Hindawi Publishing CorporationEvidence-Based Complementary and Alternative MedicineVolume 2013 Article ID 525613 17 pageshttpdxdoiorg1011552013525613

Research ArticleEvaluation of Aromatic Plants and Compounds Used toFight Multidrug Resistant Infections

Ramar Perumal Samy1 Jayapal Manikandan23 and Mohammed Al Qahtani2

1 Infectious Diseases Programme MD4 5 Science Drive 2 Department of Microbiology Yong Loo Lin School of MedicineNational University Health System (NUHS) National University of Singapore Singapore 117597

2 Center of Excellence in Genomic Medicine Research King Abdulaziz University PO Box 80216 Jeddah 21589 Saudi Arabia3 School of Anatomy Physiology and Human Biology The University of Western Australia 35 Stirling HighwayCrawley WA 6009 Australia

Correspondence should be addressed to Ramar Perumal Samy rperumalsamyyahoocoukand Mohammed Al Qahtani mhalqahtanikauedusa

Received 29 November 2012 Revised 7 May 2013 Accepted 23 May 2013

Academic Editor Rong Zeng

Copyright copy 2013 Ramar Perumal Samy et al This is an open access article distributed under the Creative Commons AttributionLicense which permits unrestricted use distribution and reproduction in any medium provided the original work is properlycited

Traditional medicine plays a vital role for primary health care in India where it is widely practiced to treat various ailments Amongthose obtained from the healers 78 medicinal plants were scientifically evaluated for antibacterial activity Methanol extract ofplants (100120583g of residue) was tested against the multidrug resistant (MDR) Gram-negative and Gram-positive bacteria Forty-seven plants showed strong activity against Burkholderia pseudomallei (strain TES and KHW) and Staphylococcus aureus of whichTragia involucrata L Citrus acida Roxb Hookf and Aegle marmelos (L) Correa ex Roxb showed powerful inhibition of bacteriaEighteen plants displayed only a moderate effect while six plants failed to provide any evidence of inhibition against the testedbacteria Purified compounds showed higher antimicrobial activity than crude extractsThe compounds showed less toxic effect tothe human skin fibroblasts (HEPK) cells than their corresponding aromatic fractions Phytochemical screening indicates that thepresence of various secondary metabolites may be responsible for this activity Most of the plant extracts contained high levels ofphenolic or polyphenolic compounds and exhibited activity against MDR pathogens In conclusion plants are promising agentsthat deserve further exploration Lead molecules available from such extracts may serve as potential antimicrobial agents for futuredrug development to combat diseases caused by the MDR bacterial strains as reported in this study

1 Introduction

Treatment of infections is compromised worldwide by theemergence of bacteria that are resistant tomultiple antibiotics[1] New and emerging drug resistance bacteria strains par-ticularlymethicillin-resistant Staphylococcus aureus (MRSA)vancomycin-resistant enterococci (VRE) Mycobacteriumtuberculosis (MTB) and multidrug resistance (MDR) Gram-negative bacteria are increasing worldwide and add to thegravity of the situation [2] S aureus cause a variety ofsyndromes such as food poisoning toxic shock syndromeskin lesions [3] hyperproliferative skin disease [4] andatopic dermatitis [5 6] Community-acquired pneumoniacaused by Streptococcus pneumoniae Klebsiella pneumonia

and S aureus accounts for significant mortality in SoutheastAsia [7] Melioidosis has been recognized as an importanthuman infection caused by Burkholderia pseudomallei inSingapore MalaysiaThailand and Northern Australia [8 9]Cases have also been reported from some other tropical andsubtropical regions like Africa and America and a number ofcases in man has recently been reported to increase in ChinaTaiwan and South India [10 11] Infection with antibioticresistant bacteria negatively impacts on public health dueto an increased incidence of treatment failure and severityof diseases Development of resistant bacteria due to thechromosomal mutations is more commonly associated withthe horizontal transfer of resistance determinants borne onmobile genetic elements [12] B pseudomallei is intrinsically

2 Evidence-Based Complementary and Alternative Medicine

resistant to many antibiotics [13 14] Considering the highercost for producing synthetic drugs and the various sideeffects associated with their use the need to search foralternative agents from medicinal plants and essential oilsused in folklore medicine is further justified to overcomethese issues In India there are about 550 tribal communitiescovered under 227 ethnic groups residing in about 5000villages throughout different forest and vegetation regions[15] India is one of the worldrsquos 12 megabiodiversity countries[16 17] Plant derived medicines have played a major rolein human societies throughout the history and prehistory ofmankind [18]The traditional healers (traditional physicians)or medicinemen have a long history of their own diagnosticand treatment system which they have acquired from theirancestors [19] Approximately 80 of the world populationstill relies on traditional medicine for the treatment of com-mon diseases [20ndash22] Medicinal plants thus offer significantpotential for the development of novel antibacterial therapiesand adjunct treatments [23] Plant derived drugs serve as aprototype to develop more effective and less toxic medicinesIn previous studies few attempts were made to confirm theantimicrobial activity of some indigenous medicinal plants[24 25] Not only extracts of variousmedicinal plants but alsoessential oils and their constituents have been investigatedfor their antimicrobial properties against bacteria and fungi[26ndash28]

The principal compounds from the leaves showed a betterantibacterial activity against P aeruginosa and B subtilisbacteria and a significant antifungal activity on C albicans[29] The essential oil from R officinalis (alpha pineneverbenonebornyl acetate) was found to be more sensitiveto the Gram-positive bacteria (MIC 25ndash4mgmL) thanto the Gram-negative bacteria [30] Several essential oilskill bacteria by damaging the cell membrane structure andinhibiting their membrane function [31] Because of theantimicrobial potency of plant extracts and oils they becomea rich source of raw materials for many biotechnological andpharmaceutical industries for the development of therapeuticdrugs The increasing trend in the use of aromatic plantsand essential oils in food cosmetic and pharmaceuticalindustries suggests that a systematic study of traditionalmedicinal plants is very important in order to find activecompounds from such sources [32ndash34] The purpose ofthis study is to survey and investigate popular medicinalaromatic plants and their essential oils with a view to fightagainst multidrug resistant human pathogens In the presentstudy 71 plant species were selected on the basis of theavailable medicinal information and screened for their invitro antimicrobial efficacy against bacteria

2 Materials and Methods

21 Ethnomedicinal Survey and Collection of Plants Eth-nomedicinal surveys were conducted duringMarch 1998 andJuly 2001 from various tribal localities (Kolli hills Kalrayanhills Pachamalai Javadi hills Mundanthurai) of Eastern andWestern Ghats Tamil Nadu India For ethnobotanical stud-ies questioners were used to collect the general informationon the tribes and the key information on medicinal details

was collected through interviews The medicinal plants wereidentified by a taxonomist using the standard Flora ofTamil Nadu Carnatic [35] and the voucher specimens weredeposited in the departmentrsquos herbarium at the EntomologyResearch Institute Loyola College Chennai India

22 Preparation of Plant Extracts Using a Soxhlet apparatusthe shade-dried and powdered plant materials (200 g of each)were extracted with 1000mL of methanol (CH

3OH) for 10 h

The collected methanol extracts were filtered (Whatman no1 filter paper) and evaporated with a rotary evaporator andfreeze dryer (lyophilized) to obtain the crude extracts (BuchiLabortechnik AG Switzerland) The dried crude extractswere stored at 4∘C for antimicrobial assays [34]

23 Culture of Microorganisms The following Gram-nega-tive Burkholderia pseudomallei (TES21) Burkholderia pseu-domallei (KHW22) Klebsiella pneumoniae (ATCC15380)Klebsiella pneumoniae Pseudomonas aeruginosa (ATCC2-7853)Vibrio damsela and Salmonella typhi (ATCC51812) andGram-positive Staphylococcus aureus (ATCC 29213) Strepto-coccus pyogenes and Streptococcus pneumoniae (ATCC49619)microorganisms were used for cultures B pseudomalleibacterial strains such as KHW and TES were isolated fromthe patient samples obtained from the Department of Micro-biology NUS Singapore The strains were subcultured on20mLTryptic Soy (TS) andMueller Hinton (MH) agar plates(pH 74) and incubated overnight at 37∘C before use

24 Antimicrobial Activity The standard bacterial cultureswere stored at minus70∘C subcultured on 20mL MH and TSagar plates (pH 74) and incubated overnight at 37∘C priorto use The antimicrobial property was tested using the disc-diffusion method [36] Five young colonies of each strain ofbacteria taken from their respective cultures grown overnighton TS agar plates (Oxoid limited Wode Road BasingstokeHants England UK) were suspended in 5mL of sterile saline(09) and the density of the suspension was adjusted toapproximately 3 times 108 colony forming unit (CFU) The swabwas used to inoculate the dried surface of TS agar plateby streaking four times over the surface of the agar androtating the plate approximately 90∘C to ensure an evendistribution of the inoculums The medium was allowed todry for about 3min before adding a 6 millimeter in diameter(mm) sterile paper disc (Becton Dickinson USA) on thesurface Each disc was tapped gently down onto the agar toprovide a uniform contact Lyophilized residue (100120583gmL)of each plant extracts and purified fractions was weighedand dissolved in 1mL of water and 20120583L of the extractsand oils (containing 100120583g of residue) were applied on eachdisc (3 replicates) while the sterile blank disc served asa normal control The antimicrobial effect of the extractson the clinical isolates was determined in comparison withthe reference antibiotics (chloramphenicol 30120583gdisc andceftazidime 30 120583gdisc) which were used as positive controlsThe plates were incubated at 37∘C for 24 h and the inhibitionzones were measured and calculated

Evidence-Based Complementary and Alternative Medicine 3

25Minimum Inhibitory Concentrations (MICs) Assay MICswere evaluated based on the in vitro screening of 16 purifiedfractions that were found to have potent antimicrobial activ-ity Broth dilution method was used for the MIC assay withsome slight modifications as recommend by the NCCLS [37]Two-fold serial dilutions of all the fractionated compoundswere made with MH and TS broth in microtiter plate wellsto adjust the final concentration from 78 to 250120583gmL whilewells containing the broth alonewithout any sample served asa controlThree replicates (119899 = 3) were used for each dilutionand culture containing approximately 1 times 105 CFUmL Theplateswere incubated at 37∘C for 24 h and the absorbancewasmeasured at 560 nm

26 Cytotoxicity Assay The cytotoxic effects of variousextracts were tested by MTT assay [38] using human skinfibroblast HEPK cells The toxic effect of plant extracts andessential oils was assayed on human skin fibroblast (HEPK)cell proliferation in 96-well microtitre plates Confluent cells(5 times 106 cells per well) were incubated with extracts and oilsfor 24 h and the percentage inhibitory concentration (IC

50)

was determined

27 Phytochemical Analysis of Plant Extracts Themost activeextracts were used for purification of antimicrobial com-pounds [39] 100 g each of the plant powder was percolatedwith 500mL of 4 aqueous HCl (adjusted to pH 2) andheated at 50∘C for 3 hThe extract waswashedwith 2times250mLof diethyl ether and the organic phase evaporated to drynessusing vacuum rotary evaporator The dark brown gummyresidues obtained by acid hydrolysis were chromatographedon silica gel column 60 times 32 cm (60ndash120 mesh pH 7 Merck)by gradually eluting with n-hexaneethyl acetate (8 2 6 43 7 and 1 1) and chloroformmethanol (3 2) The aliquotsof each fraction were subjected to thin layer chromatography(TLC) on silica gel coated TLC plate (1mm Merck) usingthe solvent system consisting of 20 (vv) n-hexaneethylacetateThe chromatograms were detected using 50H

2SO4

solution as a spray reagent [34]The individual fractions werecollected and concentrated by vacuum rotary evaporator at40∘C All the purified compounds recovered from the silicagel column were monitored by reading the absorbance at190ndash350 nm (UV spectrophotometer Hitachi Japan) Theactive fractions were further purified and the final yieldsof the compounds were recorded The lyophilized pooledconcentrated compounds were then assayed (100120583gmL)against bacteria The phytochemical screening was done onthe pure compounds using the chemical method previouslyreported for the detection of secondary metabolites [40]The different chemical constituents tested include alkaloidsflavonoids glycosides polyphenols saponin sterols triter-penes tannins reducing sugars gallic acid catechol andaglycones

28 Statistical Analysis The bacterial growth inhibitoryactivity (inhibition zones millimeter in diameter) was com-pared for significant differences within the bacterial strainsOne way analysis of variance was performed (mean plusmn SD

119899 = 3 replicates) using GraphPad Prism 4 USA lowast119875 lt 001was considered statistically significant (inhibition zones ofextractsfractions versus antibiotic drugs)

3 Results

31 Study Area for the Collection of AromaticMedicinal PlantsTheWestern and Eastern Ghats were selected for the presentstudy with the cite map showing the landmarks (Figures 1(a)-1(b)) Kalrayan hills are situated north of Attur taluk (Salemdistrict) one of the major range of hills in the Eastern Ghatsof Tamil Nadu (Figure 1(c)) Pachamalai hills are situated tothe north of Thuraiyur taluk of Tiruchirappalli district Therich biodiversity part of Eastern Ghats lies between latitudes11∘091015840 0010158401015840 to 11∘ 271015840 0010158401015840 N and longitudes 78∘281015840 0010158401015840 to78∘ 491015840 0010158401015840 E and occupies an area of about 52761 squarekm It is located near 11∘ 111015840N 78∘21E1118∘N 7835∘E11187835 (Figure 1(d)) Mundanthurai is located nearly 45 kmwest of Tirunelveli district TN between latitude 8∘ 251015840 and8∘ 531015840 N and longitude 77∘ 101015840 and 77∘ 351015840 E This is the onlyarea of Western Ghats that has the longest raining period ofabout 8 months and forms the catchment area for 14 riversand streams (Figure 1(e)) Kolli Malai is a small mountainrange located in Namakkal district The mountains are about1000ndash1300m in height and cover an area of approximately280 km The Kolli hills are part of the Eastern Ghats whichis a mountain range that runs mostly parallel to the east coastof Tamil Nadu in South India (Figure 1(f)) Javadi hills areone of the largest in the Eastern Ghats in Vellore district inthe northern part of the state of Tamil Nadu They consistof bluish gray hills with peaks averaging 3600ndash3800 feetor 1100ndash1150 meter (Figure 1(g)) Based on the vegetationtype (Figures 2(a)ndash2(d)) the study area consists of (i) drydeciduous (ii) moist deciduous and (iii) rain forests anddiverse proportion of plant parts in abundance (Figures 2(e)-2(f)) Three different types of tribes (ie Kani Malayali andPaliyan tribes) inhabit in the hill ranges The Kani tribeslocated at Mundanthurai raise different types of vegetablesin their own fields while the Malayali tribes cultivate riceThey all engage not only in the agricultural work but alsoare involved in silvicultural work assigned by the forestdepartment Government of TN India

32 Medicinal Plants Glory Western Ghats (Mundanthurai)and Eastern Ghats (Kolli hills Javadi hills Kalrayan hillsPachamalai hills) possess a rich diversity of medicinal plantsthat are used as food and drug by different groups oftribal communities Urbanization habitat degradation andfragmentation of these forests have resulted in the depletionof natural resources on which these tribes used to dependfor their livelihoods It has become increasingly difficult forthem to live in their traditional way In addition the impactof modernization and urbanization has encroached in andaround tribal settlements thus changing their lifestyles

33 Plants Valued as Edibles Various types of plant parts arecollected during different seasons cooked and eaten alongwith boiled rice (Table 1) For example Solanum nigrum leafis most commonly used in all the four regions There are

4 Evidence-Based Complementary and Alternative Medicine

Figure 1 (a) The site for collection of medicinal plants in Western and Eastern Ghats of Tamil Nadu (b) The landmark (map) of traditionalmedicine distribution and collection of different types of plants (c) District map showing the collection site of plants from Kalrayan hills(Salem) (d) Pachamalai hills (Thiruchirappalli) and (e) Mundanthurai (Tirunelveli) rich biodiversity hot-spot of the Western Ghats (f)Kolli hills (Namakkal) (g) Javadi hills (Vellore) part of the Eastern Ghats which is a mountain range that runs mostly parallel to the eastcoast of South India

a large number of wild edible fruits including yielding plantssuch as Citrus acida Ficus benghalensis Ficus microcarpaFicus racemosa Phyllanthus emblica Solanum trilobatumand Syzygium cumini are popularly used by the tribes

34 Plants Used for Snakebite Treatment Thirty-four plantsused for snakebite treatment are documented (Table 1)Snakebite is amajor health hazard that leads to highmortalityin tribal settlements The majority of the antidotes are

prepared freshly from plant materials frequently collectedfrom the leaves of A paniculata A echioides Aristolochiaindica E alba E prostrata M pudica O sanctum Tinvolucrata and Cleistanthus collinus (Oduvanthalai) thewhole plants ofAchyranthes aspera andWedelia calendulaceathe stem-barks and nuts of Strychnos nux-vomica the rootsof Hemidesmus indicus Tephrosia purpurea Rauwolfia ser-pentina C roseus and so forth and the tubers of Gloriosasuperba The tuber paste is usually applied externally on

Evidence-Based Complementary and Alternative Medicine 5

Table 1 Some of the important traditional medicinal plant species families voucher specimens parts used yield of extracts phytochemicalscreening and toxicity on human macrophage cells

Scientific name Family Voucher specimen Plant parts Yield (gm) Phytochemicalanalysis

Adhatoda vasica Nees Acanthaceae D2020 Leaf 64 VasicineAegle marmelos (L)Correa ex Roxb Rutaceae D2018 Root-bark 58 Alkaloids

Alangium salvifolium (L)f Wangerin Alangiaceae 0140 Leaf 63 Phenolic

Andrographis paniculataWallich ex Nees Acanthaceae 0061 Leaf 68 Andrographolide

Andrographis echioidesNees Acanthaceae 0116 Leaf 70 Terpenoids

Acalypha indica L Euphorbiaceae 29644 Leaf 61 AcalypheAcalypha lanceolata L Euphorbiaceae 15791 Leaf 71 AlkaloidsAchyranthes aspera L Amaranthaceae 2666 Leaf 27 BetaineAgeratum conyzoides L Asteraceae 4812 Leaf 37 Essential oilsAsteracantha longifolia L Acanthaceae 0234 Stem 74 GlycosidesAzadirachta indica A Juss Meliaceae D0204 Leaf bark 67 Tannins

Borassus flabellifer L Arecaceae D0202 Root 08 Flavonoidsphenolics

Boerhavia erecta L Nyctaginaceae 10897 Whole plant 28 PhenolicsCalotropis procera (Ait)Ait f Asclepiadaceae D073 Root-bark 13 Terpenoids

Calotropis gigantea (L)RBrex Ait Asclepiadaceae D070 Milky latex 48 Alkaloids

Cassia auriculata L Caesalpiniaceae 0141 Leaf 49 Saponins

Cassia occidentalis L Caesalpiniaceae 0111 Root 95 Flavonoidssaponins

Cassia tora L Caesalpiniaceae 0100 Stem bark 69 SaponinsCassia fistula L Caesalpiniaceae 037 Whole plant 79 SaponinsCardiospermumhalicacabum L Sapindaceae 0125 Whole plant 58 Flavonoids

Catharanthus roseus (L)GDon Apocynaceae 0029 Leaf

root1407 Alkaloids

Cinnamomum zeylanicumGarcin ex Blume Lauraceae 00209 Bark 31 Essential oil

TanninCinnamomum inersReinw ex Blume Lauraceae 043-c Leaf bark Alkaloids

Cissus quadrangularisRoxb Vitaceae D02023 Leaf 68 Glycosides

Citrus acida Roxb Hookf Rutaceae 0213 Leaf 52 SaponinsTerpenoids

Centella asiatica (L) Umbelliferae 0138 Whole plant 89 FlavonoidsAlkaloids

Clerodendrum inerme (L)Gaertn Verbenaceae D02043 Stem 78 Sterols diterpenes

Clitoria ternatea L Papilionaceae D02026 Seed 98 ProteinCleistanthus collinus(Roxb) Benth andHookf

Euphorbiaceae 0011 Whole plant 003 Cleistanthincollinusin

Cleome gynandropsis L Capparidaceae 12247 Leaf 62 GlycosidesCleome viscose L Capparidaceae 29999 Leaf 27 Phenolics

6 Evidence-Based Complementary and Alternative Medicine

Table 1 Continued

Scientific name Family Voucher specimen Plant parts Yield (gm) Phytochemicalanalysis

Coccinia grandisWampA Cucurbitaceae D02030 Leaf root 09 GlycosidesCymbopogon citratus(DC) Gramineae D012 Root 025 Essential oil

Datura metel L Solanaceae D02038 Leaf stem 39 SteroidsEucalyptus globulus Labill Myrtaceae D0220 Leaf 12 TerpenoidsEclipta alba (L) Hassk Asteraceae D028 Whole plant 07 PhenolicEuphorbia hirta Linn Euphorbiaceae 0018-c Whole plant 012 mdash

Eclipta prostrata (L) Asteraceae D210 Leaf 110 Triterpenoidsaponin

Eugenia caryophyllus(Sprengel) Bullock ampHarrison

Myrtaceae 0025 Flower buds 116 Essential oils

Elettaria cardamomumWhite et Mason Zingiberaceae 0009 Fruit pods 317 Essential oils

Gloriosa superba L Liliaceae 020-S Tuber 108 Alkaloids phenol

Jatropha curcas L Euphorbiaceae 015 Whole plant 53 Alkaloidsflavonoids

Hyptis suaveolens (L) Poit Lamiaceae 24688 Leaf 63 Essential oilHemidesmus indicus L Asclepiadaceae D-009 Roots CoumarinsIchnocarpus frutescens(L) RBr Apocynaceae 0110 Root flower 73 Terpenoids

Leucas aspera (Willd)Link Labiatae 0114 Leaf 83 Triterpenes

Lawsonia inermis L Lythraceae T261 Leaf 09 Glycosidesphenolic

Madhuca longifolia (L) JFMacbr Sapotaceae D01415 Nut 93 Sitosterol

Merremia hastate L(Desr) Hallierf Convolvulaceae 10894 Whole plant 40 Alkaloids

Mentha piperita L Lamiaceae 0217-c Whole plant 07 Essential oilsMorinda tinctoria Roxb Rubiaceae 0122 Leaf 14 GlycosidesMimosa pudica L Mimosaceae 0071 Whole plant 06 mdashOldenlandia umbellata L Rubiaceae D02047 Leaf 44 AlkaloidsOcimum sanctum L Lamiaceae 0016 Whole plant 30 AlkaloidsPiper attenuatum BuchHamex Miq Piperaceae 007 Flower 46 Alkaloids

Plumbago zeylanica (L)Cav Plumbaginaceae 0121 Root 48 Plumbagin

Plectranthus amboinicus(L) Spreng Lamiaceae 0410 Whole plant 12 Essential oils

terpenoidsPhyllanthus debilis L(Klein ex Willd) Euphorbiaceae 0120 Whole plant 49 Polyphenol

Phyllanthusmadraspatensis L Euphorbiaceae 0117 Whole plant 50 Polyphenol

Premna tomentosaWilld Verbenaceae 0129 Leaf 53 DiterpenesRosmarinus officinalis L Lamiaceae 0017 Root 023 Essential oilsRauwolfia serpentine L Apocynaceae 020-S Root 115 AlkaloidSebastiania chamaelea (L)Muell Arg Euphorbiaceae 0034 Leaf 11 Polyphenol

Solanum trilobatum L Solanaceae D02054 Leaf flower 40 Tannins

Evidence-Based Complementary and Alternative Medicine 7

Table 1 Continued

Scientific name Family Voucher specimen Plant parts Yield (gm) Phytochemicalanalysis

Sphaeranthus indicus L Asteraceae D02060 Whole plant 10 Essential oilSwertia chirata (L) Ham Gentianaceae D0540 Whole plant 16 GlycodisesStrychnos nux-vomica L Loganiaceae S-22 Nuts 036 AlkaloidsTragia involucrata L Euphorbiaceae D068 Leaves 16 ShellsolTinospora cordifolia(Willd) Miers ex Hooff ampThoms

Menispermaceae 0118 Leafroot stem 50 Glycosides tannins

Tridax procumbens L Compositae 10649 Leaf 18 FlavonoidsTerminalia arjuna (DC)W amp A Combretaceae 033-c Bark 80 Phenolics

Tephrosia purpurea (L)Pers Fabaceae S-43 Whole plant 08 Isoflavone

Vitex negundo L Verbenaceae 0031 Leaf 24 TerpineolVetiveria zizanioides L Gramineae 0051 Root 103 Essential oilWithania somnifera (L)Dunal Solanaceae D02063 Root 21 Alkaloids

Wedelia calendulacea Less Asteraceae S-24 Leaves FlavonoidsZingiber officinale Rosc Zingiberaceae 0327 Rhizome 23 TanninsZanthoxylum limonella(Dennst) Alston Rutaceae 009 Bark 19 Alkaloids essential

oilClass of chemical compounds A alkaloids S saponins T tannins St steroids G glycosides T terpenoids P polyphenol P phenolics Sh shellsol Hhydrocarbon esters

the site of snakebite and decoction is given orally fortreatment by indigenous people Besides these tribes relyon the medicinal plants as ingredients for fabricating a kindof medicated stone for health management ldquoVishakallurdquo(poison stone) is used by the indigenous groups called Kaniin Kerala India to treat a snakebiteWhen the stone is placeddirectly on the bitten area it sticks to the body to absorb thepoison and then become detached when absorption seems tobe complete The ingredients of Vishakallu stones are madewith leaves of Ocimum sanctum Anisomeles malabaricaLeucas aspera Piper betle Santalum album and the pebblescollected from the river bank

35 Survey of Medicinal Plants and Their Health Care ValuesThe present study is an attempt to provide scientific basis andobtain justification for the traditional beliefs of reliance on arich diversity of ethnomedicinal plants along with the richheritage of traditional medicine practices related to healthcare system made available by the primitive tribal commu-nities located at different settlements The native traditionalpractitioners called ldquovaidyarsrdquo have a good knowledge aboutthe traditional plants locally available for treatment of var-ious diseases (Figures 3(a)ndash3(p)) Such traditional medicalknowledge is used for preparing home remedies ill healthprevention and routine health maintenanceThis knowledgeis also applicable to cover other sectors of social life Duringthe ethnobotanical survey the wealth of 78 medicinal plantspecies used by the indigenous tribal community for varioustypes of health treatment was documented The botanical

names family names parts used chemical constituents andtheir application are provided (Table 1)

36 Antimicrobial Activity of Crude Extracts In this studywe reported the antimicrobial screening of methanolic crudeextracts of 78 medicinal plants (Table 2) Results revealedthat 68 plant extracts displayed potent activities against oneor more Gram-positive and -negative bacteria Of whichTragia involucrata Citrus acida Aegle marmelos Adha-toda vasica Calotropis procera Andrographis paniculata andMentha piperita Azadirachta indica Sphaeranthus indicusand Elettaria cardamomum showed the highest antibacte-rial activity against the multidrug resistant B pseudomallei(KHW and TES) and S aureus at 100120583gmL concentrationThe extracts showed pronounced antibacterial activity withtheir inhibitory zones ranging from 20 to 31mm in diam-eter as compared to the standard drugs chloramphenicoland ceftazidime (29ndash33mm) The majority of the plantsdemonstrated a powerful antimicrobial potency against themultidrug resistant strains of B pseudomallei (KHW andTES) K pneumonia and S aureus Approximately twenty-one plant extracts exerted only a weak or moderate effectagainst the tested bacteria while the crude extract of 13plants failed to show any effect at all Except for the plantextracts of T involucrata A lanceolata A vasica and Sindicus extracts the majority of the plant extracts wereineffective against theV damsela infection fascinatingly only11 plants exhibited activity against P aeruginosa of whichS indicus M piperita and C procera were found to havevery strong inhibition of bacteria at the tested concentrations

8 Evidence-Based Complementary and Alternative Medicine

Table 2 Antimicrobial activity of methanol extract of aromatic medicinal plants and essential oils evaluated against multidrug resistant(MDR) human pathogens at 100120583gmL concentration

Scientific name Microorganisms growth inhibition zones (6 millimeter in diameters)KHW TES Kp Kpr Pa Sa Stp Sp Vd Vd

Adhatoda vasica Nees 28 19 22 18 12 mdash 10 17 16 mdashAegle marmelos (L) Correa ex Roxb 29 17 9 10 mdash 15 mdash mdash mdash mdashAlangium salvifolium (L) f Wangerin 15 10 8 8 mdash 12 9 10 mdash mdashAndrographis echioides L 12 9 mdash mdash mdash 17 8 9 mdash mdashAndrographis paniculataWallich ex Nees 26 21 19 13 mdash 25 8 16 12 mdashAcalypha indica L mdash mdash mdash mdash mdash 18 mdash mdash mdash mdashAcalypha lanceolata L mdash mdash 12 13 mdash 10 mdash mdash 21 mdashAchyranthes aspera L mdash mdash 7 14 mdash 15 12 11 mdash mdashAgeratum conyzoides L 10 8 mdash mdash mdash mdash mdash mdash mdash mdashAsteracantha longifolia L 16 12 mdash mdash mdash 15 mdash mdash mdash mdashAzadirachta indica A Juss 15 17 21 16 14 23 12 14 mdash mdashBorassus flabellifer L 9 10 17 8 mdash mdash 9 10 mdash mdashBoerhavia erecta L mdash mdash 8 9 7 16 mdash mdash mdash mdashCalotropis procera (L) mdash mdash 15 mdash 18 28 9 mdash mdash mdashCalotropis gigantea (L) RBrex Ait 11 9 mdash mdash mdash 20 mdash mdash 9 8Cardiospermum halicacabum L 23 mdash mdash 14 mdash 9 19 mdash mdash mdashCatharanthus roseus (L) GDon 13 7 12 9 mdash 15 11 8 mdash mdashCassia auriculata L 17 13 12 mdash mdash 19 13 mdash mdash mdashCassia occidentalis L 18 mdash mdash mdash mdash mdash mdash mdash mdash mdashCassia tora L mdash mdash mdash mdash mdash mdash mdash mdash mdash mdashCassia fistula L mdash mdash mdash mdash mdash mdash mdash mdash mdash mdashCitrus acida Roxb Hookf 26 22 mdash 12 29 9 13 8 mdashCissus quadrangularis L mdash mdash mdash mdash mdash mdash mdash mdash mdash mdashCinnamomum zeylanicum Garcin exBlume 14 16 7 20 mdash 22 19 7 mdash mdash

Cinnamomum iners Reinw ex Blume 20 16 mdash 15 mdash 16 12 mdash mdash mdashRosmarinus officinalis L mdash 9 mdash 10 mdash 7 mdash 8 mdash mdashCentella asiatica (L) 9 8 mdash mdash mdash 11 mdash mdash mdash mdashClerodendrum inerme (L) Gaertn 13 7 12 mdash mdash 15 11 8 mdash mdashClitoria ternatea L mdash mdash mdash mdash mdash mdash mdash mdash mdash mdashClitoria ternatea L 16 12 mdash 13 mdash 8 10 mdash mdash mdashCleome gynandropsis L mdash mdash 12 19 11 mdash 9 15 12 mdashCleome viscose L mdash mdash 8 10 14 20 mdash mdash mdash mdashCoccinia grandisWampA 9 10 17 8 mdash mdash 9 10 mdash mdashCymbopogon citratus (DC) 16 18 mdash 17 mdash 14 mdash mdash mdash mdashDatura metel L mdash mdash mdash mdash mdash mdash mdash mdash mdash mdashEclipta alba (L) Hassk 20 mdash mdash 9 mdash mdash 10 mdash mdash mdashEuphorbia hirta Linn 11 mdash mdash mdash mdash 16 mdash mdash mdash mdashEucalyptus globulus Labill mdash mdash mdash 7 mdash mdash mdash 7 mdash mdashEugenia caryophyllus Bullock amp Harrison mdash 7 mdash 11 mdash 9 mdash 8 mdash mdashElettaria cardamomumWhite et Mason 21 20 7 14 mdash 22 12 17 mdash mdashHyptis suaveolens (L) Poit 8 10 8 7 mdash mdash mdash mdash mdash mdashIchnocarpus frutescens (L) WJ Aiton mdash mdash mdash mdash mdash mdash mdash mdash mdash mdashJatropha curcas L mdash mdash mdash 8 mdash 11 mdash 12 mdash 7Leucas aspera (Willd) Link 9 10 17 8 mdash 12 9 10 mdash mdashLawsonia inermis L mdash mdash mdash mdash mdash mdash mdash mdash mdash mdashMadhuca longifolia (L) JF Macbr 18 16 14 19 12 mdash 8 7 9 mdash

Evidence-Based Complementary and Alternative Medicine 9

Table 2 Continued

Scientific name Microorganisms growth inhibition zones (6 millimeter in diameters)KHW TES Kp Kpr Pa Sa Stp Sp Vd Vd

Merremia hastate L (Desr) Hallierf mdash mdash mdash mdash mdash mdash mdash mdash mdash mdashMorinda tinctoria Roxb mdash mdash mdash mdash mdash mdash mdash mdash mdash mdashMentha piperita L 23 17 26 12 20 25 19 mdash mdash mdashOcimum sanctum L 12 9 11 7 mdash 15 7 8 mdash mdashOldenlandia umbellata L mdash mdash mdash mdash mdash 17 mdash mdash mdash mdashPiper attenuatum Buch Hamex Miq 13 7 12 21 mdash 17 11 8 mdash mdashPlumbago zeylanica (L) Cav 9 10 17 8 mdash 12 9 10 mdash mdashPlectranthus amboinicus (L) Spreng mdash mdash mdash 8 11 15 mdash mdash mdash mdashPhyllanthus debilis L (Klein ex Willd) 7 8 mdash 9 mdash 9 18 7 mdash mdashPhyllanthus maderaspatensis L 17 mdash mdash mdash mdash mdash mdash mdash mdash mdashPremna tomentosaWilld 13 10 mdash 15 mdash 9 10 mdash mdash mdashGloriosa superba L 17 16 7 8 mdash 15 mdash mdash mdash 8Sebastiania chamaelea (L) Muell Arg 19 12 mdash 17 mdash 13 19 8 mdash mdashSolanum trilobatum L mdash mdash 13 9 mdash 8 mdash mdash mdash mdashSphaeranthus indicus L 20 18 21 7 22 mdash 11 9 mdash 16Swertia chirata (L) Ham mdash mdash mdash mdash mdash mdash mdash mdash mdash mdashTerminalia arjuna (W amp A) mdash mdash mdash mdash mdash 16 mdash mdash mdash mdashTinospora cordifolia (Willd) Miers exHooff ampThoms mdash mdash 23 16 mdash 12 15 mdash mdash mdash

Tridax procumbens L 9 8 7 mdash mdash 14 mdash mdash mdash mdashTragia involucrata L 25 23 20 mdash mdash 31 28 22 19 mdashVitex negundo L mdash mdash mdash mdash mdash 14 mdash mdash mdash mdashVetiveria zizanioides (L) 11 9 7 7 mdash 16 mdash mdash 8Withania somnifera (L) Dunal 12 20 mdash mdash mdash 15 12 mdash mdash mdashZingiber officinale Rosc 14 11 7 15 mdash 7 12 7 mdash mdashZanthoxylum limonella (Dennst) Alston 13 7 12 9 mdash mdash 11 8 mdash mdashChloramphenicol (30120583gdisc) 21 12 15 17 29 16 15 18 13 11Ceftazidime (30 120583gdisc) 33 16 22 19 16 25 21 20 12 15lowastBacteria (+minus) Results obtained in the disc diffusion assay antibacterial activity is expressed as the mean plusmn SD (119899 = 3) of the inhibition by the extract andits diameter around the discs One way analysis of variance was performed (mean plusmn SD 119899 = 3 replicates) Size of inhibition zones were including the sterileblank discs 6 millimeter (mm) in diameters Absence of bacterial inhibition indicates (mdash) antibiotic disc (30 120583gdisc)

Interestingly sixteen plants such as Andrographis echioidesC auriculata C viscose C gigantea T arjuna Oldenlandiaumbellata Boerhavia erecta and E hirta exerted a strongactivity against the Gram-positive S aureus bacteria

37 Phytochemical Screening of Plants The results obtainedfrom the phytochemical screening as shown in Table 1 indi-cate the presence of various types of secondary metabolitessuch as polyphenols tannins saponins alkaloids and gly-cosidespolysaccharides Most of the plant extracts relativelyrich in alkaloids phenols flavonoids polyphenols tanninssterols and terpenoids were found to inhibit the growth oforganisms

38 Antimicrobial Activity of Fractioned Compounds Activecomponents were purified from the most active extractsfor further testing The compound shellsol of T involucrata

and C acida exhibited the most potent action againstthe antibiotic resistant strains of B pseudomallei (KHW)S aureus B pseudomallei (TES) and K pneumoniae Amarmelos was also found to inhibit the growth of B pseudo-mallei (KHW) more effectively than other tested bacteria Avasica showed the broad spectrum growth inhibitory activityon B pseudomallei (KHW) K pneumoniae K pneumoniaeresistant B pseudomallei (TES) S pyogenes and V damselaHowever E cardamomum displayed antimicrobial activityon some of the B pseudomallei (KHW) S pyogenes Bpseudomallei (TES) and S typhi strains Similarly A indicaexerted the growth inhibition on K pneumoniae and Saureus Remarkably Sebastiania chamaelea was more activeagainst K pneumoniae K pneumoniae and S pyogenesThe compound from S indicus inhibited the growth of Kpneumoniae K pneumoniae B pseudomallei (KHW andTES) and S typhi strains as compared to the activity shown

10 Evidence-Based Complementary and Alternative Medicine

Figure 2 Diverse biodiversity richness of medicinal plants in Western and Eastern Ghats (a) Topography of plant covering area in Kollihills (Namakkal district Tamil Nadu) (b) Aerial view of occurrence of medicinal plants in Mundanthurai hills (Tirunelveli district TN) (c)Pachamalai hills (Trichy district) and its natural vegetation inhabitants for Malaiyali tribes (d) Deforestation of natural herbal resources dueto urbanization in Kalrayan hills (Salem district) in the Eastern Ghats of TN (e) Medicinal plants and its various parts used by the natives(traditional healers) for the treatment of diverse human illness with a very high percentage of leaves and whole plants often used for herbaldrug preparation by the local practitioners (f) Various category of plants like shrub herb climbers and tree and the parts used in medicine

Evidence-Based Complementary and Alternative Medicine 11

Figure 3 Medicinal aromatic herbs spices and toxic plants were collected from the tribal areas of the Western and Eastern Ghats region inTamil Nadu India (a) A vasica Nees (leaf) (b) Eclipta alba (L) Hassk (whole plant) (c)Mimosa pudica L (whole plant) (d) P amboinicus(L) Spreng (whole plant) (e) T procumbens (L) (whole plant) (f) Euphorbia hirta Linn (whole plant) (g) A paniculata Wallich ex Nees(H) C roseus (L) GDon (whole plant) used for therapy (i) Cinnamomum iners Reinw ex Blume (leaf) (j) E globulus Labill (leaf and bark)(k) Z officinale Rosc (Rhizome) (l) E caryophyllus (Sprengel) Bullock and Harrison (flower buds) (m) M piperita L (whole plant) (n) Ccitratus (DC) Clitoria ternatea L (whole plant) (o) C zeylanicum Garcin ex Blume (bark) (p) Elettaria cardamomumWhite et Mason (fruitpod) used for medicine and food preparation

by the crude extracts (Figure 4)The antimicrobial efficacy offractions collected from the oil yielding plants was also com-pared with that of the tested compounds C zeylanicum andR officinaliswere themost sensitive in controlling the growthof B pseudomallei (KHW) S aureus K pneumonia and Spneumoniae Fascinatingly all the compounds obtained fromaromatic plants except those from E globules were foundto be very effective against the multidrug resistant humanpathogen B pseudomallei (KHW) that causes melioidosis

On the other hand compounds from C citrates O sanctumE caryophyllus and Z zizanioide showed some promisingeffect only against S aureus (Figure 5) The activity of thecompounds were pronounced more than that of the oilyielding plant fractions

39 Minimum Inhibitory Concentrations (MICs) The antibi-otic potential of the purified fractions was obtained fromthe MIC determination The hydrocarbon ester shellsol

12 Evidence-Based Complementary and Alternative Medicine

(mean plusmn SD n = 3)

0

5

10

15

20

25

30

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn V

d St

Bacteria (+minus)

lowast

lowast

lowast

A vasica

In vitro assay(P

lant

frac

tion)

Inhi

bitio

n zo

nes (

mm

)

(a)

(mean plusmn SD n = 3)

0

5

10

15

20

25

30

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn V

d St

Bacteria (+minus)

lowast

A marmelos

Inhi

bitio

n zo

nes (

mm

)

In vitro assay

(b)

(mean plusmn SD n = 3)

0

5

10

15

20

25

30

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn V

d St

Bacteria (+minus)

lowastlowast

A indica

Inhi

bitio

n zo

nes (

mm

)

Incu

batio

n (37∘C

24 h

)

(c)

(mean plusmn SD n = 3)

0

5

10

15

20

25

30

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn V

d St

Bacteria (+minus)

lowast

lowast lowast

35

C acida

Inhi

bitio

n zo

nes (

mm

)(P

lant

frac

tion)

(d)

(mean plusmn SD n = 3)

0

5

10

15

20

25

30

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn V

d St

Bacteria (+minus)

lowastlowast

lowast

S chamaelea

Inhi

bitio

n zo

nes (

mm

)

(e)

(mean plusmn SD n = 3)

0

5

10

15

20

25

30

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn V

d St

Bacteria (+minus)

lowastlowast

lowastlowast

S indicus

Inhi

bitio

n zo

nes (

mm

)

Incu

batio

n (37∘C

24 h

)

(f)

(mean plusmn SD n = 3)

0

5

10

15

20

25

30

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn V

d St

Bacteria (+minus)

lowast

lowast lowast

E cardamomum

Inhi

bitio

n zo

nes (

mm

)(P

lant

frac

tion)

(g)

(mean plusmn SD n = 3)

0

5

10

15

20

25

30

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn V

d St

Bacteria (+minus)

lowast

lowast

lowast

lowast

Shellsol

35

Inhi

bitio

n zo

nes (

mm

)

Incu

batio

n (37∘C

24 h

)

(h)

Figure 4 In vitro antimicrobial activity of purified fractions from the most active plant extracts tested against bacteria Growth inhibitionzones were measured and analyzed with mean plusmn standard deviation (SD) (119899 = 3) using one way analysis of variance Level of significanceat (lowast119875 gt 001) Most of the fractions exerted a potent inhibitory effect against multidrug resistant Gram-negative bacteria (B pseudomalleistrains KHW and TES) K pneumonia and Gram-positive bacteria S aureus

(T involucrata) andC acida showed an interesting inhibitorypotential against S aureus (MIC of 78 120583gmL) and B pseu-domallei strain of KHW (MIC of 156 120583gmL) A vasicashowed an MIC of 156120583gmL against B pseudomallei(KHW) and an MIC of 3125120583gmL against K pneumo-niae K pneumoniae S pyogenes and V damsela strainsFractions from A marmelos and terpenoid from A indicaexerted bacteriostatic effect with MIC values of 3125120583gmLon some selected bacteria including B pseudomallei ofKHW S aureus and B pseudomallei of TES The MIC of3125 120583gmL was found for E cardamomum against S aureus

K pneumonia and S pyogenes S indicus displayed a verystrong inhibition against MDR K pneumoniae (MIC of156 120583gmL) and against B pseudomallei (KHW and TES)at MIC of 3125 120583gmL When the antimicrobial efficacies ofpurified fractions from aromatic plants were compared theC zeylanicum fraction displayed an important antimicrobialeffect against S aureus (MIC of 78 120583gmL) MDR B pseu-domallei of KHW (MIC of 156 120583gmL) and S pneumoniae(MIC of 3125 120583gmL) The essential oil from M piperateashowed MIC value of 3125120583gmL against K pneumoniae Saureus and B pseudomallei (KHW) respectivelyO sanctum

Evidence-Based Complementary and Alternative Medicine 13

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn

Bacteria (+minus)

0

5

10

15

20

25

30

Inhi

bitio

n zo

nes (

mm

)(P

lant

frac

tion)

lowastlowast

lowast

(mean plusmn SD n = 3)In vitro assay

C citratus

(a)

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn

Bacteria (+minus)

0

5

10

15

20

25

30

Inhi

bitio

n zo

nes (

mm

)

lowast

lowast

lowastlowast

lowast

lowast

(mean plusmn SD n = 3)In vitro assay

35

C zeylanicum

(b)

Bacteria (+minus)

0

5

10

15

20

25

30

Inhi

bitio

n zo

nes (

mm

)

lowast

lowast

lowast

(mean plusmn SD n = 3)

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn V

d St

O sanctum

Incu

batio

n (37∘C

24 h

)

(c)

Bacteria (+minus)

0

5

10

15

20

25

Inhi

bitio

n zo

nes (

mm

)

lowast

lowast

(mean plusmn SD n = 3)

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn V

d St

M piperita

(Pla

nt fr

actio

n)

(d)

Bacteria (+minus)

0

5

10

15

20

25

30

Inhi

bitio

n zo

nes (

mm

)lowast

lowast

lowast

(mean plusmn SD n = 3)

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn V

d St

E caryophyllus

(e)

0

5

10

15

20

25

30

Inhi

bitio

n zo

nes (

mm

) (mean plusmn SD n = 3)

Bacteria (+minus)

KHW

TES

Kp

KprPa Sa

Stp

Spn

E globulus

Incu

batio

n (37∘C

24 h

)

(f)

Bacteria (+minus)

0

5

10

15

20

25

30

Inhi

bitio

n zo

nes (

mm

)(P

lant

frac

tion)

lowast

lowast

lowast

lowast

(mean plusmn SD n = 3)

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn V

d

R officinalis

(g)

0

5

10

15

20

25

30

Inhi

bitio

n zo

nes (

mm

)

lowast

lowast

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn V

d St

Bacteria (+minus)V zizanioides

(mean plusmn SD n = 3)

Incu

batio

n (37∘C

24 h

)

(h)

Figure 5 Comparison of antimicrobial effect of plant compounds obtained from the most popularly used sources of essential oils as assayedby the disc-diffusion method in vitro It displayed a powerful activity against B pseudomallei and S aureus than the other bacteria strainsOther compounds showed only a moderate or weak action against the tested bacteria Values for zone of bacterial growth inhibition werepresented as mean plusmn SD (119899 = 3) with level of significance at (lowast119875 gt 001)

and C citratus fractions also showed antimicrobial activity(MICs of 3125ndash125120583gmL) only at higher concentrationsagainst the tested bacteria In addition to that higher concen-trations (gt250120583gmL) (of Vetiveria fractions) were requiredto inhibit Vibro species and others (including E globulusfractions) failed to show any effect at tested concentrations(78ndash125 120583gmL) However the purified fractions (from mostactive medicinal plants) showed strong bacteriostatic inhibi-tion against the tested organisms (Table 3)

310 Cytotoxic Effects of Plants When the components wereassayed for cytotoxicity against the normal human skinfibroblasts (HEPK) cells the compounds obtained from

E cardamomum T involucrata S indicusC acidaA vasicaA marmelos A indica and A paniculata did not showtoxicity up to 1000120583gmL (see Figures S1 and S2 in Supple-mentaryMaterial available online at httpdxdoiorg1011552013525613) A slight reduction of cell proliferation wasnoted only at higher doses (2000120583gmL) In contrast cellproliferation was markedly reduced after exposure of HEPKcells to O sanctum E globulus V zizanioides C citratusand E globulus compounds There was no gradual reductionin skin cell proliferation seen after exposure to C zeylan-icum R officinalis and M piperita (see Figures S3 and S4)compounds The toxicity was found to be concentration-dependent when the skin fibroblasts (HEPK) cells were

14 Evidence-Based Complementary and Alternative Medicine

Table 3 Minimum inhibitory concentrations (MICs) of purified plant fractions and essential oils against antibiotic resistant bacteria

Botanical name Family Parts used Gram-positive and -negative bacteria (MICs 120583gmL)KHW TES Kp KPr Sa Stp Sp Vd St

A indica Juss Meliaceae Seed (fraction) 3125 625 125 lt250 3125 mdash mdash mdash mdashA marmelos (L) Rutaceae Root-bark (F) 3125 3125 mdash mdash 3125 625 125 250 mdashA paniculata Nees Acanthaceae Leaf (fraction) 250 mdash 125 mdash 625 mdash mdash 125 mdashA vasica Nees Acanthaceae Fraction (Stem) 156 3125 3125 625 mdash lt250 3125 3125 mdashC acida Roxb Rutaceae Leaf (fraction) 156 625 mdash mdash 78 mdash 3125 mdash mdashE cardamomumWhite et Mason Acanthaceae Fraction (WP) 625 mdash 3125 mdash 3125 250 625 mdash mdashS indicus (L) Euphorbiaceae Whole plant (F) 3125 125 156 mdash 625 mdash mdash lt250 mdashT involucrata (L) Euphorbiaceae Shellosol (leaf) 156 3125 mdash mdash 78 625 mdash mdash mdashCinnamomum zeylanicum (L) Lauraceae Bark (fraction) 156 625 mdash mdash 78 3125 mdash mdash 125Cymbopogon citratus (L) Graminae Leaf (fraction) 250 lt250 625 mdash 625 125 mdash mdash mdashEugenia caryophyllus (L) Myrtaceae Flower buds (F) 625 125 mdash mdash 625 mdash mdash mdash mdashEucalyptus globulus (L) Myrtaceae Fraction (leaf) mdash mdash mdash mdash mdash mdash lt250 mdash lt250Mentha piperita (L) Labiatae Fraction (WP) mdash 625 3125 mdash 3125 mdash lt250 mdash mdashOcimum sanctum (L) Labiatae Leaf (fraction) 625 125 mdash 3125 625 mdash 125 lt250 mdashRosmarinus officinalis (L) Labiatae Rosemary oil 3125 mdash mdash mdash 3125 mdash mdash mdash mdashVetiveria zizanioides (L) Graminae Root (fraction) mdash 125 125 mdash mdash mdash gt250 gt250The bacterial growth inhibitory activity was compared for significant differences within the bacterial strains by broth-dilution method at 250 125 625 3125156 and 78 120583gmL F fractions

exposed to various compounds The cell proliferation wasincreased by the influence of the plant components at thelower concentrations Whereas the oil yielding plant com-pounds showed inhibition of cell proliferation and toxicity athigher doses 250ndash1000120583gmL

4 Discussion

The Western Ghats is considered as one of the richestbiodiversity hotspots in the world [41] In this survey wecollected nearly 78 medicinal plants from Western andEastern Ghats that are edible and popularly used for curingvarious ailments including snakebite Traditional remedieshave a long-standing history in many tribal settlements inTN India and they continue to provide useful and applicabletools for treating ailments [42] The ingredients that makeup the ldquoVishakallurdquo stone which is used as an antidote forsnakebite are different herbs and pebbles available from theriver banks Likewise aqueous paste and decoction obtainedfrom the leaves of A paniculata are widely used for snakebitetreatment by indigenous people [43] Previous studies havereported that ethnomedicine plays major roles in conservingthe disappearing knowledge of tribal communities [44ndash47]The traditional beliefs of reliance on a rich diversity ofethnomedicinal plants located at different settlements havealso been confirmed in another study [48 49] Herein weexplored the various types of traditional practices reported bythe primitive tribal communities with a view to gain furtherknowledge from such studies

In the present investigation potentially rich sources oftribal medicine (71 plants) were scientifically evaluated fortheir antibacterial activity against the MDR bacteria and theaccumulated data was disseminated for the first time to the

scientific community Out of the 71medicinal plants screenedfor the antibacterial activity 10 of them (T involucrataC acida A marmelos A vasica C procera A paniculataand M piperita A indica S indicus and E cardamomum)displayed the highest antibacterial activity against the mul-tidrug resistant B pseudomallei (KHW and TES) and Saureus strains The antibacterial activity of those crude plantextracts was as equally effective as that of the standard drugsOur findings corroborated with the previous reports madeon the antistaphylococcal activity of tribal medicinal plants[34 50 51] On the other hand isolated components from themost active extracts of T involucrata shellsol and C acidaexhibited the most potent action against the antibiotic resis-tant B pseudomallei (KHW) K pneumoniae and S aureusstrainsThese results further confirmed our previous findingson the leaves of T involucrata and its compounds hydrocar-bon ester-like shellsol which displayed a high antibacterialeffect against the different bacterial strains especially thatof S aureus [34] Eugenol and caryophyllene are the activeagents contained in the M piperita [52] and O sanctumplants which are believed to be mainly responsible for theantimicrobial properties of these plants [53] Interestinglythe compounds obtained from the aromatic plants such asC zeylanicum and R officinalis were also found to be veryeffective against the multidrug resistant human pathogen Kpneumoniae S aureus S typhi and B pseudomallei (KHW)that causes melioidosis

The inhibitory potential determined for the shellsol(T involucrata) vaseline (A vasica) C acida and C zey-lanicum indicates that the MIC of 78ndash3125120583gmL foundagainst the B pseudomallei of KHW K pneumoniae Kpneumoniae S pyogenes and V damsel and S pneumoniaewas quite low Similarly lower MIC values were found for

Evidence-Based Complementary and Alternative Medicine 15

M piperita (MIC of 113ndash225 mgmL) against the abovebacterial strains [52] MIC was found for the most activealcohol extracts of A salvifolium (MIC 0034ndash0263mgmL)on S aureus [54] and the MIC for S trilobatum aqueousextracts determined against the tested organisms rangedfrom 006 to 05mgmL [55] C zeylanicum was foundto have an effective antibacterial activity (MIC 64 120583gmL)against P aeruginosa E coli B subtilis and S aureus[56] Previously several investigators have demonstrated thatactive agents exert interesting activity against bacteria evenat lower concentrations tested [57 58] The bacteriostaticmechanism involves damage to the cell walls of bacteriafollowed by inhibition of protein synthesis that ultimatelyleads to bacterial death [59] The most active plants arewidely used by various tribes as traditional treatment (iecut wounds skin infection and scabies) thus indicatingthe potential for further development into promising drugsIn order to ascertain the safety and efficacy of the mostactive compounds their effect on human skin fibroblast cellswas evaluated Chemical constituents of E cardamomumT involucrata S indicus C acida A vasica A marmelosA indica and A paniculata plants failed to produce anynoticeable toxicity up to 1000120583gmL Although some ofthe tested compounds exhibited a slight reduction of cellproliferation and some minor morphological changes suchchanges were insignificant at lower doses and became evidentonly at higher doses However certain aromatic compoundsof O sanctum E globulus V zizanioides C citratus and Eglobulus plants showed reduction of cell proliferation againstHEPK cells

Our phytochemical screening also provides evidenceof the presence of several types of compounds that aremainly responsible for the remarkable antibacterial effectof these plants The differences noted for the bactericidalactivity of various plant extracts in this study appears tobe directly related to the diversity of compounds (shown inparentheses) that are accumulated in the following plants(eg A marmelos and O umbellate) [60ndash62] Compoundslike tannins phenol and polyphenols can bind the Gram-negative bacteria to form a heavy soluble complex on the cellsurface which subsequently disturbs the availability of recep-tor on cells and kills the bacteria [63] Several species havingwide spectra of antimicrobial activity mainly due to theactive constituents such as essential oil phenolic compoundslike thymol carvacrol in oregano and thyme eugenol inclove and cinnamon were also identified previously [64 65]Essential oils degrade the cell wall interact with the cell com-ponents and then disrupt the cytoplasmic membrane [66]The antimicrobial effect of phenolic compounds may involvemultiplemodes of action including damage to themembraneprotein interference with membrane integrated enzymes[67] causing leakage of cellular components coagulation ofcytoplasm depletion of the proton motive force alterationof fatty acid and phospholipid constituents impairmentof enzymatic mechanisms for production and metabolismalteration of nutrient uptake and electron transport [68]influencing the synthesis of DNA and RNA and destroyingprotein translation and the function of the mitochondrionin eukaryotes [69] The mode of action of antimicrobial

agents depends on the type of microorganism and is mainlyrelated to their cell wall structure and the outer membranearrangement Most of the plant spices and herbs containcomplex phenolics (ie phenolic acids flavonoids tanninslignans coumarins quinines) In addition the mechanismsof action of each phenolic compound against various bacteriaare also very complicated [26 70] Further investigation istherefore required to understand the relationship betweenthe antimicrobial action and the chemical structure of everyphenolic compound in the tested extracts The informationavailable from previous pharmacological sources combinedwith the findings herein reported on the medicinal plantextracts may serve as essential data for future drug develop-ment to combat diseases caused by theMDRbacterial strains

Abbreviations

MDR Multidrug resistant bacteriaTES Strain of Burkholderia pseudomalleiMRSA Methicillin-resistant Staphylococcus aureusVRE Vancomycin-resistant enterococciMTB Mycobacterium tuberculosisCH3OH Methanol

CFU Colony forming unitHCl Hydrochloric acidTLC Thin layer chromatographyH2SO4 Sulphuric acids

UV Ultraviolet spectrophotometerHEPK Human skin fibroblasts

Acknowledgments

Theauthors are thankful to the ERI Loyola College ChennaiIndia for his support during the survey The authors alsothank the Department of Microbiology NUHS NUS forthe bacterial cultures used in this investigation The authorsare thankful to the tribal people for sharing their healingknowledge on medicinal aromatic plants

References

[1] M N Alekshun and S B Levy ldquoMolecular mechanisms ofantibacterialmultidrug resistancerdquoCell vol 128 no 6 pp 1037ndash1050 2007

[2] A D Russell ldquoAntibiotic and biocide resistance in bacteriaintroductionrdquo Symposium Series Society for Applied Microbiol-ogy vol 31 no 92 pp 1Sndash3S 2002

[3] D R Guay ldquoTreatment of bacterial skin and skin structureinfectionsrdquo Expert Opinion on Pharmacotherapy vol 4 no 8pp 1259ndash1275 2003

[4] A J Brosnahan and P M Schlievert ldquoGram-positive bac-terial superantigen outside-in signaling causes toxic shocksyndromerdquo FEBS Journal vol 278 no 23 pp 4649ndash4667 2011

[5] K Fellermann J Wehkamp K R Herrlinger and E F StangeldquoCrohnrsquos disease a defensin deficiency syndromerdquo EuropeanJournal of Gastroenterology and Hepatology vol 15 no 6 pp627ndash634 2003

16 Evidence-Based Complementary and Alternative Medicine

[6] L Chen S Lin R Agha-Majzoub L Overbergh C Mathieuand L S Chan ldquoCCL27 is a critical factor for the developmentof atopic dermatitis in the keratin-14 IL-4 transgenic mousemodelrdquo International Immunology vol 18 no 8 pp 1233ndash12422006

[7] J H Song V Thamlikitkul and P R Hsueh ldquoClinical andeconomic burden of community-acquired pneumonia amongstadults in the Asia-Pacific regionrdquo International Journal ofAntimicrobial Agents vol 38 no 2 pp 108ndash117 2011

[8] D A B Dance ldquoMelioidosisrdquo Current Opinion in InfectiousDiseases vol 15 no 2 pp 127ndash132 2002

[9] N J White ldquoMelioidosisrdquo The Lancet vol 361 no 9370 pp1715ndash1722 2003

[10] S Yang ldquoMelioidosis research in Chinardquo Acta Tropica vol 77no 2 pp 157ndash165 2000

[11] P R Hsueh L J Teng L N Lee et al ldquoMelioidosis an emerginginfection in Taiwanrdquo Emerging Infectious Diseases vol 7 no 3pp 428ndash433 2001

[12] C Walsh and S Fanning ldquoAntimicrobial resistance in food-borne pathogensmdasha cause for concernrdquo Current Drug Targetsvol 9 no 9 pp 808ndash815 2008

[13] M Vorachit P Chongtrakool S Arkomsean and S BoonsongldquoAntimicrobial resistance in Burkholderia pseudomalleirdquo ActaTropica vol 74 no 2-3 pp 139ndash144 2000

[14] F M Thibault E Hernandez D R Vidal M Girardet and JD Cavallo ldquoAntibiotic susceptibility of 65 isolates of Burkholde-ria pseudomallei and Burkholderia mallei to 35 antimicrobialagentsrdquo Journal of Antimicrobial Chemotherapy vol 54 no 6pp 1134ndash1138 2004

[15] R L S Sikarwar ldquoEthnogynaecological uses of plants new toIndiardquo Ethnobotany vol 12 no 1-2 pp 112ndash115 2002

[16] R Siva ldquoStatus of natural dyes and dye-yielding plants in IndiardquoCurrent Science vol 92 no 7 pp 916ndash925 2007

[17] P Kumar Singh V Kumar R K Tiwari A Sharma ChV Rao and R H Singh ldquoMedico-ethnobotany of lsquochatararsquoblock of district sonebhadra Uttar Pradesh Indiardquo Advances inBiological Research vol 4 no 1 pp 65ndash80 2010

[18] W H Lewis and M P Elwin-Lewis Medical Botany PlantsAffecting Human Health John Wiley amp Sons New York NYUSA 2003

[19] R Perumal Samy P N Pushparaj and P GopalakrishnakoneldquoA compilation of bioactive compounds fromAyurvedardquo Bioin-formation vol 3 no 3 pp 100ndash110 2008

[20] WorldHealthOrganization (WHO)TraditionalMedicine Strat-egy 2002ndash2005 Genera World Health Organization GenevaSwitzerland 2002

[21] P S Haddad G A Azar S Groom and M Boivin ldquoNaturalhealth products modulation of immune function and preven-tion of chronic diseasesrdquo Evidence Based Complementary andAlternative Medicine vol 2 no 4 pp 513ndash520 2005

[22] K Poonam andG S Singh ldquoEthnobotanical study of medicinalplants used by the Taungya community in Terai Arc LandscapeIndiardquo Journal of Ethnopharmacology vol 123 no 1 pp 167ndash1762009

[23] G B Mahady ldquoMedicinal plants for the prevention and treat-ment of bacterial infectionsrdquo Current Pharmaceutical Designvol 11 no 19 pp 2405ndash2427 2005

[24] V P Kumar N S Chauhan H Padh and M Rajani ldquoSearchfor antibacterial and antifungal agents from selected Indianmedicinal plantsrdquo Journal of Ethnopharmacology vol 107 no2 pp 182ndash188 2006

[25] R Nair T Kalariya and S Chanda ldquoAntibacterial activity ofsome plant extracts used in folk medicinerdquo Journal of HerbalPharmacotherapy vol 7 no 3-4 pp 191ndash201 2007

[26] D Kalemba and A Kunicka ldquoAntibacterial and antifungalproperties of essential oilsrdquo Current Medicinal Chemistry vol10 no 10 pp 813ndash829 2003

[27] A H Ashour ldquoAntibacterial antifungal and anticancer activi-ties of volatile oils and extracts from stems leaves and flowers ofEucalyptus sideroxylon and Eucalyptus torquatardquoCancer BiologyandTherapy vol 7 no 3 pp 399ndash403 2008

[28] M J Abad L M Bedoya L Apaza and P Bermejo ldquoTheArtemisia L genus a review of bioactive essential oilsrdquoMolecules vol 17 no 3 pp 2542ndash2566 2012

[29] H N Marzoug M Romdhane A Lebrihi et al ldquoEucalyptusoleosa essential oils chemical composition and antimicrobialand antioxidant activities of the oils from different plant parts(stems leaves flowers and fruits)rdquoMolecules vol 16 no 2 pp1695ndash1709 2011

[30] C Proestos D Sereli and M Komaitis ldquoDetermination ofphenolic compounds in aromatic plants by RP-HPLC and GC-MSrdquo Food Chemistry vol 95 no 1 pp 44ndash52 2006

[31] S Chao G Young C Oberg and K Nakaoka ldquoInhibition ofmethicillin-resistant Staphylococcus aureus (MRSA) by essentialoilsrdquo Flavour and Fragrance Journal vol 23 no 6 pp 444ndash4492008

[32] M Akin D Oguz and H T Saracoglu ldquoAntibacterial effectsof some plant extracts from Labiatae (Lamiaceae) growingnaturally around Sirnak-Silopi Turkeyrdquo International Journal ofPharmaceutical and Applied Sciences vol 1 no 1 pp 4ndash47 2010

[33] K Bayoub T Baibai D Mountassif A Retmane and ASoukri ldquoAntibacterial activities of the crude ethanol extracts ofmedicinal plants against Listeriamonocytogenes and some otherpathogenic strainsrdquo African Journal of Biotechnology vol 9 no27 pp 4251ndash4258 2010

[34] R P Samy P Gopalakrishnakone M Sarumathi P Houghtonand S Ignacimuthu ldquoPurification of antibacterial agents fromTragia involucratamdasha popular tribal medicine for wound heal-ingrdquo Journal of Ethnopharmacology vol 107 no 1 pp 99ndash1062006

[35] K M Matthew The Flora of Tamil Nadu Carnatic Part IndashIIIprinted at the Diocesan Press Madras India 1981ndash1983

[36] AW BauerWMKirby J C Sherris andM Turck ldquoAntibioticsusceptibility testing by a standardized single disk methodrdquoAmerican Journal of Clinical Pathology vol 45 no 4 pp 493ndash496 1966

[37] National Committee for Clinical Laboratory Standards Per-formance Standards For Antimicrobial Susceptibility Testing12th Informational Supplement Approved standard M100-S12National Committee for Clinical Laboratory Standards WaynePa USA 2002

[38] E A Elsinghorst ldquoMeasurement of invasion by gentamicinresistancerdquoMethods in Enzymology vol 236 pp 405ndash420 1994

[39] J B Harborne Phytochemical Methods Chapman and HallNew York NY USA 1976

[40] H Wagner S Bladt and E M Zgairski Plant Drug AnalysisSpringer New York NY USA 1984

[41] N Myers R A Mittermeler C G Mittermeler G A B daFonseca and J Kent ldquoBiodiversity hotspots for conservationprioritiesrdquo Nature vol 403 no 6772 pp 853ndash858 2000

[42] R Perumal Samy and S Ignacimuthu ldquoAntibacterial activity ofsome folkloremedicinal plants used by tribals inWestern Ghats

Evidence-Based Complementary and Alternative Medicine 17

of Indiardquo Journal of Ethnopharmacology vol 69 no 1 pp 63ndash712000

[43] K Nampoothiri A Chrispal A Begum S Jasmine K GGopinath and A Zachariah ldquoA clinical study of renal tubulardysfunction inCleistanthus collinus (Oduvanthalai) poisoningrdquoClinical Toxicology vol 48 no 3 pp 193ndash197 2010

[44] M Strathern UsefulKnowledge The Isiah Berlin Lecture Uni-versity of Manchester Brunswick UK

[45] C N Sunil and A K Pradeep ldquo(2001) Another new species ofTripogon (Poaceae) from Indiardquo Sida vol 19 pp 803ndash806 2005

[46] S Biber-Klemm and T Cottier Right To Plant GeneticResources and Traditional Knowledge Basic Issues and Prospec-tive Oxfordshire Nosworthy Way UK 2006

[47] V S Ramachandran S Shijo Joseph and R Aruna ldquoEth-nobotanical studies from amaravathy range of Indira GandhiWildlife SanctuaryWesternGhats CoimbatoreDistrict South-ern Indiardquo Ethnobotanical Leaflets vol 13 pp 1069ndash1087 2009

[48] A H Rajasab and I Mohamad ldquoDocumentation of folkknowledge on edible wild plants of North Karnatakardquo IndianJournal of Traditional Knowledge vol 3 no 4 pp 419ndash4292004

[49] M Rajadurai V G Vidhya M Ramya and A Bhaskar ldquoEthno-medicinal plants used by the traditional healers of pachamalaihills Tamilnadu Indiardquo Studies on Ethno-Medicine vol 3 no 1pp 39ndash41 2009

[50] S K Adesina O Idowu A O Ogundaini et al ldquoAntimicrobialconstituents of the leaves of Acalypha wilkesiana and Acalyphahispidardquo Phytotherapy Research vol 14 no 5 pp 371ndash374 2000

[51] M T Gutierrez-Lugo M P Singh W M Maiese and BN Timmermann ldquoNew antimicrobial cycloartane triterpenesfrom Acalypha communisrdquo Journal of Natural Products vol 65no 6 pp 872ndash875 2002

[52] A K Tyagi andAMalik ldquoAntimicrobial potential and chemicalcomposition of Mentha piperita oil in liquid and vapour phaseagainst food spoiling microorganismsrdquo Food Control vol 22no 11 pp 1707ndash1714 2011

[53] G Devendran and U Balasubramanian ldquoQualitative phyto-chemical screening and GC-MS analysis of Ocimum sanctumL leavesrdquoAsian Journal of Plant Science and Research vol 1 no4 pp 44ndash48 2011

[54] V C Jain N M Patel D P Shah P K Patel and B HJoshi ldquoAntioxidant and antimicrobial activities of Alangiumsalvifolium (Lf) Wang rootrdquo Global Journal of Pharmacologyvol 4 no 1 pp 13ndash18 2010

[55] P Swapna Latha andK Kannabiran ldquoAntimicrobial activity andphytochemicals of Solanum trilobatum LinnrdquoAfrican Journal ofBiotechnology vol 5 no 23 pp 2402ndash2404 2006

[56] M Usha1 S Ragini and S M A Naqvi ldquoAntibacterial activityof acetone and ethanol extracts of Cinnamon (Cinnamomumzeylanicum) and Ajowan (Trachyspermum ammi) on four foodspoilage bacteriardquo International Research Journal of BiologicalSciences vol 1 no 4 pp 7ndash11 2012

[57] S Prabuseenivasan M Jayakumar and S Ignacimuthu ldquoInvitro antibacterial activity of some plant essential oilsrdquo BMCComplementary and Alternative Medicine vol 6 article 39 pp1ndash8 2006

[58] G A Ayoola F M Lawore T Adelowotan et al ldquoChemi-cal analysis and antimicrobial activity of the essential oil ofSyzigium aromaticum (clove)rdquo African Journal of Biotechnologyvol 7 no 13 pp 2227ndash2231 2008

[59] Y J Fu L Chen Y Zu et al ldquoThe antibacterial activity of cloveessential oil against Propionibacterium acnes and its mechanismof actionrdquo Archives of Dermatology vol 145 no 1 pp 86ndash882009

[60] D Venkatesan C M Karrunakarn S Selva Kumar and P TPalani Swamy ldquoIdentification of phytochemical constituents ofAegle marmelos responsible for antimicrobial activity againstselected pathogenic organismsrdquo Ethnobotanical Leaflets vol 13pp 1362ndash1372 2009

[61] P Arun K G Purushotham J Johnsy jayarani and V KumarildquoIn vitro Antibacterial Activity of Oldenlandia umbellata anIndian medicinal Plantrdquo Journal of Pharmaceutical Science andTechnology vol 2 no 4 pp 198ndash201 2010

[62] A Manjamalai R Sardar Sathyajith Singh C Guruvayoorap-pan and V M Berlin Grace ldquoAnalysis of phytochemicalconstituents and antimicrobial activity of some medicinalplants in Taminadu Indiardquo Global Journal of Biotechnology andBiochemistry vol 5 no 2 pp 120ndash128 2010

[63] E Haslam ldquoNatural polyphenols (vegetable tannins) as drugsPossible modes of actionrdquo Journal of Natural Products vol 59no 2 pp 205ndash215 1996

[64] Y Cai Q Luo M Sun and H Corke ldquoAntioxidant activityand phenolic compounds of 112 traditional Chinese medicinalplants associated with anticancerrdquo Life Sciences vol 74 no 17pp 2157ndash2184 2004

[65] B Shan Y Z Cai M Sun and H Corke ldquoAntioxidant capacityof 26 spice extracts and characterization of their phenolicconstituentsrdquo Journal of Agricultural and Food Chemistry vol53 no 20 pp 7749ndash7759 2005

[66] R J W Lambert P N Skandamis P J Coote and G-J ENychas ldquoA study of the minimum inhibitory concentration andmode of action of oregano essential oil thymol and carvacrolrdquoJournal of AppliedMicrobiology vol 91 no 3 pp 453ndash462 2001

[67] M Raccach ldquoThe antimicrobial activity of phenolic antioxi-dants in food a reviewrdquo Journal of Food Safety vol 6 no 3 pp141ndash170 1984

[68] M Taniguchi Y Yano E Tada et al ldquoMode of action of polygo-dial an antifungal sesquiterpene dialdehyderdquo Agricultural andBiological Chemistry vol 52 no 6 pp 1409ndash1414 1988

[69] G J E Nychas ldquoNatural antimicrobials from plantsrdquo in NewMethods of Food Preservation G W Gould Ed pp 58ndash89Blackie Academic London UK 1995

[70] S Burt ldquoEssential oils their antibacterial properties and poten-tial applications in foodsmdasha reviewrdquo International Journal ofFood Microbiology vol 94 no 3 pp 223ndash253 2004

Submit your manuscripts athttpwwwhindawicom

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Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

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Behavioural Neurology

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Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Disease Markers

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BioMed Research International

OncologyJournal of

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Oxidative Medicine and Cellular Longevity

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PPAR Research

The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014

Immunology ResearchHindawi Publishing Corporationhttpwwwhindawicom Volume 2014

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Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Parkinsonrsquos Disease

Evidence-Based Complementary and Alternative Medicine

Volume 2014Hindawi Publishing Corporationhttpwwwhindawicom

2 Evidence-Based Complementary and Alternative Medicine

resistant to many antibiotics [13 14] Considering the highercost for producing synthetic drugs and the various sideeffects associated with their use the need to search foralternative agents from medicinal plants and essential oilsused in folklore medicine is further justified to overcomethese issues In India there are about 550 tribal communitiescovered under 227 ethnic groups residing in about 5000villages throughout different forest and vegetation regions[15] India is one of the worldrsquos 12 megabiodiversity countries[16 17] Plant derived medicines have played a major rolein human societies throughout the history and prehistory ofmankind [18]The traditional healers (traditional physicians)or medicinemen have a long history of their own diagnosticand treatment system which they have acquired from theirancestors [19] Approximately 80 of the world populationstill relies on traditional medicine for the treatment of com-mon diseases [20ndash22] Medicinal plants thus offer significantpotential for the development of novel antibacterial therapiesand adjunct treatments [23] Plant derived drugs serve as aprototype to develop more effective and less toxic medicinesIn previous studies few attempts were made to confirm theantimicrobial activity of some indigenous medicinal plants[24 25] Not only extracts of variousmedicinal plants but alsoessential oils and their constituents have been investigatedfor their antimicrobial properties against bacteria and fungi[26ndash28]

The principal compounds from the leaves showed a betterantibacterial activity against P aeruginosa and B subtilisbacteria and a significant antifungal activity on C albicans[29] The essential oil from R officinalis (alpha pineneverbenonebornyl acetate) was found to be more sensitiveto the Gram-positive bacteria (MIC 25ndash4mgmL) thanto the Gram-negative bacteria [30] Several essential oilskill bacteria by damaging the cell membrane structure andinhibiting their membrane function [31] Because of theantimicrobial potency of plant extracts and oils they becomea rich source of raw materials for many biotechnological andpharmaceutical industries for the development of therapeuticdrugs The increasing trend in the use of aromatic plantsand essential oils in food cosmetic and pharmaceuticalindustries suggests that a systematic study of traditionalmedicinal plants is very important in order to find activecompounds from such sources [32ndash34] The purpose ofthis study is to survey and investigate popular medicinalaromatic plants and their essential oils with a view to fightagainst multidrug resistant human pathogens In the presentstudy 71 plant species were selected on the basis of theavailable medicinal information and screened for their invitro antimicrobial efficacy against bacteria

2 Materials and Methods

21 Ethnomedicinal Survey and Collection of Plants Eth-nomedicinal surveys were conducted duringMarch 1998 andJuly 2001 from various tribal localities (Kolli hills Kalrayanhills Pachamalai Javadi hills Mundanthurai) of Eastern andWestern Ghats Tamil Nadu India For ethnobotanical stud-ies questioners were used to collect the general informationon the tribes and the key information on medicinal details

was collected through interviews The medicinal plants wereidentified by a taxonomist using the standard Flora ofTamil Nadu Carnatic [35] and the voucher specimens weredeposited in the departmentrsquos herbarium at the EntomologyResearch Institute Loyola College Chennai India

22 Preparation of Plant Extracts Using a Soxhlet apparatusthe shade-dried and powdered plant materials (200 g of each)were extracted with 1000mL of methanol (CH

3OH) for 10 h

The collected methanol extracts were filtered (Whatman no1 filter paper) and evaporated with a rotary evaporator andfreeze dryer (lyophilized) to obtain the crude extracts (BuchiLabortechnik AG Switzerland) The dried crude extractswere stored at 4∘C for antimicrobial assays [34]

23 Culture of Microorganisms The following Gram-nega-tive Burkholderia pseudomallei (TES21) Burkholderia pseu-domallei (KHW22) Klebsiella pneumoniae (ATCC15380)Klebsiella pneumoniae Pseudomonas aeruginosa (ATCC2-7853)Vibrio damsela and Salmonella typhi (ATCC51812) andGram-positive Staphylococcus aureus (ATCC 29213) Strepto-coccus pyogenes and Streptococcus pneumoniae (ATCC49619)microorganisms were used for cultures B pseudomalleibacterial strains such as KHW and TES were isolated fromthe patient samples obtained from the Department of Micro-biology NUS Singapore The strains were subcultured on20mLTryptic Soy (TS) andMueller Hinton (MH) agar plates(pH 74) and incubated overnight at 37∘C before use

24 Antimicrobial Activity The standard bacterial cultureswere stored at minus70∘C subcultured on 20mL MH and TSagar plates (pH 74) and incubated overnight at 37∘C priorto use The antimicrobial property was tested using the disc-diffusion method [36] Five young colonies of each strain ofbacteria taken from their respective cultures grown overnighton TS agar plates (Oxoid limited Wode Road BasingstokeHants England UK) were suspended in 5mL of sterile saline(09) and the density of the suspension was adjusted toapproximately 3 times 108 colony forming unit (CFU) The swabwas used to inoculate the dried surface of TS agar plateby streaking four times over the surface of the agar androtating the plate approximately 90∘C to ensure an evendistribution of the inoculums The medium was allowed todry for about 3min before adding a 6 millimeter in diameter(mm) sterile paper disc (Becton Dickinson USA) on thesurface Each disc was tapped gently down onto the agar toprovide a uniform contact Lyophilized residue (100120583gmL)of each plant extracts and purified fractions was weighedand dissolved in 1mL of water and 20120583L of the extractsand oils (containing 100120583g of residue) were applied on eachdisc (3 replicates) while the sterile blank disc served asa normal control The antimicrobial effect of the extractson the clinical isolates was determined in comparison withthe reference antibiotics (chloramphenicol 30120583gdisc andceftazidime 30 120583gdisc) which were used as positive controlsThe plates were incubated at 37∘C for 24 h and the inhibitionzones were measured and calculated

Evidence-Based Complementary and Alternative Medicine 3

25Minimum Inhibitory Concentrations (MICs) Assay MICswere evaluated based on the in vitro screening of 16 purifiedfractions that were found to have potent antimicrobial activ-ity Broth dilution method was used for the MIC assay withsome slight modifications as recommend by the NCCLS [37]Two-fold serial dilutions of all the fractionated compoundswere made with MH and TS broth in microtiter plate wellsto adjust the final concentration from 78 to 250120583gmL whilewells containing the broth alonewithout any sample served asa controlThree replicates (119899 = 3) were used for each dilutionand culture containing approximately 1 times 105 CFUmL Theplateswere incubated at 37∘C for 24 h and the absorbancewasmeasured at 560 nm

26 Cytotoxicity Assay The cytotoxic effects of variousextracts were tested by MTT assay [38] using human skinfibroblast HEPK cells The toxic effect of plant extracts andessential oils was assayed on human skin fibroblast (HEPK)cell proliferation in 96-well microtitre plates Confluent cells(5 times 106 cells per well) were incubated with extracts and oilsfor 24 h and the percentage inhibitory concentration (IC

50)

was determined

27 Phytochemical Analysis of Plant Extracts Themost activeextracts were used for purification of antimicrobial com-pounds [39] 100 g each of the plant powder was percolatedwith 500mL of 4 aqueous HCl (adjusted to pH 2) andheated at 50∘C for 3 hThe extract waswashedwith 2times250mLof diethyl ether and the organic phase evaporated to drynessusing vacuum rotary evaporator The dark brown gummyresidues obtained by acid hydrolysis were chromatographedon silica gel column 60 times 32 cm (60ndash120 mesh pH 7 Merck)by gradually eluting with n-hexaneethyl acetate (8 2 6 43 7 and 1 1) and chloroformmethanol (3 2) The aliquotsof each fraction were subjected to thin layer chromatography(TLC) on silica gel coated TLC plate (1mm Merck) usingthe solvent system consisting of 20 (vv) n-hexaneethylacetateThe chromatograms were detected using 50H

2SO4

solution as a spray reagent [34]The individual fractions werecollected and concentrated by vacuum rotary evaporator at40∘C All the purified compounds recovered from the silicagel column were monitored by reading the absorbance at190ndash350 nm (UV spectrophotometer Hitachi Japan) Theactive fractions were further purified and the final yieldsof the compounds were recorded The lyophilized pooledconcentrated compounds were then assayed (100120583gmL)against bacteria The phytochemical screening was done onthe pure compounds using the chemical method previouslyreported for the detection of secondary metabolites [40]The different chemical constituents tested include alkaloidsflavonoids glycosides polyphenols saponin sterols triter-penes tannins reducing sugars gallic acid catechol andaglycones

28 Statistical Analysis The bacterial growth inhibitoryactivity (inhibition zones millimeter in diameter) was com-pared for significant differences within the bacterial strainsOne way analysis of variance was performed (mean plusmn SD

119899 = 3 replicates) using GraphPad Prism 4 USA lowast119875 lt 001was considered statistically significant (inhibition zones ofextractsfractions versus antibiotic drugs)

3 Results

31 Study Area for the Collection of AromaticMedicinal PlantsTheWestern and Eastern Ghats were selected for the presentstudy with the cite map showing the landmarks (Figures 1(a)-1(b)) Kalrayan hills are situated north of Attur taluk (Salemdistrict) one of the major range of hills in the Eastern Ghatsof Tamil Nadu (Figure 1(c)) Pachamalai hills are situated tothe north of Thuraiyur taluk of Tiruchirappalli district Therich biodiversity part of Eastern Ghats lies between latitudes11∘091015840 0010158401015840 to 11∘ 271015840 0010158401015840 N and longitudes 78∘281015840 0010158401015840 to78∘ 491015840 0010158401015840 E and occupies an area of about 52761 squarekm It is located near 11∘ 111015840N 78∘21E1118∘N 7835∘E11187835 (Figure 1(d)) Mundanthurai is located nearly 45 kmwest of Tirunelveli district TN between latitude 8∘ 251015840 and8∘ 531015840 N and longitude 77∘ 101015840 and 77∘ 351015840 E This is the onlyarea of Western Ghats that has the longest raining period ofabout 8 months and forms the catchment area for 14 riversand streams (Figure 1(e)) Kolli Malai is a small mountainrange located in Namakkal district The mountains are about1000ndash1300m in height and cover an area of approximately280 km The Kolli hills are part of the Eastern Ghats whichis a mountain range that runs mostly parallel to the east coastof Tamil Nadu in South India (Figure 1(f)) Javadi hills areone of the largest in the Eastern Ghats in Vellore district inthe northern part of the state of Tamil Nadu They consistof bluish gray hills with peaks averaging 3600ndash3800 feetor 1100ndash1150 meter (Figure 1(g)) Based on the vegetationtype (Figures 2(a)ndash2(d)) the study area consists of (i) drydeciduous (ii) moist deciduous and (iii) rain forests anddiverse proportion of plant parts in abundance (Figures 2(e)-2(f)) Three different types of tribes (ie Kani Malayali andPaliyan tribes) inhabit in the hill ranges The Kani tribeslocated at Mundanthurai raise different types of vegetablesin their own fields while the Malayali tribes cultivate riceThey all engage not only in the agricultural work but alsoare involved in silvicultural work assigned by the forestdepartment Government of TN India

32 Medicinal Plants Glory Western Ghats (Mundanthurai)and Eastern Ghats (Kolli hills Javadi hills Kalrayan hillsPachamalai hills) possess a rich diversity of medicinal plantsthat are used as food and drug by different groups oftribal communities Urbanization habitat degradation andfragmentation of these forests have resulted in the depletionof natural resources on which these tribes used to dependfor their livelihoods It has become increasingly difficult forthem to live in their traditional way In addition the impactof modernization and urbanization has encroached in andaround tribal settlements thus changing their lifestyles

33 Plants Valued as Edibles Various types of plant parts arecollected during different seasons cooked and eaten alongwith boiled rice (Table 1) For example Solanum nigrum leafis most commonly used in all the four regions There are

4 Evidence-Based Complementary and Alternative Medicine

Figure 1 (a) The site for collection of medicinal plants in Western and Eastern Ghats of Tamil Nadu (b) The landmark (map) of traditionalmedicine distribution and collection of different types of plants (c) District map showing the collection site of plants from Kalrayan hills(Salem) (d) Pachamalai hills (Thiruchirappalli) and (e) Mundanthurai (Tirunelveli) rich biodiversity hot-spot of the Western Ghats (f)Kolli hills (Namakkal) (g) Javadi hills (Vellore) part of the Eastern Ghats which is a mountain range that runs mostly parallel to the eastcoast of South India

a large number of wild edible fruits including yielding plantssuch as Citrus acida Ficus benghalensis Ficus microcarpaFicus racemosa Phyllanthus emblica Solanum trilobatumand Syzygium cumini are popularly used by the tribes

34 Plants Used for Snakebite Treatment Thirty-four plantsused for snakebite treatment are documented (Table 1)Snakebite is amajor health hazard that leads to highmortalityin tribal settlements The majority of the antidotes are

prepared freshly from plant materials frequently collectedfrom the leaves of A paniculata A echioides Aristolochiaindica E alba E prostrata M pudica O sanctum Tinvolucrata and Cleistanthus collinus (Oduvanthalai) thewhole plants ofAchyranthes aspera andWedelia calendulaceathe stem-barks and nuts of Strychnos nux-vomica the rootsof Hemidesmus indicus Tephrosia purpurea Rauwolfia ser-pentina C roseus and so forth and the tubers of Gloriosasuperba The tuber paste is usually applied externally on

Evidence-Based Complementary and Alternative Medicine 5

Table 1 Some of the important traditional medicinal plant species families voucher specimens parts used yield of extracts phytochemicalscreening and toxicity on human macrophage cells

Scientific name Family Voucher specimen Plant parts Yield (gm) Phytochemicalanalysis

Adhatoda vasica Nees Acanthaceae D2020 Leaf 64 VasicineAegle marmelos (L)Correa ex Roxb Rutaceae D2018 Root-bark 58 Alkaloids

Alangium salvifolium (L)f Wangerin Alangiaceae 0140 Leaf 63 Phenolic

Andrographis paniculataWallich ex Nees Acanthaceae 0061 Leaf 68 Andrographolide

Andrographis echioidesNees Acanthaceae 0116 Leaf 70 Terpenoids

Acalypha indica L Euphorbiaceae 29644 Leaf 61 AcalypheAcalypha lanceolata L Euphorbiaceae 15791 Leaf 71 AlkaloidsAchyranthes aspera L Amaranthaceae 2666 Leaf 27 BetaineAgeratum conyzoides L Asteraceae 4812 Leaf 37 Essential oilsAsteracantha longifolia L Acanthaceae 0234 Stem 74 GlycosidesAzadirachta indica A Juss Meliaceae D0204 Leaf bark 67 Tannins

Borassus flabellifer L Arecaceae D0202 Root 08 Flavonoidsphenolics

Boerhavia erecta L Nyctaginaceae 10897 Whole plant 28 PhenolicsCalotropis procera (Ait)Ait f Asclepiadaceae D073 Root-bark 13 Terpenoids

Calotropis gigantea (L)RBrex Ait Asclepiadaceae D070 Milky latex 48 Alkaloids

Cassia auriculata L Caesalpiniaceae 0141 Leaf 49 Saponins

Cassia occidentalis L Caesalpiniaceae 0111 Root 95 Flavonoidssaponins

Cassia tora L Caesalpiniaceae 0100 Stem bark 69 SaponinsCassia fistula L Caesalpiniaceae 037 Whole plant 79 SaponinsCardiospermumhalicacabum L Sapindaceae 0125 Whole plant 58 Flavonoids

Catharanthus roseus (L)GDon Apocynaceae 0029 Leaf

root1407 Alkaloids

Cinnamomum zeylanicumGarcin ex Blume Lauraceae 00209 Bark 31 Essential oil

TanninCinnamomum inersReinw ex Blume Lauraceae 043-c Leaf bark Alkaloids

Cissus quadrangularisRoxb Vitaceae D02023 Leaf 68 Glycosides

Citrus acida Roxb Hookf Rutaceae 0213 Leaf 52 SaponinsTerpenoids

Centella asiatica (L) Umbelliferae 0138 Whole plant 89 FlavonoidsAlkaloids

Clerodendrum inerme (L)Gaertn Verbenaceae D02043 Stem 78 Sterols diterpenes

Clitoria ternatea L Papilionaceae D02026 Seed 98 ProteinCleistanthus collinus(Roxb) Benth andHookf

Euphorbiaceae 0011 Whole plant 003 Cleistanthincollinusin

Cleome gynandropsis L Capparidaceae 12247 Leaf 62 GlycosidesCleome viscose L Capparidaceae 29999 Leaf 27 Phenolics

6 Evidence-Based Complementary and Alternative Medicine

Table 1 Continued

Scientific name Family Voucher specimen Plant parts Yield (gm) Phytochemicalanalysis

Coccinia grandisWampA Cucurbitaceae D02030 Leaf root 09 GlycosidesCymbopogon citratus(DC) Gramineae D012 Root 025 Essential oil

Datura metel L Solanaceae D02038 Leaf stem 39 SteroidsEucalyptus globulus Labill Myrtaceae D0220 Leaf 12 TerpenoidsEclipta alba (L) Hassk Asteraceae D028 Whole plant 07 PhenolicEuphorbia hirta Linn Euphorbiaceae 0018-c Whole plant 012 mdash

Eclipta prostrata (L) Asteraceae D210 Leaf 110 Triterpenoidsaponin

Eugenia caryophyllus(Sprengel) Bullock ampHarrison

Myrtaceae 0025 Flower buds 116 Essential oils

Elettaria cardamomumWhite et Mason Zingiberaceae 0009 Fruit pods 317 Essential oils

Gloriosa superba L Liliaceae 020-S Tuber 108 Alkaloids phenol

Jatropha curcas L Euphorbiaceae 015 Whole plant 53 Alkaloidsflavonoids

Hyptis suaveolens (L) Poit Lamiaceae 24688 Leaf 63 Essential oilHemidesmus indicus L Asclepiadaceae D-009 Roots CoumarinsIchnocarpus frutescens(L) RBr Apocynaceae 0110 Root flower 73 Terpenoids

Leucas aspera (Willd)Link Labiatae 0114 Leaf 83 Triterpenes

Lawsonia inermis L Lythraceae T261 Leaf 09 Glycosidesphenolic

Madhuca longifolia (L) JFMacbr Sapotaceae D01415 Nut 93 Sitosterol

Merremia hastate L(Desr) Hallierf Convolvulaceae 10894 Whole plant 40 Alkaloids

Mentha piperita L Lamiaceae 0217-c Whole plant 07 Essential oilsMorinda tinctoria Roxb Rubiaceae 0122 Leaf 14 GlycosidesMimosa pudica L Mimosaceae 0071 Whole plant 06 mdashOldenlandia umbellata L Rubiaceae D02047 Leaf 44 AlkaloidsOcimum sanctum L Lamiaceae 0016 Whole plant 30 AlkaloidsPiper attenuatum BuchHamex Miq Piperaceae 007 Flower 46 Alkaloids

Plumbago zeylanica (L)Cav Plumbaginaceae 0121 Root 48 Plumbagin

Plectranthus amboinicus(L) Spreng Lamiaceae 0410 Whole plant 12 Essential oils

terpenoidsPhyllanthus debilis L(Klein ex Willd) Euphorbiaceae 0120 Whole plant 49 Polyphenol

Phyllanthusmadraspatensis L Euphorbiaceae 0117 Whole plant 50 Polyphenol

Premna tomentosaWilld Verbenaceae 0129 Leaf 53 DiterpenesRosmarinus officinalis L Lamiaceae 0017 Root 023 Essential oilsRauwolfia serpentine L Apocynaceae 020-S Root 115 AlkaloidSebastiania chamaelea (L)Muell Arg Euphorbiaceae 0034 Leaf 11 Polyphenol

Solanum trilobatum L Solanaceae D02054 Leaf flower 40 Tannins

Evidence-Based Complementary and Alternative Medicine 7

Table 1 Continued

Scientific name Family Voucher specimen Plant parts Yield (gm) Phytochemicalanalysis

Sphaeranthus indicus L Asteraceae D02060 Whole plant 10 Essential oilSwertia chirata (L) Ham Gentianaceae D0540 Whole plant 16 GlycodisesStrychnos nux-vomica L Loganiaceae S-22 Nuts 036 AlkaloidsTragia involucrata L Euphorbiaceae D068 Leaves 16 ShellsolTinospora cordifolia(Willd) Miers ex Hooff ampThoms

Menispermaceae 0118 Leafroot stem 50 Glycosides tannins

Tridax procumbens L Compositae 10649 Leaf 18 FlavonoidsTerminalia arjuna (DC)W amp A Combretaceae 033-c Bark 80 Phenolics

Tephrosia purpurea (L)Pers Fabaceae S-43 Whole plant 08 Isoflavone

Vitex negundo L Verbenaceae 0031 Leaf 24 TerpineolVetiveria zizanioides L Gramineae 0051 Root 103 Essential oilWithania somnifera (L)Dunal Solanaceae D02063 Root 21 Alkaloids

Wedelia calendulacea Less Asteraceae S-24 Leaves FlavonoidsZingiber officinale Rosc Zingiberaceae 0327 Rhizome 23 TanninsZanthoxylum limonella(Dennst) Alston Rutaceae 009 Bark 19 Alkaloids essential

oilClass of chemical compounds A alkaloids S saponins T tannins St steroids G glycosides T terpenoids P polyphenol P phenolics Sh shellsol Hhydrocarbon esters

the site of snakebite and decoction is given orally fortreatment by indigenous people Besides these tribes relyon the medicinal plants as ingredients for fabricating a kindof medicated stone for health management ldquoVishakallurdquo(poison stone) is used by the indigenous groups called Kaniin Kerala India to treat a snakebiteWhen the stone is placeddirectly on the bitten area it sticks to the body to absorb thepoison and then become detached when absorption seems tobe complete The ingredients of Vishakallu stones are madewith leaves of Ocimum sanctum Anisomeles malabaricaLeucas aspera Piper betle Santalum album and the pebblescollected from the river bank

35 Survey of Medicinal Plants and Their Health Care ValuesThe present study is an attempt to provide scientific basis andobtain justification for the traditional beliefs of reliance on arich diversity of ethnomedicinal plants along with the richheritage of traditional medicine practices related to healthcare system made available by the primitive tribal commu-nities located at different settlements The native traditionalpractitioners called ldquovaidyarsrdquo have a good knowledge aboutthe traditional plants locally available for treatment of var-ious diseases (Figures 3(a)ndash3(p)) Such traditional medicalknowledge is used for preparing home remedies ill healthprevention and routine health maintenanceThis knowledgeis also applicable to cover other sectors of social life Duringthe ethnobotanical survey the wealth of 78 medicinal plantspecies used by the indigenous tribal community for varioustypes of health treatment was documented The botanical

names family names parts used chemical constituents andtheir application are provided (Table 1)

36 Antimicrobial Activity of Crude Extracts In this studywe reported the antimicrobial screening of methanolic crudeextracts of 78 medicinal plants (Table 2) Results revealedthat 68 plant extracts displayed potent activities against oneor more Gram-positive and -negative bacteria Of whichTragia involucrata Citrus acida Aegle marmelos Adha-toda vasica Calotropis procera Andrographis paniculata andMentha piperita Azadirachta indica Sphaeranthus indicusand Elettaria cardamomum showed the highest antibacte-rial activity against the multidrug resistant B pseudomallei(KHW and TES) and S aureus at 100120583gmL concentrationThe extracts showed pronounced antibacterial activity withtheir inhibitory zones ranging from 20 to 31mm in diam-eter as compared to the standard drugs chloramphenicoland ceftazidime (29ndash33mm) The majority of the plantsdemonstrated a powerful antimicrobial potency against themultidrug resistant strains of B pseudomallei (KHW andTES) K pneumonia and S aureus Approximately twenty-one plant extracts exerted only a weak or moderate effectagainst the tested bacteria while the crude extract of 13plants failed to show any effect at all Except for the plantextracts of T involucrata A lanceolata A vasica and Sindicus extracts the majority of the plant extracts wereineffective against theV damsela infection fascinatingly only11 plants exhibited activity against P aeruginosa of whichS indicus M piperita and C procera were found to havevery strong inhibition of bacteria at the tested concentrations

8 Evidence-Based Complementary and Alternative Medicine

Table 2 Antimicrobial activity of methanol extract of aromatic medicinal plants and essential oils evaluated against multidrug resistant(MDR) human pathogens at 100120583gmL concentration

Scientific name Microorganisms growth inhibition zones (6 millimeter in diameters)KHW TES Kp Kpr Pa Sa Stp Sp Vd Vd

Adhatoda vasica Nees 28 19 22 18 12 mdash 10 17 16 mdashAegle marmelos (L) Correa ex Roxb 29 17 9 10 mdash 15 mdash mdash mdash mdashAlangium salvifolium (L) f Wangerin 15 10 8 8 mdash 12 9 10 mdash mdashAndrographis echioides L 12 9 mdash mdash mdash 17 8 9 mdash mdashAndrographis paniculataWallich ex Nees 26 21 19 13 mdash 25 8 16 12 mdashAcalypha indica L mdash mdash mdash mdash mdash 18 mdash mdash mdash mdashAcalypha lanceolata L mdash mdash 12 13 mdash 10 mdash mdash 21 mdashAchyranthes aspera L mdash mdash 7 14 mdash 15 12 11 mdash mdashAgeratum conyzoides L 10 8 mdash mdash mdash mdash mdash mdash mdash mdashAsteracantha longifolia L 16 12 mdash mdash mdash 15 mdash mdash mdash mdashAzadirachta indica A Juss 15 17 21 16 14 23 12 14 mdash mdashBorassus flabellifer L 9 10 17 8 mdash mdash 9 10 mdash mdashBoerhavia erecta L mdash mdash 8 9 7 16 mdash mdash mdash mdashCalotropis procera (L) mdash mdash 15 mdash 18 28 9 mdash mdash mdashCalotropis gigantea (L) RBrex Ait 11 9 mdash mdash mdash 20 mdash mdash 9 8Cardiospermum halicacabum L 23 mdash mdash 14 mdash 9 19 mdash mdash mdashCatharanthus roseus (L) GDon 13 7 12 9 mdash 15 11 8 mdash mdashCassia auriculata L 17 13 12 mdash mdash 19 13 mdash mdash mdashCassia occidentalis L 18 mdash mdash mdash mdash mdash mdash mdash mdash mdashCassia tora L mdash mdash mdash mdash mdash mdash mdash mdash mdash mdashCassia fistula L mdash mdash mdash mdash mdash mdash mdash mdash mdash mdashCitrus acida Roxb Hookf 26 22 mdash 12 29 9 13 8 mdashCissus quadrangularis L mdash mdash mdash mdash mdash mdash mdash mdash mdash mdashCinnamomum zeylanicum Garcin exBlume 14 16 7 20 mdash 22 19 7 mdash mdash

Cinnamomum iners Reinw ex Blume 20 16 mdash 15 mdash 16 12 mdash mdash mdashRosmarinus officinalis L mdash 9 mdash 10 mdash 7 mdash 8 mdash mdashCentella asiatica (L) 9 8 mdash mdash mdash 11 mdash mdash mdash mdashClerodendrum inerme (L) Gaertn 13 7 12 mdash mdash 15 11 8 mdash mdashClitoria ternatea L mdash mdash mdash mdash mdash mdash mdash mdash mdash mdashClitoria ternatea L 16 12 mdash 13 mdash 8 10 mdash mdash mdashCleome gynandropsis L mdash mdash 12 19 11 mdash 9 15 12 mdashCleome viscose L mdash mdash 8 10 14 20 mdash mdash mdash mdashCoccinia grandisWampA 9 10 17 8 mdash mdash 9 10 mdash mdashCymbopogon citratus (DC) 16 18 mdash 17 mdash 14 mdash mdash mdash mdashDatura metel L mdash mdash mdash mdash mdash mdash mdash mdash mdash mdashEclipta alba (L) Hassk 20 mdash mdash 9 mdash mdash 10 mdash mdash mdashEuphorbia hirta Linn 11 mdash mdash mdash mdash 16 mdash mdash mdash mdashEucalyptus globulus Labill mdash mdash mdash 7 mdash mdash mdash 7 mdash mdashEugenia caryophyllus Bullock amp Harrison mdash 7 mdash 11 mdash 9 mdash 8 mdash mdashElettaria cardamomumWhite et Mason 21 20 7 14 mdash 22 12 17 mdash mdashHyptis suaveolens (L) Poit 8 10 8 7 mdash mdash mdash mdash mdash mdashIchnocarpus frutescens (L) WJ Aiton mdash mdash mdash mdash mdash mdash mdash mdash mdash mdashJatropha curcas L mdash mdash mdash 8 mdash 11 mdash 12 mdash 7Leucas aspera (Willd) Link 9 10 17 8 mdash 12 9 10 mdash mdashLawsonia inermis L mdash mdash mdash mdash mdash mdash mdash mdash mdash mdashMadhuca longifolia (L) JF Macbr 18 16 14 19 12 mdash 8 7 9 mdash

Evidence-Based Complementary and Alternative Medicine 9

Table 2 Continued

Scientific name Microorganisms growth inhibition zones (6 millimeter in diameters)KHW TES Kp Kpr Pa Sa Stp Sp Vd Vd

Merremia hastate L (Desr) Hallierf mdash mdash mdash mdash mdash mdash mdash mdash mdash mdashMorinda tinctoria Roxb mdash mdash mdash mdash mdash mdash mdash mdash mdash mdashMentha piperita L 23 17 26 12 20 25 19 mdash mdash mdashOcimum sanctum L 12 9 11 7 mdash 15 7 8 mdash mdashOldenlandia umbellata L mdash mdash mdash mdash mdash 17 mdash mdash mdash mdashPiper attenuatum Buch Hamex Miq 13 7 12 21 mdash 17 11 8 mdash mdashPlumbago zeylanica (L) Cav 9 10 17 8 mdash 12 9 10 mdash mdashPlectranthus amboinicus (L) Spreng mdash mdash mdash 8 11 15 mdash mdash mdash mdashPhyllanthus debilis L (Klein ex Willd) 7 8 mdash 9 mdash 9 18 7 mdash mdashPhyllanthus maderaspatensis L 17 mdash mdash mdash mdash mdash mdash mdash mdash mdashPremna tomentosaWilld 13 10 mdash 15 mdash 9 10 mdash mdash mdashGloriosa superba L 17 16 7 8 mdash 15 mdash mdash mdash 8Sebastiania chamaelea (L) Muell Arg 19 12 mdash 17 mdash 13 19 8 mdash mdashSolanum trilobatum L mdash mdash 13 9 mdash 8 mdash mdash mdash mdashSphaeranthus indicus L 20 18 21 7 22 mdash 11 9 mdash 16Swertia chirata (L) Ham mdash mdash mdash mdash mdash mdash mdash mdash mdash mdashTerminalia arjuna (W amp A) mdash mdash mdash mdash mdash 16 mdash mdash mdash mdashTinospora cordifolia (Willd) Miers exHooff ampThoms mdash mdash 23 16 mdash 12 15 mdash mdash mdash

Tridax procumbens L 9 8 7 mdash mdash 14 mdash mdash mdash mdashTragia involucrata L 25 23 20 mdash mdash 31 28 22 19 mdashVitex negundo L mdash mdash mdash mdash mdash 14 mdash mdash mdash mdashVetiveria zizanioides (L) 11 9 7 7 mdash 16 mdash mdash 8Withania somnifera (L) Dunal 12 20 mdash mdash mdash 15 12 mdash mdash mdashZingiber officinale Rosc 14 11 7 15 mdash 7 12 7 mdash mdashZanthoxylum limonella (Dennst) Alston 13 7 12 9 mdash mdash 11 8 mdash mdashChloramphenicol (30120583gdisc) 21 12 15 17 29 16 15 18 13 11Ceftazidime (30 120583gdisc) 33 16 22 19 16 25 21 20 12 15lowastBacteria (+minus) Results obtained in the disc diffusion assay antibacterial activity is expressed as the mean plusmn SD (119899 = 3) of the inhibition by the extract andits diameter around the discs One way analysis of variance was performed (mean plusmn SD 119899 = 3 replicates) Size of inhibition zones were including the sterileblank discs 6 millimeter (mm) in diameters Absence of bacterial inhibition indicates (mdash) antibiotic disc (30 120583gdisc)

Interestingly sixteen plants such as Andrographis echioidesC auriculata C viscose C gigantea T arjuna Oldenlandiaumbellata Boerhavia erecta and E hirta exerted a strongactivity against the Gram-positive S aureus bacteria

37 Phytochemical Screening of Plants The results obtainedfrom the phytochemical screening as shown in Table 1 indi-cate the presence of various types of secondary metabolitessuch as polyphenols tannins saponins alkaloids and gly-cosidespolysaccharides Most of the plant extracts relativelyrich in alkaloids phenols flavonoids polyphenols tanninssterols and terpenoids were found to inhibit the growth oforganisms

38 Antimicrobial Activity of Fractioned Compounds Activecomponents were purified from the most active extractsfor further testing The compound shellsol of T involucrata

and C acida exhibited the most potent action againstthe antibiotic resistant strains of B pseudomallei (KHW)S aureus B pseudomallei (TES) and K pneumoniae Amarmelos was also found to inhibit the growth of B pseudo-mallei (KHW) more effectively than other tested bacteria Avasica showed the broad spectrum growth inhibitory activityon B pseudomallei (KHW) K pneumoniae K pneumoniaeresistant B pseudomallei (TES) S pyogenes and V damselaHowever E cardamomum displayed antimicrobial activityon some of the B pseudomallei (KHW) S pyogenes Bpseudomallei (TES) and S typhi strains Similarly A indicaexerted the growth inhibition on K pneumoniae and Saureus Remarkably Sebastiania chamaelea was more activeagainst K pneumoniae K pneumoniae and S pyogenesThe compound from S indicus inhibited the growth of Kpneumoniae K pneumoniae B pseudomallei (KHW andTES) and S typhi strains as compared to the activity shown

10 Evidence-Based Complementary and Alternative Medicine

Figure 2 Diverse biodiversity richness of medicinal plants in Western and Eastern Ghats (a) Topography of plant covering area in Kollihills (Namakkal district Tamil Nadu) (b) Aerial view of occurrence of medicinal plants in Mundanthurai hills (Tirunelveli district TN) (c)Pachamalai hills (Trichy district) and its natural vegetation inhabitants for Malaiyali tribes (d) Deforestation of natural herbal resources dueto urbanization in Kalrayan hills (Salem district) in the Eastern Ghats of TN (e) Medicinal plants and its various parts used by the natives(traditional healers) for the treatment of diverse human illness with a very high percentage of leaves and whole plants often used for herbaldrug preparation by the local practitioners (f) Various category of plants like shrub herb climbers and tree and the parts used in medicine

Evidence-Based Complementary and Alternative Medicine 11

Figure 3 Medicinal aromatic herbs spices and toxic plants were collected from the tribal areas of the Western and Eastern Ghats region inTamil Nadu India (a) A vasica Nees (leaf) (b) Eclipta alba (L) Hassk (whole plant) (c)Mimosa pudica L (whole plant) (d) P amboinicus(L) Spreng (whole plant) (e) T procumbens (L) (whole plant) (f) Euphorbia hirta Linn (whole plant) (g) A paniculata Wallich ex Nees(H) C roseus (L) GDon (whole plant) used for therapy (i) Cinnamomum iners Reinw ex Blume (leaf) (j) E globulus Labill (leaf and bark)(k) Z officinale Rosc (Rhizome) (l) E caryophyllus (Sprengel) Bullock and Harrison (flower buds) (m) M piperita L (whole plant) (n) Ccitratus (DC) Clitoria ternatea L (whole plant) (o) C zeylanicum Garcin ex Blume (bark) (p) Elettaria cardamomumWhite et Mason (fruitpod) used for medicine and food preparation

by the crude extracts (Figure 4)The antimicrobial efficacy offractions collected from the oil yielding plants was also com-pared with that of the tested compounds C zeylanicum andR officinaliswere themost sensitive in controlling the growthof B pseudomallei (KHW) S aureus K pneumonia and Spneumoniae Fascinatingly all the compounds obtained fromaromatic plants except those from E globules were foundto be very effective against the multidrug resistant humanpathogen B pseudomallei (KHW) that causes melioidosis

On the other hand compounds from C citrates O sanctumE caryophyllus and Z zizanioide showed some promisingeffect only against S aureus (Figure 5) The activity of thecompounds were pronounced more than that of the oilyielding plant fractions

39 Minimum Inhibitory Concentrations (MICs) The antibi-otic potential of the purified fractions was obtained fromthe MIC determination The hydrocarbon ester shellsol

12 Evidence-Based Complementary and Alternative Medicine

(mean plusmn SD n = 3)

0

5

10

15

20

25

30

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn V

d St

Bacteria (+minus)

lowast

lowast

lowast

A vasica

In vitro assay(P

lant

frac

tion)

Inhi

bitio

n zo

nes (

mm

)

(a)

(mean plusmn SD n = 3)

0

5

10

15

20

25

30

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn V

d St

Bacteria (+minus)

lowast

A marmelos

Inhi

bitio

n zo

nes (

mm

)

In vitro assay

(b)

(mean plusmn SD n = 3)

0

5

10

15

20

25

30

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn V

d St

Bacteria (+minus)

lowastlowast

A indica

Inhi

bitio

n zo

nes (

mm

)

Incu

batio

n (37∘C

24 h

)

(c)

(mean plusmn SD n = 3)

0

5

10

15

20

25

30

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn V

d St

Bacteria (+minus)

lowast

lowast lowast

35

C acida

Inhi

bitio

n zo

nes (

mm

)(P

lant

frac

tion)

(d)

(mean plusmn SD n = 3)

0

5

10

15

20

25

30

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn V

d St

Bacteria (+minus)

lowastlowast

lowast

S chamaelea

Inhi

bitio

n zo

nes (

mm

)

(e)

(mean plusmn SD n = 3)

0

5

10

15

20

25

30

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn V

d St

Bacteria (+minus)

lowastlowast

lowastlowast

S indicus

Inhi

bitio

n zo

nes (

mm

)

Incu

batio

n (37∘C

24 h

)

(f)

(mean plusmn SD n = 3)

0

5

10

15

20

25

30

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn V

d St

Bacteria (+minus)

lowast

lowast lowast

E cardamomum

Inhi

bitio

n zo

nes (

mm

)(P

lant

frac

tion)

(g)

(mean plusmn SD n = 3)

0

5

10

15

20

25

30

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn V

d St

Bacteria (+minus)

lowast

lowast

lowast

lowast

Shellsol

35

Inhi

bitio

n zo

nes (

mm

)

Incu

batio

n (37∘C

24 h

)

(h)

Figure 4 In vitro antimicrobial activity of purified fractions from the most active plant extracts tested against bacteria Growth inhibitionzones were measured and analyzed with mean plusmn standard deviation (SD) (119899 = 3) using one way analysis of variance Level of significanceat (lowast119875 gt 001) Most of the fractions exerted a potent inhibitory effect against multidrug resistant Gram-negative bacteria (B pseudomalleistrains KHW and TES) K pneumonia and Gram-positive bacteria S aureus

(T involucrata) andC acida showed an interesting inhibitorypotential against S aureus (MIC of 78 120583gmL) and B pseu-domallei strain of KHW (MIC of 156 120583gmL) A vasicashowed an MIC of 156120583gmL against B pseudomallei(KHW) and an MIC of 3125120583gmL against K pneumo-niae K pneumoniae S pyogenes and V damsela strainsFractions from A marmelos and terpenoid from A indicaexerted bacteriostatic effect with MIC values of 3125120583gmLon some selected bacteria including B pseudomallei ofKHW S aureus and B pseudomallei of TES The MIC of3125 120583gmL was found for E cardamomum against S aureus

K pneumonia and S pyogenes S indicus displayed a verystrong inhibition against MDR K pneumoniae (MIC of156 120583gmL) and against B pseudomallei (KHW and TES)at MIC of 3125 120583gmL When the antimicrobial efficacies ofpurified fractions from aromatic plants were compared theC zeylanicum fraction displayed an important antimicrobialeffect against S aureus (MIC of 78 120583gmL) MDR B pseu-domallei of KHW (MIC of 156 120583gmL) and S pneumoniae(MIC of 3125 120583gmL) The essential oil from M piperateashowed MIC value of 3125120583gmL against K pneumoniae Saureus and B pseudomallei (KHW) respectivelyO sanctum

Evidence-Based Complementary and Alternative Medicine 13

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn

Bacteria (+minus)

0

5

10

15

20

25

30

Inhi

bitio

n zo

nes (

mm

)(P

lant

frac

tion)

lowastlowast

lowast

(mean plusmn SD n = 3)In vitro assay

C citratus

(a)

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn

Bacteria (+minus)

0

5

10

15

20

25

30

Inhi

bitio

n zo

nes (

mm

)

lowast

lowast

lowastlowast

lowast

lowast

(mean plusmn SD n = 3)In vitro assay

35

C zeylanicum

(b)

Bacteria (+minus)

0

5

10

15

20

25

30

Inhi

bitio

n zo

nes (

mm

)

lowast

lowast

lowast

(mean plusmn SD n = 3)

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn V

d St

O sanctum

Incu

batio

n (37∘C

24 h

)

(c)

Bacteria (+minus)

0

5

10

15

20

25

Inhi

bitio

n zo

nes (

mm

)

lowast

lowast

(mean plusmn SD n = 3)

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn V

d St

M piperita

(Pla

nt fr

actio

n)

(d)

Bacteria (+minus)

0

5

10

15

20

25

30

Inhi

bitio

n zo

nes (

mm

)lowast

lowast

lowast

(mean plusmn SD n = 3)

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn V

d St

E caryophyllus

(e)

0

5

10

15

20

25

30

Inhi

bitio

n zo

nes (

mm

) (mean plusmn SD n = 3)

Bacteria (+minus)

KHW

TES

Kp

KprPa Sa

Stp

Spn

E globulus

Incu

batio

n (37∘C

24 h

)

(f)

Bacteria (+minus)

0

5

10

15

20

25

30

Inhi

bitio

n zo

nes (

mm

)(P

lant

frac

tion)

lowast

lowast

lowast

lowast

(mean plusmn SD n = 3)

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn V

d

R officinalis

(g)

0

5

10

15

20

25

30

Inhi

bitio

n zo

nes (

mm

)

lowast

lowast

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn V

d St

Bacteria (+minus)V zizanioides

(mean plusmn SD n = 3)

Incu

batio

n (37∘C

24 h

)

(h)

Figure 5 Comparison of antimicrobial effect of plant compounds obtained from the most popularly used sources of essential oils as assayedby the disc-diffusion method in vitro It displayed a powerful activity against B pseudomallei and S aureus than the other bacteria strainsOther compounds showed only a moderate or weak action against the tested bacteria Values for zone of bacterial growth inhibition werepresented as mean plusmn SD (119899 = 3) with level of significance at (lowast119875 gt 001)

and C citratus fractions also showed antimicrobial activity(MICs of 3125ndash125120583gmL) only at higher concentrationsagainst the tested bacteria In addition to that higher concen-trations (gt250120583gmL) (of Vetiveria fractions) were requiredto inhibit Vibro species and others (including E globulusfractions) failed to show any effect at tested concentrations(78ndash125 120583gmL) However the purified fractions (from mostactive medicinal plants) showed strong bacteriostatic inhibi-tion against the tested organisms (Table 3)

310 Cytotoxic Effects of Plants When the components wereassayed for cytotoxicity against the normal human skinfibroblasts (HEPK) cells the compounds obtained from

E cardamomum T involucrata S indicusC acidaA vasicaA marmelos A indica and A paniculata did not showtoxicity up to 1000120583gmL (see Figures S1 and S2 in Supple-mentaryMaterial available online at httpdxdoiorg1011552013525613) A slight reduction of cell proliferation wasnoted only at higher doses (2000120583gmL) In contrast cellproliferation was markedly reduced after exposure of HEPKcells to O sanctum E globulus V zizanioides C citratusand E globulus compounds There was no gradual reductionin skin cell proliferation seen after exposure to C zeylan-icum R officinalis and M piperita (see Figures S3 and S4)compounds The toxicity was found to be concentration-dependent when the skin fibroblasts (HEPK) cells were

14 Evidence-Based Complementary and Alternative Medicine

Table 3 Minimum inhibitory concentrations (MICs) of purified plant fractions and essential oils against antibiotic resistant bacteria

Botanical name Family Parts used Gram-positive and -negative bacteria (MICs 120583gmL)KHW TES Kp KPr Sa Stp Sp Vd St

A indica Juss Meliaceae Seed (fraction) 3125 625 125 lt250 3125 mdash mdash mdash mdashA marmelos (L) Rutaceae Root-bark (F) 3125 3125 mdash mdash 3125 625 125 250 mdashA paniculata Nees Acanthaceae Leaf (fraction) 250 mdash 125 mdash 625 mdash mdash 125 mdashA vasica Nees Acanthaceae Fraction (Stem) 156 3125 3125 625 mdash lt250 3125 3125 mdashC acida Roxb Rutaceae Leaf (fraction) 156 625 mdash mdash 78 mdash 3125 mdash mdashE cardamomumWhite et Mason Acanthaceae Fraction (WP) 625 mdash 3125 mdash 3125 250 625 mdash mdashS indicus (L) Euphorbiaceae Whole plant (F) 3125 125 156 mdash 625 mdash mdash lt250 mdashT involucrata (L) Euphorbiaceae Shellosol (leaf) 156 3125 mdash mdash 78 625 mdash mdash mdashCinnamomum zeylanicum (L) Lauraceae Bark (fraction) 156 625 mdash mdash 78 3125 mdash mdash 125Cymbopogon citratus (L) Graminae Leaf (fraction) 250 lt250 625 mdash 625 125 mdash mdash mdashEugenia caryophyllus (L) Myrtaceae Flower buds (F) 625 125 mdash mdash 625 mdash mdash mdash mdashEucalyptus globulus (L) Myrtaceae Fraction (leaf) mdash mdash mdash mdash mdash mdash lt250 mdash lt250Mentha piperita (L) Labiatae Fraction (WP) mdash 625 3125 mdash 3125 mdash lt250 mdash mdashOcimum sanctum (L) Labiatae Leaf (fraction) 625 125 mdash 3125 625 mdash 125 lt250 mdashRosmarinus officinalis (L) Labiatae Rosemary oil 3125 mdash mdash mdash 3125 mdash mdash mdash mdashVetiveria zizanioides (L) Graminae Root (fraction) mdash 125 125 mdash mdash mdash gt250 gt250The bacterial growth inhibitory activity was compared for significant differences within the bacterial strains by broth-dilution method at 250 125 625 3125156 and 78 120583gmL F fractions

exposed to various compounds The cell proliferation wasincreased by the influence of the plant components at thelower concentrations Whereas the oil yielding plant com-pounds showed inhibition of cell proliferation and toxicity athigher doses 250ndash1000120583gmL

4 Discussion

The Western Ghats is considered as one of the richestbiodiversity hotspots in the world [41] In this survey wecollected nearly 78 medicinal plants from Western andEastern Ghats that are edible and popularly used for curingvarious ailments including snakebite Traditional remedieshave a long-standing history in many tribal settlements inTN India and they continue to provide useful and applicabletools for treating ailments [42] The ingredients that makeup the ldquoVishakallurdquo stone which is used as an antidote forsnakebite are different herbs and pebbles available from theriver banks Likewise aqueous paste and decoction obtainedfrom the leaves of A paniculata are widely used for snakebitetreatment by indigenous people [43] Previous studies havereported that ethnomedicine plays major roles in conservingthe disappearing knowledge of tribal communities [44ndash47]The traditional beliefs of reliance on a rich diversity ofethnomedicinal plants located at different settlements havealso been confirmed in another study [48 49] Herein weexplored the various types of traditional practices reported bythe primitive tribal communities with a view to gain furtherknowledge from such studies

In the present investigation potentially rich sources oftribal medicine (71 plants) were scientifically evaluated fortheir antibacterial activity against the MDR bacteria and theaccumulated data was disseminated for the first time to the

scientific community Out of the 71medicinal plants screenedfor the antibacterial activity 10 of them (T involucrataC acida A marmelos A vasica C procera A paniculataand M piperita A indica S indicus and E cardamomum)displayed the highest antibacterial activity against the mul-tidrug resistant B pseudomallei (KHW and TES) and Saureus strains The antibacterial activity of those crude plantextracts was as equally effective as that of the standard drugsOur findings corroborated with the previous reports madeon the antistaphylococcal activity of tribal medicinal plants[34 50 51] On the other hand isolated components from themost active extracts of T involucrata shellsol and C acidaexhibited the most potent action against the antibiotic resis-tant B pseudomallei (KHW) K pneumoniae and S aureusstrainsThese results further confirmed our previous findingson the leaves of T involucrata and its compounds hydrocar-bon ester-like shellsol which displayed a high antibacterialeffect against the different bacterial strains especially thatof S aureus [34] Eugenol and caryophyllene are the activeagents contained in the M piperita [52] and O sanctumplants which are believed to be mainly responsible for theantimicrobial properties of these plants [53] Interestinglythe compounds obtained from the aromatic plants such asC zeylanicum and R officinalis were also found to be veryeffective against the multidrug resistant human pathogen Kpneumoniae S aureus S typhi and B pseudomallei (KHW)that causes melioidosis

The inhibitory potential determined for the shellsol(T involucrata) vaseline (A vasica) C acida and C zey-lanicum indicates that the MIC of 78ndash3125120583gmL foundagainst the B pseudomallei of KHW K pneumoniae Kpneumoniae S pyogenes and V damsel and S pneumoniaewas quite low Similarly lower MIC values were found for

Evidence-Based Complementary and Alternative Medicine 15

M piperita (MIC of 113ndash225 mgmL) against the abovebacterial strains [52] MIC was found for the most activealcohol extracts of A salvifolium (MIC 0034ndash0263mgmL)on S aureus [54] and the MIC for S trilobatum aqueousextracts determined against the tested organisms rangedfrom 006 to 05mgmL [55] C zeylanicum was foundto have an effective antibacterial activity (MIC 64 120583gmL)against P aeruginosa E coli B subtilis and S aureus[56] Previously several investigators have demonstrated thatactive agents exert interesting activity against bacteria evenat lower concentrations tested [57 58] The bacteriostaticmechanism involves damage to the cell walls of bacteriafollowed by inhibition of protein synthesis that ultimatelyleads to bacterial death [59] The most active plants arewidely used by various tribes as traditional treatment (iecut wounds skin infection and scabies) thus indicatingthe potential for further development into promising drugsIn order to ascertain the safety and efficacy of the mostactive compounds their effect on human skin fibroblast cellswas evaluated Chemical constituents of E cardamomumT involucrata S indicus C acida A vasica A marmelosA indica and A paniculata plants failed to produce anynoticeable toxicity up to 1000120583gmL Although some ofthe tested compounds exhibited a slight reduction of cellproliferation and some minor morphological changes suchchanges were insignificant at lower doses and became evidentonly at higher doses However certain aromatic compoundsof O sanctum E globulus V zizanioides C citratus and Eglobulus plants showed reduction of cell proliferation againstHEPK cells

Our phytochemical screening also provides evidenceof the presence of several types of compounds that aremainly responsible for the remarkable antibacterial effectof these plants The differences noted for the bactericidalactivity of various plant extracts in this study appears tobe directly related to the diversity of compounds (shown inparentheses) that are accumulated in the following plants(eg A marmelos and O umbellate) [60ndash62] Compoundslike tannins phenol and polyphenols can bind the Gram-negative bacteria to form a heavy soluble complex on the cellsurface which subsequently disturbs the availability of recep-tor on cells and kills the bacteria [63] Several species havingwide spectra of antimicrobial activity mainly due to theactive constituents such as essential oil phenolic compoundslike thymol carvacrol in oregano and thyme eugenol inclove and cinnamon were also identified previously [64 65]Essential oils degrade the cell wall interact with the cell com-ponents and then disrupt the cytoplasmic membrane [66]The antimicrobial effect of phenolic compounds may involvemultiplemodes of action including damage to themembraneprotein interference with membrane integrated enzymes[67] causing leakage of cellular components coagulation ofcytoplasm depletion of the proton motive force alterationof fatty acid and phospholipid constituents impairmentof enzymatic mechanisms for production and metabolismalteration of nutrient uptake and electron transport [68]influencing the synthesis of DNA and RNA and destroyingprotein translation and the function of the mitochondrionin eukaryotes [69] The mode of action of antimicrobial

agents depends on the type of microorganism and is mainlyrelated to their cell wall structure and the outer membranearrangement Most of the plant spices and herbs containcomplex phenolics (ie phenolic acids flavonoids tanninslignans coumarins quinines) In addition the mechanismsof action of each phenolic compound against various bacteriaare also very complicated [26 70] Further investigation istherefore required to understand the relationship betweenthe antimicrobial action and the chemical structure of everyphenolic compound in the tested extracts The informationavailable from previous pharmacological sources combinedwith the findings herein reported on the medicinal plantextracts may serve as essential data for future drug develop-ment to combat diseases caused by theMDRbacterial strains

Abbreviations

MDR Multidrug resistant bacteriaTES Strain of Burkholderia pseudomalleiMRSA Methicillin-resistant Staphylococcus aureusVRE Vancomycin-resistant enterococciMTB Mycobacterium tuberculosisCH3OH Methanol

CFU Colony forming unitHCl Hydrochloric acidTLC Thin layer chromatographyH2SO4 Sulphuric acids

UV Ultraviolet spectrophotometerHEPK Human skin fibroblasts

Acknowledgments

Theauthors are thankful to the ERI Loyola College ChennaiIndia for his support during the survey The authors alsothank the Department of Microbiology NUHS NUS forthe bacterial cultures used in this investigation The authorsare thankful to the tribal people for sharing their healingknowledge on medicinal aromatic plants

References

[1] M N Alekshun and S B Levy ldquoMolecular mechanisms ofantibacterialmultidrug resistancerdquoCell vol 128 no 6 pp 1037ndash1050 2007

[2] A D Russell ldquoAntibiotic and biocide resistance in bacteriaintroductionrdquo Symposium Series Society for Applied Microbiol-ogy vol 31 no 92 pp 1Sndash3S 2002

[3] D R Guay ldquoTreatment of bacterial skin and skin structureinfectionsrdquo Expert Opinion on Pharmacotherapy vol 4 no 8pp 1259ndash1275 2003

[4] A J Brosnahan and P M Schlievert ldquoGram-positive bac-terial superantigen outside-in signaling causes toxic shocksyndromerdquo FEBS Journal vol 278 no 23 pp 4649ndash4667 2011

[5] K Fellermann J Wehkamp K R Herrlinger and E F StangeldquoCrohnrsquos disease a defensin deficiency syndromerdquo EuropeanJournal of Gastroenterology and Hepatology vol 15 no 6 pp627ndash634 2003

16 Evidence-Based Complementary and Alternative Medicine

[6] L Chen S Lin R Agha-Majzoub L Overbergh C Mathieuand L S Chan ldquoCCL27 is a critical factor for the developmentof atopic dermatitis in the keratin-14 IL-4 transgenic mousemodelrdquo International Immunology vol 18 no 8 pp 1233ndash12422006

[7] J H Song V Thamlikitkul and P R Hsueh ldquoClinical andeconomic burden of community-acquired pneumonia amongstadults in the Asia-Pacific regionrdquo International Journal ofAntimicrobial Agents vol 38 no 2 pp 108ndash117 2011

[8] D A B Dance ldquoMelioidosisrdquo Current Opinion in InfectiousDiseases vol 15 no 2 pp 127ndash132 2002

[9] N J White ldquoMelioidosisrdquo The Lancet vol 361 no 9370 pp1715ndash1722 2003

[10] S Yang ldquoMelioidosis research in Chinardquo Acta Tropica vol 77no 2 pp 157ndash165 2000

[11] P R Hsueh L J Teng L N Lee et al ldquoMelioidosis an emerginginfection in Taiwanrdquo Emerging Infectious Diseases vol 7 no 3pp 428ndash433 2001

[12] C Walsh and S Fanning ldquoAntimicrobial resistance in food-borne pathogensmdasha cause for concernrdquo Current Drug Targetsvol 9 no 9 pp 808ndash815 2008

[13] M Vorachit P Chongtrakool S Arkomsean and S BoonsongldquoAntimicrobial resistance in Burkholderia pseudomalleirdquo ActaTropica vol 74 no 2-3 pp 139ndash144 2000

[14] F M Thibault E Hernandez D R Vidal M Girardet and JD Cavallo ldquoAntibiotic susceptibility of 65 isolates of Burkholde-ria pseudomallei and Burkholderia mallei to 35 antimicrobialagentsrdquo Journal of Antimicrobial Chemotherapy vol 54 no 6pp 1134ndash1138 2004

[15] R L S Sikarwar ldquoEthnogynaecological uses of plants new toIndiardquo Ethnobotany vol 12 no 1-2 pp 112ndash115 2002

[16] R Siva ldquoStatus of natural dyes and dye-yielding plants in IndiardquoCurrent Science vol 92 no 7 pp 916ndash925 2007

[17] P Kumar Singh V Kumar R K Tiwari A Sharma ChV Rao and R H Singh ldquoMedico-ethnobotany of lsquochatararsquoblock of district sonebhadra Uttar Pradesh Indiardquo Advances inBiological Research vol 4 no 1 pp 65ndash80 2010

[18] W H Lewis and M P Elwin-Lewis Medical Botany PlantsAffecting Human Health John Wiley amp Sons New York NYUSA 2003

[19] R Perumal Samy P N Pushparaj and P GopalakrishnakoneldquoA compilation of bioactive compounds fromAyurvedardquo Bioin-formation vol 3 no 3 pp 100ndash110 2008

[20] WorldHealthOrganization (WHO)TraditionalMedicine Strat-egy 2002ndash2005 Genera World Health Organization GenevaSwitzerland 2002

[21] P S Haddad G A Azar S Groom and M Boivin ldquoNaturalhealth products modulation of immune function and preven-tion of chronic diseasesrdquo Evidence Based Complementary andAlternative Medicine vol 2 no 4 pp 513ndash520 2005

[22] K Poonam andG S Singh ldquoEthnobotanical study of medicinalplants used by the Taungya community in Terai Arc LandscapeIndiardquo Journal of Ethnopharmacology vol 123 no 1 pp 167ndash1762009

[23] G B Mahady ldquoMedicinal plants for the prevention and treat-ment of bacterial infectionsrdquo Current Pharmaceutical Designvol 11 no 19 pp 2405ndash2427 2005

[24] V P Kumar N S Chauhan H Padh and M Rajani ldquoSearchfor antibacterial and antifungal agents from selected Indianmedicinal plantsrdquo Journal of Ethnopharmacology vol 107 no2 pp 182ndash188 2006

[25] R Nair T Kalariya and S Chanda ldquoAntibacterial activity ofsome plant extracts used in folk medicinerdquo Journal of HerbalPharmacotherapy vol 7 no 3-4 pp 191ndash201 2007

[26] D Kalemba and A Kunicka ldquoAntibacterial and antifungalproperties of essential oilsrdquo Current Medicinal Chemistry vol10 no 10 pp 813ndash829 2003

[27] A H Ashour ldquoAntibacterial antifungal and anticancer activi-ties of volatile oils and extracts from stems leaves and flowers ofEucalyptus sideroxylon and Eucalyptus torquatardquoCancer BiologyandTherapy vol 7 no 3 pp 399ndash403 2008

[28] M J Abad L M Bedoya L Apaza and P Bermejo ldquoTheArtemisia L genus a review of bioactive essential oilsrdquoMolecules vol 17 no 3 pp 2542ndash2566 2012

[29] H N Marzoug M Romdhane A Lebrihi et al ldquoEucalyptusoleosa essential oils chemical composition and antimicrobialand antioxidant activities of the oils from different plant parts(stems leaves flowers and fruits)rdquoMolecules vol 16 no 2 pp1695ndash1709 2011

[30] C Proestos D Sereli and M Komaitis ldquoDetermination ofphenolic compounds in aromatic plants by RP-HPLC and GC-MSrdquo Food Chemistry vol 95 no 1 pp 44ndash52 2006

[31] S Chao G Young C Oberg and K Nakaoka ldquoInhibition ofmethicillin-resistant Staphylococcus aureus (MRSA) by essentialoilsrdquo Flavour and Fragrance Journal vol 23 no 6 pp 444ndash4492008

[32] M Akin D Oguz and H T Saracoglu ldquoAntibacterial effectsof some plant extracts from Labiatae (Lamiaceae) growingnaturally around Sirnak-Silopi Turkeyrdquo International Journal ofPharmaceutical and Applied Sciences vol 1 no 1 pp 4ndash47 2010

[33] K Bayoub T Baibai D Mountassif A Retmane and ASoukri ldquoAntibacterial activities of the crude ethanol extracts ofmedicinal plants against Listeriamonocytogenes and some otherpathogenic strainsrdquo African Journal of Biotechnology vol 9 no27 pp 4251ndash4258 2010

[34] R P Samy P Gopalakrishnakone M Sarumathi P Houghtonand S Ignacimuthu ldquoPurification of antibacterial agents fromTragia involucratamdasha popular tribal medicine for wound heal-ingrdquo Journal of Ethnopharmacology vol 107 no 1 pp 99ndash1062006

[35] K M Matthew The Flora of Tamil Nadu Carnatic Part IndashIIIprinted at the Diocesan Press Madras India 1981ndash1983

[36] AW BauerWMKirby J C Sherris andM Turck ldquoAntibioticsusceptibility testing by a standardized single disk methodrdquoAmerican Journal of Clinical Pathology vol 45 no 4 pp 493ndash496 1966

[37] National Committee for Clinical Laboratory Standards Per-formance Standards For Antimicrobial Susceptibility Testing12th Informational Supplement Approved standard M100-S12National Committee for Clinical Laboratory Standards WaynePa USA 2002

[38] E A Elsinghorst ldquoMeasurement of invasion by gentamicinresistancerdquoMethods in Enzymology vol 236 pp 405ndash420 1994

[39] J B Harborne Phytochemical Methods Chapman and HallNew York NY USA 1976

[40] H Wagner S Bladt and E M Zgairski Plant Drug AnalysisSpringer New York NY USA 1984

[41] N Myers R A Mittermeler C G Mittermeler G A B daFonseca and J Kent ldquoBiodiversity hotspots for conservationprioritiesrdquo Nature vol 403 no 6772 pp 853ndash858 2000

[42] R Perumal Samy and S Ignacimuthu ldquoAntibacterial activity ofsome folkloremedicinal plants used by tribals inWestern Ghats

Evidence-Based Complementary and Alternative Medicine 17

of Indiardquo Journal of Ethnopharmacology vol 69 no 1 pp 63ndash712000

[43] K Nampoothiri A Chrispal A Begum S Jasmine K GGopinath and A Zachariah ldquoA clinical study of renal tubulardysfunction inCleistanthus collinus (Oduvanthalai) poisoningrdquoClinical Toxicology vol 48 no 3 pp 193ndash197 2010

[44] M Strathern UsefulKnowledge The Isiah Berlin Lecture Uni-versity of Manchester Brunswick UK

[45] C N Sunil and A K Pradeep ldquo(2001) Another new species ofTripogon (Poaceae) from Indiardquo Sida vol 19 pp 803ndash806 2005

[46] S Biber-Klemm and T Cottier Right To Plant GeneticResources and Traditional Knowledge Basic Issues and Prospec-tive Oxfordshire Nosworthy Way UK 2006

[47] V S Ramachandran S Shijo Joseph and R Aruna ldquoEth-nobotanical studies from amaravathy range of Indira GandhiWildlife SanctuaryWesternGhats CoimbatoreDistrict South-ern Indiardquo Ethnobotanical Leaflets vol 13 pp 1069ndash1087 2009

[48] A H Rajasab and I Mohamad ldquoDocumentation of folkknowledge on edible wild plants of North Karnatakardquo IndianJournal of Traditional Knowledge vol 3 no 4 pp 419ndash4292004

[49] M Rajadurai V G Vidhya M Ramya and A Bhaskar ldquoEthno-medicinal plants used by the traditional healers of pachamalaihills Tamilnadu Indiardquo Studies on Ethno-Medicine vol 3 no 1pp 39ndash41 2009

[50] S K Adesina O Idowu A O Ogundaini et al ldquoAntimicrobialconstituents of the leaves of Acalypha wilkesiana and Acalyphahispidardquo Phytotherapy Research vol 14 no 5 pp 371ndash374 2000

[51] M T Gutierrez-Lugo M P Singh W M Maiese and BN Timmermann ldquoNew antimicrobial cycloartane triterpenesfrom Acalypha communisrdquo Journal of Natural Products vol 65no 6 pp 872ndash875 2002

[52] A K Tyagi andAMalik ldquoAntimicrobial potential and chemicalcomposition of Mentha piperita oil in liquid and vapour phaseagainst food spoiling microorganismsrdquo Food Control vol 22no 11 pp 1707ndash1714 2011

[53] G Devendran and U Balasubramanian ldquoQualitative phyto-chemical screening and GC-MS analysis of Ocimum sanctumL leavesrdquoAsian Journal of Plant Science and Research vol 1 no4 pp 44ndash48 2011

[54] V C Jain N M Patel D P Shah P K Patel and B HJoshi ldquoAntioxidant and antimicrobial activities of Alangiumsalvifolium (Lf) Wang rootrdquo Global Journal of Pharmacologyvol 4 no 1 pp 13ndash18 2010

[55] P Swapna Latha andK Kannabiran ldquoAntimicrobial activity andphytochemicals of Solanum trilobatum LinnrdquoAfrican Journal ofBiotechnology vol 5 no 23 pp 2402ndash2404 2006

[56] M Usha1 S Ragini and S M A Naqvi ldquoAntibacterial activityof acetone and ethanol extracts of Cinnamon (Cinnamomumzeylanicum) and Ajowan (Trachyspermum ammi) on four foodspoilage bacteriardquo International Research Journal of BiologicalSciences vol 1 no 4 pp 7ndash11 2012

[57] S Prabuseenivasan M Jayakumar and S Ignacimuthu ldquoInvitro antibacterial activity of some plant essential oilsrdquo BMCComplementary and Alternative Medicine vol 6 article 39 pp1ndash8 2006

[58] G A Ayoola F M Lawore T Adelowotan et al ldquoChemi-cal analysis and antimicrobial activity of the essential oil ofSyzigium aromaticum (clove)rdquo African Journal of Biotechnologyvol 7 no 13 pp 2227ndash2231 2008

[59] Y J Fu L Chen Y Zu et al ldquoThe antibacterial activity of cloveessential oil against Propionibacterium acnes and its mechanismof actionrdquo Archives of Dermatology vol 145 no 1 pp 86ndash882009

[60] D Venkatesan C M Karrunakarn S Selva Kumar and P TPalani Swamy ldquoIdentification of phytochemical constituents ofAegle marmelos responsible for antimicrobial activity againstselected pathogenic organismsrdquo Ethnobotanical Leaflets vol 13pp 1362ndash1372 2009

[61] P Arun K G Purushotham J Johnsy jayarani and V KumarildquoIn vitro Antibacterial Activity of Oldenlandia umbellata anIndian medicinal Plantrdquo Journal of Pharmaceutical Science andTechnology vol 2 no 4 pp 198ndash201 2010

[62] A Manjamalai R Sardar Sathyajith Singh C Guruvayoorap-pan and V M Berlin Grace ldquoAnalysis of phytochemicalconstituents and antimicrobial activity of some medicinalplants in Taminadu Indiardquo Global Journal of Biotechnology andBiochemistry vol 5 no 2 pp 120ndash128 2010

[63] E Haslam ldquoNatural polyphenols (vegetable tannins) as drugsPossible modes of actionrdquo Journal of Natural Products vol 59no 2 pp 205ndash215 1996

[64] Y Cai Q Luo M Sun and H Corke ldquoAntioxidant activityand phenolic compounds of 112 traditional Chinese medicinalplants associated with anticancerrdquo Life Sciences vol 74 no 17pp 2157ndash2184 2004

[65] B Shan Y Z Cai M Sun and H Corke ldquoAntioxidant capacityof 26 spice extracts and characterization of their phenolicconstituentsrdquo Journal of Agricultural and Food Chemistry vol53 no 20 pp 7749ndash7759 2005

[66] R J W Lambert P N Skandamis P J Coote and G-J ENychas ldquoA study of the minimum inhibitory concentration andmode of action of oregano essential oil thymol and carvacrolrdquoJournal of AppliedMicrobiology vol 91 no 3 pp 453ndash462 2001

[67] M Raccach ldquoThe antimicrobial activity of phenolic antioxi-dants in food a reviewrdquo Journal of Food Safety vol 6 no 3 pp141ndash170 1984

[68] M Taniguchi Y Yano E Tada et al ldquoMode of action of polygo-dial an antifungal sesquiterpene dialdehyderdquo Agricultural andBiological Chemistry vol 52 no 6 pp 1409ndash1414 1988

[69] G J E Nychas ldquoNatural antimicrobials from plantsrdquo in NewMethods of Food Preservation G W Gould Ed pp 58ndash89Blackie Academic London UK 1995

[70] S Burt ldquoEssential oils their antibacterial properties and poten-tial applications in foodsmdasha reviewrdquo International Journal ofFood Microbiology vol 94 no 3 pp 223ndash253 2004

Submit your manuscripts athttpwwwhindawicom

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Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

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Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Behavioural Neurology

EndocrinologyInternational Journal of

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Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Disease Markers

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

BioMed Research International

OncologyJournal of

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Oxidative Medicine and Cellular Longevity

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PPAR Research

The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014

Immunology ResearchHindawi Publishing Corporationhttpwwwhindawicom Volume 2014

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Parkinsonrsquos Disease

Evidence-Based Complementary and Alternative Medicine

Volume 2014Hindawi Publishing Corporationhttpwwwhindawicom

Evidence-Based Complementary and Alternative Medicine 3

25Minimum Inhibitory Concentrations (MICs) Assay MICswere evaluated based on the in vitro screening of 16 purifiedfractions that were found to have potent antimicrobial activ-ity Broth dilution method was used for the MIC assay withsome slight modifications as recommend by the NCCLS [37]Two-fold serial dilutions of all the fractionated compoundswere made with MH and TS broth in microtiter plate wellsto adjust the final concentration from 78 to 250120583gmL whilewells containing the broth alonewithout any sample served asa controlThree replicates (119899 = 3) were used for each dilutionand culture containing approximately 1 times 105 CFUmL Theplateswere incubated at 37∘C for 24 h and the absorbancewasmeasured at 560 nm

26 Cytotoxicity Assay The cytotoxic effects of variousextracts were tested by MTT assay [38] using human skinfibroblast HEPK cells The toxic effect of plant extracts andessential oils was assayed on human skin fibroblast (HEPK)cell proliferation in 96-well microtitre plates Confluent cells(5 times 106 cells per well) were incubated with extracts and oilsfor 24 h and the percentage inhibitory concentration (IC

50)

was determined

27 Phytochemical Analysis of Plant Extracts Themost activeextracts were used for purification of antimicrobial com-pounds [39] 100 g each of the plant powder was percolatedwith 500mL of 4 aqueous HCl (adjusted to pH 2) andheated at 50∘C for 3 hThe extract waswashedwith 2times250mLof diethyl ether and the organic phase evaporated to drynessusing vacuum rotary evaporator The dark brown gummyresidues obtained by acid hydrolysis were chromatographedon silica gel column 60 times 32 cm (60ndash120 mesh pH 7 Merck)by gradually eluting with n-hexaneethyl acetate (8 2 6 43 7 and 1 1) and chloroformmethanol (3 2) The aliquotsof each fraction were subjected to thin layer chromatography(TLC) on silica gel coated TLC plate (1mm Merck) usingthe solvent system consisting of 20 (vv) n-hexaneethylacetateThe chromatograms were detected using 50H

2SO4

solution as a spray reagent [34]The individual fractions werecollected and concentrated by vacuum rotary evaporator at40∘C All the purified compounds recovered from the silicagel column were monitored by reading the absorbance at190ndash350 nm (UV spectrophotometer Hitachi Japan) Theactive fractions were further purified and the final yieldsof the compounds were recorded The lyophilized pooledconcentrated compounds were then assayed (100120583gmL)against bacteria The phytochemical screening was done onthe pure compounds using the chemical method previouslyreported for the detection of secondary metabolites [40]The different chemical constituents tested include alkaloidsflavonoids glycosides polyphenols saponin sterols triter-penes tannins reducing sugars gallic acid catechol andaglycones

28 Statistical Analysis The bacterial growth inhibitoryactivity (inhibition zones millimeter in diameter) was com-pared for significant differences within the bacterial strainsOne way analysis of variance was performed (mean plusmn SD

119899 = 3 replicates) using GraphPad Prism 4 USA lowast119875 lt 001was considered statistically significant (inhibition zones ofextractsfractions versus antibiotic drugs)

3 Results

31 Study Area for the Collection of AromaticMedicinal PlantsTheWestern and Eastern Ghats were selected for the presentstudy with the cite map showing the landmarks (Figures 1(a)-1(b)) Kalrayan hills are situated north of Attur taluk (Salemdistrict) one of the major range of hills in the Eastern Ghatsof Tamil Nadu (Figure 1(c)) Pachamalai hills are situated tothe north of Thuraiyur taluk of Tiruchirappalli district Therich biodiversity part of Eastern Ghats lies between latitudes11∘091015840 0010158401015840 to 11∘ 271015840 0010158401015840 N and longitudes 78∘281015840 0010158401015840 to78∘ 491015840 0010158401015840 E and occupies an area of about 52761 squarekm It is located near 11∘ 111015840N 78∘21E1118∘N 7835∘E11187835 (Figure 1(d)) Mundanthurai is located nearly 45 kmwest of Tirunelveli district TN between latitude 8∘ 251015840 and8∘ 531015840 N and longitude 77∘ 101015840 and 77∘ 351015840 E This is the onlyarea of Western Ghats that has the longest raining period ofabout 8 months and forms the catchment area for 14 riversand streams (Figure 1(e)) Kolli Malai is a small mountainrange located in Namakkal district The mountains are about1000ndash1300m in height and cover an area of approximately280 km The Kolli hills are part of the Eastern Ghats whichis a mountain range that runs mostly parallel to the east coastof Tamil Nadu in South India (Figure 1(f)) Javadi hills areone of the largest in the Eastern Ghats in Vellore district inthe northern part of the state of Tamil Nadu They consistof bluish gray hills with peaks averaging 3600ndash3800 feetor 1100ndash1150 meter (Figure 1(g)) Based on the vegetationtype (Figures 2(a)ndash2(d)) the study area consists of (i) drydeciduous (ii) moist deciduous and (iii) rain forests anddiverse proportion of plant parts in abundance (Figures 2(e)-2(f)) Three different types of tribes (ie Kani Malayali andPaliyan tribes) inhabit in the hill ranges The Kani tribeslocated at Mundanthurai raise different types of vegetablesin their own fields while the Malayali tribes cultivate riceThey all engage not only in the agricultural work but alsoare involved in silvicultural work assigned by the forestdepartment Government of TN India

32 Medicinal Plants Glory Western Ghats (Mundanthurai)and Eastern Ghats (Kolli hills Javadi hills Kalrayan hillsPachamalai hills) possess a rich diversity of medicinal plantsthat are used as food and drug by different groups oftribal communities Urbanization habitat degradation andfragmentation of these forests have resulted in the depletionof natural resources on which these tribes used to dependfor their livelihoods It has become increasingly difficult forthem to live in their traditional way In addition the impactof modernization and urbanization has encroached in andaround tribal settlements thus changing their lifestyles

33 Plants Valued as Edibles Various types of plant parts arecollected during different seasons cooked and eaten alongwith boiled rice (Table 1) For example Solanum nigrum leafis most commonly used in all the four regions There are

4 Evidence-Based Complementary and Alternative Medicine

Figure 1 (a) The site for collection of medicinal plants in Western and Eastern Ghats of Tamil Nadu (b) The landmark (map) of traditionalmedicine distribution and collection of different types of plants (c) District map showing the collection site of plants from Kalrayan hills(Salem) (d) Pachamalai hills (Thiruchirappalli) and (e) Mundanthurai (Tirunelveli) rich biodiversity hot-spot of the Western Ghats (f)Kolli hills (Namakkal) (g) Javadi hills (Vellore) part of the Eastern Ghats which is a mountain range that runs mostly parallel to the eastcoast of South India

a large number of wild edible fruits including yielding plantssuch as Citrus acida Ficus benghalensis Ficus microcarpaFicus racemosa Phyllanthus emblica Solanum trilobatumand Syzygium cumini are popularly used by the tribes

34 Plants Used for Snakebite Treatment Thirty-four plantsused for snakebite treatment are documented (Table 1)Snakebite is amajor health hazard that leads to highmortalityin tribal settlements The majority of the antidotes are

prepared freshly from plant materials frequently collectedfrom the leaves of A paniculata A echioides Aristolochiaindica E alba E prostrata M pudica O sanctum Tinvolucrata and Cleistanthus collinus (Oduvanthalai) thewhole plants ofAchyranthes aspera andWedelia calendulaceathe stem-barks and nuts of Strychnos nux-vomica the rootsof Hemidesmus indicus Tephrosia purpurea Rauwolfia ser-pentina C roseus and so forth and the tubers of Gloriosasuperba The tuber paste is usually applied externally on

Evidence-Based Complementary and Alternative Medicine 5

Table 1 Some of the important traditional medicinal plant species families voucher specimens parts used yield of extracts phytochemicalscreening and toxicity on human macrophage cells

Scientific name Family Voucher specimen Plant parts Yield (gm) Phytochemicalanalysis

Adhatoda vasica Nees Acanthaceae D2020 Leaf 64 VasicineAegle marmelos (L)Correa ex Roxb Rutaceae D2018 Root-bark 58 Alkaloids

Alangium salvifolium (L)f Wangerin Alangiaceae 0140 Leaf 63 Phenolic

Andrographis paniculataWallich ex Nees Acanthaceae 0061 Leaf 68 Andrographolide

Andrographis echioidesNees Acanthaceae 0116 Leaf 70 Terpenoids

Acalypha indica L Euphorbiaceae 29644 Leaf 61 AcalypheAcalypha lanceolata L Euphorbiaceae 15791 Leaf 71 AlkaloidsAchyranthes aspera L Amaranthaceae 2666 Leaf 27 BetaineAgeratum conyzoides L Asteraceae 4812 Leaf 37 Essential oilsAsteracantha longifolia L Acanthaceae 0234 Stem 74 GlycosidesAzadirachta indica A Juss Meliaceae D0204 Leaf bark 67 Tannins

Borassus flabellifer L Arecaceae D0202 Root 08 Flavonoidsphenolics

Boerhavia erecta L Nyctaginaceae 10897 Whole plant 28 PhenolicsCalotropis procera (Ait)Ait f Asclepiadaceae D073 Root-bark 13 Terpenoids

Calotropis gigantea (L)RBrex Ait Asclepiadaceae D070 Milky latex 48 Alkaloids

Cassia auriculata L Caesalpiniaceae 0141 Leaf 49 Saponins

Cassia occidentalis L Caesalpiniaceae 0111 Root 95 Flavonoidssaponins

Cassia tora L Caesalpiniaceae 0100 Stem bark 69 SaponinsCassia fistula L Caesalpiniaceae 037 Whole plant 79 SaponinsCardiospermumhalicacabum L Sapindaceae 0125 Whole plant 58 Flavonoids

Catharanthus roseus (L)GDon Apocynaceae 0029 Leaf

root1407 Alkaloids

Cinnamomum zeylanicumGarcin ex Blume Lauraceae 00209 Bark 31 Essential oil

TanninCinnamomum inersReinw ex Blume Lauraceae 043-c Leaf bark Alkaloids

Cissus quadrangularisRoxb Vitaceae D02023 Leaf 68 Glycosides

Citrus acida Roxb Hookf Rutaceae 0213 Leaf 52 SaponinsTerpenoids

Centella asiatica (L) Umbelliferae 0138 Whole plant 89 FlavonoidsAlkaloids

Clerodendrum inerme (L)Gaertn Verbenaceae D02043 Stem 78 Sterols diterpenes

Clitoria ternatea L Papilionaceae D02026 Seed 98 ProteinCleistanthus collinus(Roxb) Benth andHookf

Euphorbiaceae 0011 Whole plant 003 Cleistanthincollinusin

Cleome gynandropsis L Capparidaceae 12247 Leaf 62 GlycosidesCleome viscose L Capparidaceae 29999 Leaf 27 Phenolics

6 Evidence-Based Complementary and Alternative Medicine

Table 1 Continued

Scientific name Family Voucher specimen Plant parts Yield (gm) Phytochemicalanalysis

Coccinia grandisWampA Cucurbitaceae D02030 Leaf root 09 GlycosidesCymbopogon citratus(DC) Gramineae D012 Root 025 Essential oil

Datura metel L Solanaceae D02038 Leaf stem 39 SteroidsEucalyptus globulus Labill Myrtaceae D0220 Leaf 12 TerpenoidsEclipta alba (L) Hassk Asteraceae D028 Whole plant 07 PhenolicEuphorbia hirta Linn Euphorbiaceae 0018-c Whole plant 012 mdash

Eclipta prostrata (L) Asteraceae D210 Leaf 110 Triterpenoidsaponin

Eugenia caryophyllus(Sprengel) Bullock ampHarrison

Myrtaceae 0025 Flower buds 116 Essential oils

Elettaria cardamomumWhite et Mason Zingiberaceae 0009 Fruit pods 317 Essential oils

Gloriosa superba L Liliaceae 020-S Tuber 108 Alkaloids phenol

Jatropha curcas L Euphorbiaceae 015 Whole plant 53 Alkaloidsflavonoids

Hyptis suaveolens (L) Poit Lamiaceae 24688 Leaf 63 Essential oilHemidesmus indicus L Asclepiadaceae D-009 Roots CoumarinsIchnocarpus frutescens(L) RBr Apocynaceae 0110 Root flower 73 Terpenoids

Leucas aspera (Willd)Link Labiatae 0114 Leaf 83 Triterpenes

Lawsonia inermis L Lythraceae T261 Leaf 09 Glycosidesphenolic

Madhuca longifolia (L) JFMacbr Sapotaceae D01415 Nut 93 Sitosterol

Merremia hastate L(Desr) Hallierf Convolvulaceae 10894 Whole plant 40 Alkaloids

Mentha piperita L Lamiaceae 0217-c Whole plant 07 Essential oilsMorinda tinctoria Roxb Rubiaceae 0122 Leaf 14 GlycosidesMimosa pudica L Mimosaceae 0071 Whole plant 06 mdashOldenlandia umbellata L Rubiaceae D02047 Leaf 44 AlkaloidsOcimum sanctum L Lamiaceae 0016 Whole plant 30 AlkaloidsPiper attenuatum BuchHamex Miq Piperaceae 007 Flower 46 Alkaloids

Plumbago zeylanica (L)Cav Plumbaginaceae 0121 Root 48 Plumbagin

Plectranthus amboinicus(L) Spreng Lamiaceae 0410 Whole plant 12 Essential oils

terpenoidsPhyllanthus debilis L(Klein ex Willd) Euphorbiaceae 0120 Whole plant 49 Polyphenol

Phyllanthusmadraspatensis L Euphorbiaceae 0117 Whole plant 50 Polyphenol

Premna tomentosaWilld Verbenaceae 0129 Leaf 53 DiterpenesRosmarinus officinalis L Lamiaceae 0017 Root 023 Essential oilsRauwolfia serpentine L Apocynaceae 020-S Root 115 AlkaloidSebastiania chamaelea (L)Muell Arg Euphorbiaceae 0034 Leaf 11 Polyphenol

Solanum trilobatum L Solanaceae D02054 Leaf flower 40 Tannins

Evidence-Based Complementary and Alternative Medicine 7

Table 1 Continued

Scientific name Family Voucher specimen Plant parts Yield (gm) Phytochemicalanalysis

Sphaeranthus indicus L Asteraceae D02060 Whole plant 10 Essential oilSwertia chirata (L) Ham Gentianaceae D0540 Whole plant 16 GlycodisesStrychnos nux-vomica L Loganiaceae S-22 Nuts 036 AlkaloidsTragia involucrata L Euphorbiaceae D068 Leaves 16 ShellsolTinospora cordifolia(Willd) Miers ex Hooff ampThoms

Menispermaceae 0118 Leafroot stem 50 Glycosides tannins

Tridax procumbens L Compositae 10649 Leaf 18 FlavonoidsTerminalia arjuna (DC)W amp A Combretaceae 033-c Bark 80 Phenolics

Tephrosia purpurea (L)Pers Fabaceae S-43 Whole plant 08 Isoflavone

Vitex negundo L Verbenaceae 0031 Leaf 24 TerpineolVetiveria zizanioides L Gramineae 0051 Root 103 Essential oilWithania somnifera (L)Dunal Solanaceae D02063 Root 21 Alkaloids

Wedelia calendulacea Less Asteraceae S-24 Leaves FlavonoidsZingiber officinale Rosc Zingiberaceae 0327 Rhizome 23 TanninsZanthoxylum limonella(Dennst) Alston Rutaceae 009 Bark 19 Alkaloids essential

oilClass of chemical compounds A alkaloids S saponins T tannins St steroids G glycosides T terpenoids P polyphenol P phenolics Sh shellsol Hhydrocarbon esters

the site of snakebite and decoction is given orally fortreatment by indigenous people Besides these tribes relyon the medicinal plants as ingredients for fabricating a kindof medicated stone for health management ldquoVishakallurdquo(poison stone) is used by the indigenous groups called Kaniin Kerala India to treat a snakebiteWhen the stone is placeddirectly on the bitten area it sticks to the body to absorb thepoison and then become detached when absorption seems tobe complete The ingredients of Vishakallu stones are madewith leaves of Ocimum sanctum Anisomeles malabaricaLeucas aspera Piper betle Santalum album and the pebblescollected from the river bank

35 Survey of Medicinal Plants and Their Health Care ValuesThe present study is an attempt to provide scientific basis andobtain justification for the traditional beliefs of reliance on arich diversity of ethnomedicinal plants along with the richheritage of traditional medicine practices related to healthcare system made available by the primitive tribal commu-nities located at different settlements The native traditionalpractitioners called ldquovaidyarsrdquo have a good knowledge aboutthe traditional plants locally available for treatment of var-ious diseases (Figures 3(a)ndash3(p)) Such traditional medicalknowledge is used for preparing home remedies ill healthprevention and routine health maintenanceThis knowledgeis also applicable to cover other sectors of social life Duringthe ethnobotanical survey the wealth of 78 medicinal plantspecies used by the indigenous tribal community for varioustypes of health treatment was documented The botanical

names family names parts used chemical constituents andtheir application are provided (Table 1)

36 Antimicrobial Activity of Crude Extracts In this studywe reported the antimicrobial screening of methanolic crudeextracts of 78 medicinal plants (Table 2) Results revealedthat 68 plant extracts displayed potent activities against oneor more Gram-positive and -negative bacteria Of whichTragia involucrata Citrus acida Aegle marmelos Adha-toda vasica Calotropis procera Andrographis paniculata andMentha piperita Azadirachta indica Sphaeranthus indicusand Elettaria cardamomum showed the highest antibacte-rial activity against the multidrug resistant B pseudomallei(KHW and TES) and S aureus at 100120583gmL concentrationThe extracts showed pronounced antibacterial activity withtheir inhibitory zones ranging from 20 to 31mm in diam-eter as compared to the standard drugs chloramphenicoland ceftazidime (29ndash33mm) The majority of the plantsdemonstrated a powerful antimicrobial potency against themultidrug resistant strains of B pseudomallei (KHW andTES) K pneumonia and S aureus Approximately twenty-one plant extracts exerted only a weak or moderate effectagainst the tested bacteria while the crude extract of 13plants failed to show any effect at all Except for the plantextracts of T involucrata A lanceolata A vasica and Sindicus extracts the majority of the plant extracts wereineffective against theV damsela infection fascinatingly only11 plants exhibited activity against P aeruginosa of whichS indicus M piperita and C procera were found to havevery strong inhibition of bacteria at the tested concentrations

8 Evidence-Based Complementary and Alternative Medicine

Table 2 Antimicrobial activity of methanol extract of aromatic medicinal plants and essential oils evaluated against multidrug resistant(MDR) human pathogens at 100120583gmL concentration

Scientific name Microorganisms growth inhibition zones (6 millimeter in diameters)KHW TES Kp Kpr Pa Sa Stp Sp Vd Vd

Adhatoda vasica Nees 28 19 22 18 12 mdash 10 17 16 mdashAegle marmelos (L) Correa ex Roxb 29 17 9 10 mdash 15 mdash mdash mdash mdashAlangium salvifolium (L) f Wangerin 15 10 8 8 mdash 12 9 10 mdash mdashAndrographis echioides L 12 9 mdash mdash mdash 17 8 9 mdash mdashAndrographis paniculataWallich ex Nees 26 21 19 13 mdash 25 8 16 12 mdashAcalypha indica L mdash mdash mdash mdash mdash 18 mdash mdash mdash mdashAcalypha lanceolata L mdash mdash 12 13 mdash 10 mdash mdash 21 mdashAchyranthes aspera L mdash mdash 7 14 mdash 15 12 11 mdash mdashAgeratum conyzoides L 10 8 mdash mdash mdash mdash mdash mdash mdash mdashAsteracantha longifolia L 16 12 mdash mdash mdash 15 mdash mdash mdash mdashAzadirachta indica A Juss 15 17 21 16 14 23 12 14 mdash mdashBorassus flabellifer L 9 10 17 8 mdash mdash 9 10 mdash mdashBoerhavia erecta L mdash mdash 8 9 7 16 mdash mdash mdash mdashCalotropis procera (L) mdash mdash 15 mdash 18 28 9 mdash mdash mdashCalotropis gigantea (L) RBrex Ait 11 9 mdash mdash mdash 20 mdash mdash 9 8Cardiospermum halicacabum L 23 mdash mdash 14 mdash 9 19 mdash mdash mdashCatharanthus roseus (L) GDon 13 7 12 9 mdash 15 11 8 mdash mdashCassia auriculata L 17 13 12 mdash mdash 19 13 mdash mdash mdashCassia occidentalis L 18 mdash mdash mdash mdash mdash mdash mdash mdash mdashCassia tora L mdash mdash mdash mdash mdash mdash mdash mdash mdash mdashCassia fistula L mdash mdash mdash mdash mdash mdash mdash mdash mdash mdashCitrus acida Roxb Hookf 26 22 mdash 12 29 9 13 8 mdashCissus quadrangularis L mdash mdash mdash mdash mdash mdash mdash mdash mdash mdashCinnamomum zeylanicum Garcin exBlume 14 16 7 20 mdash 22 19 7 mdash mdash

Cinnamomum iners Reinw ex Blume 20 16 mdash 15 mdash 16 12 mdash mdash mdashRosmarinus officinalis L mdash 9 mdash 10 mdash 7 mdash 8 mdash mdashCentella asiatica (L) 9 8 mdash mdash mdash 11 mdash mdash mdash mdashClerodendrum inerme (L) Gaertn 13 7 12 mdash mdash 15 11 8 mdash mdashClitoria ternatea L mdash mdash mdash mdash mdash mdash mdash mdash mdash mdashClitoria ternatea L 16 12 mdash 13 mdash 8 10 mdash mdash mdashCleome gynandropsis L mdash mdash 12 19 11 mdash 9 15 12 mdashCleome viscose L mdash mdash 8 10 14 20 mdash mdash mdash mdashCoccinia grandisWampA 9 10 17 8 mdash mdash 9 10 mdash mdashCymbopogon citratus (DC) 16 18 mdash 17 mdash 14 mdash mdash mdash mdashDatura metel L mdash mdash mdash mdash mdash mdash mdash mdash mdash mdashEclipta alba (L) Hassk 20 mdash mdash 9 mdash mdash 10 mdash mdash mdashEuphorbia hirta Linn 11 mdash mdash mdash mdash 16 mdash mdash mdash mdashEucalyptus globulus Labill mdash mdash mdash 7 mdash mdash mdash 7 mdash mdashEugenia caryophyllus Bullock amp Harrison mdash 7 mdash 11 mdash 9 mdash 8 mdash mdashElettaria cardamomumWhite et Mason 21 20 7 14 mdash 22 12 17 mdash mdashHyptis suaveolens (L) Poit 8 10 8 7 mdash mdash mdash mdash mdash mdashIchnocarpus frutescens (L) WJ Aiton mdash mdash mdash mdash mdash mdash mdash mdash mdash mdashJatropha curcas L mdash mdash mdash 8 mdash 11 mdash 12 mdash 7Leucas aspera (Willd) Link 9 10 17 8 mdash 12 9 10 mdash mdashLawsonia inermis L mdash mdash mdash mdash mdash mdash mdash mdash mdash mdashMadhuca longifolia (L) JF Macbr 18 16 14 19 12 mdash 8 7 9 mdash

Evidence-Based Complementary and Alternative Medicine 9

Table 2 Continued

Scientific name Microorganisms growth inhibition zones (6 millimeter in diameters)KHW TES Kp Kpr Pa Sa Stp Sp Vd Vd

Merremia hastate L (Desr) Hallierf mdash mdash mdash mdash mdash mdash mdash mdash mdash mdashMorinda tinctoria Roxb mdash mdash mdash mdash mdash mdash mdash mdash mdash mdashMentha piperita L 23 17 26 12 20 25 19 mdash mdash mdashOcimum sanctum L 12 9 11 7 mdash 15 7 8 mdash mdashOldenlandia umbellata L mdash mdash mdash mdash mdash 17 mdash mdash mdash mdashPiper attenuatum Buch Hamex Miq 13 7 12 21 mdash 17 11 8 mdash mdashPlumbago zeylanica (L) Cav 9 10 17 8 mdash 12 9 10 mdash mdashPlectranthus amboinicus (L) Spreng mdash mdash mdash 8 11 15 mdash mdash mdash mdashPhyllanthus debilis L (Klein ex Willd) 7 8 mdash 9 mdash 9 18 7 mdash mdashPhyllanthus maderaspatensis L 17 mdash mdash mdash mdash mdash mdash mdash mdash mdashPremna tomentosaWilld 13 10 mdash 15 mdash 9 10 mdash mdash mdashGloriosa superba L 17 16 7 8 mdash 15 mdash mdash mdash 8Sebastiania chamaelea (L) Muell Arg 19 12 mdash 17 mdash 13 19 8 mdash mdashSolanum trilobatum L mdash mdash 13 9 mdash 8 mdash mdash mdash mdashSphaeranthus indicus L 20 18 21 7 22 mdash 11 9 mdash 16Swertia chirata (L) Ham mdash mdash mdash mdash mdash mdash mdash mdash mdash mdashTerminalia arjuna (W amp A) mdash mdash mdash mdash mdash 16 mdash mdash mdash mdashTinospora cordifolia (Willd) Miers exHooff ampThoms mdash mdash 23 16 mdash 12 15 mdash mdash mdash

Tridax procumbens L 9 8 7 mdash mdash 14 mdash mdash mdash mdashTragia involucrata L 25 23 20 mdash mdash 31 28 22 19 mdashVitex negundo L mdash mdash mdash mdash mdash 14 mdash mdash mdash mdashVetiveria zizanioides (L) 11 9 7 7 mdash 16 mdash mdash 8Withania somnifera (L) Dunal 12 20 mdash mdash mdash 15 12 mdash mdash mdashZingiber officinale Rosc 14 11 7 15 mdash 7 12 7 mdash mdashZanthoxylum limonella (Dennst) Alston 13 7 12 9 mdash mdash 11 8 mdash mdashChloramphenicol (30120583gdisc) 21 12 15 17 29 16 15 18 13 11Ceftazidime (30 120583gdisc) 33 16 22 19 16 25 21 20 12 15lowastBacteria (+minus) Results obtained in the disc diffusion assay antibacterial activity is expressed as the mean plusmn SD (119899 = 3) of the inhibition by the extract andits diameter around the discs One way analysis of variance was performed (mean plusmn SD 119899 = 3 replicates) Size of inhibition zones were including the sterileblank discs 6 millimeter (mm) in diameters Absence of bacterial inhibition indicates (mdash) antibiotic disc (30 120583gdisc)

Interestingly sixteen plants such as Andrographis echioidesC auriculata C viscose C gigantea T arjuna Oldenlandiaumbellata Boerhavia erecta and E hirta exerted a strongactivity against the Gram-positive S aureus bacteria

37 Phytochemical Screening of Plants The results obtainedfrom the phytochemical screening as shown in Table 1 indi-cate the presence of various types of secondary metabolitessuch as polyphenols tannins saponins alkaloids and gly-cosidespolysaccharides Most of the plant extracts relativelyrich in alkaloids phenols flavonoids polyphenols tanninssterols and terpenoids were found to inhibit the growth oforganisms

38 Antimicrobial Activity of Fractioned Compounds Activecomponents were purified from the most active extractsfor further testing The compound shellsol of T involucrata

and C acida exhibited the most potent action againstthe antibiotic resistant strains of B pseudomallei (KHW)S aureus B pseudomallei (TES) and K pneumoniae Amarmelos was also found to inhibit the growth of B pseudo-mallei (KHW) more effectively than other tested bacteria Avasica showed the broad spectrum growth inhibitory activityon B pseudomallei (KHW) K pneumoniae K pneumoniaeresistant B pseudomallei (TES) S pyogenes and V damselaHowever E cardamomum displayed antimicrobial activityon some of the B pseudomallei (KHW) S pyogenes Bpseudomallei (TES) and S typhi strains Similarly A indicaexerted the growth inhibition on K pneumoniae and Saureus Remarkably Sebastiania chamaelea was more activeagainst K pneumoniae K pneumoniae and S pyogenesThe compound from S indicus inhibited the growth of Kpneumoniae K pneumoniae B pseudomallei (KHW andTES) and S typhi strains as compared to the activity shown

10 Evidence-Based Complementary and Alternative Medicine

Figure 2 Diverse biodiversity richness of medicinal plants in Western and Eastern Ghats (a) Topography of plant covering area in Kollihills (Namakkal district Tamil Nadu) (b) Aerial view of occurrence of medicinal plants in Mundanthurai hills (Tirunelveli district TN) (c)Pachamalai hills (Trichy district) and its natural vegetation inhabitants for Malaiyali tribes (d) Deforestation of natural herbal resources dueto urbanization in Kalrayan hills (Salem district) in the Eastern Ghats of TN (e) Medicinal plants and its various parts used by the natives(traditional healers) for the treatment of diverse human illness with a very high percentage of leaves and whole plants often used for herbaldrug preparation by the local practitioners (f) Various category of plants like shrub herb climbers and tree and the parts used in medicine

Evidence-Based Complementary and Alternative Medicine 11

Figure 3 Medicinal aromatic herbs spices and toxic plants were collected from the tribal areas of the Western and Eastern Ghats region inTamil Nadu India (a) A vasica Nees (leaf) (b) Eclipta alba (L) Hassk (whole plant) (c)Mimosa pudica L (whole plant) (d) P amboinicus(L) Spreng (whole plant) (e) T procumbens (L) (whole plant) (f) Euphorbia hirta Linn (whole plant) (g) A paniculata Wallich ex Nees(H) C roseus (L) GDon (whole plant) used for therapy (i) Cinnamomum iners Reinw ex Blume (leaf) (j) E globulus Labill (leaf and bark)(k) Z officinale Rosc (Rhizome) (l) E caryophyllus (Sprengel) Bullock and Harrison (flower buds) (m) M piperita L (whole plant) (n) Ccitratus (DC) Clitoria ternatea L (whole plant) (o) C zeylanicum Garcin ex Blume (bark) (p) Elettaria cardamomumWhite et Mason (fruitpod) used for medicine and food preparation

by the crude extracts (Figure 4)The antimicrobial efficacy offractions collected from the oil yielding plants was also com-pared with that of the tested compounds C zeylanicum andR officinaliswere themost sensitive in controlling the growthof B pseudomallei (KHW) S aureus K pneumonia and Spneumoniae Fascinatingly all the compounds obtained fromaromatic plants except those from E globules were foundto be very effective against the multidrug resistant humanpathogen B pseudomallei (KHW) that causes melioidosis

On the other hand compounds from C citrates O sanctumE caryophyllus and Z zizanioide showed some promisingeffect only against S aureus (Figure 5) The activity of thecompounds were pronounced more than that of the oilyielding plant fractions

39 Minimum Inhibitory Concentrations (MICs) The antibi-otic potential of the purified fractions was obtained fromthe MIC determination The hydrocarbon ester shellsol

12 Evidence-Based Complementary and Alternative Medicine

(mean plusmn SD n = 3)

0

5

10

15

20

25

30

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn V

d St

Bacteria (+minus)

lowast

lowast

lowast

A vasica

In vitro assay(P

lant

frac

tion)

Inhi

bitio

n zo

nes (

mm

)

(a)

(mean plusmn SD n = 3)

0

5

10

15

20

25

30

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn V

d St

Bacteria (+minus)

lowast

A marmelos

Inhi

bitio

n zo

nes (

mm

)

In vitro assay

(b)

(mean plusmn SD n = 3)

0

5

10

15

20

25

30

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn V

d St

Bacteria (+minus)

lowastlowast

A indica

Inhi

bitio

n zo

nes (

mm

)

Incu

batio

n (37∘C

24 h

)

(c)

(mean plusmn SD n = 3)

0

5

10

15

20

25

30

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn V

d St

Bacteria (+minus)

lowast

lowast lowast

35

C acida

Inhi

bitio

n zo

nes (

mm

)(P

lant

frac

tion)

(d)

(mean plusmn SD n = 3)

0

5

10

15

20

25

30

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn V

d St

Bacteria (+minus)

lowastlowast

lowast

S chamaelea

Inhi

bitio

n zo

nes (

mm

)

(e)

(mean plusmn SD n = 3)

0

5

10

15

20

25

30

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn V

d St

Bacteria (+minus)

lowastlowast

lowastlowast

S indicus

Inhi

bitio

n zo

nes (

mm

)

Incu

batio

n (37∘C

24 h

)

(f)

(mean plusmn SD n = 3)

0

5

10

15

20

25

30

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn V

d St

Bacteria (+minus)

lowast

lowast lowast

E cardamomum

Inhi

bitio

n zo

nes (

mm

)(P

lant

frac

tion)

(g)

(mean plusmn SD n = 3)

0

5

10

15

20

25

30

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn V

d St

Bacteria (+minus)

lowast

lowast

lowast

lowast

Shellsol

35

Inhi

bitio

n zo

nes (

mm

)

Incu

batio

n (37∘C

24 h

)

(h)

Figure 4 In vitro antimicrobial activity of purified fractions from the most active plant extracts tested against bacteria Growth inhibitionzones were measured and analyzed with mean plusmn standard deviation (SD) (119899 = 3) using one way analysis of variance Level of significanceat (lowast119875 gt 001) Most of the fractions exerted a potent inhibitory effect against multidrug resistant Gram-negative bacteria (B pseudomalleistrains KHW and TES) K pneumonia and Gram-positive bacteria S aureus

(T involucrata) andC acida showed an interesting inhibitorypotential against S aureus (MIC of 78 120583gmL) and B pseu-domallei strain of KHW (MIC of 156 120583gmL) A vasicashowed an MIC of 156120583gmL against B pseudomallei(KHW) and an MIC of 3125120583gmL against K pneumo-niae K pneumoniae S pyogenes and V damsela strainsFractions from A marmelos and terpenoid from A indicaexerted bacteriostatic effect with MIC values of 3125120583gmLon some selected bacteria including B pseudomallei ofKHW S aureus and B pseudomallei of TES The MIC of3125 120583gmL was found for E cardamomum against S aureus

K pneumonia and S pyogenes S indicus displayed a verystrong inhibition against MDR K pneumoniae (MIC of156 120583gmL) and against B pseudomallei (KHW and TES)at MIC of 3125 120583gmL When the antimicrobial efficacies ofpurified fractions from aromatic plants were compared theC zeylanicum fraction displayed an important antimicrobialeffect against S aureus (MIC of 78 120583gmL) MDR B pseu-domallei of KHW (MIC of 156 120583gmL) and S pneumoniae(MIC of 3125 120583gmL) The essential oil from M piperateashowed MIC value of 3125120583gmL against K pneumoniae Saureus and B pseudomallei (KHW) respectivelyO sanctum

Evidence-Based Complementary and Alternative Medicine 13

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn

Bacteria (+minus)

0

5

10

15

20

25

30

Inhi

bitio

n zo

nes (

mm

)(P

lant

frac

tion)

lowastlowast

lowast

(mean plusmn SD n = 3)In vitro assay

C citratus

(a)

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn

Bacteria (+minus)

0

5

10

15

20

25

30

Inhi

bitio

n zo

nes (

mm

)

lowast

lowast

lowastlowast

lowast

lowast

(mean plusmn SD n = 3)In vitro assay

35

C zeylanicum

(b)

Bacteria (+minus)

0

5

10

15

20

25

30

Inhi

bitio

n zo

nes (

mm

)

lowast

lowast

lowast

(mean plusmn SD n = 3)

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn V

d St

O sanctum

Incu

batio

n (37∘C

24 h

)

(c)

Bacteria (+minus)

0

5

10

15

20

25

Inhi

bitio

n zo

nes (

mm

)

lowast

lowast

(mean plusmn SD n = 3)

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn V

d St

M piperita

(Pla

nt fr

actio

n)

(d)

Bacteria (+minus)

0

5

10

15

20

25

30

Inhi

bitio

n zo

nes (

mm

)lowast

lowast

lowast

(mean plusmn SD n = 3)

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn V

d St

E caryophyllus

(e)

0

5

10

15

20

25

30

Inhi

bitio

n zo

nes (

mm

) (mean plusmn SD n = 3)

Bacteria (+minus)

KHW

TES

Kp

KprPa Sa

Stp

Spn

E globulus

Incu

batio

n (37∘C

24 h

)

(f)

Bacteria (+minus)

0

5

10

15

20

25

30

Inhi

bitio

n zo

nes (

mm

)(P

lant

frac

tion)

lowast

lowast

lowast

lowast

(mean plusmn SD n = 3)

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn V

d

R officinalis

(g)

0

5

10

15

20

25

30

Inhi

bitio

n zo

nes (

mm

)

lowast

lowast

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn V

d St

Bacteria (+minus)V zizanioides

(mean plusmn SD n = 3)

Incu

batio

n (37∘C

24 h

)

(h)

Figure 5 Comparison of antimicrobial effect of plant compounds obtained from the most popularly used sources of essential oils as assayedby the disc-diffusion method in vitro It displayed a powerful activity against B pseudomallei and S aureus than the other bacteria strainsOther compounds showed only a moderate or weak action against the tested bacteria Values for zone of bacterial growth inhibition werepresented as mean plusmn SD (119899 = 3) with level of significance at (lowast119875 gt 001)

and C citratus fractions also showed antimicrobial activity(MICs of 3125ndash125120583gmL) only at higher concentrationsagainst the tested bacteria In addition to that higher concen-trations (gt250120583gmL) (of Vetiveria fractions) were requiredto inhibit Vibro species and others (including E globulusfractions) failed to show any effect at tested concentrations(78ndash125 120583gmL) However the purified fractions (from mostactive medicinal plants) showed strong bacteriostatic inhibi-tion against the tested organisms (Table 3)

310 Cytotoxic Effects of Plants When the components wereassayed for cytotoxicity against the normal human skinfibroblasts (HEPK) cells the compounds obtained from

E cardamomum T involucrata S indicusC acidaA vasicaA marmelos A indica and A paniculata did not showtoxicity up to 1000120583gmL (see Figures S1 and S2 in Supple-mentaryMaterial available online at httpdxdoiorg1011552013525613) A slight reduction of cell proliferation wasnoted only at higher doses (2000120583gmL) In contrast cellproliferation was markedly reduced after exposure of HEPKcells to O sanctum E globulus V zizanioides C citratusand E globulus compounds There was no gradual reductionin skin cell proliferation seen after exposure to C zeylan-icum R officinalis and M piperita (see Figures S3 and S4)compounds The toxicity was found to be concentration-dependent when the skin fibroblasts (HEPK) cells were

14 Evidence-Based Complementary and Alternative Medicine

Table 3 Minimum inhibitory concentrations (MICs) of purified plant fractions and essential oils against antibiotic resistant bacteria

Botanical name Family Parts used Gram-positive and -negative bacteria (MICs 120583gmL)KHW TES Kp KPr Sa Stp Sp Vd St

A indica Juss Meliaceae Seed (fraction) 3125 625 125 lt250 3125 mdash mdash mdash mdashA marmelos (L) Rutaceae Root-bark (F) 3125 3125 mdash mdash 3125 625 125 250 mdashA paniculata Nees Acanthaceae Leaf (fraction) 250 mdash 125 mdash 625 mdash mdash 125 mdashA vasica Nees Acanthaceae Fraction (Stem) 156 3125 3125 625 mdash lt250 3125 3125 mdashC acida Roxb Rutaceae Leaf (fraction) 156 625 mdash mdash 78 mdash 3125 mdash mdashE cardamomumWhite et Mason Acanthaceae Fraction (WP) 625 mdash 3125 mdash 3125 250 625 mdash mdashS indicus (L) Euphorbiaceae Whole plant (F) 3125 125 156 mdash 625 mdash mdash lt250 mdashT involucrata (L) Euphorbiaceae Shellosol (leaf) 156 3125 mdash mdash 78 625 mdash mdash mdashCinnamomum zeylanicum (L) Lauraceae Bark (fraction) 156 625 mdash mdash 78 3125 mdash mdash 125Cymbopogon citratus (L) Graminae Leaf (fraction) 250 lt250 625 mdash 625 125 mdash mdash mdashEugenia caryophyllus (L) Myrtaceae Flower buds (F) 625 125 mdash mdash 625 mdash mdash mdash mdashEucalyptus globulus (L) Myrtaceae Fraction (leaf) mdash mdash mdash mdash mdash mdash lt250 mdash lt250Mentha piperita (L) Labiatae Fraction (WP) mdash 625 3125 mdash 3125 mdash lt250 mdash mdashOcimum sanctum (L) Labiatae Leaf (fraction) 625 125 mdash 3125 625 mdash 125 lt250 mdashRosmarinus officinalis (L) Labiatae Rosemary oil 3125 mdash mdash mdash 3125 mdash mdash mdash mdashVetiveria zizanioides (L) Graminae Root (fraction) mdash 125 125 mdash mdash mdash gt250 gt250The bacterial growth inhibitory activity was compared for significant differences within the bacterial strains by broth-dilution method at 250 125 625 3125156 and 78 120583gmL F fractions

exposed to various compounds The cell proliferation wasincreased by the influence of the plant components at thelower concentrations Whereas the oil yielding plant com-pounds showed inhibition of cell proliferation and toxicity athigher doses 250ndash1000120583gmL

4 Discussion

The Western Ghats is considered as one of the richestbiodiversity hotspots in the world [41] In this survey wecollected nearly 78 medicinal plants from Western andEastern Ghats that are edible and popularly used for curingvarious ailments including snakebite Traditional remedieshave a long-standing history in many tribal settlements inTN India and they continue to provide useful and applicabletools for treating ailments [42] The ingredients that makeup the ldquoVishakallurdquo stone which is used as an antidote forsnakebite are different herbs and pebbles available from theriver banks Likewise aqueous paste and decoction obtainedfrom the leaves of A paniculata are widely used for snakebitetreatment by indigenous people [43] Previous studies havereported that ethnomedicine plays major roles in conservingthe disappearing knowledge of tribal communities [44ndash47]The traditional beliefs of reliance on a rich diversity ofethnomedicinal plants located at different settlements havealso been confirmed in another study [48 49] Herein weexplored the various types of traditional practices reported bythe primitive tribal communities with a view to gain furtherknowledge from such studies

In the present investigation potentially rich sources oftribal medicine (71 plants) were scientifically evaluated fortheir antibacterial activity against the MDR bacteria and theaccumulated data was disseminated for the first time to the

scientific community Out of the 71medicinal plants screenedfor the antibacterial activity 10 of them (T involucrataC acida A marmelos A vasica C procera A paniculataand M piperita A indica S indicus and E cardamomum)displayed the highest antibacterial activity against the mul-tidrug resistant B pseudomallei (KHW and TES) and Saureus strains The antibacterial activity of those crude plantextracts was as equally effective as that of the standard drugsOur findings corroborated with the previous reports madeon the antistaphylococcal activity of tribal medicinal plants[34 50 51] On the other hand isolated components from themost active extracts of T involucrata shellsol and C acidaexhibited the most potent action against the antibiotic resis-tant B pseudomallei (KHW) K pneumoniae and S aureusstrainsThese results further confirmed our previous findingson the leaves of T involucrata and its compounds hydrocar-bon ester-like shellsol which displayed a high antibacterialeffect against the different bacterial strains especially thatof S aureus [34] Eugenol and caryophyllene are the activeagents contained in the M piperita [52] and O sanctumplants which are believed to be mainly responsible for theantimicrobial properties of these plants [53] Interestinglythe compounds obtained from the aromatic plants such asC zeylanicum and R officinalis were also found to be veryeffective against the multidrug resistant human pathogen Kpneumoniae S aureus S typhi and B pseudomallei (KHW)that causes melioidosis

The inhibitory potential determined for the shellsol(T involucrata) vaseline (A vasica) C acida and C zey-lanicum indicates that the MIC of 78ndash3125120583gmL foundagainst the B pseudomallei of KHW K pneumoniae Kpneumoniae S pyogenes and V damsel and S pneumoniaewas quite low Similarly lower MIC values were found for

Evidence-Based Complementary and Alternative Medicine 15

M piperita (MIC of 113ndash225 mgmL) against the abovebacterial strains [52] MIC was found for the most activealcohol extracts of A salvifolium (MIC 0034ndash0263mgmL)on S aureus [54] and the MIC for S trilobatum aqueousextracts determined against the tested organisms rangedfrom 006 to 05mgmL [55] C zeylanicum was foundto have an effective antibacterial activity (MIC 64 120583gmL)against P aeruginosa E coli B subtilis and S aureus[56] Previously several investigators have demonstrated thatactive agents exert interesting activity against bacteria evenat lower concentrations tested [57 58] The bacteriostaticmechanism involves damage to the cell walls of bacteriafollowed by inhibition of protein synthesis that ultimatelyleads to bacterial death [59] The most active plants arewidely used by various tribes as traditional treatment (iecut wounds skin infection and scabies) thus indicatingthe potential for further development into promising drugsIn order to ascertain the safety and efficacy of the mostactive compounds their effect on human skin fibroblast cellswas evaluated Chemical constituents of E cardamomumT involucrata S indicus C acida A vasica A marmelosA indica and A paniculata plants failed to produce anynoticeable toxicity up to 1000120583gmL Although some ofthe tested compounds exhibited a slight reduction of cellproliferation and some minor morphological changes suchchanges were insignificant at lower doses and became evidentonly at higher doses However certain aromatic compoundsof O sanctum E globulus V zizanioides C citratus and Eglobulus plants showed reduction of cell proliferation againstHEPK cells

Our phytochemical screening also provides evidenceof the presence of several types of compounds that aremainly responsible for the remarkable antibacterial effectof these plants The differences noted for the bactericidalactivity of various plant extracts in this study appears tobe directly related to the diversity of compounds (shown inparentheses) that are accumulated in the following plants(eg A marmelos and O umbellate) [60ndash62] Compoundslike tannins phenol and polyphenols can bind the Gram-negative bacteria to form a heavy soluble complex on the cellsurface which subsequently disturbs the availability of recep-tor on cells and kills the bacteria [63] Several species havingwide spectra of antimicrobial activity mainly due to theactive constituents such as essential oil phenolic compoundslike thymol carvacrol in oregano and thyme eugenol inclove and cinnamon were also identified previously [64 65]Essential oils degrade the cell wall interact with the cell com-ponents and then disrupt the cytoplasmic membrane [66]The antimicrobial effect of phenolic compounds may involvemultiplemodes of action including damage to themembraneprotein interference with membrane integrated enzymes[67] causing leakage of cellular components coagulation ofcytoplasm depletion of the proton motive force alterationof fatty acid and phospholipid constituents impairmentof enzymatic mechanisms for production and metabolismalteration of nutrient uptake and electron transport [68]influencing the synthesis of DNA and RNA and destroyingprotein translation and the function of the mitochondrionin eukaryotes [69] The mode of action of antimicrobial

agents depends on the type of microorganism and is mainlyrelated to their cell wall structure and the outer membranearrangement Most of the plant spices and herbs containcomplex phenolics (ie phenolic acids flavonoids tanninslignans coumarins quinines) In addition the mechanismsof action of each phenolic compound against various bacteriaare also very complicated [26 70] Further investigation istherefore required to understand the relationship betweenthe antimicrobial action and the chemical structure of everyphenolic compound in the tested extracts The informationavailable from previous pharmacological sources combinedwith the findings herein reported on the medicinal plantextracts may serve as essential data for future drug develop-ment to combat diseases caused by theMDRbacterial strains

Abbreviations

MDR Multidrug resistant bacteriaTES Strain of Burkholderia pseudomalleiMRSA Methicillin-resistant Staphylococcus aureusVRE Vancomycin-resistant enterococciMTB Mycobacterium tuberculosisCH3OH Methanol

CFU Colony forming unitHCl Hydrochloric acidTLC Thin layer chromatographyH2SO4 Sulphuric acids

UV Ultraviolet spectrophotometerHEPK Human skin fibroblasts

Acknowledgments

Theauthors are thankful to the ERI Loyola College ChennaiIndia for his support during the survey The authors alsothank the Department of Microbiology NUHS NUS forthe bacterial cultures used in this investigation The authorsare thankful to the tribal people for sharing their healingknowledge on medicinal aromatic plants

References

[1] M N Alekshun and S B Levy ldquoMolecular mechanisms ofantibacterialmultidrug resistancerdquoCell vol 128 no 6 pp 1037ndash1050 2007

[2] A D Russell ldquoAntibiotic and biocide resistance in bacteriaintroductionrdquo Symposium Series Society for Applied Microbiol-ogy vol 31 no 92 pp 1Sndash3S 2002

[3] D R Guay ldquoTreatment of bacterial skin and skin structureinfectionsrdquo Expert Opinion on Pharmacotherapy vol 4 no 8pp 1259ndash1275 2003

[4] A J Brosnahan and P M Schlievert ldquoGram-positive bac-terial superantigen outside-in signaling causes toxic shocksyndromerdquo FEBS Journal vol 278 no 23 pp 4649ndash4667 2011

[5] K Fellermann J Wehkamp K R Herrlinger and E F StangeldquoCrohnrsquos disease a defensin deficiency syndromerdquo EuropeanJournal of Gastroenterology and Hepatology vol 15 no 6 pp627ndash634 2003

16 Evidence-Based Complementary and Alternative Medicine

[6] L Chen S Lin R Agha-Majzoub L Overbergh C Mathieuand L S Chan ldquoCCL27 is a critical factor for the developmentof atopic dermatitis in the keratin-14 IL-4 transgenic mousemodelrdquo International Immunology vol 18 no 8 pp 1233ndash12422006

[7] J H Song V Thamlikitkul and P R Hsueh ldquoClinical andeconomic burden of community-acquired pneumonia amongstadults in the Asia-Pacific regionrdquo International Journal ofAntimicrobial Agents vol 38 no 2 pp 108ndash117 2011

[8] D A B Dance ldquoMelioidosisrdquo Current Opinion in InfectiousDiseases vol 15 no 2 pp 127ndash132 2002

[9] N J White ldquoMelioidosisrdquo The Lancet vol 361 no 9370 pp1715ndash1722 2003

[10] S Yang ldquoMelioidosis research in Chinardquo Acta Tropica vol 77no 2 pp 157ndash165 2000

[11] P R Hsueh L J Teng L N Lee et al ldquoMelioidosis an emerginginfection in Taiwanrdquo Emerging Infectious Diseases vol 7 no 3pp 428ndash433 2001

[12] C Walsh and S Fanning ldquoAntimicrobial resistance in food-borne pathogensmdasha cause for concernrdquo Current Drug Targetsvol 9 no 9 pp 808ndash815 2008

[13] M Vorachit P Chongtrakool S Arkomsean and S BoonsongldquoAntimicrobial resistance in Burkholderia pseudomalleirdquo ActaTropica vol 74 no 2-3 pp 139ndash144 2000

[14] F M Thibault E Hernandez D R Vidal M Girardet and JD Cavallo ldquoAntibiotic susceptibility of 65 isolates of Burkholde-ria pseudomallei and Burkholderia mallei to 35 antimicrobialagentsrdquo Journal of Antimicrobial Chemotherapy vol 54 no 6pp 1134ndash1138 2004

[15] R L S Sikarwar ldquoEthnogynaecological uses of plants new toIndiardquo Ethnobotany vol 12 no 1-2 pp 112ndash115 2002

[16] R Siva ldquoStatus of natural dyes and dye-yielding plants in IndiardquoCurrent Science vol 92 no 7 pp 916ndash925 2007

[17] P Kumar Singh V Kumar R K Tiwari A Sharma ChV Rao and R H Singh ldquoMedico-ethnobotany of lsquochatararsquoblock of district sonebhadra Uttar Pradesh Indiardquo Advances inBiological Research vol 4 no 1 pp 65ndash80 2010

[18] W H Lewis and M P Elwin-Lewis Medical Botany PlantsAffecting Human Health John Wiley amp Sons New York NYUSA 2003

[19] R Perumal Samy P N Pushparaj and P GopalakrishnakoneldquoA compilation of bioactive compounds fromAyurvedardquo Bioin-formation vol 3 no 3 pp 100ndash110 2008

[20] WorldHealthOrganization (WHO)TraditionalMedicine Strat-egy 2002ndash2005 Genera World Health Organization GenevaSwitzerland 2002

[21] P S Haddad G A Azar S Groom and M Boivin ldquoNaturalhealth products modulation of immune function and preven-tion of chronic diseasesrdquo Evidence Based Complementary andAlternative Medicine vol 2 no 4 pp 513ndash520 2005

[22] K Poonam andG S Singh ldquoEthnobotanical study of medicinalplants used by the Taungya community in Terai Arc LandscapeIndiardquo Journal of Ethnopharmacology vol 123 no 1 pp 167ndash1762009

[23] G B Mahady ldquoMedicinal plants for the prevention and treat-ment of bacterial infectionsrdquo Current Pharmaceutical Designvol 11 no 19 pp 2405ndash2427 2005

[24] V P Kumar N S Chauhan H Padh and M Rajani ldquoSearchfor antibacterial and antifungal agents from selected Indianmedicinal plantsrdquo Journal of Ethnopharmacology vol 107 no2 pp 182ndash188 2006

[25] R Nair T Kalariya and S Chanda ldquoAntibacterial activity ofsome plant extracts used in folk medicinerdquo Journal of HerbalPharmacotherapy vol 7 no 3-4 pp 191ndash201 2007

[26] D Kalemba and A Kunicka ldquoAntibacterial and antifungalproperties of essential oilsrdquo Current Medicinal Chemistry vol10 no 10 pp 813ndash829 2003

[27] A H Ashour ldquoAntibacterial antifungal and anticancer activi-ties of volatile oils and extracts from stems leaves and flowers ofEucalyptus sideroxylon and Eucalyptus torquatardquoCancer BiologyandTherapy vol 7 no 3 pp 399ndash403 2008

[28] M J Abad L M Bedoya L Apaza and P Bermejo ldquoTheArtemisia L genus a review of bioactive essential oilsrdquoMolecules vol 17 no 3 pp 2542ndash2566 2012

[29] H N Marzoug M Romdhane A Lebrihi et al ldquoEucalyptusoleosa essential oils chemical composition and antimicrobialand antioxidant activities of the oils from different plant parts(stems leaves flowers and fruits)rdquoMolecules vol 16 no 2 pp1695ndash1709 2011

[30] C Proestos D Sereli and M Komaitis ldquoDetermination ofphenolic compounds in aromatic plants by RP-HPLC and GC-MSrdquo Food Chemistry vol 95 no 1 pp 44ndash52 2006

[31] S Chao G Young C Oberg and K Nakaoka ldquoInhibition ofmethicillin-resistant Staphylococcus aureus (MRSA) by essentialoilsrdquo Flavour and Fragrance Journal vol 23 no 6 pp 444ndash4492008

[32] M Akin D Oguz and H T Saracoglu ldquoAntibacterial effectsof some plant extracts from Labiatae (Lamiaceae) growingnaturally around Sirnak-Silopi Turkeyrdquo International Journal ofPharmaceutical and Applied Sciences vol 1 no 1 pp 4ndash47 2010

[33] K Bayoub T Baibai D Mountassif A Retmane and ASoukri ldquoAntibacterial activities of the crude ethanol extracts ofmedicinal plants against Listeriamonocytogenes and some otherpathogenic strainsrdquo African Journal of Biotechnology vol 9 no27 pp 4251ndash4258 2010

[34] R P Samy P Gopalakrishnakone M Sarumathi P Houghtonand S Ignacimuthu ldquoPurification of antibacterial agents fromTragia involucratamdasha popular tribal medicine for wound heal-ingrdquo Journal of Ethnopharmacology vol 107 no 1 pp 99ndash1062006

[35] K M Matthew The Flora of Tamil Nadu Carnatic Part IndashIIIprinted at the Diocesan Press Madras India 1981ndash1983

[36] AW BauerWMKirby J C Sherris andM Turck ldquoAntibioticsusceptibility testing by a standardized single disk methodrdquoAmerican Journal of Clinical Pathology vol 45 no 4 pp 493ndash496 1966

[37] National Committee for Clinical Laboratory Standards Per-formance Standards For Antimicrobial Susceptibility Testing12th Informational Supplement Approved standard M100-S12National Committee for Clinical Laboratory Standards WaynePa USA 2002

[38] E A Elsinghorst ldquoMeasurement of invasion by gentamicinresistancerdquoMethods in Enzymology vol 236 pp 405ndash420 1994

[39] J B Harborne Phytochemical Methods Chapman and HallNew York NY USA 1976

[40] H Wagner S Bladt and E M Zgairski Plant Drug AnalysisSpringer New York NY USA 1984

[41] N Myers R A Mittermeler C G Mittermeler G A B daFonseca and J Kent ldquoBiodiversity hotspots for conservationprioritiesrdquo Nature vol 403 no 6772 pp 853ndash858 2000

[42] R Perumal Samy and S Ignacimuthu ldquoAntibacterial activity ofsome folkloremedicinal plants used by tribals inWestern Ghats

Evidence-Based Complementary and Alternative Medicine 17

of Indiardquo Journal of Ethnopharmacology vol 69 no 1 pp 63ndash712000

[43] K Nampoothiri A Chrispal A Begum S Jasmine K GGopinath and A Zachariah ldquoA clinical study of renal tubulardysfunction inCleistanthus collinus (Oduvanthalai) poisoningrdquoClinical Toxicology vol 48 no 3 pp 193ndash197 2010

[44] M Strathern UsefulKnowledge The Isiah Berlin Lecture Uni-versity of Manchester Brunswick UK

[45] C N Sunil and A K Pradeep ldquo(2001) Another new species ofTripogon (Poaceae) from Indiardquo Sida vol 19 pp 803ndash806 2005

[46] S Biber-Klemm and T Cottier Right To Plant GeneticResources and Traditional Knowledge Basic Issues and Prospec-tive Oxfordshire Nosworthy Way UK 2006

[47] V S Ramachandran S Shijo Joseph and R Aruna ldquoEth-nobotanical studies from amaravathy range of Indira GandhiWildlife SanctuaryWesternGhats CoimbatoreDistrict South-ern Indiardquo Ethnobotanical Leaflets vol 13 pp 1069ndash1087 2009

[48] A H Rajasab and I Mohamad ldquoDocumentation of folkknowledge on edible wild plants of North Karnatakardquo IndianJournal of Traditional Knowledge vol 3 no 4 pp 419ndash4292004

[49] M Rajadurai V G Vidhya M Ramya and A Bhaskar ldquoEthno-medicinal plants used by the traditional healers of pachamalaihills Tamilnadu Indiardquo Studies on Ethno-Medicine vol 3 no 1pp 39ndash41 2009

[50] S K Adesina O Idowu A O Ogundaini et al ldquoAntimicrobialconstituents of the leaves of Acalypha wilkesiana and Acalyphahispidardquo Phytotherapy Research vol 14 no 5 pp 371ndash374 2000

[51] M T Gutierrez-Lugo M P Singh W M Maiese and BN Timmermann ldquoNew antimicrobial cycloartane triterpenesfrom Acalypha communisrdquo Journal of Natural Products vol 65no 6 pp 872ndash875 2002

[52] A K Tyagi andAMalik ldquoAntimicrobial potential and chemicalcomposition of Mentha piperita oil in liquid and vapour phaseagainst food spoiling microorganismsrdquo Food Control vol 22no 11 pp 1707ndash1714 2011

[53] G Devendran and U Balasubramanian ldquoQualitative phyto-chemical screening and GC-MS analysis of Ocimum sanctumL leavesrdquoAsian Journal of Plant Science and Research vol 1 no4 pp 44ndash48 2011

[54] V C Jain N M Patel D P Shah P K Patel and B HJoshi ldquoAntioxidant and antimicrobial activities of Alangiumsalvifolium (Lf) Wang rootrdquo Global Journal of Pharmacologyvol 4 no 1 pp 13ndash18 2010

[55] P Swapna Latha andK Kannabiran ldquoAntimicrobial activity andphytochemicals of Solanum trilobatum LinnrdquoAfrican Journal ofBiotechnology vol 5 no 23 pp 2402ndash2404 2006

[56] M Usha1 S Ragini and S M A Naqvi ldquoAntibacterial activityof acetone and ethanol extracts of Cinnamon (Cinnamomumzeylanicum) and Ajowan (Trachyspermum ammi) on four foodspoilage bacteriardquo International Research Journal of BiologicalSciences vol 1 no 4 pp 7ndash11 2012

[57] S Prabuseenivasan M Jayakumar and S Ignacimuthu ldquoInvitro antibacterial activity of some plant essential oilsrdquo BMCComplementary and Alternative Medicine vol 6 article 39 pp1ndash8 2006

[58] G A Ayoola F M Lawore T Adelowotan et al ldquoChemi-cal analysis and antimicrobial activity of the essential oil ofSyzigium aromaticum (clove)rdquo African Journal of Biotechnologyvol 7 no 13 pp 2227ndash2231 2008

[59] Y J Fu L Chen Y Zu et al ldquoThe antibacterial activity of cloveessential oil against Propionibacterium acnes and its mechanismof actionrdquo Archives of Dermatology vol 145 no 1 pp 86ndash882009

[60] D Venkatesan C M Karrunakarn S Selva Kumar and P TPalani Swamy ldquoIdentification of phytochemical constituents ofAegle marmelos responsible for antimicrobial activity againstselected pathogenic organismsrdquo Ethnobotanical Leaflets vol 13pp 1362ndash1372 2009

[61] P Arun K G Purushotham J Johnsy jayarani and V KumarildquoIn vitro Antibacterial Activity of Oldenlandia umbellata anIndian medicinal Plantrdquo Journal of Pharmaceutical Science andTechnology vol 2 no 4 pp 198ndash201 2010

[62] A Manjamalai R Sardar Sathyajith Singh C Guruvayoorap-pan and V M Berlin Grace ldquoAnalysis of phytochemicalconstituents and antimicrobial activity of some medicinalplants in Taminadu Indiardquo Global Journal of Biotechnology andBiochemistry vol 5 no 2 pp 120ndash128 2010

[63] E Haslam ldquoNatural polyphenols (vegetable tannins) as drugsPossible modes of actionrdquo Journal of Natural Products vol 59no 2 pp 205ndash215 1996

[64] Y Cai Q Luo M Sun and H Corke ldquoAntioxidant activityand phenolic compounds of 112 traditional Chinese medicinalplants associated with anticancerrdquo Life Sciences vol 74 no 17pp 2157ndash2184 2004

[65] B Shan Y Z Cai M Sun and H Corke ldquoAntioxidant capacityof 26 spice extracts and characterization of their phenolicconstituentsrdquo Journal of Agricultural and Food Chemistry vol53 no 20 pp 7749ndash7759 2005

[66] R J W Lambert P N Skandamis P J Coote and G-J ENychas ldquoA study of the minimum inhibitory concentration andmode of action of oregano essential oil thymol and carvacrolrdquoJournal of AppliedMicrobiology vol 91 no 3 pp 453ndash462 2001

[67] M Raccach ldquoThe antimicrobial activity of phenolic antioxi-dants in food a reviewrdquo Journal of Food Safety vol 6 no 3 pp141ndash170 1984

[68] M Taniguchi Y Yano E Tada et al ldquoMode of action of polygo-dial an antifungal sesquiterpene dialdehyderdquo Agricultural andBiological Chemistry vol 52 no 6 pp 1409ndash1414 1988

[69] G J E Nychas ldquoNatural antimicrobials from plantsrdquo in NewMethods of Food Preservation G W Gould Ed pp 58ndash89Blackie Academic London UK 1995

[70] S Burt ldquoEssential oils their antibacterial properties and poten-tial applications in foodsmdasha reviewrdquo International Journal ofFood Microbiology vol 94 no 3 pp 223ndash253 2004

Submit your manuscripts athttpwwwhindawicom

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Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

MEDIATORSINFLAMMATION

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Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Behavioural Neurology

EndocrinologyInternational Journal of

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Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Disease Markers

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

BioMed Research International

OncologyJournal of

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Oxidative Medicine and Cellular Longevity

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PPAR Research

The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014

Immunology ResearchHindawi Publishing Corporationhttpwwwhindawicom Volume 2014

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ObesityJournal of

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Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Parkinsonrsquos Disease

Evidence-Based Complementary and Alternative Medicine

Volume 2014Hindawi Publishing Corporationhttpwwwhindawicom

4 Evidence-Based Complementary and Alternative Medicine

Figure 1 (a) The site for collection of medicinal plants in Western and Eastern Ghats of Tamil Nadu (b) The landmark (map) of traditionalmedicine distribution and collection of different types of plants (c) District map showing the collection site of plants from Kalrayan hills(Salem) (d) Pachamalai hills (Thiruchirappalli) and (e) Mundanthurai (Tirunelveli) rich biodiversity hot-spot of the Western Ghats (f)Kolli hills (Namakkal) (g) Javadi hills (Vellore) part of the Eastern Ghats which is a mountain range that runs mostly parallel to the eastcoast of South India

a large number of wild edible fruits including yielding plantssuch as Citrus acida Ficus benghalensis Ficus microcarpaFicus racemosa Phyllanthus emblica Solanum trilobatumand Syzygium cumini are popularly used by the tribes

34 Plants Used for Snakebite Treatment Thirty-four plantsused for snakebite treatment are documented (Table 1)Snakebite is amajor health hazard that leads to highmortalityin tribal settlements The majority of the antidotes are

prepared freshly from plant materials frequently collectedfrom the leaves of A paniculata A echioides Aristolochiaindica E alba E prostrata M pudica O sanctum Tinvolucrata and Cleistanthus collinus (Oduvanthalai) thewhole plants ofAchyranthes aspera andWedelia calendulaceathe stem-barks and nuts of Strychnos nux-vomica the rootsof Hemidesmus indicus Tephrosia purpurea Rauwolfia ser-pentina C roseus and so forth and the tubers of Gloriosasuperba The tuber paste is usually applied externally on

Evidence-Based Complementary and Alternative Medicine 5

Table 1 Some of the important traditional medicinal plant species families voucher specimens parts used yield of extracts phytochemicalscreening and toxicity on human macrophage cells

Scientific name Family Voucher specimen Plant parts Yield (gm) Phytochemicalanalysis

Adhatoda vasica Nees Acanthaceae D2020 Leaf 64 VasicineAegle marmelos (L)Correa ex Roxb Rutaceae D2018 Root-bark 58 Alkaloids

Alangium salvifolium (L)f Wangerin Alangiaceae 0140 Leaf 63 Phenolic

Andrographis paniculataWallich ex Nees Acanthaceae 0061 Leaf 68 Andrographolide

Andrographis echioidesNees Acanthaceae 0116 Leaf 70 Terpenoids

Acalypha indica L Euphorbiaceae 29644 Leaf 61 AcalypheAcalypha lanceolata L Euphorbiaceae 15791 Leaf 71 AlkaloidsAchyranthes aspera L Amaranthaceae 2666 Leaf 27 BetaineAgeratum conyzoides L Asteraceae 4812 Leaf 37 Essential oilsAsteracantha longifolia L Acanthaceae 0234 Stem 74 GlycosidesAzadirachta indica A Juss Meliaceae D0204 Leaf bark 67 Tannins

Borassus flabellifer L Arecaceae D0202 Root 08 Flavonoidsphenolics

Boerhavia erecta L Nyctaginaceae 10897 Whole plant 28 PhenolicsCalotropis procera (Ait)Ait f Asclepiadaceae D073 Root-bark 13 Terpenoids

Calotropis gigantea (L)RBrex Ait Asclepiadaceae D070 Milky latex 48 Alkaloids

Cassia auriculata L Caesalpiniaceae 0141 Leaf 49 Saponins

Cassia occidentalis L Caesalpiniaceae 0111 Root 95 Flavonoidssaponins

Cassia tora L Caesalpiniaceae 0100 Stem bark 69 SaponinsCassia fistula L Caesalpiniaceae 037 Whole plant 79 SaponinsCardiospermumhalicacabum L Sapindaceae 0125 Whole plant 58 Flavonoids

Catharanthus roseus (L)GDon Apocynaceae 0029 Leaf

root1407 Alkaloids

Cinnamomum zeylanicumGarcin ex Blume Lauraceae 00209 Bark 31 Essential oil

TanninCinnamomum inersReinw ex Blume Lauraceae 043-c Leaf bark Alkaloids

Cissus quadrangularisRoxb Vitaceae D02023 Leaf 68 Glycosides

Citrus acida Roxb Hookf Rutaceae 0213 Leaf 52 SaponinsTerpenoids

Centella asiatica (L) Umbelliferae 0138 Whole plant 89 FlavonoidsAlkaloids

Clerodendrum inerme (L)Gaertn Verbenaceae D02043 Stem 78 Sterols diterpenes

Clitoria ternatea L Papilionaceae D02026 Seed 98 ProteinCleistanthus collinus(Roxb) Benth andHookf

Euphorbiaceae 0011 Whole plant 003 Cleistanthincollinusin

Cleome gynandropsis L Capparidaceae 12247 Leaf 62 GlycosidesCleome viscose L Capparidaceae 29999 Leaf 27 Phenolics

6 Evidence-Based Complementary and Alternative Medicine

Table 1 Continued

Scientific name Family Voucher specimen Plant parts Yield (gm) Phytochemicalanalysis

Coccinia grandisWampA Cucurbitaceae D02030 Leaf root 09 GlycosidesCymbopogon citratus(DC) Gramineae D012 Root 025 Essential oil

Datura metel L Solanaceae D02038 Leaf stem 39 SteroidsEucalyptus globulus Labill Myrtaceae D0220 Leaf 12 TerpenoidsEclipta alba (L) Hassk Asteraceae D028 Whole plant 07 PhenolicEuphorbia hirta Linn Euphorbiaceae 0018-c Whole plant 012 mdash

Eclipta prostrata (L) Asteraceae D210 Leaf 110 Triterpenoidsaponin

Eugenia caryophyllus(Sprengel) Bullock ampHarrison

Myrtaceae 0025 Flower buds 116 Essential oils

Elettaria cardamomumWhite et Mason Zingiberaceae 0009 Fruit pods 317 Essential oils

Gloriosa superba L Liliaceae 020-S Tuber 108 Alkaloids phenol

Jatropha curcas L Euphorbiaceae 015 Whole plant 53 Alkaloidsflavonoids

Hyptis suaveolens (L) Poit Lamiaceae 24688 Leaf 63 Essential oilHemidesmus indicus L Asclepiadaceae D-009 Roots CoumarinsIchnocarpus frutescens(L) RBr Apocynaceae 0110 Root flower 73 Terpenoids

Leucas aspera (Willd)Link Labiatae 0114 Leaf 83 Triterpenes

Lawsonia inermis L Lythraceae T261 Leaf 09 Glycosidesphenolic

Madhuca longifolia (L) JFMacbr Sapotaceae D01415 Nut 93 Sitosterol

Merremia hastate L(Desr) Hallierf Convolvulaceae 10894 Whole plant 40 Alkaloids

Mentha piperita L Lamiaceae 0217-c Whole plant 07 Essential oilsMorinda tinctoria Roxb Rubiaceae 0122 Leaf 14 GlycosidesMimosa pudica L Mimosaceae 0071 Whole plant 06 mdashOldenlandia umbellata L Rubiaceae D02047 Leaf 44 AlkaloidsOcimum sanctum L Lamiaceae 0016 Whole plant 30 AlkaloidsPiper attenuatum BuchHamex Miq Piperaceae 007 Flower 46 Alkaloids

Plumbago zeylanica (L)Cav Plumbaginaceae 0121 Root 48 Plumbagin

Plectranthus amboinicus(L) Spreng Lamiaceae 0410 Whole plant 12 Essential oils

terpenoidsPhyllanthus debilis L(Klein ex Willd) Euphorbiaceae 0120 Whole plant 49 Polyphenol

Phyllanthusmadraspatensis L Euphorbiaceae 0117 Whole plant 50 Polyphenol

Premna tomentosaWilld Verbenaceae 0129 Leaf 53 DiterpenesRosmarinus officinalis L Lamiaceae 0017 Root 023 Essential oilsRauwolfia serpentine L Apocynaceae 020-S Root 115 AlkaloidSebastiania chamaelea (L)Muell Arg Euphorbiaceae 0034 Leaf 11 Polyphenol

Solanum trilobatum L Solanaceae D02054 Leaf flower 40 Tannins

Evidence-Based Complementary and Alternative Medicine 7

Table 1 Continued

Scientific name Family Voucher specimen Plant parts Yield (gm) Phytochemicalanalysis

Sphaeranthus indicus L Asteraceae D02060 Whole plant 10 Essential oilSwertia chirata (L) Ham Gentianaceae D0540 Whole plant 16 GlycodisesStrychnos nux-vomica L Loganiaceae S-22 Nuts 036 AlkaloidsTragia involucrata L Euphorbiaceae D068 Leaves 16 ShellsolTinospora cordifolia(Willd) Miers ex Hooff ampThoms

Menispermaceae 0118 Leafroot stem 50 Glycosides tannins

Tridax procumbens L Compositae 10649 Leaf 18 FlavonoidsTerminalia arjuna (DC)W amp A Combretaceae 033-c Bark 80 Phenolics

Tephrosia purpurea (L)Pers Fabaceae S-43 Whole plant 08 Isoflavone

Vitex negundo L Verbenaceae 0031 Leaf 24 TerpineolVetiveria zizanioides L Gramineae 0051 Root 103 Essential oilWithania somnifera (L)Dunal Solanaceae D02063 Root 21 Alkaloids

Wedelia calendulacea Less Asteraceae S-24 Leaves FlavonoidsZingiber officinale Rosc Zingiberaceae 0327 Rhizome 23 TanninsZanthoxylum limonella(Dennst) Alston Rutaceae 009 Bark 19 Alkaloids essential

oilClass of chemical compounds A alkaloids S saponins T tannins St steroids G glycosides T terpenoids P polyphenol P phenolics Sh shellsol Hhydrocarbon esters

the site of snakebite and decoction is given orally fortreatment by indigenous people Besides these tribes relyon the medicinal plants as ingredients for fabricating a kindof medicated stone for health management ldquoVishakallurdquo(poison stone) is used by the indigenous groups called Kaniin Kerala India to treat a snakebiteWhen the stone is placeddirectly on the bitten area it sticks to the body to absorb thepoison and then become detached when absorption seems tobe complete The ingredients of Vishakallu stones are madewith leaves of Ocimum sanctum Anisomeles malabaricaLeucas aspera Piper betle Santalum album and the pebblescollected from the river bank

35 Survey of Medicinal Plants and Their Health Care ValuesThe present study is an attempt to provide scientific basis andobtain justification for the traditional beliefs of reliance on arich diversity of ethnomedicinal plants along with the richheritage of traditional medicine practices related to healthcare system made available by the primitive tribal commu-nities located at different settlements The native traditionalpractitioners called ldquovaidyarsrdquo have a good knowledge aboutthe traditional plants locally available for treatment of var-ious diseases (Figures 3(a)ndash3(p)) Such traditional medicalknowledge is used for preparing home remedies ill healthprevention and routine health maintenanceThis knowledgeis also applicable to cover other sectors of social life Duringthe ethnobotanical survey the wealth of 78 medicinal plantspecies used by the indigenous tribal community for varioustypes of health treatment was documented The botanical

names family names parts used chemical constituents andtheir application are provided (Table 1)

36 Antimicrobial Activity of Crude Extracts In this studywe reported the antimicrobial screening of methanolic crudeextracts of 78 medicinal plants (Table 2) Results revealedthat 68 plant extracts displayed potent activities against oneor more Gram-positive and -negative bacteria Of whichTragia involucrata Citrus acida Aegle marmelos Adha-toda vasica Calotropis procera Andrographis paniculata andMentha piperita Azadirachta indica Sphaeranthus indicusand Elettaria cardamomum showed the highest antibacte-rial activity against the multidrug resistant B pseudomallei(KHW and TES) and S aureus at 100120583gmL concentrationThe extracts showed pronounced antibacterial activity withtheir inhibitory zones ranging from 20 to 31mm in diam-eter as compared to the standard drugs chloramphenicoland ceftazidime (29ndash33mm) The majority of the plantsdemonstrated a powerful antimicrobial potency against themultidrug resistant strains of B pseudomallei (KHW andTES) K pneumonia and S aureus Approximately twenty-one plant extracts exerted only a weak or moderate effectagainst the tested bacteria while the crude extract of 13plants failed to show any effect at all Except for the plantextracts of T involucrata A lanceolata A vasica and Sindicus extracts the majority of the plant extracts wereineffective against theV damsela infection fascinatingly only11 plants exhibited activity against P aeruginosa of whichS indicus M piperita and C procera were found to havevery strong inhibition of bacteria at the tested concentrations

8 Evidence-Based Complementary and Alternative Medicine

Table 2 Antimicrobial activity of methanol extract of aromatic medicinal plants and essential oils evaluated against multidrug resistant(MDR) human pathogens at 100120583gmL concentration

Scientific name Microorganisms growth inhibition zones (6 millimeter in diameters)KHW TES Kp Kpr Pa Sa Stp Sp Vd Vd

Adhatoda vasica Nees 28 19 22 18 12 mdash 10 17 16 mdashAegle marmelos (L) Correa ex Roxb 29 17 9 10 mdash 15 mdash mdash mdash mdashAlangium salvifolium (L) f Wangerin 15 10 8 8 mdash 12 9 10 mdash mdashAndrographis echioides L 12 9 mdash mdash mdash 17 8 9 mdash mdashAndrographis paniculataWallich ex Nees 26 21 19 13 mdash 25 8 16 12 mdashAcalypha indica L mdash mdash mdash mdash mdash 18 mdash mdash mdash mdashAcalypha lanceolata L mdash mdash 12 13 mdash 10 mdash mdash 21 mdashAchyranthes aspera L mdash mdash 7 14 mdash 15 12 11 mdash mdashAgeratum conyzoides L 10 8 mdash mdash mdash mdash mdash mdash mdash mdashAsteracantha longifolia L 16 12 mdash mdash mdash 15 mdash mdash mdash mdashAzadirachta indica A Juss 15 17 21 16 14 23 12 14 mdash mdashBorassus flabellifer L 9 10 17 8 mdash mdash 9 10 mdash mdashBoerhavia erecta L mdash mdash 8 9 7 16 mdash mdash mdash mdashCalotropis procera (L) mdash mdash 15 mdash 18 28 9 mdash mdash mdashCalotropis gigantea (L) RBrex Ait 11 9 mdash mdash mdash 20 mdash mdash 9 8Cardiospermum halicacabum L 23 mdash mdash 14 mdash 9 19 mdash mdash mdashCatharanthus roseus (L) GDon 13 7 12 9 mdash 15 11 8 mdash mdashCassia auriculata L 17 13 12 mdash mdash 19 13 mdash mdash mdashCassia occidentalis L 18 mdash mdash mdash mdash mdash mdash mdash mdash mdashCassia tora L mdash mdash mdash mdash mdash mdash mdash mdash mdash mdashCassia fistula L mdash mdash mdash mdash mdash mdash mdash mdash mdash mdashCitrus acida Roxb Hookf 26 22 mdash 12 29 9 13 8 mdashCissus quadrangularis L mdash mdash mdash mdash mdash mdash mdash mdash mdash mdashCinnamomum zeylanicum Garcin exBlume 14 16 7 20 mdash 22 19 7 mdash mdash

Cinnamomum iners Reinw ex Blume 20 16 mdash 15 mdash 16 12 mdash mdash mdashRosmarinus officinalis L mdash 9 mdash 10 mdash 7 mdash 8 mdash mdashCentella asiatica (L) 9 8 mdash mdash mdash 11 mdash mdash mdash mdashClerodendrum inerme (L) Gaertn 13 7 12 mdash mdash 15 11 8 mdash mdashClitoria ternatea L mdash mdash mdash mdash mdash mdash mdash mdash mdash mdashClitoria ternatea L 16 12 mdash 13 mdash 8 10 mdash mdash mdashCleome gynandropsis L mdash mdash 12 19 11 mdash 9 15 12 mdashCleome viscose L mdash mdash 8 10 14 20 mdash mdash mdash mdashCoccinia grandisWampA 9 10 17 8 mdash mdash 9 10 mdash mdashCymbopogon citratus (DC) 16 18 mdash 17 mdash 14 mdash mdash mdash mdashDatura metel L mdash mdash mdash mdash mdash mdash mdash mdash mdash mdashEclipta alba (L) Hassk 20 mdash mdash 9 mdash mdash 10 mdash mdash mdashEuphorbia hirta Linn 11 mdash mdash mdash mdash 16 mdash mdash mdash mdashEucalyptus globulus Labill mdash mdash mdash 7 mdash mdash mdash 7 mdash mdashEugenia caryophyllus Bullock amp Harrison mdash 7 mdash 11 mdash 9 mdash 8 mdash mdashElettaria cardamomumWhite et Mason 21 20 7 14 mdash 22 12 17 mdash mdashHyptis suaveolens (L) Poit 8 10 8 7 mdash mdash mdash mdash mdash mdashIchnocarpus frutescens (L) WJ Aiton mdash mdash mdash mdash mdash mdash mdash mdash mdash mdashJatropha curcas L mdash mdash mdash 8 mdash 11 mdash 12 mdash 7Leucas aspera (Willd) Link 9 10 17 8 mdash 12 9 10 mdash mdashLawsonia inermis L mdash mdash mdash mdash mdash mdash mdash mdash mdash mdashMadhuca longifolia (L) JF Macbr 18 16 14 19 12 mdash 8 7 9 mdash

Evidence-Based Complementary and Alternative Medicine 9

Table 2 Continued

Scientific name Microorganisms growth inhibition zones (6 millimeter in diameters)KHW TES Kp Kpr Pa Sa Stp Sp Vd Vd

Merremia hastate L (Desr) Hallierf mdash mdash mdash mdash mdash mdash mdash mdash mdash mdashMorinda tinctoria Roxb mdash mdash mdash mdash mdash mdash mdash mdash mdash mdashMentha piperita L 23 17 26 12 20 25 19 mdash mdash mdashOcimum sanctum L 12 9 11 7 mdash 15 7 8 mdash mdashOldenlandia umbellata L mdash mdash mdash mdash mdash 17 mdash mdash mdash mdashPiper attenuatum Buch Hamex Miq 13 7 12 21 mdash 17 11 8 mdash mdashPlumbago zeylanica (L) Cav 9 10 17 8 mdash 12 9 10 mdash mdashPlectranthus amboinicus (L) Spreng mdash mdash mdash 8 11 15 mdash mdash mdash mdashPhyllanthus debilis L (Klein ex Willd) 7 8 mdash 9 mdash 9 18 7 mdash mdashPhyllanthus maderaspatensis L 17 mdash mdash mdash mdash mdash mdash mdash mdash mdashPremna tomentosaWilld 13 10 mdash 15 mdash 9 10 mdash mdash mdashGloriosa superba L 17 16 7 8 mdash 15 mdash mdash mdash 8Sebastiania chamaelea (L) Muell Arg 19 12 mdash 17 mdash 13 19 8 mdash mdashSolanum trilobatum L mdash mdash 13 9 mdash 8 mdash mdash mdash mdashSphaeranthus indicus L 20 18 21 7 22 mdash 11 9 mdash 16Swertia chirata (L) Ham mdash mdash mdash mdash mdash mdash mdash mdash mdash mdashTerminalia arjuna (W amp A) mdash mdash mdash mdash mdash 16 mdash mdash mdash mdashTinospora cordifolia (Willd) Miers exHooff ampThoms mdash mdash 23 16 mdash 12 15 mdash mdash mdash

Tridax procumbens L 9 8 7 mdash mdash 14 mdash mdash mdash mdashTragia involucrata L 25 23 20 mdash mdash 31 28 22 19 mdashVitex negundo L mdash mdash mdash mdash mdash 14 mdash mdash mdash mdashVetiveria zizanioides (L) 11 9 7 7 mdash 16 mdash mdash 8Withania somnifera (L) Dunal 12 20 mdash mdash mdash 15 12 mdash mdash mdashZingiber officinale Rosc 14 11 7 15 mdash 7 12 7 mdash mdashZanthoxylum limonella (Dennst) Alston 13 7 12 9 mdash mdash 11 8 mdash mdashChloramphenicol (30120583gdisc) 21 12 15 17 29 16 15 18 13 11Ceftazidime (30 120583gdisc) 33 16 22 19 16 25 21 20 12 15lowastBacteria (+minus) Results obtained in the disc diffusion assay antibacterial activity is expressed as the mean plusmn SD (119899 = 3) of the inhibition by the extract andits diameter around the discs One way analysis of variance was performed (mean plusmn SD 119899 = 3 replicates) Size of inhibition zones were including the sterileblank discs 6 millimeter (mm) in diameters Absence of bacterial inhibition indicates (mdash) antibiotic disc (30 120583gdisc)

Interestingly sixteen plants such as Andrographis echioidesC auriculata C viscose C gigantea T arjuna Oldenlandiaumbellata Boerhavia erecta and E hirta exerted a strongactivity against the Gram-positive S aureus bacteria

37 Phytochemical Screening of Plants The results obtainedfrom the phytochemical screening as shown in Table 1 indi-cate the presence of various types of secondary metabolitessuch as polyphenols tannins saponins alkaloids and gly-cosidespolysaccharides Most of the plant extracts relativelyrich in alkaloids phenols flavonoids polyphenols tanninssterols and terpenoids were found to inhibit the growth oforganisms

38 Antimicrobial Activity of Fractioned Compounds Activecomponents were purified from the most active extractsfor further testing The compound shellsol of T involucrata

and C acida exhibited the most potent action againstthe antibiotic resistant strains of B pseudomallei (KHW)S aureus B pseudomallei (TES) and K pneumoniae Amarmelos was also found to inhibit the growth of B pseudo-mallei (KHW) more effectively than other tested bacteria Avasica showed the broad spectrum growth inhibitory activityon B pseudomallei (KHW) K pneumoniae K pneumoniaeresistant B pseudomallei (TES) S pyogenes and V damselaHowever E cardamomum displayed antimicrobial activityon some of the B pseudomallei (KHW) S pyogenes Bpseudomallei (TES) and S typhi strains Similarly A indicaexerted the growth inhibition on K pneumoniae and Saureus Remarkably Sebastiania chamaelea was more activeagainst K pneumoniae K pneumoniae and S pyogenesThe compound from S indicus inhibited the growth of Kpneumoniae K pneumoniae B pseudomallei (KHW andTES) and S typhi strains as compared to the activity shown

10 Evidence-Based Complementary and Alternative Medicine

Figure 2 Diverse biodiversity richness of medicinal plants in Western and Eastern Ghats (a) Topography of plant covering area in Kollihills (Namakkal district Tamil Nadu) (b) Aerial view of occurrence of medicinal plants in Mundanthurai hills (Tirunelveli district TN) (c)Pachamalai hills (Trichy district) and its natural vegetation inhabitants for Malaiyali tribes (d) Deforestation of natural herbal resources dueto urbanization in Kalrayan hills (Salem district) in the Eastern Ghats of TN (e) Medicinal plants and its various parts used by the natives(traditional healers) for the treatment of diverse human illness with a very high percentage of leaves and whole plants often used for herbaldrug preparation by the local practitioners (f) Various category of plants like shrub herb climbers and tree and the parts used in medicine

Evidence-Based Complementary and Alternative Medicine 11

Figure 3 Medicinal aromatic herbs spices and toxic plants were collected from the tribal areas of the Western and Eastern Ghats region inTamil Nadu India (a) A vasica Nees (leaf) (b) Eclipta alba (L) Hassk (whole plant) (c)Mimosa pudica L (whole plant) (d) P amboinicus(L) Spreng (whole plant) (e) T procumbens (L) (whole plant) (f) Euphorbia hirta Linn (whole plant) (g) A paniculata Wallich ex Nees(H) C roseus (L) GDon (whole plant) used for therapy (i) Cinnamomum iners Reinw ex Blume (leaf) (j) E globulus Labill (leaf and bark)(k) Z officinale Rosc (Rhizome) (l) E caryophyllus (Sprengel) Bullock and Harrison (flower buds) (m) M piperita L (whole plant) (n) Ccitratus (DC) Clitoria ternatea L (whole plant) (o) C zeylanicum Garcin ex Blume (bark) (p) Elettaria cardamomumWhite et Mason (fruitpod) used for medicine and food preparation

by the crude extracts (Figure 4)The antimicrobial efficacy offractions collected from the oil yielding plants was also com-pared with that of the tested compounds C zeylanicum andR officinaliswere themost sensitive in controlling the growthof B pseudomallei (KHW) S aureus K pneumonia and Spneumoniae Fascinatingly all the compounds obtained fromaromatic plants except those from E globules were foundto be very effective against the multidrug resistant humanpathogen B pseudomallei (KHW) that causes melioidosis

On the other hand compounds from C citrates O sanctumE caryophyllus and Z zizanioide showed some promisingeffect only against S aureus (Figure 5) The activity of thecompounds were pronounced more than that of the oilyielding plant fractions

39 Minimum Inhibitory Concentrations (MICs) The antibi-otic potential of the purified fractions was obtained fromthe MIC determination The hydrocarbon ester shellsol

12 Evidence-Based Complementary and Alternative Medicine

(mean plusmn SD n = 3)

0

5

10

15

20

25

30

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn V

d St

Bacteria (+minus)

lowast

lowast

lowast

A vasica

In vitro assay(P

lant

frac

tion)

Inhi

bitio

n zo

nes (

mm

)

(a)

(mean plusmn SD n = 3)

0

5

10

15

20

25

30

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn V

d St

Bacteria (+minus)

lowast

A marmelos

Inhi

bitio

n zo

nes (

mm

)

In vitro assay

(b)

(mean plusmn SD n = 3)

0

5

10

15

20

25

30

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn V

d St

Bacteria (+minus)

lowastlowast

A indica

Inhi

bitio

n zo

nes (

mm

)

Incu

batio

n (37∘C

24 h

)

(c)

(mean plusmn SD n = 3)

0

5

10

15

20

25

30

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn V

d St

Bacteria (+minus)

lowast

lowast lowast

35

C acida

Inhi

bitio

n zo

nes (

mm

)(P

lant

frac

tion)

(d)

(mean plusmn SD n = 3)

0

5

10

15

20

25

30

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn V

d St

Bacteria (+minus)

lowastlowast

lowast

S chamaelea

Inhi

bitio

n zo

nes (

mm

)

(e)

(mean plusmn SD n = 3)

0

5

10

15

20

25

30

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn V

d St

Bacteria (+minus)

lowastlowast

lowastlowast

S indicus

Inhi

bitio

n zo

nes (

mm

)

Incu

batio

n (37∘C

24 h

)

(f)

(mean plusmn SD n = 3)

0

5

10

15

20

25

30

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn V

d St

Bacteria (+minus)

lowast

lowast lowast

E cardamomum

Inhi

bitio

n zo

nes (

mm

)(P

lant

frac

tion)

(g)

(mean plusmn SD n = 3)

0

5

10

15

20

25

30

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn V

d St

Bacteria (+minus)

lowast

lowast

lowast

lowast

Shellsol

35

Inhi

bitio

n zo

nes (

mm

)

Incu

batio

n (37∘C

24 h

)

(h)

Figure 4 In vitro antimicrobial activity of purified fractions from the most active plant extracts tested against bacteria Growth inhibitionzones were measured and analyzed with mean plusmn standard deviation (SD) (119899 = 3) using one way analysis of variance Level of significanceat (lowast119875 gt 001) Most of the fractions exerted a potent inhibitory effect against multidrug resistant Gram-negative bacteria (B pseudomalleistrains KHW and TES) K pneumonia and Gram-positive bacteria S aureus

(T involucrata) andC acida showed an interesting inhibitorypotential against S aureus (MIC of 78 120583gmL) and B pseu-domallei strain of KHW (MIC of 156 120583gmL) A vasicashowed an MIC of 156120583gmL against B pseudomallei(KHW) and an MIC of 3125120583gmL against K pneumo-niae K pneumoniae S pyogenes and V damsela strainsFractions from A marmelos and terpenoid from A indicaexerted bacteriostatic effect with MIC values of 3125120583gmLon some selected bacteria including B pseudomallei ofKHW S aureus and B pseudomallei of TES The MIC of3125 120583gmL was found for E cardamomum against S aureus

K pneumonia and S pyogenes S indicus displayed a verystrong inhibition against MDR K pneumoniae (MIC of156 120583gmL) and against B pseudomallei (KHW and TES)at MIC of 3125 120583gmL When the antimicrobial efficacies ofpurified fractions from aromatic plants were compared theC zeylanicum fraction displayed an important antimicrobialeffect against S aureus (MIC of 78 120583gmL) MDR B pseu-domallei of KHW (MIC of 156 120583gmL) and S pneumoniae(MIC of 3125 120583gmL) The essential oil from M piperateashowed MIC value of 3125120583gmL against K pneumoniae Saureus and B pseudomallei (KHW) respectivelyO sanctum

Evidence-Based Complementary and Alternative Medicine 13

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn

Bacteria (+minus)

0

5

10

15

20

25

30

Inhi

bitio

n zo

nes (

mm

)(P

lant

frac

tion)

lowastlowast

lowast

(mean plusmn SD n = 3)In vitro assay

C citratus

(a)

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn

Bacteria (+minus)

0

5

10

15

20

25

30

Inhi

bitio

n zo

nes (

mm

)

lowast

lowast

lowastlowast

lowast

lowast

(mean plusmn SD n = 3)In vitro assay

35

C zeylanicum

(b)

Bacteria (+minus)

0

5

10

15

20

25

30

Inhi

bitio

n zo

nes (

mm

)

lowast

lowast

lowast

(mean plusmn SD n = 3)

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn V

d St

O sanctum

Incu

batio

n (37∘C

24 h

)

(c)

Bacteria (+minus)

0

5

10

15

20

25

Inhi

bitio

n zo

nes (

mm

)

lowast

lowast

(mean plusmn SD n = 3)

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn V

d St

M piperita

(Pla

nt fr

actio

n)

(d)

Bacteria (+minus)

0

5

10

15

20

25

30

Inhi

bitio

n zo

nes (

mm

)lowast

lowast

lowast

(mean plusmn SD n = 3)

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn V

d St

E caryophyllus

(e)

0

5

10

15

20

25

30

Inhi

bitio

n zo

nes (

mm

) (mean plusmn SD n = 3)

Bacteria (+minus)

KHW

TES

Kp

KprPa Sa

Stp

Spn

E globulus

Incu

batio

n (37∘C

24 h

)

(f)

Bacteria (+minus)

0

5

10

15

20

25

30

Inhi

bitio

n zo

nes (

mm

)(P

lant

frac

tion)

lowast

lowast

lowast

lowast

(mean plusmn SD n = 3)

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn V

d

R officinalis

(g)

0

5

10

15

20

25

30

Inhi

bitio

n zo

nes (

mm

)

lowast

lowast

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn V

d St

Bacteria (+minus)V zizanioides

(mean plusmn SD n = 3)

Incu

batio

n (37∘C

24 h

)

(h)

Figure 5 Comparison of antimicrobial effect of plant compounds obtained from the most popularly used sources of essential oils as assayedby the disc-diffusion method in vitro It displayed a powerful activity against B pseudomallei and S aureus than the other bacteria strainsOther compounds showed only a moderate or weak action against the tested bacteria Values for zone of bacterial growth inhibition werepresented as mean plusmn SD (119899 = 3) with level of significance at (lowast119875 gt 001)

and C citratus fractions also showed antimicrobial activity(MICs of 3125ndash125120583gmL) only at higher concentrationsagainst the tested bacteria In addition to that higher concen-trations (gt250120583gmL) (of Vetiveria fractions) were requiredto inhibit Vibro species and others (including E globulusfractions) failed to show any effect at tested concentrations(78ndash125 120583gmL) However the purified fractions (from mostactive medicinal plants) showed strong bacteriostatic inhibi-tion against the tested organisms (Table 3)

310 Cytotoxic Effects of Plants When the components wereassayed for cytotoxicity against the normal human skinfibroblasts (HEPK) cells the compounds obtained from

E cardamomum T involucrata S indicusC acidaA vasicaA marmelos A indica and A paniculata did not showtoxicity up to 1000120583gmL (see Figures S1 and S2 in Supple-mentaryMaterial available online at httpdxdoiorg1011552013525613) A slight reduction of cell proliferation wasnoted only at higher doses (2000120583gmL) In contrast cellproliferation was markedly reduced after exposure of HEPKcells to O sanctum E globulus V zizanioides C citratusand E globulus compounds There was no gradual reductionin skin cell proliferation seen after exposure to C zeylan-icum R officinalis and M piperita (see Figures S3 and S4)compounds The toxicity was found to be concentration-dependent when the skin fibroblasts (HEPK) cells were

14 Evidence-Based Complementary and Alternative Medicine

Table 3 Minimum inhibitory concentrations (MICs) of purified plant fractions and essential oils against antibiotic resistant bacteria

Botanical name Family Parts used Gram-positive and -negative bacteria (MICs 120583gmL)KHW TES Kp KPr Sa Stp Sp Vd St

A indica Juss Meliaceae Seed (fraction) 3125 625 125 lt250 3125 mdash mdash mdash mdashA marmelos (L) Rutaceae Root-bark (F) 3125 3125 mdash mdash 3125 625 125 250 mdashA paniculata Nees Acanthaceae Leaf (fraction) 250 mdash 125 mdash 625 mdash mdash 125 mdashA vasica Nees Acanthaceae Fraction (Stem) 156 3125 3125 625 mdash lt250 3125 3125 mdashC acida Roxb Rutaceae Leaf (fraction) 156 625 mdash mdash 78 mdash 3125 mdash mdashE cardamomumWhite et Mason Acanthaceae Fraction (WP) 625 mdash 3125 mdash 3125 250 625 mdash mdashS indicus (L) Euphorbiaceae Whole plant (F) 3125 125 156 mdash 625 mdash mdash lt250 mdashT involucrata (L) Euphorbiaceae Shellosol (leaf) 156 3125 mdash mdash 78 625 mdash mdash mdashCinnamomum zeylanicum (L) Lauraceae Bark (fraction) 156 625 mdash mdash 78 3125 mdash mdash 125Cymbopogon citratus (L) Graminae Leaf (fraction) 250 lt250 625 mdash 625 125 mdash mdash mdashEugenia caryophyllus (L) Myrtaceae Flower buds (F) 625 125 mdash mdash 625 mdash mdash mdash mdashEucalyptus globulus (L) Myrtaceae Fraction (leaf) mdash mdash mdash mdash mdash mdash lt250 mdash lt250Mentha piperita (L) Labiatae Fraction (WP) mdash 625 3125 mdash 3125 mdash lt250 mdash mdashOcimum sanctum (L) Labiatae Leaf (fraction) 625 125 mdash 3125 625 mdash 125 lt250 mdashRosmarinus officinalis (L) Labiatae Rosemary oil 3125 mdash mdash mdash 3125 mdash mdash mdash mdashVetiveria zizanioides (L) Graminae Root (fraction) mdash 125 125 mdash mdash mdash gt250 gt250The bacterial growth inhibitory activity was compared for significant differences within the bacterial strains by broth-dilution method at 250 125 625 3125156 and 78 120583gmL F fractions

exposed to various compounds The cell proliferation wasincreased by the influence of the plant components at thelower concentrations Whereas the oil yielding plant com-pounds showed inhibition of cell proliferation and toxicity athigher doses 250ndash1000120583gmL

4 Discussion

The Western Ghats is considered as one of the richestbiodiversity hotspots in the world [41] In this survey wecollected nearly 78 medicinal plants from Western andEastern Ghats that are edible and popularly used for curingvarious ailments including snakebite Traditional remedieshave a long-standing history in many tribal settlements inTN India and they continue to provide useful and applicabletools for treating ailments [42] The ingredients that makeup the ldquoVishakallurdquo stone which is used as an antidote forsnakebite are different herbs and pebbles available from theriver banks Likewise aqueous paste and decoction obtainedfrom the leaves of A paniculata are widely used for snakebitetreatment by indigenous people [43] Previous studies havereported that ethnomedicine plays major roles in conservingthe disappearing knowledge of tribal communities [44ndash47]The traditional beliefs of reliance on a rich diversity ofethnomedicinal plants located at different settlements havealso been confirmed in another study [48 49] Herein weexplored the various types of traditional practices reported bythe primitive tribal communities with a view to gain furtherknowledge from such studies

In the present investigation potentially rich sources oftribal medicine (71 plants) were scientifically evaluated fortheir antibacterial activity against the MDR bacteria and theaccumulated data was disseminated for the first time to the

scientific community Out of the 71medicinal plants screenedfor the antibacterial activity 10 of them (T involucrataC acida A marmelos A vasica C procera A paniculataand M piperita A indica S indicus and E cardamomum)displayed the highest antibacterial activity against the mul-tidrug resistant B pseudomallei (KHW and TES) and Saureus strains The antibacterial activity of those crude plantextracts was as equally effective as that of the standard drugsOur findings corroborated with the previous reports madeon the antistaphylococcal activity of tribal medicinal plants[34 50 51] On the other hand isolated components from themost active extracts of T involucrata shellsol and C acidaexhibited the most potent action against the antibiotic resis-tant B pseudomallei (KHW) K pneumoniae and S aureusstrainsThese results further confirmed our previous findingson the leaves of T involucrata and its compounds hydrocar-bon ester-like shellsol which displayed a high antibacterialeffect against the different bacterial strains especially thatof S aureus [34] Eugenol and caryophyllene are the activeagents contained in the M piperita [52] and O sanctumplants which are believed to be mainly responsible for theantimicrobial properties of these plants [53] Interestinglythe compounds obtained from the aromatic plants such asC zeylanicum and R officinalis were also found to be veryeffective against the multidrug resistant human pathogen Kpneumoniae S aureus S typhi and B pseudomallei (KHW)that causes melioidosis

The inhibitory potential determined for the shellsol(T involucrata) vaseline (A vasica) C acida and C zey-lanicum indicates that the MIC of 78ndash3125120583gmL foundagainst the B pseudomallei of KHW K pneumoniae Kpneumoniae S pyogenes and V damsel and S pneumoniaewas quite low Similarly lower MIC values were found for

Evidence-Based Complementary and Alternative Medicine 15

M piperita (MIC of 113ndash225 mgmL) against the abovebacterial strains [52] MIC was found for the most activealcohol extracts of A salvifolium (MIC 0034ndash0263mgmL)on S aureus [54] and the MIC for S trilobatum aqueousextracts determined against the tested organisms rangedfrom 006 to 05mgmL [55] C zeylanicum was foundto have an effective antibacterial activity (MIC 64 120583gmL)against P aeruginosa E coli B subtilis and S aureus[56] Previously several investigators have demonstrated thatactive agents exert interesting activity against bacteria evenat lower concentrations tested [57 58] The bacteriostaticmechanism involves damage to the cell walls of bacteriafollowed by inhibition of protein synthesis that ultimatelyleads to bacterial death [59] The most active plants arewidely used by various tribes as traditional treatment (iecut wounds skin infection and scabies) thus indicatingthe potential for further development into promising drugsIn order to ascertain the safety and efficacy of the mostactive compounds their effect on human skin fibroblast cellswas evaluated Chemical constituents of E cardamomumT involucrata S indicus C acida A vasica A marmelosA indica and A paniculata plants failed to produce anynoticeable toxicity up to 1000120583gmL Although some ofthe tested compounds exhibited a slight reduction of cellproliferation and some minor morphological changes suchchanges were insignificant at lower doses and became evidentonly at higher doses However certain aromatic compoundsof O sanctum E globulus V zizanioides C citratus and Eglobulus plants showed reduction of cell proliferation againstHEPK cells

Our phytochemical screening also provides evidenceof the presence of several types of compounds that aremainly responsible for the remarkable antibacterial effectof these plants The differences noted for the bactericidalactivity of various plant extracts in this study appears tobe directly related to the diversity of compounds (shown inparentheses) that are accumulated in the following plants(eg A marmelos and O umbellate) [60ndash62] Compoundslike tannins phenol and polyphenols can bind the Gram-negative bacteria to form a heavy soluble complex on the cellsurface which subsequently disturbs the availability of recep-tor on cells and kills the bacteria [63] Several species havingwide spectra of antimicrobial activity mainly due to theactive constituents such as essential oil phenolic compoundslike thymol carvacrol in oregano and thyme eugenol inclove and cinnamon were also identified previously [64 65]Essential oils degrade the cell wall interact with the cell com-ponents and then disrupt the cytoplasmic membrane [66]The antimicrobial effect of phenolic compounds may involvemultiplemodes of action including damage to themembraneprotein interference with membrane integrated enzymes[67] causing leakage of cellular components coagulation ofcytoplasm depletion of the proton motive force alterationof fatty acid and phospholipid constituents impairmentof enzymatic mechanisms for production and metabolismalteration of nutrient uptake and electron transport [68]influencing the synthesis of DNA and RNA and destroyingprotein translation and the function of the mitochondrionin eukaryotes [69] The mode of action of antimicrobial

agents depends on the type of microorganism and is mainlyrelated to their cell wall structure and the outer membranearrangement Most of the plant spices and herbs containcomplex phenolics (ie phenolic acids flavonoids tanninslignans coumarins quinines) In addition the mechanismsof action of each phenolic compound against various bacteriaare also very complicated [26 70] Further investigation istherefore required to understand the relationship betweenthe antimicrobial action and the chemical structure of everyphenolic compound in the tested extracts The informationavailable from previous pharmacological sources combinedwith the findings herein reported on the medicinal plantextracts may serve as essential data for future drug develop-ment to combat diseases caused by theMDRbacterial strains

Abbreviations

MDR Multidrug resistant bacteriaTES Strain of Burkholderia pseudomalleiMRSA Methicillin-resistant Staphylococcus aureusVRE Vancomycin-resistant enterococciMTB Mycobacterium tuberculosisCH3OH Methanol

CFU Colony forming unitHCl Hydrochloric acidTLC Thin layer chromatographyH2SO4 Sulphuric acids

UV Ultraviolet spectrophotometerHEPK Human skin fibroblasts

Acknowledgments

Theauthors are thankful to the ERI Loyola College ChennaiIndia for his support during the survey The authors alsothank the Department of Microbiology NUHS NUS forthe bacterial cultures used in this investigation The authorsare thankful to the tribal people for sharing their healingknowledge on medicinal aromatic plants

References

[1] M N Alekshun and S B Levy ldquoMolecular mechanisms ofantibacterialmultidrug resistancerdquoCell vol 128 no 6 pp 1037ndash1050 2007

[2] A D Russell ldquoAntibiotic and biocide resistance in bacteriaintroductionrdquo Symposium Series Society for Applied Microbiol-ogy vol 31 no 92 pp 1Sndash3S 2002

[3] D R Guay ldquoTreatment of bacterial skin and skin structureinfectionsrdquo Expert Opinion on Pharmacotherapy vol 4 no 8pp 1259ndash1275 2003

[4] A J Brosnahan and P M Schlievert ldquoGram-positive bac-terial superantigen outside-in signaling causes toxic shocksyndromerdquo FEBS Journal vol 278 no 23 pp 4649ndash4667 2011

[5] K Fellermann J Wehkamp K R Herrlinger and E F StangeldquoCrohnrsquos disease a defensin deficiency syndromerdquo EuropeanJournal of Gastroenterology and Hepatology vol 15 no 6 pp627ndash634 2003

16 Evidence-Based Complementary and Alternative Medicine

[6] L Chen S Lin R Agha-Majzoub L Overbergh C Mathieuand L S Chan ldquoCCL27 is a critical factor for the developmentof atopic dermatitis in the keratin-14 IL-4 transgenic mousemodelrdquo International Immunology vol 18 no 8 pp 1233ndash12422006

[7] J H Song V Thamlikitkul and P R Hsueh ldquoClinical andeconomic burden of community-acquired pneumonia amongstadults in the Asia-Pacific regionrdquo International Journal ofAntimicrobial Agents vol 38 no 2 pp 108ndash117 2011

[8] D A B Dance ldquoMelioidosisrdquo Current Opinion in InfectiousDiseases vol 15 no 2 pp 127ndash132 2002

[9] N J White ldquoMelioidosisrdquo The Lancet vol 361 no 9370 pp1715ndash1722 2003

[10] S Yang ldquoMelioidosis research in Chinardquo Acta Tropica vol 77no 2 pp 157ndash165 2000

[11] P R Hsueh L J Teng L N Lee et al ldquoMelioidosis an emerginginfection in Taiwanrdquo Emerging Infectious Diseases vol 7 no 3pp 428ndash433 2001

[12] C Walsh and S Fanning ldquoAntimicrobial resistance in food-borne pathogensmdasha cause for concernrdquo Current Drug Targetsvol 9 no 9 pp 808ndash815 2008

[13] M Vorachit P Chongtrakool S Arkomsean and S BoonsongldquoAntimicrobial resistance in Burkholderia pseudomalleirdquo ActaTropica vol 74 no 2-3 pp 139ndash144 2000

[14] F M Thibault E Hernandez D R Vidal M Girardet and JD Cavallo ldquoAntibiotic susceptibility of 65 isolates of Burkholde-ria pseudomallei and Burkholderia mallei to 35 antimicrobialagentsrdquo Journal of Antimicrobial Chemotherapy vol 54 no 6pp 1134ndash1138 2004

[15] R L S Sikarwar ldquoEthnogynaecological uses of plants new toIndiardquo Ethnobotany vol 12 no 1-2 pp 112ndash115 2002

[16] R Siva ldquoStatus of natural dyes and dye-yielding plants in IndiardquoCurrent Science vol 92 no 7 pp 916ndash925 2007

[17] P Kumar Singh V Kumar R K Tiwari A Sharma ChV Rao and R H Singh ldquoMedico-ethnobotany of lsquochatararsquoblock of district sonebhadra Uttar Pradesh Indiardquo Advances inBiological Research vol 4 no 1 pp 65ndash80 2010

[18] W H Lewis and M P Elwin-Lewis Medical Botany PlantsAffecting Human Health John Wiley amp Sons New York NYUSA 2003

[19] R Perumal Samy P N Pushparaj and P GopalakrishnakoneldquoA compilation of bioactive compounds fromAyurvedardquo Bioin-formation vol 3 no 3 pp 100ndash110 2008

[20] WorldHealthOrganization (WHO)TraditionalMedicine Strat-egy 2002ndash2005 Genera World Health Organization GenevaSwitzerland 2002

[21] P S Haddad G A Azar S Groom and M Boivin ldquoNaturalhealth products modulation of immune function and preven-tion of chronic diseasesrdquo Evidence Based Complementary andAlternative Medicine vol 2 no 4 pp 513ndash520 2005

[22] K Poonam andG S Singh ldquoEthnobotanical study of medicinalplants used by the Taungya community in Terai Arc LandscapeIndiardquo Journal of Ethnopharmacology vol 123 no 1 pp 167ndash1762009

[23] G B Mahady ldquoMedicinal plants for the prevention and treat-ment of bacterial infectionsrdquo Current Pharmaceutical Designvol 11 no 19 pp 2405ndash2427 2005

[24] V P Kumar N S Chauhan H Padh and M Rajani ldquoSearchfor antibacterial and antifungal agents from selected Indianmedicinal plantsrdquo Journal of Ethnopharmacology vol 107 no2 pp 182ndash188 2006

[25] R Nair T Kalariya and S Chanda ldquoAntibacterial activity ofsome plant extracts used in folk medicinerdquo Journal of HerbalPharmacotherapy vol 7 no 3-4 pp 191ndash201 2007

[26] D Kalemba and A Kunicka ldquoAntibacterial and antifungalproperties of essential oilsrdquo Current Medicinal Chemistry vol10 no 10 pp 813ndash829 2003

[27] A H Ashour ldquoAntibacterial antifungal and anticancer activi-ties of volatile oils and extracts from stems leaves and flowers ofEucalyptus sideroxylon and Eucalyptus torquatardquoCancer BiologyandTherapy vol 7 no 3 pp 399ndash403 2008

[28] M J Abad L M Bedoya L Apaza and P Bermejo ldquoTheArtemisia L genus a review of bioactive essential oilsrdquoMolecules vol 17 no 3 pp 2542ndash2566 2012

[29] H N Marzoug M Romdhane A Lebrihi et al ldquoEucalyptusoleosa essential oils chemical composition and antimicrobialand antioxidant activities of the oils from different plant parts(stems leaves flowers and fruits)rdquoMolecules vol 16 no 2 pp1695ndash1709 2011

[30] C Proestos D Sereli and M Komaitis ldquoDetermination ofphenolic compounds in aromatic plants by RP-HPLC and GC-MSrdquo Food Chemistry vol 95 no 1 pp 44ndash52 2006

[31] S Chao G Young C Oberg and K Nakaoka ldquoInhibition ofmethicillin-resistant Staphylococcus aureus (MRSA) by essentialoilsrdquo Flavour and Fragrance Journal vol 23 no 6 pp 444ndash4492008

[32] M Akin D Oguz and H T Saracoglu ldquoAntibacterial effectsof some plant extracts from Labiatae (Lamiaceae) growingnaturally around Sirnak-Silopi Turkeyrdquo International Journal ofPharmaceutical and Applied Sciences vol 1 no 1 pp 4ndash47 2010

[33] K Bayoub T Baibai D Mountassif A Retmane and ASoukri ldquoAntibacterial activities of the crude ethanol extracts ofmedicinal plants against Listeriamonocytogenes and some otherpathogenic strainsrdquo African Journal of Biotechnology vol 9 no27 pp 4251ndash4258 2010

[34] R P Samy P Gopalakrishnakone M Sarumathi P Houghtonand S Ignacimuthu ldquoPurification of antibacterial agents fromTragia involucratamdasha popular tribal medicine for wound heal-ingrdquo Journal of Ethnopharmacology vol 107 no 1 pp 99ndash1062006

[35] K M Matthew The Flora of Tamil Nadu Carnatic Part IndashIIIprinted at the Diocesan Press Madras India 1981ndash1983

[36] AW BauerWMKirby J C Sherris andM Turck ldquoAntibioticsusceptibility testing by a standardized single disk methodrdquoAmerican Journal of Clinical Pathology vol 45 no 4 pp 493ndash496 1966

[37] National Committee for Clinical Laboratory Standards Per-formance Standards For Antimicrobial Susceptibility Testing12th Informational Supplement Approved standard M100-S12National Committee for Clinical Laboratory Standards WaynePa USA 2002

[38] E A Elsinghorst ldquoMeasurement of invasion by gentamicinresistancerdquoMethods in Enzymology vol 236 pp 405ndash420 1994

[39] J B Harborne Phytochemical Methods Chapman and HallNew York NY USA 1976

[40] H Wagner S Bladt and E M Zgairski Plant Drug AnalysisSpringer New York NY USA 1984

[41] N Myers R A Mittermeler C G Mittermeler G A B daFonseca and J Kent ldquoBiodiversity hotspots for conservationprioritiesrdquo Nature vol 403 no 6772 pp 853ndash858 2000

[42] R Perumal Samy and S Ignacimuthu ldquoAntibacterial activity ofsome folkloremedicinal plants used by tribals inWestern Ghats

Evidence-Based Complementary and Alternative Medicine 17

of Indiardquo Journal of Ethnopharmacology vol 69 no 1 pp 63ndash712000

[43] K Nampoothiri A Chrispal A Begum S Jasmine K GGopinath and A Zachariah ldquoA clinical study of renal tubulardysfunction inCleistanthus collinus (Oduvanthalai) poisoningrdquoClinical Toxicology vol 48 no 3 pp 193ndash197 2010

[44] M Strathern UsefulKnowledge The Isiah Berlin Lecture Uni-versity of Manchester Brunswick UK

[45] C N Sunil and A K Pradeep ldquo(2001) Another new species ofTripogon (Poaceae) from Indiardquo Sida vol 19 pp 803ndash806 2005

[46] S Biber-Klemm and T Cottier Right To Plant GeneticResources and Traditional Knowledge Basic Issues and Prospec-tive Oxfordshire Nosworthy Way UK 2006

[47] V S Ramachandran S Shijo Joseph and R Aruna ldquoEth-nobotanical studies from amaravathy range of Indira GandhiWildlife SanctuaryWesternGhats CoimbatoreDistrict South-ern Indiardquo Ethnobotanical Leaflets vol 13 pp 1069ndash1087 2009

[48] A H Rajasab and I Mohamad ldquoDocumentation of folkknowledge on edible wild plants of North Karnatakardquo IndianJournal of Traditional Knowledge vol 3 no 4 pp 419ndash4292004

[49] M Rajadurai V G Vidhya M Ramya and A Bhaskar ldquoEthno-medicinal plants used by the traditional healers of pachamalaihills Tamilnadu Indiardquo Studies on Ethno-Medicine vol 3 no 1pp 39ndash41 2009

[50] S K Adesina O Idowu A O Ogundaini et al ldquoAntimicrobialconstituents of the leaves of Acalypha wilkesiana and Acalyphahispidardquo Phytotherapy Research vol 14 no 5 pp 371ndash374 2000

[51] M T Gutierrez-Lugo M P Singh W M Maiese and BN Timmermann ldquoNew antimicrobial cycloartane triterpenesfrom Acalypha communisrdquo Journal of Natural Products vol 65no 6 pp 872ndash875 2002

[52] A K Tyagi andAMalik ldquoAntimicrobial potential and chemicalcomposition of Mentha piperita oil in liquid and vapour phaseagainst food spoiling microorganismsrdquo Food Control vol 22no 11 pp 1707ndash1714 2011

[53] G Devendran and U Balasubramanian ldquoQualitative phyto-chemical screening and GC-MS analysis of Ocimum sanctumL leavesrdquoAsian Journal of Plant Science and Research vol 1 no4 pp 44ndash48 2011

[54] V C Jain N M Patel D P Shah P K Patel and B HJoshi ldquoAntioxidant and antimicrobial activities of Alangiumsalvifolium (Lf) Wang rootrdquo Global Journal of Pharmacologyvol 4 no 1 pp 13ndash18 2010

[55] P Swapna Latha andK Kannabiran ldquoAntimicrobial activity andphytochemicals of Solanum trilobatum LinnrdquoAfrican Journal ofBiotechnology vol 5 no 23 pp 2402ndash2404 2006

[56] M Usha1 S Ragini and S M A Naqvi ldquoAntibacterial activityof acetone and ethanol extracts of Cinnamon (Cinnamomumzeylanicum) and Ajowan (Trachyspermum ammi) on four foodspoilage bacteriardquo International Research Journal of BiologicalSciences vol 1 no 4 pp 7ndash11 2012

[57] S Prabuseenivasan M Jayakumar and S Ignacimuthu ldquoInvitro antibacterial activity of some plant essential oilsrdquo BMCComplementary and Alternative Medicine vol 6 article 39 pp1ndash8 2006

[58] G A Ayoola F M Lawore T Adelowotan et al ldquoChemi-cal analysis and antimicrobial activity of the essential oil ofSyzigium aromaticum (clove)rdquo African Journal of Biotechnologyvol 7 no 13 pp 2227ndash2231 2008

[59] Y J Fu L Chen Y Zu et al ldquoThe antibacterial activity of cloveessential oil against Propionibacterium acnes and its mechanismof actionrdquo Archives of Dermatology vol 145 no 1 pp 86ndash882009

[60] D Venkatesan C M Karrunakarn S Selva Kumar and P TPalani Swamy ldquoIdentification of phytochemical constituents ofAegle marmelos responsible for antimicrobial activity againstselected pathogenic organismsrdquo Ethnobotanical Leaflets vol 13pp 1362ndash1372 2009

[61] P Arun K G Purushotham J Johnsy jayarani and V KumarildquoIn vitro Antibacterial Activity of Oldenlandia umbellata anIndian medicinal Plantrdquo Journal of Pharmaceutical Science andTechnology vol 2 no 4 pp 198ndash201 2010

[62] A Manjamalai R Sardar Sathyajith Singh C Guruvayoorap-pan and V M Berlin Grace ldquoAnalysis of phytochemicalconstituents and antimicrobial activity of some medicinalplants in Taminadu Indiardquo Global Journal of Biotechnology andBiochemistry vol 5 no 2 pp 120ndash128 2010

[63] E Haslam ldquoNatural polyphenols (vegetable tannins) as drugsPossible modes of actionrdquo Journal of Natural Products vol 59no 2 pp 205ndash215 1996

[64] Y Cai Q Luo M Sun and H Corke ldquoAntioxidant activityand phenolic compounds of 112 traditional Chinese medicinalplants associated with anticancerrdquo Life Sciences vol 74 no 17pp 2157ndash2184 2004

[65] B Shan Y Z Cai M Sun and H Corke ldquoAntioxidant capacityof 26 spice extracts and characterization of their phenolicconstituentsrdquo Journal of Agricultural and Food Chemistry vol53 no 20 pp 7749ndash7759 2005

[66] R J W Lambert P N Skandamis P J Coote and G-J ENychas ldquoA study of the minimum inhibitory concentration andmode of action of oregano essential oil thymol and carvacrolrdquoJournal of AppliedMicrobiology vol 91 no 3 pp 453ndash462 2001

[67] M Raccach ldquoThe antimicrobial activity of phenolic antioxi-dants in food a reviewrdquo Journal of Food Safety vol 6 no 3 pp141ndash170 1984

[68] M Taniguchi Y Yano E Tada et al ldquoMode of action of polygo-dial an antifungal sesquiterpene dialdehyderdquo Agricultural andBiological Chemistry vol 52 no 6 pp 1409ndash1414 1988

[69] G J E Nychas ldquoNatural antimicrobials from plantsrdquo in NewMethods of Food Preservation G W Gould Ed pp 58ndash89Blackie Academic London UK 1995

[70] S Burt ldquoEssential oils their antibacterial properties and poten-tial applications in foodsmdasha reviewrdquo International Journal ofFood Microbiology vol 94 no 3 pp 223ndash253 2004

Submit your manuscripts athttpwwwhindawicom

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Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

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Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Behavioural Neurology

EndocrinologyInternational Journal of

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Disease Markers

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BioMed Research International

OncologyJournal of

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Oxidative Medicine and Cellular Longevity

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PPAR Research

The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014

Immunology ResearchHindawi Publishing Corporationhttpwwwhindawicom Volume 2014

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Parkinsonrsquos Disease

Evidence-Based Complementary and Alternative Medicine

Volume 2014Hindawi Publishing Corporationhttpwwwhindawicom

Evidence-Based Complementary and Alternative Medicine 5

Table 1 Some of the important traditional medicinal plant species families voucher specimens parts used yield of extracts phytochemicalscreening and toxicity on human macrophage cells

Scientific name Family Voucher specimen Plant parts Yield (gm) Phytochemicalanalysis

Adhatoda vasica Nees Acanthaceae D2020 Leaf 64 VasicineAegle marmelos (L)Correa ex Roxb Rutaceae D2018 Root-bark 58 Alkaloids

Alangium salvifolium (L)f Wangerin Alangiaceae 0140 Leaf 63 Phenolic

Andrographis paniculataWallich ex Nees Acanthaceae 0061 Leaf 68 Andrographolide

Andrographis echioidesNees Acanthaceae 0116 Leaf 70 Terpenoids

Acalypha indica L Euphorbiaceae 29644 Leaf 61 AcalypheAcalypha lanceolata L Euphorbiaceae 15791 Leaf 71 AlkaloidsAchyranthes aspera L Amaranthaceae 2666 Leaf 27 BetaineAgeratum conyzoides L Asteraceae 4812 Leaf 37 Essential oilsAsteracantha longifolia L Acanthaceae 0234 Stem 74 GlycosidesAzadirachta indica A Juss Meliaceae D0204 Leaf bark 67 Tannins

Borassus flabellifer L Arecaceae D0202 Root 08 Flavonoidsphenolics

Boerhavia erecta L Nyctaginaceae 10897 Whole plant 28 PhenolicsCalotropis procera (Ait)Ait f Asclepiadaceae D073 Root-bark 13 Terpenoids

Calotropis gigantea (L)RBrex Ait Asclepiadaceae D070 Milky latex 48 Alkaloids

Cassia auriculata L Caesalpiniaceae 0141 Leaf 49 Saponins

Cassia occidentalis L Caesalpiniaceae 0111 Root 95 Flavonoidssaponins

Cassia tora L Caesalpiniaceae 0100 Stem bark 69 SaponinsCassia fistula L Caesalpiniaceae 037 Whole plant 79 SaponinsCardiospermumhalicacabum L Sapindaceae 0125 Whole plant 58 Flavonoids

Catharanthus roseus (L)GDon Apocynaceae 0029 Leaf

root1407 Alkaloids

Cinnamomum zeylanicumGarcin ex Blume Lauraceae 00209 Bark 31 Essential oil

TanninCinnamomum inersReinw ex Blume Lauraceae 043-c Leaf bark Alkaloids

Cissus quadrangularisRoxb Vitaceae D02023 Leaf 68 Glycosides

Citrus acida Roxb Hookf Rutaceae 0213 Leaf 52 SaponinsTerpenoids

Centella asiatica (L) Umbelliferae 0138 Whole plant 89 FlavonoidsAlkaloids

Clerodendrum inerme (L)Gaertn Verbenaceae D02043 Stem 78 Sterols diterpenes

Clitoria ternatea L Papilionaceae D02026 Seed 98 ProteinCleistanthus collinus(Roxb) Benth andHookf

Euphorbiaceae 0011 Whole plant 003 Cleistanthincollinusin

Cleome gynandropsis L Capparidaceae 12247 Leaf 62 GlycosidesCleome viscose L Capparidaceae 29999 Leaf 27 Phenolics

6 Evidence-Based Complementary and Alternative Medicine

Table 1 Continued

Scientific name Family Voucher specimen Plant parts Yield (gm) Phytochemicalanalysis

Coccinia grandisWampA Cucurbitaceae D02030 Leaf root 09 GlycosidesCymbopogon citratus(DC) Gramineae D012 Root 025 Essential oil

Datura metel L Solanaceae D02038 Leaf stem 39 SteroidsEucalyptus globulus Labill Myrtaceae D0220 Leaf 12 TerpenoidsEclipta alba (L) Hassk Asteraceae D028 Whole plant 07 PhenolicEuphorbia hirta Linn Euphorbiaceae 0018-c Whole plant 012 mdash

Eclipta prostrata (L) Asteraceae D210 Leaf 110 Triterpenoidsaponin

Eugenia caryophyllus(Sprengel) Bullock ampHarrison

Myrtaceae 0025 Flower buds 116 Essential oils

Elettaria cardamomumWhite et Mason Zingiberaceae 0009 Fruit pods 317 Essential oils

Gloriosa superba L Liliaceae 020-S Tuber 108 Alkaloids phenol

Jatropha curcas L Euphorbiaceae 015 Whole plant 53 Alkaloidsflavonoids

Hyptis suaveolens (L) Poit Lamiaceae 24688 Leaf 63 Essential oilHemidesmus indicus L Asclepiadaceae D-009 Roots CoumarinsIchnocarpus frutescens(L) RBr Apocynaceae 0110 Root flower 73 Terpenoids

Leucas aspera (Willd)Link Labiatae 0114 Leaf 83 Triterpenes

Lawsonia inermis L Lythraceae T261 Leaf 09 Glycosidesphenolic

Madhuca longifolia (L) JFMacbr Sapotaceae D01415 Nut 93 Sitosterol

Merremia hastate L(Desr) Hallierf Convolvulaceae 10894 Whole plant 40 Alkaloids

Mentha piperita L Lamiaceae 0217-c Whole plant 07 Essential oilsMorinda tinctoria Roxb Rubiaceae 0122 Leaf 14 GlycosidesMimosa pudica L Mimosaceae 0071 Whole plant 06 mdashOldenlandia umbellata L Rubiaceae D02047 Leaf 44 AlkaloidsOcimum sanctum L Lamiaceae 0016 Whole plant 30 AlkaloidsPiper attenuatum BuchHamex Miq Piperaceae 007 Flower 46 Alkaloids

Plumbago zeylanica (L)Cav Plumbaginaceae 0121 Root 48 Plumbagin

Plectranthus amboinicus(L) Spreng Lamiaceae 0410 Whole plant 12 Essential oils

terpenoidsPhyllanthus debilis L(Klein ex Willd) Euphorbiaceae 0120 Whole plant 49 Polyphenol

Phyllanthusmadraspatensis L Euphorbiaceae 0117 Whole plant 50 Polyphenol

Premna tomentosaWilld Verbenaceae 0129 Leaf 53 DiterpenesRosmarinus officinalis L Lamiaceae 0017 Root 023 Essential oilsRauwolfia serpentine L Apocynaceae 020-S Root 115 AlkaloidSebastiania chamaelea (L)Muell Arg Euphorbiaceae 0034 Leaf 11 Polyphenol

Solanum trilobatum L Solanaceae D02054 Leaf flower 40 Tannins

Evidence-Based Complementary and Alternative Medicine 7

Table 1 Continued

Scientific name Family Voucher specimen Plant parts Yield (gm) Phytochemicalanalysis

Sphaeranthus indicus L Asteraceae D02060 Whole plant 10 Essential oilSwertia chirata (L) Ham Gentianaceae D0540 Whole plant 16 GlycodisesStrychnos nux-vomica L Loganiaceae S-22 Nuts 036 AlkaloidsTragia involucrata L Euphorbiaceae D068 Leaves 16 ShellsolTinospora cordifolia(Willd) Miers ex Hooff ampThoms

Menispermaceae 0118 Leafroot stem 50 Glycosides tannins

Tridax procumbens L Compositae 10649 Leaf 18 FlavonoidsTerminalia arjuna (DC)W amp A Combretaceae 033-c Bark 80 Phenolics

Tephrosia purpurea (L)Pers Fabaceae S-43 Whole plant 08 Isoflavone

Vitex negundo L Verbenaceae 0031 Leaf 24 TerpineolVetiveria zizanioides L Gramineae 0051 Root 103 Essential oilWithania somnifera (L)Dunal Solanaceae D02063 Root 21 Alkaloids

Wedelia calendulacea Less Asteraceae S-24 Leaves FlavonoidsZingiber officinale Rosc Zingiberaceae 0327 Rhizome 23 TanninsZanthoxylum limonella(Dennst) Alston Rutaceae 009 Bark 19 Alkaloids essential

oilClass of chemical compounds A alkaloids S saponins T tannins St steroids G glycosides T terpenoids P polyphenol P phenolics Sh shellsol Hhydrocarbon esters

the site of snakebite and decoction is given orally fortreatment by indigenous people Besides these tribes relyon the medicinal plants as ingredients for fabricating a kindof medicated stone for health management ldquoVishakallurdquo(poison stone) is used by the indigenous groups called Kaniin Kerala India to treat a snakebiteWhen the stone is placeddirectly on the bitten area it sticks to the body to absorb thepoison and then become detached when absorption seems tobe complete The ingredients of Vishakallu stones are madewith leaves of Ocimum sanctum Anisomeles malabaricaLeucas aspera Piper betle Santalum album and the pebblescollected from the river bank

35 Survey of Medicinal Plants and Their Health Care ValuesThe present study is an attempt to provide scientific basis andobtain justification for the traditional beliefs of reliance on arich diversity of ethnomedicinal plants along with the richheritage of traditional medicine practices related to healthcare system made available by the primitive tribal commu-nities located at different settlements The native traditionalpractitioners called ldquovaidyarsrdquo have a good knowledge aboutthe traditional plants locally available for treatment of var-ious diseases (Figures 3(a)ndash3(p)) Such traditional medicalknowledge is used for preparing home remedies ill healthprevention and routine health maintenanceThis knowledgeis also applicable to cover other sectors of social life Duringthe ethnobotanical survey the wealth of 78 medicinal plantspecies used by the indigenous tribal community for varioustypes of health treatment was documented The botanical

names family names parts used chemical constituents andtheir application are provided (Table 1)

36 Antimicrobial Activity of Crude Extracts In this studywe reported the antimicrobial screening of methanolic crudeextracts of 78 medicinal plants (Table 2) Results revealedthat 68 plant extracts displayed potent activities against oneor more Gram-positive and -negative bacteria Of whichTragia involucrata Citrus acida Aegle marmelos Adha-toda vasica Calotropis procera Andrographis paniculata andMentha piperita Azadirachta indica Sphaeranthus indicusand Elettaria cardamomum showed the highest antibacte-rial activity against the multidrug resistant B pseudomallei(KHW and TES) and S aureus at 100120583gmL concentrationThe extracts showed pronounced antibacterial activity withtheir inhibitory zones ranging from 20 to 31mm in diam-eter as compared to the standard drugs chloramphenicoland ceftazidime (29ndash33mm) The majority of the plantsdemonstrated a powerful antimicrobial potency against themultidrug resistant strains of B pseudomallei (KHW andTES) K pneumonia and S aureus Approximately twenty-one plant extracts exerted only a weak or moderate effectagainst the tested bacteria while the crude extract of 13plants failed to show any effect at all Except for the plantextracts of T involucrata A lanceolata A vasica and Sindicus extracts the majority of the plant extracts wereineffective against theV damsela infection fascinatingly only11 plants exhibited activity against P aeruginosa of whichS indicus M piperita and C procera were found to havevery strong inhibition of bacteria at the tested concentrations

8 Evidence-Based Complementary and Alternative Medicine

Table 2 Antimicrobial activity of methanol extract of aromatic medicinal plants and essential oils evaluated against multidrug resistant(MDR) human pathogens at 100120583gmL concentration

Scientific name Microorganisms growth inhibition zones (6 millimeter in diameters)KHW TES Kp Kpr Pa Sa Stp Sp Vd Vd

Adhatoda vasica Nees 28 19 22 18 12 mdash 10 17 16 mdashAegle marmelos (L) Correa ex Roxb 29 17 9 10 mdash 15 mdash mdash mdash mdashAlangium salvifolium (L) f Wangerin 15 10 8 8 mdash 12 9 10 mdash mdashAndrographis echioides L 12 9 mdash mdash mdash 17 8 9 mdash mdashAndrographis paniculataWallich ex Nees 26 21 19 13 mdash 25 8 16 12 mdashAcalypha indica L mdash mdash mdash mdash mdash 18 mdash mdash mdash mdashAcalypha lanceolata L mdash mdash 12 13 mdash 10 mdash mdash 21 mdashAchyranthes aspera L mdash mdash 7 14 mdash 15 12 11 mdash mdashAgeratum conyzoides L 10 8 mdash mdash mdash mdash mdash mdash mdash mdashAsteracantha longifolia L 16 12 mdash mdash mdash 15 mdash mdash mdash mdashAzadirachta indica A Juss 15 17 21 16 14 23 12 14 mdash mdashBorassus flabellifer L 9 10 17 8 mdash mdash 9 10 mdash mdashBoerhavia erecta L mdash mdash 8 9 7 16 mdash mdash mdash mdashCalotropis procera (L) mdash mdash 15 mdash 18 28 9 mdash mdash mdashCalotropis gigantea (L) RBrex Ait 11 9 mdash mdash mdash 20 mdash mdash 9 8Cardiospermum halicacabum L 23 mdash mdash 14 mdash 9 19 mdash mdash mdashCatharanthus roseus (L) GDon 13 7 12 9 mdash 15 11 8 mdash mdashCassia auriculata L 17 13 12 mdash mdash 19 13 mdash mdash mdashCassia occidentalis L 18 mdash mdash mdash mdash mdash mdash mdash mdash mdashCassia tora L mdash mdash mdash mdash mdash mdash mdash mdash mdash mdashCassia fistula L mdash mdash mdash mdash mdash mdash mdash mdash mdash mdashCitrus acida Roxb Hookf 26 22 mdash 12 29 9 13 8 mdashCissus quadrangularis L mdash mdash mdash mdash mdash mdash mdash mdash mdash mdashCinnamomum zeylanicum Garcin exBlume 14 16 7 20 mdash 22 19 7 mdash mdash

Cinnamomum iners Reinw ex Blume 20 16 mdash 15 mdash 16 12 mdash mdash mdashRosmarinus officinalis L mdash 9 mdash 10 mdash 7 mdash 8 mdash mdashCentella asiatica (L) 9 8 mdash mdash mdash 11 mdash mdash mdash mdashClerodendrum inerme (L) Gaertn 13 7 12 mdash mdash 15 11 8 mdash mdashClitoria ternatea L mdash mdash mdash mdash mdash mdash mdash mdash mdash mdashClitoria ternatea L 16 12 mdash 13 mdash 8 10 mdash mdash mdashCleome gynandropsis L mdash mdash 12 19 11 mdash 9 15 12 mdashCleome viscose L mdash mdash 8 10 14 20 mdash mdash mdash mdashCoccinia grandisWampA 9 10 17 8 mdash mdash 9 10 mdash mdashCymbopogon citratus (DC) 16 18 mdash 17 mdash 14 mdash mdash mdash mdashDatura metel L mdash mdash mdash mdash mdash mdash mdash mdash mdash mdashEclipta alba (L) Hassk 20 mdash mdash 9 mdash mdash 10 mdash mdash mdashEuphorbia hirta Linn 11 mdash mdash mdash mdash 16 mdash mdash mdash mdashEucalyptus globulus Labill mdash mdash mdash 7 mdash mdash mdash 7 mdash mdashEugenia caryophyllus Bullock amp Harrison mdash 7 mdash 11 mdash 9 mdash 8 mdash mdashElettaria cardamomumWhite et Mason 21 20 7 14 mdash 22 12 17 mdash mdashHyptis suaveolens (L) Poit 8 10 8 7 mdash mdash mdash mdash mdash mdashIchnocarpus frutescens (L) WJ Aiton mdash mdash mdash mdash mdash mdash mdash mdash mdash mdashJatropha curcas L mdash mdash mdash 8 mdash 11 mdash 12 mdash 7Leucas aspera (Willd) Link 9 10 17 8 mdash 12 9 10 mdash mdashLawsonia inermis L mdash mdash mdash mdash mdash mdash mdash mdash mdash mdashMadhuca longifolia (L) JF Macbr 18 16 14 19 12 mdash 8 7 9 mdash

Evidence-Based Complementary and Alternative Medicine 9

Table 2 Continued

Scientific name Microorganisms growth inhibition zones (6 millimeter in diameters)KHW TES Kp Kpr Pa Sa Stp Sp Vd Vd

Merremia hastate L (Desr) Hallierf mdash mdash mdash mdash mdash mdash mdash mdash mdash mdashMorinda tinctoria Roxb mdash mdash mdash mdash mdash mdash mdash mdash mdash mdashMentha piperita L 23 17 26 12 20 25 19 mdash mdash mdashOcimum sanctum L 12 9 11 7 mdash 15 7 8 mdash mdashOldenlandia umbellata L mdash mdash mdash mdash mdash 17 mdash mdash mdash mdashPiper attenuatum Buch Hamex Miq 13 7 12 21 mdash 17 11 8 mdash mdashPlumbago zeylanica (L) Cav 9 10 17 8 mdash 12 9 10 mdash mdashPlectranthus amboinicus (L) Spreng mdash mdash mdash 8 11 15 mdash mdash mdash mdashPhyllanthus debilis L (Klein ex Willd) 7 8 mdash 9 mdash 9 18 7 mdash mdashPhyllanthus maderaspatensis L 17 mdash mdash mdash mdash mdash mdash mdash mdash mdashPremna tomentosaWilld 13 10 mdash 15 mdash 9 10 mdash mdash mdashGloriosa superba L 17 16 7 8 mdash 15 mdash mdash mdash 8Sebastiania chamaelea (L) Muell Arg 19 12 mdash 17 mdash 13 19 8 mdash mdashSolanum trilobatum L mdash mdash 13 9 mdash 8 mdash mdash mdash mdashSphaeranthus indicus L 20 18 21 7 22 mdash 11 9 mdash 16Swertia chirata (L) Ham mdash mdash mdash mdash mdash mdash mdash mdash mdash mdashTerminalia arjuna (W amp A) mdash mdash mdash mdash mdash 16 mdash mdash mdash mdashTinospora cordifolia (Willd) Miers exHooff ampThoms mdash mdash 23 16 mdash 12 15 mdash mdash mdash

Tridax procumbens L 9 8 7 mdash mdash 14 mdash mdash mdash mdashTragia involucrata L 25 23 20 mdash mdash 31 28 22 19 mdashVitex negundo L mdash mdash mdash mdash mdash 14 mdash mdash mdash mdashVetiveria zizanioides (L) 11 9 7 7 mdash 16 mdash mdash 8Withania somnifera (L) Dunal 12 20 mdash mdash mdash 15 12 mdash mdash mdashZingiber officinale Rosc 14 11 7 15 mdash 7 12 7 mdash mdashZanthoxylum limonella (Dennst) Alston 13 7 12 9 mdash mdash 11 8 mdash mdashChloramphenicol (30120583gdisc) 21 12 15 17 29 16 15 18 13 11Ceftazidime (30 120583gdisc) 33 16 22 19 16 25 21 20 12 15lowastBacteria (+minus) Results obtained in the disc diffusion assay antibacterial activity is expressed as the mean plusmn SD (119899 = 3) of the inhibition by the extract andits diameter around the discs One way analysis of variance was performed (mean plusmn SD 119899 = 3 replicates) Size of inhibition zones were including the sterileblank discs 6 millimeter (mm) in diameters Absence of bacterial inhibition indicates (mdash) antibiotic disc (30 120583gdisc)

Interestingly sixteen plants such as Andrographis echioidesC auriculata C viscose C gigantea T arjuna Oldenlandiaumbellata Boerhavia erecta and E hirta exerted a strongactivity against the Gram-positive S aureus bacteria

37 Phytochemical Screening of Plants The results obtainedfrom the phytochemical screening as shown in Table 1 indi-cate the presence of various types of secondary metabolitessuch as polyphenols tannins saponins alkaloids and gly-cosidespolysaccharides Most of the plant extracts relativelyrich in alkaloids phenols flavonoids polyphenols tanninssterols and terpenoids were found to inhibit the growth oforganisms

38 Antimicrobial Activity of Fractioned Compounds Activecomponents were purified from the most active extractsfor further testing The compound shellsol of T involucrata

and C acida exhibited the most potent action againstthe antibiotic resistant strains of B pseudomallei (KHW)S aureus B pseudomallei (TES) and K pneumoniae Amarmelos was also found to inhibit the growth of B pseudo-mallei (KHW) more effectively than other tested bacteria Avasica showed the broad spectrum growth inhibitory activityon B pseudomallei (KHW) K pneumoniae K pneumoniaeresistant B pseudomallei (TES) S pyogenes and V damselaHowever E cardamomum displayed antimicrobial activityon some of the B pseudomallei (KHW) S pyogenes Bpseudomallei (TES) and S typhi strains Similarly A indicaexerted the growth inhibition on K pneumoniae and Saureus Remarkably Sebastiania chamaelea was more activeagainst K pneumoniae K pneumoniae and S pyogenesThe compound from S indicus inhibited the growth of Kpneumoniae K pneumoniae B pseudomallei (KHW andTES) and S typhi strains as compared to the activity shown

10 Evidence-Based Complementary and Alternative Medicine

Figure 2 Diverse biodiversity richness of medicinal plants in Western and Eastern Ghats (a) Topography of plant covering area in Kollihills (Namakkal district Tamil Nadu) (b) Aerial view of occurrence of medicinal plants in Mundanthurai hills (Tirunelveli district TN) (c)Pachamalai hills (Trichy district) and its natural vegetation inhabitants for Malaiyali tribes (d) Deforestation of natural herbal resources dueto urbanization in Kalrayan hills (Salem district) in the Eastern Ghats of TN (e) Medicinal plants and its various parts used by the natives(traditional healers) for the treatment of diverse human illness with a very high percentage of leaves and whole plants often used for herbaldrug preparation by the local practitioners (f) Various category of plants like shrub herb climbers and tree and the parts used in medicine

Evidence-Based Complementary and Alternative Medicine 11

Figure 3 Medicinal aromatic herbs spices and toxic plants were collected from the tribal areas of the Western and Eastern Ghats region inTamil Nadu India (a) A vasica Nees (leaf) (b) Eclipta alba (L) Hassk (whole plant) (c)Mimosa pudica L (whole plant) (d) P amboinicus(L) Spreng (whole plant) (e) T procumbens (L) (whole plant) (f) Euphorbia hirta Linn (whole plant) (g) A paniculata Wallich ex Nees(H) C roseus (L) GDon (whole plant) used for therapy (i) Cinnamomum iners Reinw ex Blume (leaf) (j) E globulus Labill (leaf and bark)(k) Z officinale Rosc (Rhizome) (l) E caryophyllus (Sprengel) Bullock and Harrison (flower buds) (m) M piperita L (whole plant) (n) Ccitratus (DC) Clitoria ternatea L (whole plant) (o) C zeylanicum Garcin ex Blume (bark) (p) Elettaria cardamomumWhite et Mason (fruitpod) used for medicine and food preparation

by the crude extracts (Figure 4)The antimicrobial efficacy offractions collected from the oil yielding plants was also com-pared with that of the tested compounds C zeylanicum andR officinaliswere themost sensitive in controlling the growthof B pseudomallei (KHW) S aureus K pneumonia and Spneumoniae Fascinatingly all the compounds obtained fromaromatic plants except those from E globules were foundto be very effective against the multidrug resistant humanpathogen B pseudomallei (KHW) that causes melioidosis

On the other hand compounds from C citrates O sanctumE caryophyllus and Z zizanioide showed some promisingeffect only against S aureus (Figure 5) The activity of thecompounds were pronounced more than that of the oilyielding plant fractions

39 Minimum Inhibitory Concentrations (MICs) The antibi-otic potential of the purified fractions was obtained fromthe MIC determination The hydrocarbon ester shellsol

12 Evidence-Based Complementary and Alternative Medicine

(mean plusmn SD n = 3)

0

5

10

15

20

25

30

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn V

d St

Bacteria (+minus)

lowast

lowast

lowast

A vasica

In vitro assay(P

lant

frac

tion)

Inhi

bitio

n zo

nes (

mm

)

(a)

(mean plusmn SD n = 3)

0

5

10

15

20

25

30

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn V

d St

Bacteria (+minus)

lowast

A marmelos

Inhi

bitio

n zo

nes (

mm

)

In vitro assay

(b)

(mean plusmn SD n = 3)

0

5

10

15

20

25

30

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn V

d St

Bacteria (+minus)

lowastlowast

A indica

Inhi

bitio

n zo

nes (

mm

)

Incu

batio

n (37∘C

24 h

)

(c)

(mean plusmn SD n = 3)

0

5

10

15

20

25

30

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn V

d St

Bacteria (+minus)

lowast

lowast lowast

35

C acida

Inhi

bitio

n zo

nes (

mm

)(P

lant

frac

tion)

(d)

(mean plusmn SD n = 3)

0

5

10

15

20

25

30

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn V

d St

Bacteria (+minus)

lowastlowast

lowast

S chamaelea

Inhi

bitio

n zo

nes (

mm

)

(e)

(mean plusmn SD n = 3)

0

5

10

15

20

25

30

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn V

d St

Bacteria (+minus)

lowastlowast

lowastlowast

S indicus

Inhi

bitio

n zo

nes (

mm

)

Incu

batio

n (37∘C

24 h

)

(f)

(mean plusmn SD n = 3)

0

5

10

15

20

25

30

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn V

d St

Bacteria (+minus)

lowast

lowast lowast

E cardamomum

Inhi

bitio

n zo

nes (

mm

)(P

lant

frac

tion)

(g)

(mean plusmn SD n = 3)

0

5

10

15

20

25

30

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn V

d St

Bacteria (+minus)

lowast

lowast

lowast

lowast

Shellsol

35

Inhi

bitio

n zo

nes (

mm

)

Incu

batio

n (37∘C

24 h

)

(h)

Figure 4 In vitro antimicrobial activity of purified fractions from the most active plant extracts tested against bacteria Growth inhibitionzones were measured and analyzed with mean plusmn standard deviation (SD) (119899 = 3) using one way analysis of variance Level of significanceat (lowast119875 gt 001) Most of the fractions exerted a potent inhibitory effect against multidrug resistant Gram-negative bacteria (B pseudomalleistrains KHW and TES) K pneumonia and Gram-positive bacteria S aureus

(T involucrata) andC acida showed an interesting inhibitorypotential against S aureus (MIC of 78 120583gmL) and B pseu-domallei strain of KHW (MIC of 156 120583gmL) A vasicashowed an MIC of 156120583gmL against B pseudomallei(KHW) and an MIC of 3125120583gmL against K pneumo-niae K pneumoniae S pyogenes and V damsela strainsFractions from A marmelos and terpenoid from A indicaexerted bacteriostatic effect with MIC values of 3125120583gmLon some selected bacteria including B pseudomallei ofKHW S aureus and B pseudomallei of TES The MIC of3125 120583gmL was found for E cardamomum against S aureus

K pneumonia and S pyogenes S indicus displayed a verystrong inhibition against MDR K pneumoniae (MIC of156 120583gmL) and against B pseudomallei (KHW and TES)at MIC of 3125 120583gmL When the antimicrobial efficacies ofpurified fractions from aromatic plants were compared theC zeylanicum fraction displayed an important antimicrobialeffect against S aureus (MIC of 78 120583gmL) MDR B pseu-domallei of KHW (MIC of 156 120583gmL) and S pneumoniae(MIC of 3125 120583gmL) The essential oil from M piperateashowed MIC value of 3125120583gmL against K pneumoniae Saureus and B pseudomallei (KHW) respectivelyO sanctum

Evidence-Based Complementary and Alternative Medicine 13

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn

Bacteria (+minus)

0

5

10

15

20

25

30

Inhi

bitio

n zo

nes (

mm

)(P

lant

frac

tion)

lowastlowast

lowast

(mean plusmn SD n = 3)In vitro assay

C citratus

(a)

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn

Bacteria (+minus)

0

5

10

15

20

25

30

Inhi

bitio

n zo

nes (

mm

)

lowast

lowast

lowastlowast

lowast

lowast

(mean plusmn SD n = 3)In vitro assay

35

C zeylanicum

(b)

Bacteria (+minus)

0

5

10

15

20

25

30

Inhi

bitio

n zo

nes (

mm

)

lowast

lowast

lowast

(mean plusmn SD n = 3)

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn V

d St

O sanctum

Incu

batio

n (37∘C

24 h

)

(c)

Bacteria (+minus)

0

5

10

15

20

25

Inhi

bitio

n zo

nes (

mm

)

lowast

lowast

(mean plusmn SD n = 3)

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn V

d St

M piperita

(Pla

nt fr

actio

n)

(d)

Bacteria (+minus)

0

5

10

15

20

25

30

Inhi

bitio

n zo

nes (

mm

)lowast

lowast

lowast

(mean plusmn SD n = 3)

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn V

d St

E caryophyllus

(e)

0

5

10

15

20

25

30

Inhi

bitio

n zo

nes (

mm

) (mean plusmn SD n = 3)

Bacteria (+minus)

KHW

TES

Kp

KprPa Sa

Stp

Spn

E globulus

Incu

batio

n (37∘C

24 h

)

(f)

Bacteria (+minus)

0

5

10

15

20

25

30

Inhi

bitio

n zo

nes (

mm

)(P

lant

frac

tion)

lowast

lowast

lowast

lowast

(mean plusmn SD n = 3)

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn V

d

R officinalis

(g)

0

5

10

15

20

25

30

Inhi

bitio

n zo

nes (

mm

)

lowast

lowast

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn V

d St

Bacteria (+minus)V zizanioides

(mean plusmn SD n = 3)

Incu

batio

n (37∘C

24 h

)

(h)

Figure 5 Comparison of antimicrobial effect of plant compounds obtained from the most popularly used sources of essential oils as assayedby the disc-diffusion method in vitro It displayed a powerful activity against B pseudomallei and S aureus than the other bacteria strainsOther compounds showed only a moderate or weak action against the tested bacteria Values for zone of bacterial growth inhibition werepresented as mean plusmn SD (119899 = 3) with level of significance at (lowast119875 gt 001)

and C citratus fractions also showed antimicrobial activity(MICs of 3125ndash125120583gmL) only at higher concentrationsagainst the tested bacteria In addition to that higher concen-trations (gt250120583gmL) (of Vetiveria fractions) were requiredto inhibit Vibro species and others (including E globulusfractions) failed to show any effect at tested concentrations(78ndash125 120583gmL) However the purified fractions (from mostactive medicinal plants) showed strong bacteriostatic inhibi-tion against the tested organisms (Table 3)

310 Cytotoxic Effects of Plants When the components wereassayed for cytotoxicity against the normal human skinfibroblasts (HEPK) cells the compounds obtained from

E cardamomum T involucrata S indicusC acidaA vasicaA marmelos A indica and A paniculata did not showtoxicity up to 1000120583gmL (see Figures S1 and S2 in Supple-mentaryMaterial available online at httpdxdoiorg1011552013525613) A slight reduction of cell proliferation wasnoted only at higher doses (2000120583gmL) In contrast cellproliferation was markedly reduced after exposure of HEPKcells to O sanctum E globulus V zizanioides C citratusand E globulus compounds There was no gradual reductionin skin cell proliferation seen after exposure to C zeylan-icum R officinalis and M piperita (see Figures S3 and S4)compounds The toxicity was found to be concentration-dependent when the skin fibroblasts (HEPK) cells were

14 Evidence-Based Complementary and Alternative Medicine

Table 3 Minimum inhibitory concentrations (MICs) of purified plant fractions and essential oils against antibiotic resistant bacteria

Botanical name Family Parts used Gram-positive and -negative bacteria (MICs 120583gmL)KHW TES Kp KPr Sa Stp Sp Vd St

A indica Juss Meliaceae Seed (fraction) 3125 625 125 lt250 3125 mdash mdash mdash mdashA marmelos (L) Rutaceae Root-bark (F) 3125 3125 mdash mdash 3125 625 125 250 mdashA paniculata Nees Acanthaceae Leaf (fraction) 250 mdash 125 mdash 625 mdash mdash 125 mdashA vasica Nees Acanthaceae Fraction (Stem) 156 3125 3125 625 mdash lt250 3125 3125 mdashC acida Roxb Rutaceae Leaf (fraction) 156 625 mdash mdash 78 mdash 3125 mdash mdashE cardamomumWhite et Mason Acanthaceae Fraction (WP) 625 mdash 3125 mdash 3125 250 625 mdash mdashS indicus (L) Euphorbiaceae Whole plant (F) 3125 125 156 mdash 625 mdash mdash lt250 mdashT involucrata (L) Euphorbiaceae Shellosol (leaf) 156 3125 mdash mdash 78 625 mdash mdash mdashCinnamomum zeylanicum (L) Lauraceae Bark (fraction) 156 625 mdash mdash 78 3125 mdash mdash 125Cymbopogon citratus (L) Graminae Leaf (fraction) 250 lt250 625 mdash 625 125 mdash mdash mdashEugenia caryophyllus (L) Myrtaceae Flower buds (F) 625 125 mdash mdash 625 mdash mdash mdash mdashEucalyptus globulus (L) Myrtaceae Fraction (leaf) mdash mdash mdash mdash mdash mdash lt250 mdash lt250Mentha piperita (L) Labiatae Fraction (WP) mdash 625 3125 mdash 3125 mdash lt250 mdash mdashOcimum sanctum (L) Labiatae Leaf (fraction) 625 125 mdash 3125 625 mdash 125 lt250 mdashRosmarinus officinalis (L) Labiatae Rosemary oil 3125 mdash mdash mdash 3125 mdash mdash mdash mdashVetiveria zizanioides (L) Graminae Root (fraction) mdash 125 125 mdash mdash mdash gt250 gt250The bacterial growth inhibitory activity was compared for significant differences within the bacterial strains by broth-dilution method at 250 125 625 3125156 and 78 120583gmL F fractions

exposed to various compounds The cell proliferation wasincreased by the influence of the plant components at thelower concentrations Whereas the oil yielding plant com-pounds showed inhibition of cell proliferation and toxicity athigher doses 250ndash1000120583gmL

4 Discussion

The Western Ghats is considered as one of the richestbiodiversity hotspots in the world [41] In this survey wecollected nearly 78 medicinal plants from Western andEastern Ghats that are edible and popularly used for curingvarious ailments including snakebite Traditional remedieshave a long-standing history in many tribal settlements inTN India and they continue to provide useful and applicabletools for treating ailments [42] The ingredients that makeup the ldquoVishakallurdquo stone which is used as an antidote forsnakebite are different herbs and pebbles available from theriver banks Likewise aqueous paste and decoction obtainedfrom the leaves of A paniculata are widely used for snakebitetreatment by indigenous people [43] Previous studies havereported that ethnomedicine plays major roles in conservingthe disappearing knowledge of tribal communities [44ndash47]The traditional beliefs of reliance on a rich diversity ofethnomedicinal plants located at different settlements havealso been confirmed in another study [48 49] Herein weexplored the various types of traditional practices reported bythe primitive tribal communities with a view to gain furtherknowledge from such studies

In the present investigation potentially rich sources oftribal medicine (71 plants) were scientifically evaluated fortheir antibacterial activity against the MDR bacteria and theaccumulated data was disseminated for the first time to the

scientific community Out of the 71medicinal plants screenedfor the antibacterial activity 10 of them (T involucrataC acida A marmelos A vasica C procera A paniculataand M piperita A indica S indicus and E cardamomum)displayed the highest antibacterial activity against the mul-tidrug resistant B pseudomallei (KHW and TES) and Saureus strains The antibacterial activity of those crude plantextracts was as equally effective as that of the standard drugsOur findings corroborated with the previous reports madeon the antistaphylococcal activity of tribal medicinal plants[34 50 51] On the other hand isolated components from themost active extracts of T involucrata shellsol and C acidaexhibited the most potent action against the antibiotic resis-tant B pseudomallei (KHW) K pneumoniae and S aureusstrainsThese results further confirmed our previous findingson the leaves of T involucrata and its compounds hydrocar-bon ester-like shellsol which displayed a high antibacterialeffect against the different bacterial strains especially thatof S aureus [34] Eugenol and caryophyllene are the activeagents contained in the M piperita [52] and O sanctumplants which are believed to be mainly responsible for theantimicrobial properties of these plants [53] Interestinglythe compounds obtained from the aromatic plants such asC zeylanicum and R officinalis were also found to be veryeffective against the multidrug resistant human pathogen Kpneumoniae S aureus S typhi and B pseudomallei (KHW)that causes melioidosis

The inhibitory potential determined for the shellsol(T involucrata) vaseline (A vasica) C acida and C zey-lanicum indicates that the MIC of 78ndash3125120583gmL foundagainst the B pseudomallei of KHW K pneumoniae Kpneumoniae S pyogenes and V damsel and S pneumoniaewas quite low Similarly lower MIC values were found for

Evidence-Based Complementary and Alternative Medicine 15

M piperita (MIC of 113ndash225 mgmL) against the abovebacterial strains [52] MIC was found for the most activealcohol extracts of A salvifolium (MIC 0034ndash0263mgmL)on S aureus [54] and the MIC for S trilobatum aqueousextracts determined against the tested organisms rangedfrom 006 to 05mgmL [55] C zeylanicum was foundto have an effective antibacterial activity (MIC 64 120583gmL)against P aeruginosa E coli B subtilis and S aureus[56] Previously several investigators have demonstrated thatactive agents exert interesting activity against bacteria evenat lower concentrations tested [57 58] The bacteriostaticmechanism involves damage to the cell walls of bacteriafollowed by inhibition of protein synthesis that ultimatelyleads to bacterial death [59] The most active plants arewidely used by various tribes as traditional treatment (iecut wounds skin infection and scabies) thus indicatingthe potential for further development into promising drugsIn order to ascertain the safety and efficacy of the mostactive compounds their effect on human skin fibroblast cellswas evaluated Chemical constituents of E cardamomumT involucrata S indicus C acida A vasica A marmelosA indica and A paniculata plants failed to produce anynoticeable toxicity up to 1000120583gmL Although some ofthe tested compounds exhibited a slight reduction of cellproliferation and some minor morphological changes suchchanges were insignificant at lower doses and became evidentonly at higher doses However certain aromatic compoundsof O sanctum E globulus V zizanioides C citratus and Eglobulus plants showed reduction of cell proliferation againstHEPK cells

Our phytochemical screening also provides evidenceof the presence of several types of compounds that aremainly responsible for the remarkable antibacterial effectof these plants The differences noted for the bactericidalactivity of various plant extracts in this study appears tobe directly related to the diversity of compounds (shown inparentheses) that are accumulated in the following plants(eg A marmelos and O umbellate) [60ndash62] Compoundslike tannins phenol and polyphenols can bind the Gram-negative bacteria to form a heavy soluble complex on the cellsurface which subsequently disturbs the availability of recep-tor on cells and kills the bacteria [63] Several species havingwide spectra of antimicrobial activity mainly due to theactive constituents such as essential oil phenolic compoundslike thymol carvacrol in oregano and thyme eugenol inclove and cinnamon were also identified previously [64 65]Essential oils degrade the cell wall interact with the cell com-ponents and then disrupt the cytoplasmic membrane [66]The antimicrobial effect of phenolic compounds may involvemultiplemodes of action including damage to themembraneprotein interference with membrane integrated enzymes[67] causing leakage of cellular components coagulation ofcytoplasm depletion of the proton motive force alterationof fatty acid and phospholipid constituents impairmentof enzymatic mechanisms for production and metabolismalteration of nutrient uptake and electron transport [68]influencing the synthesis of DNA and RNA and destroyingprotein translation and the function of the mitochondrionin eukaryotes [69] The mode of action of antimicrobial

agents depends on the type of microorganism and is mainlyrelated to their cell wall structure and the outer membranearrangement Most of the plant spices and herbs containcomplex phenolics (ie phenolic acids flavonoids tanninslignans coumarins quinines) In addition the mechanismsof action of each phenolic compound against various bacteriaare also very complicated [26 70] Further investigation istherefore required to understand the relationship betweenthe antimicrobial action and the chemical structure of everyphenolic compound in the tested extracts The informationavailable from previous pharmacological sources combinedwith the findings herein reported on the medicinal plantextracts may serve as essential data for future drug develop-ment to combat diseases caused by theMDRbacterial strains

Abbreviations

MDR Multidrug resistant bacteriaTES Strain of Burkholderia pseudomalleiMRSA Methicillin-resistant Staphylococcus aureusVRE Vancomycin-resistant enterococciMTB Mycobacterium tuberculosisCH3OH Methanol

CFU Colony forming unitHCl Hydrochloric acidTLC Thin layer chromatographyH2SO4 Sulphuric acids

UV Ultraviolet spectrophotometerHEPK Human skin fibroblasts

Acknowledgments

Theauthors are thankful to the ERI Loyola College ChennaiIndia for his support during the survey The authors alsothank the Department of Microbiology NUHS NUS forthe bacterial cultures used in this investigation The authorsare thankful to the tribal people for sharing their healingknowledge on medicinal aromatic plants

References

[1] M N Alekshun and S B Levy ldquoMolecular mechanisms ofantibacterialmultidrug resistancerdquoCell vol 128 no 6 pp 1037ndash1050 2007

[2] A D Russell ldquoAntibiotic and biocide resistance in bacteriaintroductionrdquo Symposium Series Society for Applied Microbiol-ogy vol 31 no 92 pp 1Sndash3S 2002

[3] D R Guay ldquoTreatment of bacterial skin and skin structureinfectionsrdquo Expert Opinion on Pharmacotherapy vol 4 no 8pp 1259ndash1275 2003

[4] A J Brosnahan and P M Schlievert ldquoGram-positive bac-terial superantigen outside-in signaling causes toxic shocksyndromerdquo FEBS Journal vol 278 no 23 pp 4649ndash4667 2011

[5] K Fellermann J Wehkamp K R Herrlinger and E F StangeldquoCrohnrsquos disease a defensin deficiency syndromerdquo EuropeanJournal of Gastroenterology and Hepatology vol 15 no 6 pp627ndash634 2003

16 Evidence-Based Complementary and Alternative Medicine

[6] L Chen S Lin R Agha-Majzoub L Overbergh C Mathieuand L S Chan ldquoCCL27 is a critical factor for the developmentof atopic dermatitis in the keratin-14 IL-4 transgenic mousemodelrdquo International Immunology vol 18 no 8 pp 1233ndash12422006

[7] J H Song V Thamlikitkul and P R Hsueh ldquoClinical andeconomic burden of community-acquired pneumonia amongstadults in the Asia-Pacific regionrdquo International Journal ofAntimicrobial Agents vol 38 no 2 pp 108ndash117 2011

[8] D A B Dance ldquoMelioidosisrdquo Current Opinion in InfectiousDiseases vol 15 no 2 pp 127ndash132 2002

[9] N J White ldquoMelioidosisrdquo The Lancet vol 361 no 9370 pp1715ndash1722 2003

[10] S Yang ldquoMelioidosis research in Chinardquo Acta Tropica vol 77no 2 pp 157ndash165 2000

[11] P R Hsueh L J Teng L N Lee et al ldquoMelioidosis an emerginginfection in Taiwanrdquo Emerging Infectious Diseases vol 7 no 3pp 428ndash433 2001

[12] C Walsh and S Fanning ldquoAntimicrobial resistance in food-borne pathogensmdasha cause for concernrdquo Current Drug Targetsvol 9 no 9 pp 808ndash815 2008

[13] M Vorachit P Chongtrakool S Arkomsean and S BoonsongldquoAntimicrobial resistance in Burkholderia pseudomalleirdquo ActaTropica vol 74 no 2-3 pp 139ndash144 2000

[14] F M Thibault E Hernandez D R Vidal M Girardet and JD Cavallo ldquoAntibiotic susceptibility of 65 isolates of Burkholde-ria pseudomallei and Burkholderia mallei to 35 antimicrobialagentsrdquo Journal of Antimicrobial Chemotherapy vol 54 no 6pp 1134ndash1138 2004

[15] R L S Sikarwar ldquoEthnogynaecological uses of plants new toIndiardquo Ethnobotany vol 12 no 1-2 pp 112ndash115 2002

[16] R Siva ldquoStatus of natural dyes and dye-yielding plants in IndiardquoCurrent Science vol 92 no 7 pp 916ndash925 2007

[17] P Kumar Singh V Kumar R K Tiwari A Sharma ChV Rao and R H Singh ldquoMedico-ethnobotany of lsquochatararsquoblock of district sonebhadra Uttar Pradesh Indiardquo Advances inBiological Research vol 4 no 1 pp 65ndash80 2010

[18] W H Lewis and M P Elwin-Lewis Medical Botany PlantsAffecting Human Health John Wiley amp Sons New York NYUSA 2003

[19] R Perumal Samy P N Pushparaj and P GopalakrishnakoneldquoA compilation of bioactive compounds fromAyurvedardquo Bioin-formation vol 3 no 3 pp 100ndash110 2008

[20] WorldHealthOrganization (WHO)TraditionalMedicine Strat-egy 2002ndash2005 Genera World Health Organization GenevaSwitzerland 2002

[21] P S Haddad G A Azar S Groom and M Boivin ldquoNaturalhealth products modulation of immune function and preven-tion of chronic diseasesrdquo Evidence Based Complementary andAlternative Medicine vol 2 no 4 pp 513ndash520 2005

[22] K Poonam andG S Singh ldquoEthnobotanical study of medicinalplants used by the Taungya community in Terai Arc LandscapeIndiardquo Journal of Ethnopharmacology vol 123 no 1 pp 167ndash1762009

[23] G B Mahady ldquoMedicinal plants for the prevention and treat-ment of bacterial infectionsrdquo Current Pharmaceutical Designvol 11 no 19 pp 2405ndash2427 2005

[24] V P Kumar N S Chauhan H Padh and M Rajani ldquoSearchfor antibacterial and antifungal agents from selected Indianmedicinal plantsrdquo Journal of Ethnopharmacology vol 107 no2 pp 182ndash188 2006

[25] R Nair T Kalariya and S Chanda ldquoAntibacterial activity ofsome plant extracts used in folk medicinerdquo Journal of HerbalPharmacotherapy vol 7 no 3-4 pp 191ndash201 2007

[26] D Kalemba and A Kunicka ldquoAntibacterial and antifungalproperties of essential oilsrdquo Current Medicinal Chemistry vol10 no 10 pp 813ndash829 2003

[27] A H Ashour ldquoAntibacterial antifungal and anticancer activi-ties of volatile oils and extracts from stems leaves and flowers ofEucalyptus sideroxylon and Eucalyptus torquatardquoCancer BiologyandTherapy vol 7 no 3 pp 399ndash403 2008

[28] M J Abad L M Bedoya L Apaza and P Bermejo ldquoTheArtemisia L genus a review of bioactive essential oilsrdquoMolecules vol 17 no 3 pp 2542ndash2566 2012

[29] H N Marzoug M Romdhane A Lebrihi et al ldquoEucalyptusoleosa essential oils chemical composition and antimicrobialand antioxidant activities of the oils from different plant parts(stems leaves flowers and fruits)rdquoMolecules vol 16 no 2 pp1695ndash1709 2011

[30] C Proestos D Sereli and M Komaitis ldquoDetermination ofphenolic compounds in aromatic plants by RP-HPLC and GC-MSrdquo Food Chemistry vol 95 no 1 pp 44ndash52 2006

[31] S Chao G Young C Oberg and K Nakaoka ldquoInhibition ofmethicillin-resistant Staphylococcus aureus (MRSA) by essentialoilsrdquo Flavour and Fragrance Journal vol 23 no 6 pp 444ndash4492008

[32] M Akin D Oguz and H T Saracoglu ldquoAntibacterial effectsof some plant extracts from Labiatae (Lamiaceae) growingnaturally around Sirnak-Silopi Turkeyrdquo International Journal ofPharmaceutical and Applied Sciences vol 1 no 1 pp 4ndash47 2010

[33] K Bayoub T Baibai D Mountassif A Retmane and ASoukri ldquoAntibacterial activities of the crude ethanol extracts ofmedicinal plants against Listeriamonocytogenes and some otherpathogenic strainsrdquo African Journal of Biotechnology vol 9 no27 pp 4251ndash4258 2010

[34] R P Samy P Gopalakrishnakone M Sarumathi P Houghtonand S Ignacimuthu ldquoPurification of antibacterial agents fromTragia involucratamdasha popular tribal medicine for wound heal-ingrdquo Journal of Ethnopharmacology vol 107 no 1 pp 99ndash1062006

[35] K M Matthew The Flora of Tamil Nadu Carnatic Part IndashIIIprinted at the Diocesan Press Madras India 1981ndash1983

[36] AW BauerWMKirby J C Sherris andM Turck ldquoAntibioticsusceptibility testing by a standardized single disk methodrdquoAmerican Journal of Clinical Pathology vol 45 no 4 pp 493ndash496 1966

[37] National Committee for Clinical Laboratory Standards Per-formance Standards For Antimicrobial Susceptibility Testing12th Informational Supplement Approved standard M100-S12National Committee for Clinical Laboratory Standards WaynePa USA 2002

[38] E A Elsinghorst ldquoMeasurement of invasion by gentamicinresistancerdquoMethods in Enzymology vol 236 pp 405ndash420 1994

[39] J B Harborne Phytochemical Methods Chapman and HallNew York NY USA 1976

[40] H Wagner S Bladt and E M Zgairski Plant Drug AnalysisSpringer New York NY USA 1984

[41] N Myers R A Mittermeler C G Mittermeler G A B daFonseca and J Kent ldquoBiodiversity hotspots for conservationprioritiesrdquo Nature vol 403 no 6772 pp 853ndash858 2000

[42] R Perumal Samy and S Ignacimuthu ldquoAntibacterial activity ofsome folkloremedicinal plants used by tribals inWestern Ghats

Evidence-Based Complementary and Alternative Medicine 17

of Indiardquo Journal of Ethnopharmacology vol 69 no 1 pp 63ndash712000

[43] K Nampoothiri A Chrispal A Begum S Jasmine K GGopinath and A Zachariah ldquoA clinical study of renal tubulardysfunction inCleistanthus collinus (Oduvanthalai) poisoningrdquoClinical Toxicology vol 48 no 3 pp 193ndash197 2010

[44] M Strathern UsefulKnowledge The Isiah Berlin Lecture Uni-versity of Manchester Brunswick UK

[45] C N Sunil and A K Pradeep ldquo(2001) Another new species ofTripogon (Poaceae) from Indiardquo Sida vol 19 pp 803ndash806 2005

[46] S Biber-Klemm and T Cottier Right To Plant GeneticResources and Traditional Knowledge Basic Issues and Prospec-tive Oxfordshire Nosworthy Way UK 2006

[47] V S Ramachandran S Shijo Joseph and R Aruna ldquoEth-nobotanical studies from amaravathy range of Indira GandhiWildlife SanctuaryWesternGhats CoimbatoreDistrict South-ern Indiardquo Ethnobotanical Leaflets vol 13 pp 1069ndash1087 2009

[48] A H Rajasab and I Mohamad ldquoDocumentation of folkknowledge on edible wild plants of North Karnatakardquo IndianJournal of Traditional Knowledge vol 3 no 4 pp 419ndash4292004

[49] M Rajadurai V G Vidhya M Ramya and A Bhaskar ldquoEthno-medicinal plants used by the traditional healers of pachamalaihills Tamilnadu Indiardquo Studies on Ethno-Medicine vol 3 no 1pp 39ndash41 2009

[50] S K Adesina O Idowu A O Ogundaini et al ldquoAntimicrobialconstituents of the leaves of Acalypha wilkesiana and Acalyphahispidardquo Phytotherapy Research vol 14 no 5 pp 371ndash374 2000

[51] M T Gutierrez-Lugo M P Singh W M Maiese and BN Timmermann ldquoNew antimicrobial cycloartane triterpenesfrom Acalypha communisrdquo Journal of Natural Products vol 65no 6 pp 872ndash875 2002

[52] A K Tyagi andAMalik ldquoAntimicrobial potential and chemicalcomposition of Mentha piperita oil in liquid and vapour phaseagainst food spoiling microorganismsrdquo Food Control vol 22no 11 pp 1707ndash1714 2011

[53] G Devendran and U Balasubramanian ldquoQualitative phyto-chemical screening and GC-MS analysis of Ocimum sanctumL leavesrdquoAsian Journal of Plant Science and Research vol 1 no4 pp 44ndash48 2011

[54] V C Jain N M Patel D P Shah P K Patel and B HJoshi ldquoAntioxidant and antimicrobial activities of Alangiumsalvifolium (Lf) Wang rootrdquo Global Journal of Pharmacologyvol 4 no 1 pp 13ndash18 2010

[55] P Swapna Latha andK Kannabiran ldquoAntimicrobial activity andphytochemicals of Solanum trilobatum LinnrdquoAfrican Journal ofBiotechnology vol 5 no 23 pp 2402ndash2404 2006

[56] M Usha1 S Ragini and S M A Naqvi ldquoAntibacterial activityof acetone and ethanol extracts of Cinnamon (Cinnamomumzeylanicum) and Ajowan (Trachyspermum ammi) on four foodspoilage bacteriardquo International Research Journal of BiologicalSciences vol 1 no 4 pp 7ndash11 2012

[57] S Prabuseenivasan M Jayakumar and S Ignacimuthu ldquoInvitro antibacterial activity of some plant essential oilsrdquo BMCComplementary and Alternative Medicine vol 6 article 39 pp1ndash8 2006

[58] G A Ayoola F M Lawore T Adelowotan et al ldquoChemi-cal analysis and antimicrobial activity of the essential oil ofSyzigium aromaticum (clove)rdquo African Journal of Biotechnologyvol 7 no 13 pp 2227ndash2231 2008

[59] Y J Fu L Chen Y Zu et al ldquoThe antibacterial activity of cloveessential oil against Propionibacterium acnes and its mechanismof actionrdquo Archives of Dermatology vol 145 no 1 pp 86ndash882009

[60] D Venkatesan C M Karrunakarn S Selva Kumar and P TPalani Swamy ldquoIdentification of phytochemical constituents ofAegle marmelos responsible for antimicrobial activity againstselected pathogenic organismsrdquo Ethnobotanical Leaflets vol 13pp 1362ndash1372 2009

[61] P Arun K G Purushotham J Johnsy jayarani and V KumarildquoIn vitro Antibacterial Activity of Oldenlandia umbellata anIndian medicinal Plantrdquo Journal of Pharmaceutical Science andTechnology vol 2 no 4 pp 198ndash201 2010

[62] A Manjamalai R Sardar Sathyajith Singh C Guruvayoorap-pan and V M Berlin Grace ldquoAnalysis of phytochemicalconstituents and antimicrobial activity of some medicinalplants in Taminadu Indiardquo Global Journal of Biotechnology andBiochemistry vol 5 no 2 pp 120ndash128 2010

[63] E Haslam ldquoNatural polyphenols (vegetable tannins) as drugsPossible modes of actionrdquo Journal of Natural Products vol 59no 2 pp 205ndash215 1996

[64] Y Cai Q Luo M Sun and H Corke ldquoAntioxidant activityand phenolic compounds of 112 traditional Chinese medicinalplants associated with anticancerrdquo Life Sciences vol 74 no 17pp 2157ndash2184 2004

[65] B Shan Y Z Cai M Sun and H Corke ldquoAntioxidant capacityof 26 spice extracts and characterization of their phenolicconstituentsrdquo Journal of Agricultural and Food Chemistry vol53 no 20 pp 7749ndash7759 2005

[66] R J W Lambert P N Skandamis P J Coote and G-J ENychas ldquoA study of the minimum inhibitory concentration andmode of action of oregano essential oil thymol and carvacrolrdquoJournal of AppliedMicrobiology vol 91 no 3 pp 453ndash462 2001

[67] M Raccach ldquoThe antimicrobial activity of phenolic antioxi-dants in food a reviewrdquo Journal of Food Safety vol 6 no 3 pp141ndash170 1984

[68] M Taniguchi Y Yano E Tada et al ldquoMode of action of polygo-dial an antifungal sesquiterpene dialdehyderdquo Agricultural andBiological Chemistry vol 52 no 6 pp 1409ndash1414 1988

[69] G J E Nychas ldquoNatural antimicrobials from plantsrdquo in NewMethods of Food Preservation G W Gould Ed pp 58ndash89Blackie Academic London UK 1995

[70] S Burt ldquoEssential oils their antibacterial properties and poten-tial applications in foodsmdasha reviewrdquo International Journal ofFood Microbiology vol 94 no 3 pp 223ndash253 2004

Submit your manuscripts athttpwwwhindawicom

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Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

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Behavioural Neurology

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Disease Markers

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OncologyJournal of

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Oxidative Medicine and Cellular Longevity

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PPAR Research

The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014

Immunology ResearchHindawi Publishing Corporationhttpwwwhindawicom Volume 2014

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Parkinsonrsquos Disease

Evidence-Based Complementary and Alternative Medicine

Volume 2014Hindawi Publishing Corporationhttpwwwhindawicom

6 Evidence-Based Complementary and Alternative Medicine

Table 1 Continued

Scientific name Family Voucher specimen Plant parts Yield (gm) Phytochemicalanalysis

Coccinia grandisWampA Cucurbitaceae D02030 Leaf root 09 GlycosidesCymbopogon citratus(DC) Gramineae D012 Root 025 Essential oil

Datura metel L Solanaceae D02038 Leaf stem 39 SteroidsEucalyptus globulus Labill Myrtaceae D0220 Leaf 12 TerpenoidsEclipta alba (L) Hassk Asteraceae D028 Whole plant 07 PhenolicEuphorbia hirta Linn Euphorbiaceae 0018-c Whole plant 012 mdash

Eclipta prostrata (L) Asteraceae D210 Leaf 110 Triterpenoidsaponin

Eugenia caryophyllus(Sprengel) Bullock ampHarrison

Myrtaceae 0025 Flower buds 116 Essential oils

Elettaria cardamomumWhite et Mason Zingiberaceae 0009 Fruit pods 317 Essential oils

Gloriosa superba L Liliaceae 020-S Tuber 108 Alkaloids phenol

Jatropha curcas L Euphorbiaceae 015 Whole plant 53 Alkaloidsflavonoids

Hyptis suaveolens (L) Poit Lamiaceae 24688 Leaf 63 Essential oilHemidesmus indicus L Asclepiadaceae D-009 Roots CoumarinsIchnocarpus frutescens(L) RBr Apocynaceae 0110 Root flower 73 Terpenoids

Leucas aspera (Willd)Link Labiatae 0114 Leaf 83 Triterpenes

Lawsonia inermis L Lythraceae T261 Leaf 09 Glycosidesphenolic

Madhuca longifolia (L) JFMacbr Sapotaceae D01415 Nut 93 Sitosterol

Merremia hastate L(Desr) Hallierf Convolvulaceae 10894 Whole plant 40 Alkaloids

Mentha piperita L Lamiaceae 0217-c Whole plant 07 Essential oilsMorinda tinctoria Roxb Rubiaceae 0122 Leaf 14 GlycosidesMimosa pudica L Mimosaceae 0071 Whole plant 06 mdashOldenlandia umbellata L Rubiaceae D02047 Leaf 44 AlkaloidsOcimum sanctum L Lamiaceae 0016 Whole plant 30 AlkaloidsPiper attenuatum BuchHamex Miq Piperaceae 007 Flower 46 Alkaloids

Plumbago zeylanica (L)Cav Plumbaginaceae 0121 Root 48 Plumbagin

Plectranthus amboinicus(L) Spreng Lamiaceae 0410 Whole plant 12 Essential oils

terpenoidsPhyllanthus debilis L(Klein ex Willd) Euphorbiaceae 0120 Whole plant 49 Polyphenol

Phyllanthusmadraspatensis L Euphorbiaceae 0117 Whole plant 50 Polyphenol

Premna tomentosaWilld Verbenaceae 0129 Leaf 53 DiterpenesRosmarinus officinalis L Lamiaceae 0017 Root 023 Essential oilsRauwolfia serpentine L Apocynaceae 020-S Root 115 AlkaloidSebastiania chamaelea (L)Muell Arg Euphorbiaceae 0034 Leaf 11 Polyphenol

Solanum trilobatum L Solanaceae D02054 Leaf flower 40 Tannins

Evidence-Based Complementary and Alternative Medicine 7

Table 1 Continued

Scientific name Family Voucher specimen Plant parts Yield (gm) Phytochemicalanalysis

Sphaeranthus indicus L Asteraceae D02060 Whole plant 10 Essential oilSwertia chirata (L) Ham Gentianaceae D0540 Whole plant 16 GlycodisesStrychnos nux-vomica L Loganiaceae S-22 Nuts 036 AlkaloidsTragia involucrata L Euphorbiaceae D068 Leaves 16 ShellsolTinospora cordifolia(Willd) Miers ex Hooff ampThoms

Menispermaceae 0118 Leafroot stem 50 Glycosides tannins

Tridax procumbens L Compositae 10649 Leaf 18 FlavonoidsTerminalia arjuna (DC)W amp A Combretaceae 033-c Bark 80 Phenolics

Tephrosia purpurea (L)Pers Fabaceae S-43 Whole plant 08 Isoflavone

Vitex negundo L Verbenaceae 0031 Leaf 24 TerpineolVetiveria zizanioides L Gramineae 0051 Root 103 Essential oilWithania somnifera (L)Dunal Solanaceae D02063 Root 21 Alkaloids

Wedelia calendulacea Less Asteraceae S-24 Leaves FlavonoidsZingiber officinale Rosc Zingiberaceae 0327 Rhizome 23 TanninsZanthoxylum limonella(Dennst) Alston Rutaceae 009 Bark 19 Alkaloids essential

oilClass of chemical compounds A alkaloids S saponins T tannins St steroids G glycosides T terpenoids P polyphenol P phenolics Sh shellsol Hhydrocarbon esters

the site of snakebite and decoction is given orally fortreatment by indigenous people Besides these tribes relyon the medicinal plants as ingredients for fabricating a kindof medicated stone for health management ldquoVishakallurdquo(poison stone) is used by the indigenous groups called Kaniin Kerala India to treat a snakebiteWhen the stone is placeddirectly on the bitten area it sticks to the body to absorb thepoison and then become detached when absorption seems tobe complete The ingredients of Vishakallu stones are madewith leaves of Ocimum sanctum Anisomeles malabaricaLeucas aspera Piper betle Santalum album and the pebblescollected from the river bank

35 Survey of Medicinal Plants and Their Health Care ValuesThe present study is an attempt to provide scientific basis andobtain justification for the traditional beliefs of reliance on arich diversity of ethnomedicinal plants along with the richheritage of traditional medicine practices related to healthcare system made available by the primitive tribal commu-nities located at different settlements The native traditionalpractitioners called ldquovaidyarsrdquo have a good knowledge aboutthe traditional plants locally available for treatment of var-ious diseases (Figures 3(a)ndash3(p)) Such traditional medicalknowledge is used for preparing home remedies ill healthprevention and routine health maintenanceThis knowledgeis also applicable to cover other sectors of social life Duringthe ethnobotanical survey the wealth of 78 medicinal plantspecies used by the indigenous tribal community for varioustypes of health treatment was documented The botanical

names family names parts used chemical constituents andtheir application are provided (Table 1)

36 Antimicrobial Activity of Crude Extracts In this studywe reported the antimicrobial screening of methanolic crudeextracts of 78 medicinal plants (Table 2) Results revealedthat 68 plant extracts displayed potent activities against oneor more Gram-positive and -negative bacteria Of whichTragia involucrata Citrus acida Aegle marmelos Adha-toda vasica Calotropis procera Andrographis paniculata andMentha piperita Azadirachta indica Sphaeranthus indicusand Elettaria cardamomum showed the highest antibacte-rial activity against the multidrug resistant B pseudomallei(KHW and TES) and S aureus at 100120583gmL concentrationThe extracts showed pronounced antibacterial activity withtheir inhibitory zones ranging from 20 to 31mm in diam-eter as compared to the standard drugs chloramphenicoland ceftazidime (29ndash33mm) The majority of the plantsdemonstrated a powerful antimicrobial potency against themultidrug resistant strains of B pseudomallei (KHW andTES) K pneumonia and S aureus Approximately twenty-one plant extracts exerted only a weak or moderate effectagainst the tested bacteria while the crude extract of 13plants failed to show any effect at all Except for the plantextracts of T involucrata A lanceolata A vasica and Sindicus extracts the majority of the plant extracts wereineffective against theV damsela infection fascinatingly only11 plants exhibited activity against P aeruginosa of whichS indicus M piperita and C procera were found to havevery strong inhibition of bacteria at the tested concentrations

8 Evidence-Based Complementary and Alternative Medicine

Table 2 Antimicrobial activity of methanol extract of aromatic medicinal plants and essential oils evaluated against multidrug resistant(MDR) human pathogens at 100120583gmL concentration

Scientific name Microorganisms growth inhibition zones (6 millimeter in diameters)KHW TES Kp Kpr Pa Sa Stp Sp Vd Vd

Adhatoda vasica Nees 28 19 22 18 12 mdash 10 17 16 mdashAegle marmelos (L) Correa ex Roxb 29 17 9 10 mdash 15 mdash mdash mdash mdashAlangium salvifolium (L) f Wangerin 15 10 8 8 mdash 12 9 10 mdash mdashAndrographis echioides L 12 9 mdash mdash mdash 17 8 9 mdash mdashAndrographis paniculataWallich ex Nees 26 21 19 13 mdash 25 8 16 12 mdashAcalypha indica L mdash mdash mdash mdash mdash 18 mdash mdash mdash mdashAcalypha lanceolata L mdash mdash 12 13 mdash 10 mdash mdash 21 mdashAchyranthes aspera L mdash mdash 7 14 mdash 15 12 11 mdash mdashAgeratum conyzoides L 10 8 mdash mdash mdash mdash mdash mdash mdash mdashAsteracantha longifolia L 16 12 mdash mdash mdash 15 mdash mdash mdash mdashAzadirachta indica A Juss 15 17 21 16 14 23 12 14 mdash mdashBorassus flabellifer L 9 10 17 8 mdash mdash 9 10 mdash mdashBoerhavia erecta L mdash mdash 8 9 7 16 mdash mdash mdash mdashCalotropis procera (L) mdash mdash 15 mdash 18 28 9 mdash mdash mdashCalotropis gigantea (L) RBrex Ait 11 9 mdash mdash mdash 20 mdash mdash 9 8Cardiospermum halicacabum L 23 mdash mdash 14 mdash 9 19 mdash mdash mdashCatharanthus roseus (L) GDon 13 7 12 9 mdash 15 11 8 mdash mdashCassia auriculata L 17 13 12 mdash mdash 19 13 mdash mdash mdashCassia occidentalis L 18 mdash mdash mdash mdash mdash mdash mdash mdash mdashCassia tora L mdash mdash mdash mdash mdash mdash mdash mdash mdash mdashCassia fistula L mdash mdash mdash mdash mdash mdash mdash mdash mdash mdashCitrus acida Roxb Hookf 26 22 mdash 12 29 9 13 8 mdashCissus quadrangularis L mdash mdash mdash mdash mdash mdash mdash mdash mdash mdashCinnamomum zeylanicum Garcin exBlume 14 16 7 20 mdash 22 19 7 mdash mdash

Cinnamomum iners Reinw ex Blume 20 16 mdash 15 mdash 16 12 mdash mdash mdashRosmarinus officinalis L mdash 9 mdash 10 mdash 7 mdash 8 mdash mdashCentella asiatica (L) 9 8 mdash mdash mdash 11 mdash mdash mdash mdashClerodendrum inerme (L) Gaertn 13 7 12 mdash mdash 15 11 8 mdash mdashClitoria ternatea L mdash mdash mdash mdash mdash mdash mdash mdash mdash mdashClitoria ternatea L 16 12 mdash 13 mdash 8 10 mdash mdash mdashCleome gynandropsis L mdash mdash 12 19 11 mdash 9 15 12 mdashCleome viscose L mdash mdash 8 10 14 20 mdash mdash mdash mdashCoccinia grandisWampA 9 10 17 8 mdash mdash 9 10 mdash mdashCymbopogon citratus (DC) 16 18 mdash 17 mdash 14 mdash mdash mdash mdashDatura metel L mdash mdash mdash mdash mdash mdash mdash mdash mdash mdashEclipta alba (L) Hassk 20 mdash mdash 9 mdash mdash 10 mdash mdash mdashEuphorbia hirta Linn 11 mdash mdash mdash mdash 16 mdash mdash mdash mdashEucalyptus globulus Labill mdash mdash mdash 7 mdash mdash mdash 7 mdash mdashEugenia caryophyllus Bullock amp Harrison mdash 7 mdash 11 mdash 9 mdash 8 mdash mdashElettaria cardamomumWhite et Mason 21 20 7 14 mdash 22 12 17 mdash mdashHyptis suaveolens (L) Poit 8 10 8 7 mdash mdash mdash mdash mdash mdashIchnocarpus frutescens (L) WJ Aiton mdash mdash mdash mdash mdash mdash mdash mdash mdash mdashJatropha curcas L mdash mdash mdash 8 mdash 11 mdash 12 mdash 7Leucas aspera (Willd) Link 9 10 17 8 mdash 12 9 10 mdash mdashLawsonia inermis L mdash mdash mdash mdash mdash mdash mdash mdash mdash mdashMadhuca longifolia (L) JF Macbr 18 16 14 19 12 mdash 8 7 9 mdash

Evidence-Based Complementary and Alternative Medicine 9

Table 2 Continued

Scientific name Microorganisms growth inhibition zones (6 millimeter in diameters)KHW TES Kp Kpr Pa Sa Stp Sp Vd Vd

Merremia hastate L (Desr) Hallierf mdash mdash mdash mdash mdash mdash mdash mdash mdash mdashMorinda tinctoria Roxb mdash mdash mdash mdash mdash mdash mdash mdash mdash mdashMentha piperita L 23 17 26 12 20 25 19 mdash mdash mdashOcimum sanctum L 12 9 11 7 mdash 15 7 8 mdash mdashOldenlandia umbellata L mdash mdash mdash mdash mdash 17 mdash mdash mdash mdashPiper attenuatum Buch Hamex Miq 13 7 12 21 mdash 17 11 8 mdash mdashPlumbago zeylanica (L) Cav 9 10 17 8 mdash 12 9 10 mdash mdashPlectranthus amboinicus (L) Spreng mdash mdash mdash 8 11 15 mdash mdash mdash mdashPhyllanthus debilis L (Klein ex Willd) 7 8 mdash 9 mdash 9 18 7 mdash mdashPhyllanthus maderaspatensis L 17 mdash mdash mdash mdash mdash mdash mdash mdash mdashPremna tomentosaWilld 13 10 mdash 15 mdash 9 10 mdash mdash mdashGloriosa superba L 17 16 7 8 mdash 15 mdash mdash mdash 8Sebastiania chamaelea (L) Muell Arg 19 12 mdash 17 mdash 13 19 8 mdash mdashSolanum trilobatum L mdash mdash 13 9 mdash 8 mdash mdash mdash mdashSphaeranthus indicus L 20 18 21 7 22 mdash 11 9 mdash 16Swertia chirata (L) Ham mdash mdash mdash mdash mdash mdash mdash mdash mdash mdashTerminalia arjuna (W amp A) mdash mdash mdash mdash mdash 16 mdash mdash mdash mdashTinospora cordifolia (Willd) Miers exHooff ampThoms mdash mdash 23 16 mdash 12 15 mdash mdash mdash

Tridax procumbens L 9 8 7 mdash mdash 14 mdash mdash mdash mdashTragia involucrata L 25 23 20 mdash mdash 31 28 22 19 mdashVitex negundo L mdash mdash mdash mdash mdash 14 mdash mdash mdash mdashVetiveria zizanioides (L) 11 9 7 7 mdash 16 mdash mdash 8Withania somnifera (L) Dunal 12 20 mdash mdash mdash 15 12 mdash mdash mdashZingiber officinale Rosc 14 11 7 15 mdash 7 12 7 mdash mdashZanthoxylum limonella (Dennst) Alston 13 7 12 9 mdash mdash 11 8 mdash mdashChloramphenicol (30120583gdisc) 21 12 15 17 29 16 15 18 13 11Ceftazidime (30 120583gdisc) 33 16 22 19 16 25 21 20 12 15lowastBacteria (+minus) Results obtained in the disc diffusion assay antibacterial activity is expressed as the mean plusmn SD (119899 = 3) of the inhibition by the extract andits diameter around the discs One way analysis of variance was performed (mean plusmn SD 119899 = 3 replicates) Size of inhibition zones were including the sterileblank discs 6 millimeter (mm) in diameters Absence of bacterial inhibition indicates (mdash) antibiotic disc (30 120583gdisc)

Interestingly sixteen plants such as Andrographis echioidesC auriculata C viscose C gigantea T arjuna Oldenlandiaumbellata Boerhavia erecta and E hirta exerted a strongactivity against the Gram-positive S aureus bacteria

37 Phytochemical Screening of Plants The results obtainedfrom the phytochemical screening as shown in Table 1 indi-cate the presence of various types of secondary metabolitessuch as polyphenols tannins saponins alkaloids and gly-cosidespolysaccharides Most of the plant extracts relativelyrich in alkaloids phenols flavonoids polyphenols tanninssterols and terpenoids were found to inhibit the growth oforganisms

38 Antimicrobial Activity of Fractioned Compounds Activecomponents were purified from the most active extractsfor further testing The compound shellsol of T involucrata

and C acida exhibited the most potent action againstthe antibiotic resistant strains of B pseudomallei (KHW)S aureus B pseudomallei (TES) and K pneumoniae Amarmelos was also found to inhibit the growth of B pseudo-mallei (KHW) more effectively than other tested bacteria Avasica showed the broad spectrum growth inhibitory activityon B pseudomallei (KHW) K pneumoniae K pneumoniaeresistant B pseudomallei (TES) S pyogenes and V damselaHowever E cardamomum displayed antimicrobial activityon some of the B pseudomallei (KHW) S pyogenes Bpseudomallei (TES) and S typhi strains Similarly A indicaexerted the growth inhibition on K pneumoniae and Saureus Remarkably Sebastiania chamaelea was more activeagainst K pneumoniae K pneumoniae and S pyogenesThe compound from S indicus inhibited the growth of Kpneumoniae K pneumoniae B pseudomallei (KHW andTES) and S typhi strains as compared to the activity shown

10 Evidence-Based Complementary and Alternative Medicine

Figure 2 Diverse biodiversity richness of medicinal plants in Western and Eastern Ghats (a) Topography of plant covering area in Kollihills (Namakkal district Tamil Nadu) (b) Aerial view of occurrence of medicinal plants in Mundanthurai hills (Tirunelveli district TN) (c)Pachamalai hills (Trichy district) and its natural vegetation inhabitants for Malaiyali tribes (d) Deforestation of natural herbal resources dueto urbanization in Kalrayan hills (Salem district) in the Eastern Ghats of TN (e) Medicinal plants and its various parts used by the natives(traditional healers) for the treatment of diverse human illness with a very high percentage of leaves and whole plants often used for herbaldrug preparation by the local practitioners (f) Various category of plants like shrub herb climbers and tree and the parts used in medicine

Evidence-Based Complementary and Alternative Medicine 11

Figure 3 Medicinal aromatic herbs spices and toxic plants were collected from the tribal areas of the Western and Eastern Ghats region inTamil Nadu India (a) A vasica Nees (leaf) (b) Eclipta alba (L) Hassk (whole plant) (c)Mimosa pudica L (whole plant) (d) P amboinicus(L) Spreng (whole plant) (e) T procumbens (L) (whole plant) (f) Euphorbia hirta Linn (whole plant) (g) A paniculata Wallich ex Nees(H) C roseus (L) GDon (whole plant) used for therapy (i) Cinnamomum iners Reinw ex Blume (leaf) (j) E globulus Labill (leaf and bark)(k) Z officinale Rosc (Rhizome) (l) E caryophyllus (Sprengel) Bullock and Harrison (flower buds) (m) M piperita L (whole plant) (n) Ccitratus (DC) Clitoria ternatea L (whole plant) (o) C zeylanicum Garcin ex Blume (bark) (p) Elettaria cardamomumWhite et Mason (fruitpod) used for medicine and food preparation

by the crude extracts (Figure 4)The antimicrobial efficacy offractions collected from the oil yielding plants was also com-pared with that of the tested compounds C zeylanicum andR officinaliswere themost sensitive in controlling the growthof B pseudomallei (KHW) S aureus K pneumonia and Spneumoniae Fascinatingly all the compounds obtained fromaromatic plants except those from E globules were foundto be very effective against the multidrug resistant humanpathogen B pseudomallei (KHW) that causes melioidosis

On the other hand compounds from C citrates O sanctumE caryophyllus and Z zizanioide showed some promisingeffect only against S aureus (Figure 5) The activity of thecompounds were pronounced more than that of the oilyielding plant fractions

39 Minimum Inhibitory Concentrations (MICs) The antibi-otic potential of the purified fractions was obtained fromthe MIC determination The hydrocarbon ester shellsol

12 Evidence-Based Complementary and Alternative Medicine

(mean plusmn SD n = 3)

0

5

10

15

20

25

30

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn V

d St

Bacteria (+minus)

lowast

lowast

lowast

A vasica

In vitro assay(P

lant

frac

tion)

Inhi

bitio

n zo

nes (

mm

)

(a)

(mean plusmn SD n = 3)

0

5

10

15

20

25

30

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn V

d St

Bacteria (+minus)

lowast

A marmelos

Inhi

bitio

n zo

nes (

mm

)

In vitro assay

(b)

(mean plusmn SD n = 3)

0

5

10

15

20

25

30

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn V

d St

Bacteria (+minus)

lowastlowast

A indica

Inhi

bitio

n zo

nes (

mm

)

Incu

batio

n (37∘C

24 h

)

(c)

(mean plusmn SD n = 3)

0

5

10

15

20

25

30

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn V

d St

Bacteria (+minus)

lowast

lowast lowast

35

C acida

Inhi

bitio

n zo

nes (

mm

)(P

lant

frac

tion)

(d)

(mean plusmn SD n = 3)

0

5

10

15

20

25

30

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn V

d St

Bacteria (+minus)

lowastlowast

lowast

S chamaelea

Inhi

bitio

n zo

nes (

mm

)

(e)

(mean plusmn SD n = 3)

0

5

10

15

20

25

30

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn V

d St

Bacteria (+minus)

lowastlowast

lowastlowast

S indicus

Inhi

bitio

n zo

nes (

mm

)

Incu

batio

n (37∘C

24 h

)

(f)

(mean plusmn SD n = 3)

0

5

10

15

20

25

30

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn V

d St

Bacteria (+minus)

lowast

lowast lowast

E cardamomum

Inhi

bitio

n zo

nes (

mm

)(P

lant

frac

tion)

(g)

(mean plusmn SD n = 3)

0

5

10

15

20

25

30

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn V

d St

Bacteria (+minus)

lowast

lowast

lowast

lowast

Shellsol

35

Inhi

bitio

n zo

nes (

mm

)

Incu

batio

n (37∘C

24 h

)

(h)

Figure 4 In vitro antimicrobial activity of purified fractions from the most active plant extracts tested against bacteria Growth inhibitionzones were measured and analyzed with mean plusmn standard deviation (SD) (119899 = 3) using one way analysis of variance Level of significanceat (lowast119875 gt 001) Most of the fractions exerted a potent inhibitory effect against multidrug resistant Gram-negative bacteria (B pseudomalleistrains KHW and TES) K pneumonia and Gram-positive bacteria S aureus

(T involucrata) andC acida showed an interesting inhibitorypotential against S aureus (MIC of 78 120583gmL) and B pseu-domallei strain of KHW (MIC of 156 120583gmL) A vasicashowed an MIC of 156120583gmL against B pseudomallei(KHW) and an MIC of 3125120583gmL against K pneumo-niae K pneumoniae S pyogenes and V damsela strainsFractions from A marmelos and terpenoid from A indicaexerted bacteriostatic effect with MIC values of 3125120583gmLon some selected bacteria including B pseudomallei ofKHW S aureus and B pseudomallei of TES The MIC of3125 120583gmL was found for E cardamomum against S aureus

K pneumonia and S pyogenes S indicus displayed a verystrong inhibition against MDR K pneumoniae (MIC of156 120583gmL) and against B pseudomallei (KHW and TES)at MIC of 3125 120583gmL When the antimicrobial efficacies ofpurified fractions from aromatic plants were compared theC zeylanicum fraction displayed an important antimicrobialeffect against S aureus (MIC of 78 120583gmL) MDR B pseu-domallei of KHW (MIC of 156 120583gmL) and S pneumoniae(MIC of 3125 120583gmL) The essential oil from M piperateashowed MIC value of 3125120583gmL against K pneumoniae Saureus and B pseudomallei (KHW) respectivelyO sanctum

Evidence-Based Complementary and Alternative Medicine 13

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn

Bacteria (+minus)

0

5

10

15

20

25

30

Inhi

bitio

n zo

nes (

mm

)(P

lant

frac

tion)

lowastlowast

lowast

(mean plusmn SD n = 3)In vitro assay

C citratus

(a)

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn

Bacteria (+minus)

0

5

10

15

20

25

30

Inhi

bitio

n zo

nes (

mm

)

lowast

lowast

lowastlowast

lowast

lowast

(mean plusmn SD n = 3)In vitro assay

35

C zeylanicum

(b)

Bacteria (+minus)

0

5

10

15

20

25

30

Inhi

bitio

n zo

nes (

mm

)

lowast

lowast

lowast

(mean plusmn SD n = 3)

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn V

d St

O sanctum

Incu

batio

n (37∘C

24 h

)

(c)

Bacteria (+minus)

0

5

10

15

20

25

Inhi

bitio

n zo

nes (

mm

)

lowast

lowast

(mean plusmn SD n = 3)

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn V

d St

M piperita

(Pla

nt fr

actio

n)

(d)

Bacteria (+minus)

0

5

10

15

20

25

30

Inhi

bitio

n zo

nes (

mm

)lowast

lowast

lowast

(mean plusmn SD n = 3)

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn V

d St

E caryophyllus

(e)

0

5

10

15

20

25

30

Inhi

bitio

n zo

nes (

mm

) (mean plusmn SD n = 3)

Bacteria (+minus)

KHW

TES

Kp

KprPa Sa

Stp

Spn

E globulus

Incu

batio

n (37∘C

24 h

)

(f)

Bacteria (+minus)

0

5

10

15

20

25

30

Inhi

bitio

n zo

nes (

mm

)(P

lant

frac

tion)

lowast

lowast

lowast

lowast

(mean plusmn SD n = 3)

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn V

d

R officinalis

(g)

0

5

10

15

20

25

30

Inhi

bitio

n zo

nes (

mm

)

lowast

lowast

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn V

d St

Bacteria (+minus)V zizanioides

(mean plusmn SD n = 3)

Incu

batio

n (37∘C

24 h

)

(h)

Figure 5 Comparison of antimicrobial effect of plant compounds obtained from the most popularly used sources of essential oils as assayedby the disc-diffusion method in vitro It displayed a powerful activity against B pseudomallei and S aureus than the other bacteria strainsOther compounds showed only a moderate or weak action against the tested bacteria Values for zone of bacterial growth inhibition werepresented as mean plusmn SD (119899 = 3) with level of significance at (lowast119875 gt 001)

and C citratus fractions also showed antimicrobial activity(MICs of 3125ndash125120583gmL) only at higher concentrationsagainst the tested bacteria In addition to that higher concen-trations (gt250120583gmL) (of Vetiveria fractions) were requiredto inhibit Vibro species and others (including E globulusfractions) failed to show any effect at tested concentrations(78ndash125 120583gmL) However the purified fractions (from mostactive medicinal plants) showed strong bacteriostatic inhibi-tion against the tested organisms (Table 3)

310 Cytotoxic Effects of Plants When the components wereassayed for cytotoxicity against the normal human skinfibroblasts (HEPK) cells the compounds obtained from

E cardamomum T involucrata S indicusC acidaA vasicaA marmelos A indica and A paniculata did not showtoxicity up to 1000120583gmL (see Figures S1 and S2 in Supple-mentaryMaterial available online at httpdxdoiorg1011552013525613) A slight reduction of cell proliferation wasnoted only at higher doses (2000120583gmL) In contrast cellproliferation was markedly reduced after exposure of HEPKcells to O sanctum E globulus V zizanioides C citratusand E globulus compounds There was no gradual reductionin skin cell proliferation seen after exposure to C zeylan-icum R officinalis and M piperita (see Figures S3 and S4)compounds The toxicity was found to be concentration-dependent when the skin fibroblasts (HEPK) cells were

14 Evidence-Based Complementary and Alternative Medicine

Table 3 Minimum inhibitory concentrations (MICs) of purified plant fractions and essential oils against antibiotic resistant bacteria

Botanical name Family Parts used Gram-positive and -negative bacteria (MICs 120583gmL)KHW TES Kp KPr Sa Stp Sp Vd St

A indica Juss Meliaceae Seed (fraction) 3125 625 125 lt250 3125 mdash mdash mdash mdashA marmelos (L) Rutaceae Root-bark (F) 3125 3125 mdash mdash 3125 625 125 250 mdashA paniculata Nees Acanthaceae Leaf (fraction) 250 mdash 125 mdash 625 mdash mdash 125 mdashA vasica Nees Acanthaceae Fraction (Stem) 156 3125 3125 625 mdash lt250 3125 3125 mdashC acida Roxb Rutaceae Leaf (fraction) 156 625 mdash mdash 78 mdash 3125 mdash mdashE cardamomumWhite et Mason Acanthaceae Fraction (WP) 625 mdash 3125 mdash 3125 250 625 mdash mdashS indicus (L) Euphorbiaceae Whole plant (F) 3125 125 156 mdash 625 mdash mdash lt250 mdashT involucrata (L) Euphorbiaceae Shellosol (leaf) 156 3125 mdash mdash 78 625 mdash mdash mdashCinnamomum zeylanicum (L) Lauraceae Bark (fraction) 156 625 mdash mdash 78 3125 mdash mdash 125Cymbopogon citratus (L) Graminae Leaf (fraction) 250 lt250 625 mdash 625 125 mdash mdash mdashEugenia caryophyllus (L) Myrtaceae Flower buds (F) 625 125 mdash mdash 625 mdash mdash mdash mdashEucalyptus globulus (L) Myrtaceae Fraction (leaf) mdash mdash mdash mdash mdash mdash lt250 mdash lt250Mentha piperita (L) Labiatae Fraction (WP) mdash 625 3125 mdash 3125 mdash lt250 mdash mdashOcimum sanctum (L) Labiatae Leaf (fraction) 625 125 mdash 3125 625 mdash 125 lt250 mdashRosmarinus officinalis (L) Labiatae Rosemary oil 3125 mdash mdash mdash 3125 mdash mdash mdash mdashVetiveria zizanioides (L) Graminae Root (fraction) mdash 125 125 mdash mdash mdash gt250 gt250The bacterial growth inhibitory activity was compared for significant differences within the bacterial strains by broth-dilution method at 250 125 625 3125156 and 78 120583gmL F fractions

exposed to various compounds The cell proliferation wasincreased by the influence of the plant components at thelower concentrations Whereas the oil yielding plant com-pounds showed inhibition of cell proliferation and toxicity athigher doses 250ndash1000120583gmL

4 Discussion

The Western Ghats is considered as one of the richestbiodiversity hotspots in the world [41] In this survey wecollected nearly 78 medicinal plants from Western andEastern Ghats that are edible and popularly used for curingvarious ailments including snakebite Traditional remedieshave a long-standing history in many tribal settlements inTN India and they continue to provide useful and applicabletools for treating ailments [42] The ingredients that makeup the ldquoVishakallurdquo stone which is used as an antidote forsnakebite are different herbs and pebbles available from theriver banks Likewise aqueous paste and decoction obtainedfrom the leaves of A paniculata are widely used for snakebitetreatment by indigenous people [43] Previous studies havereported that ethnomedicine plays major roles in conservingthe disappearing knowledge of tribal communities [44ndash47]The traditional beliefs of reliance on a rich diversity ofethnomedicinal plants located at different settlements havealso been confirmed in another study [48 49] Herein weexplored the various types of traditional practices reported bythe primitive tribal communities with a view to gain furtherknowledge from such studies

In the present investigation potentially rich sources oftribal medicine (71 plants) were scientifically evaluated fortheir antibacterial activity against the MDR bacteria and theaccumulated data was disseminated for the first time to the

scientific community Out of the 71medicinal plants screenedfor the antibacterial activity 10 of them (T involucrataC acida A marmelos A vasica C procera A paniculataand M piperita A indica S indicus and E cardamomum)displayed the highest antibacterial activity against the mul-tidrug resistant B pseudomallei (KHW and TES) and Saureus strains The antibacterial activity of those crude plantextracts was as equally effective as that of the standard drugsOur findings corroborated with the previous reports madeon the antistaphylococcal activity of tribal medicinal plants[34 50 51] On the other hand isolated components from themost active extracts of T involucrata shellsol and C acidaexhibited the most potent action against the antibiotic resis-tant B pseudomallei (KHW) K pneumoniae and S aureusstrainsThese results further confirmed our previous findingson the leaves of T involucrata and its compounds hydrocar-bon ester-like shellsol which displayed a high antibacterialeffect against the different bacterial strains especially thatof S aureus [34] Eugenol and caryophyllene are the activeagents contained in the M piperita [52] and O sanctumplants which are believed to be mainly responsible for theantimicrobial properties of these plants [53] Interestinglythe compounds obtained from the aromatic plants such asC zeylanicum and R officinalis were also found to be veryeffective against the multidrug resistant human pathogen Kpneumoniae S aureus S typhi and B pseudomallei (KHW)that causes melioidosis

The inhibitory potential determined for the shellsol(T involucrata) vaseline (A vasica) C acida and C zey-lanicum indicates that the MIC of 78ndash3125120583gmL foundagainst the B pseudomallei of KHW K pneumoniae Kpneumoniae S pyogenes and V damsel and S pneumoniaewas quite low Similarly lower MIC values were found for

Evidence-Based Complementary and Alternative Medicine 15

M piperita (MIC of 113ndash225 mgmL) against the abovebacterial strains [52] MIC was found for the most activealcohol extracts of A salvifolium (MIC 0034ndash0263mgmL)on S aureus [54] and the MIC for S trilobatum aqueousextracts determined against the tested organisms rangedfrom 006 to 05mgmL [55] C zeylanicum was foundto have an effective antibacterial activity (MIC 64 120583gmL)against P aeruginosa E coli B subtilis and S aureus[56] Previously several investigators have demonstrated thatactive agents exert interesting activity against bacteria evenat lower concentrations tested [57 58] The bacteriostaticmechanism involves damage to the cell walls of bacteriafollowed by inhibition of protein synthesis that ultimatelyleads to bacterial death [59] The most active plants arewidely used by various tribes as traditional treatment (iecut wounds skin infection and scabies) thus indicatingthe potential for further development into promising drugsIn order to ascertain the safety and efficacy of the mostactive compounds their effect on human skin fibroblast cellswas evaluated Chemical constituents of E cardamomumT involucrata S indicus C acida A vasica A marmelosA indica and A paniculata plants failed to produce anynoticeable toxicity up to 1000120583gmL Although some ofthe tested compounds exhibited a slight reduction of cellproliferation and some minor morphological changes suchchanges were insignificant at lower doses and became evidentonly at higher doses However certain aromatic compoundsof O sanctum E globulus V zizanioides C citratus and Eglobulus plants showed reduction of cell proliferation againstHEPK cells

Our phytochemical screening also provides evidenceof the presence of several types of compounds that aremainly responsible for the remarkable antibacterial effectof these plants The differences noted for the bactericidalactivity of various plant extracts in this study appears tobe directly related to the diversity of compounds (shown inparentheses) that are accumulated in the following plants(eg A marmelos and O umbellate) [60ndash62] Compoundslike tannins phenol and polyphenols can bind the Gram-negative bacteria to form a heavy soluble complex on the cellsurface which subsequently disturbs the availability of recep-tor on cells and kills the bacteria [63] Several species havingwide spectra of antimicrobial activity mainly due to theactive constituents such as essential oil phenolic compoundslike thymol carvacrol in oregano and thyme eugenol inclove and cinnamon were also identified previously [64 65]Essential oils degrade the cell wall interact with the cell com-ponents and then disrupt the cytoplasmic membrane [66]The antimicrobial effect of phenolic compounds may involvemultiplemodes of action including damage to themembraneprotein interference with membrane integrated enzymes[67] causing leakage of cellular components coagulation ofcytoplasm depletion of the proton motive force alterationof fatty acid and phospholipid constituents impairmentof enzymatic mechanisms for production and metabolismalteration of nutrient uptake and electron transport [68]influencing the synthesis of DNA and RNA and destroyingprotein translation and the function of the mitochondrionin eukaryotes [69] The mode of action of antimicrobial

agents depends on the type of microorganism and is mainlyrelated to their cell wall structure and the outer membranearrangement Most of the plant spices and herbs containcomplex phenolics (ie phenolic acids flavonoids tanninslignans coumarins quinines) In addition the mechanismsof action of each phenolic compound against various bacteriaare also very complicated [26 70] Further investigation istherefore required to understand the relationship betweenthe antimicrobial action and the chemical structure of everyphenolic compound in the tested extracts The informationavailable from previous pharmacological sources combinedwith the findings herein reported on the medicinal plantextracts may serve as essential data for future drug develop-ment to combat diseases caused by theMDRbacterial strains

Abbreviations

MDR Multidrug resistant bacteriaTES Strain of Burkholderia pseudomalleiMRSA Methicillin-resistant Staphylococcus aureusVRE Vancomycin-resistant enterococciMTB Mycobacterium tuberculosisCH3OH Methanol

CFU Colony forming unitHCl Hydrochloric acidTLC Thin layer chromatographyH2SO4 Sulphuric acids

UV Ultraviolet spectrophotometerHEPK Human skin fibroblasts

Acknowledgments

Theauthors are thankful to the ERI Loyola College ChennaiIndia for his support during the survey The authors alsothank the Department of Microbiology NUHS NUS forthe bacterial cultures used in this investigation The authorsare thankful to the tribal people for sharing their healingknowledge on medicinal aromatic plants

References

[1] M N Alekshun and S B Levy ldquoMolecular mechanisms ofantibacterialmultidrug resistancerdquoCell vol 128 no 6 pp 1037ndash1050 2007

[2] A D Russell ldquoAntibiotic and biocide resistance in bacteriaintroductionrdquo Symposium Series Society for Applied Microbiol-ogy vol 31 no 92 pp 1Sndash3S 2002

[3] D R Guay ldquoTreatment of bacterial skin and skin structureinfectionsrdquo Expert Opinion on Pharmacotherapy vol 4 no 8pp 1259ndash1275 2003

[4] A J Brosnahan and P M Schlievert ldquoGram-positive bac-terial superantigen outside-in signaling causes toxic shocksyndromerdquo FEBS Journal vol 278 no 23 pp 4649ndash4667 2011

[5] K Fellermann J Wehkamp K R Herrlinger and E F StangeldquoCrohnrsquos disease a defensin deficiency syndromerdquo EuropeanJournal of Gastroenterology and Hepatology vol 15 no 6 pp627ndash634 2003

16 Evidence-Based Complementary and Alternative Medicine

[6] L Chen S Lin R Agha-Majzoub L Overbergh C Mathieuand L S Chan ldquoCCL27 is a critical factor for the developmentof atopic dermatitis in the keratin-14 IL-4 transgenic mousemodelrdquo International Immunology vol 18 no 8 pp 1233ndash12422006

[7] J H Song V Thamlikitkul and P R Hsueh ldquoClinical andeconomic burden of community-acquired pneumonia amongstadults in the Asia-Pacific regionrdquo International Journal ofAntimicrobial Agents vol 38 no 2 pp 108ndash117 2011

[8] D A B Dance ldquoMelioidosisrdquo Current Opinion in InfectiousDiseases vol 15 no 2 pp 127ndash132 2002

[9] N J White ldquoMelioidosisrdquo The Lancet vol 361 no 9370 pp1715ndash1722 2003

[10] S Yang ldquoMelioidosis research in Chinardquo Acta Tropica vol 77no 2 pp 157ndash165 2000

[11] P R Hsueh L J Teng L N Lee et al ldquoMelioidosis an emerginginfection in Taiwanrdquo Emerging Infectious Diseases vol 7 no 3pp 428ndash433 2001

[12] C Walsh and S Fanning ldquoAntimicrobial resistance in food-borne pathogensmdasha cause for concernrdquo Current Drug Targetsvol 9 no 9 pp 808ndash815 2008

[13] M Vorachit P Chongtrakool S Arkomsean and S BoonsongldquoAntimicrobial resistance in Burkholderia pseudomalleirdquo ActaTropica vol 74 no 2-3 pp 139ndash144 2000

[14] F M Thibault E Hernandez D R Vidal M Girardet and JD Cavallo ldquoAntibiotic susceptibility of 65 isolates of Burkholde-ria pseudomallei and Burkholderia mallei to 35 antimicrobialagentsrdquo Journal of Antimicrobial Chemotherapy vol 54 no 6pp 1134ndash1138 2004

[15] R L S Sikarwar ldquoEthnogynaecological uses of plants new toIndiardquo Ethnobotany vol 12 no 1-2 pp 112ndash115 2002

[16] R Siva ldquoStatus of natural dyes and dye-yielding plants in IndiardquoCurrent Science vol 92 no 7 pp 916ndash925 2007

[17] P Kumar Singh V Kumar R K Tiwari A Sharma ChV Rao and R H Singh ldquoMedico-ethnobotany of lsquochatararsquoblock of district sonebhadra Uttar Pradesh Indiardquo Advances inBiological Research vol 4 no 1 pp 65ndash80 2010

[18] W H Lewis and M P Elwin-Lewis Medical Botany PlantsAffecting Human Health John Wiley amp Sons New York NYUSA 2003

[19] R Perumal Samy P N Pushparaj and P GopalakrishnakoneldquoA compilation of bioactive compounds fromAyurvedardquo Bioin-formation vol 3 no 3 pp 100ndash110 2008

[20] WorldHealthOrganization (WHO)TraditionalMedicine Strat-egy 2002ndash2005 Genera World Health Organization GenevaSwitzerland 2002

[21] P S Haddad G A Azar S Groom and M Boivin ldquoNaturalhealth products modulation of immune function and preven-tion of chronic diseasesrdquo Evidence Based Complementary andAlternative Medicine vol 2 no 4 pp 513ndash520 2005

[22] K Poonam andG S Singh ldquoEthnobotanical study of medicinalplants used by the Taungya community in Terai Arc LandscapeIndiardquo Journal of Ethnopharmacology vol 123 no 1 pp 167ndash1762009

[23] G B Mahady ldquoMedicinal plants for the prevention and treat-ment of bacterial infectionsrdquo Current Pharmaceutical Designvol 11 no 19 pp 2405ndash2427 2005

[24] V P Kumar N S Chauhan H Padh and M Rajani ldquoSearchfor antibacterial and antifungal agents from selected Indianmedicinal plantsrdquo Journal of Ethnopharmacology vol 107 no2 pp 182ndash188 2006

[25] R Nair T Kalariya and S Chanda ldquoAntibacterial activity ofsome plant extracts used in folk medicinerdquo Journal of HerbalPharmacotherapy vol 7 no 3-4 pp 191ndash201 2007

[26] D Kalemba and A Kunicka ldquoAntibacterial and antifungalproperties of essential oilsrdquo Current Medicinal Chemistry vol10 no 10 pp 813ndash829 2003

[27] A H Ashour ldquoAntibacterial antifungal and anticancer activi-ties of volatile oils and extracts from stems leaves and flowers ofEucalyptus sideroxylon and Eucalyptus torquatardquoCancer BiologyandTherapy vol 7 no 3 pp 399ndash403 2008

[28] M J Abad L M Bedoya L Apaza and P Bermejo ldquoTheArtemisia L genus a review of bioactive essential oilsrdquoMolecules vol 17 no 3 pp 2542ndash2566 2012

[29] H N Marzoug M Romdhane A Lebrihi et al ldquoEucalyptusoleosa essential oils chemical composition and antimicrobialand antioxidant activities of the oils from different plant parts(stems leaves flowers and fruits)rdquoMolecules vol 16 no 2 pp1695ndash1709 2011

[30] C Proestos D Sereli and M Komaitis ldquoDetermination ofphenolic compounds in aromatic plants by RP-HPLC and GC-MSrdquo Food Chemistry vol 95 no 1 pp 44ndash52 2006

[31] S Chao G Young C Oberg and K Nakaoka ldquoInhibition ofmethicillin-resistant Staphylococcus aureus (MRSA) by essentialoilsrdquo Flavour and Fragrance Journal vol 23 no 6 pp 444ndash4492008

[32] M Akin D Oguz and H T Saracoglu ldquoAntibacterial effectsof some plant extracts from Labiatae (Lamiaceae) growingnaturally around Sirnak-Silopi Turkeyrdquo International Journal ofPharmaceutical and Applied Sciences vol 1 no 1 pp 4ndash47 2010

[33] K Bayoub T Baibai D Mountassif A Retmane and ASoukri ldquoAntibacterial activities of the crude ethanol extracts ofmedicinal plants against Listeriamonocytogenes and some otherpathogenic strainsrdquo African Journal of Biotechnology vol 9 no27 pp 4251ndash4258 2010

[34] R P Samy P Gopalakrishnakone M Sarumathi P Houghtonand S Ignacimuthu ldquoPurification of antibacterial agents fromTragia involucratamdasha popular tribal medicine for wound heal-ingrdquo Journal of Ethnopharmacology vol 107 no 1 pp 99ndash1062006

[35] K M Matthew The Flora of Tamil Nadu Carnatic Part IndashIIIprinted at the Diocesan Press Madras India 1981ndash1983

[36] AW BauerWMKirby J C Sherris andM Turck ldquoAntibioticsusceptibility testing by a standardized single disk methodrdquoAmerican Journal of Clinical Pathology vol 45 no 4 pp 493ndash496 1966

[37] National Committee for Clinical Laboratory Standards Per-formance Standards For Antimicrobial Susceptibility Testing12th Informational Supplement Approved standard M100-S12National Committee for Clinical Laboratory Standards WaynePa USA 2002

[38] E A Elsinghorst ldquoMeasurement of invasion by gentamicinresistancerdquoMethods in Enzymology vol 236 pp 405ndash420 1994

[39] J B Harborne Phytochemical Methods Chapman and HallNew York NY USA 1976

[40] H Wagner S Bladt and E M Zgairski Plant Drug AnalysisSpringer New York NY USA 1984

[41] N Myers R A Mittermeler C G Mittermeler G A B daFonseca and J Kent ldquoBiodiversity hotspots for conservationprioritiesrdquo Nature vol 403 no 6772 pp 853ndash858 2000

[42] R Perumal Samy and S Ignacimuthu ldquoAntibacterial activity ofsome folkloremedicinal plants used by tribals inWestern Ghats

Evidence-Based Complementary and Alternative Medicine 17

of Indiardquo Journal of Ethnopharmacology vol 69 no 1 pp 63ndash712000

[43] K Nampoothiri A Chrispal A Begum S Jasmine K GGopinath and A Zachariah ldquoA clinical study of renal tubulardysfunction inCleistanthus collinus (Oduvanthalai) poisoningrdquoClinical Toxicology vol 48 no 3 pp 193ndash197 2010

[44] M Strathern UsefulKnowledge The Isiah Berlin Lecture Uni-versity of Manchester Brunswick UK

[45] C N Sunil and A K Pradeep ldquo(2001) Another new species ofTripogon (Poaceae) from Indiardquo Sida vol 19 pp 803ndash806 2005

[46] S Biber-Klemm and T Cottier Right To Plant GeneticResources and Traditional Knowledge Basic Issues and Prospec-tive Oxfordshire Nosworthy Way UK 2006

[47] V S Ramachandran S Shijo Joseph and R Aruna ldquoEth-nobotanical studies from amaravathy range of Indira GandhiWildlife SanctuaryWesternGhats CoimbatoreDistrict South-ern Indiardquo Ethnobotanical Leaflets vol 13 pp 1069ndash1087 2009

[48] A H Rajasab and I Mohamad ldquoDocumentation of folkknowledge on edible wild plants of North Karnatakardquo IndianJournal of Traditional Knowledge vol 3 no 4 pp 419ndash4292004

[49] M Rajadurai V G Vidhya M Ramya and A Bhaskar ldquoEthno-medicinal plants used by the traditional healers of pachamalaihills Tamilnadu Indiardquo Studies on Ethno-Medicine vol 3 no 1pp 39ndash41 2009

[50] S K Adesina O Idowu A O Ogundaini et al ldquoAntimicrobialconstituents of the leaves of Acalypha wilkesiana and Acalyphahispidardquo Phytotherapy Research vol 14 no 5 pp 371ndash374 2000

[51] M T Gutierrez-Lugo M P Singh W M Maiese and BN Timmermann ldquoNew antimicrobial cycloartane triterpenesfrom Acalypha communisrdquo Journal of Natural Products vol 65no 6 pp 872ndash875 2002

[52] A K Tyagi andAMalik ldquoAntimicrobial potential and chemicalcomposition of Mentha piperita oil in liquid and vapour phaseagainst food spoiling microorganismsrdquo Food Control vol 22no 11 pp 1707ndash1714 2011

[53] G Devendran and U Balasubramanian ldquoQualitative phyto-chemical screening and GC-MS analysis of Ocimum sanctumL leavesrdquoAsian Journal of Plant Science and Research vol 1 no4 pp 44ndash48 2011

[54] V C Jain N M Patel D P Shah P K Patel and B HJoshi ldquoAntioxidant and antimicrobial activities of Alangiumsalvifolium (Lf) Wang rootrdquo Global Journal of Pharmacologyvol 4 no 1 pp 13ndash18 2010

[55] P Swapna Latha andK Kannabiran ldquoAntimicrobial activity andphytochemicals of Solanum trilobatum LinnrdquoAfrican Journal ofBiotechnology vol 5 no 23 pp 2402ndash2404 2006

[56] M Usha1 S Ragini and S M A Naqvi ldquoAntibacterial activityof acetone and ethanol extracts of Cinnamon (Cinnamomumzeylanicum) and Ajowan (Trachyspermum ammi) on four foodspoilage bacteriardquo International Research Journal of BiologicalSciences vol 1 no 4 pp 7ndash11 2012

[57] S Prabuseenivasan M Jayakumar and S Ignacimuthu ldquoInvitro antibacterial activity of some plant essential oilsrdquo BMCComplementary and Alternative Medicine vol 6 article 39 pp1ndash8 2006

[58] G A Ayoola F M Lawore T Adelowotan et al ldquoChemi-cal analysis and antimicrobial activity of the essential oil ofSyzigium aromaticum (clove)rdquo African Journal of Biotechnologyvol 7 no 13 pp 2227ndash2231 2008

[59] Y J Fu L Chen Y Zu et al ldquoThe antibacterial activity of cloveessential oil against Propionibacterium acnes and its mechanismof actionrdquo Archives of Dermatology vol 145 no 1 pp 86ndash882009

[60] D Venkatesan C M Karrunakarn S Selva Kumar and P TPalani Swamy ldquoIdentification of phytochemical constituents ofAegle marmelos responsible for antimicrobial activity againstselected pathogenic organismsrdquo Ethnobotanical Leaflets vol 13pp 1362ndash1372 2009

[61] P Arun K G Purushotham J Johnsy jayarani and V KumarildquoIn vitro Antibacterial Activity of Oldenlandia umbellata anIndian medicinal Plantrdquo Journal of Pharmaceutical Science andTechnology vol 2 no 4 pp 198ndash201 2010

[62] A Manjamalai R Sardar Sathyajith Singh C Guruvayoorap-pan and V M Berlin Grace ldquoAnalysis of phytochemicalconstituents and antimicrobial activity of some medicinalplants in Taminadu Indiardquo Global Journal of Biotechnology andBiochemistry vol 5 no 2 pp 120ndash128 2010

[63] E Haslam ldquoNatural polyphenols (vegetable tannins) as drugsPossible modes of actionrdquo Journal of Natural Products vol 59no 2 pp 205ndash215 1996

[64] Y Cai Q Luo M Sun and H Corke ldquoAntioxidant activityand phenolic compounds of 112 traditional Chinese medicinalplants associated with anticancerrdquo Life Sciences vol 74 no 17pp 2157ndash2184 2004

[65] B Shan Y Z Cai M Sun and H Corke ldquoAntioxidant capacityof 26 spice extracts and characterization of their phenolicconstituentsrdquo Journal of Agricultural and Food Chemistry vol53 no 20 pp 7749ndash7759 2005

[66] R J W Lambert P N Skandamis P J Coote and G-J ENychas ldquoA study of the minimum inhibitory concentration andmode of action of oregano essential oil thymol and carvacrolrdquoJournal of AppliedMicrobiology vol 91 no 3 pp 453ndash462 2001

[67] M Raccach ldquoThe antimicrobial activity of phenolic antioxi-dants in food a reviewrdquo Journal of Food Safety vol 6 no 3 pp141ndash170 1984

[68] M Taniguchi Y Yano E Tada et al ldquoMode of action of polygo-dial an antifungal sesquiterpene dialdehyderdquo Agricultural andBiological Chemistry vol 52 no 6 pp 1409ndash1414 1988

[69] G J E Nychas ldquoNatural antimicrobials from plantsrdquo in NewMethods of Food Preservation G W Gould Ed pp 58ndash89Blackie Academic London UK 1995

[70] S Burt ldquoEssential oils their antibacterial properties and poten-tial applications in foodsmdasha reviewrdquo International Journal ofFood Microbiology vol 94 no 3 pp 223ndash253 2004

Submit your manuscripts athttpwwwhindawicom

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Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

MEDIATORSINFLAMMATION

of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Behavioural Neurology

EndocrinologyInternational Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Disease Markers

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

BioMed Research International

OncologyJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Oxidative Medicine and Cellular Longevity

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

PPAR Research

The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014

Immunology ResearchHindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Journal of

ObesityJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Computational and Mathematical Methods in Medicine

OphthalmologyJournal of

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Diabetes ResearchJournal of

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Research and TreatmentAIDS

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Gastroenterology Research and Practice

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Parkinsonrsquos Disease

Evidence-Based Complementary and Alternative Medicine

Volume 2014Hindawi Publishing Corporationhttpwwwhindawicom

Evidence-Based Complementary and Alternative Medicine 7

Table 1 Continued

Scientific name Family Voucher specimen Plant parts Yield (gm) Phytochemicalanalysis

Sphaeranthus indicus L Asteraceae D02060 Whole plant 10 Essential oilSwertia chirata (L) Ham Gentianaceae D0540 Whole plant 16 GlycodisesStrychnos nux-vomica L Loganiaceae S-22 Nuts 036 AlkaloidsTragia involucrata L Euphorbiaceae D068 Leaves 16 ShellsolTinospora cordifolia(Willd) Miers ex Hooff ampThoms

Menispermaceae 0118 Leafroot stem 50 Glycosides tannins

Tridax procumbens L Compositae 10649 Leaf 18 FlavonoidsTerminalia arjuna (DC)W amp A Combretaceae 033-c Bark 80 Phenolics

Tephrosia purpurea (L)Pers Fabaceae S-43 Whole plant 08 Isoflavone

Vitex negundo L Verbenaceae 0031 Leaf 24 TerpineolVetiveria zizanioides L Gramineae 0051 Root 103 Essential oilWithania somnifera (L)Dunal Solanaceae D02063 Root 21 Alkaloids

Wedelia calendulacea Less Asteraceae S-24 Leaves FlavonoidsZingiber officinale Rosc Zingiberaceae 0327 Rhizome 23 TanninsZanthoxylum limonella(Dennst) Alston Rutaceae 009 Bark 19 Alkaloids essential

oilClass of chemical compounds A alkaloids S saponins T tannins St steroids G glycosides T terpenoids P polyphenol P phenolics Sh shellsol Hhydrocarbon esters

the site of snakebite and decoction is given orally fortreatment by indigenous people Besides these tribes relyon the medicinal plants as ingredients for fabricating a kindof medicated stone for health management ldquoVishakallurdquo(poison stone) is used by the indigenous groups called Kaniin Kerala India to treat a snakebiteWhen the stone is placeddirectly on the bitten area it sticks to the body to absorb thepoison and then become detached when absorption seems tobe complete The ingredients of Vishakallu stones are madewith leaves of Ocimum sanctum Anisomeles malabaricaLeucas aspera Piper betle Santalum album and the pebblescollected from the river bank

35 Survey of Medicinal Plants and Their Health Care ValuesThe present study is an attempt to provide scientific basis andobtain justification for the traditional beliefs of reliance on arich diversity of ethnomedicinal plants along with the richheritage of traditional medicine practices related to healthcare system made available by the primitive tribal commu-nities located at different settlements The native traditionalpractitioners called ldquovaidyarsrdquo have a good knowledge aboutthe traditional plants locally available for treatment of var-ious diseases (Figures 3(a)ndash3(p)) Such traditional medicalknowledge is used for preparing home remedies ill healthprevention and routine health maintenanceThis knowledgeis also applicable to cover other sectors of social life Duringthe ethnobotanical survey the wealth of 78 medicinal plantspecies used by the indigenous tribal community for varioustypes of health treatment was documented The botanical

names family names parts used chemical constituents andtheir application are provided (Table 1)

36 Antimicrobial Activity of Crude Extracts In this studywe reported the antimicrobial screening of methanolic crudeextracts of 78 medicinal plants (Table 2) Results revealedthat 68 plant extracts displayed potent activities against oneor more Gram-positive and -negative bacteria Of whichTragia involucrata Citrus acida Aegle marmelos Adha-toda vasica Calotropis procera Andrographis paniculata andMentha piperita Azadirachta indica Sphaeranthus indicusand Elettaria cardamomum showed the highest antibacte-rial activity against the multidrug resistant B pseudomallei(KHW and TES) and S aureus at 100120583gmL concentrationThe extracts showed pronounced antibacterial activity withtheir inhibitory zones ranging from 20 to 31mm in diam-eter as compared to the standard drugs chloramphenicoland ceftazidime (29ndash33mm) The majority of the plantsdemonstrated a powerful antimicrobial potency against themultidrug resistant strains of B pseudomallei (KHW andTES) K pneumonia and S aureus Approximately twenty-one plant extracts exerted only a weak or moderate effectagainst the tested bacteria while the crude extract of 13plants failed to show any effect at all Except for the plantextracts of T involucrata A lanceolata A vasica and Sindicus extracts the majority of the plant extracts wereineffective against theV damsela infection fascinatingly only11 plants exhibited activity against P aeruginosa of whichS indicus M piperita and C procera were found to havevery strong inhibition of bacteria at the tested concentrations

8 Evidence-Based Complementary and Alternative Medicine

Table 2 Antimicrobial activity of methanol extract of aromatic medicinal plants and essential oils evaluated against multidrug resistant(MDR) human pathogens at 100120583gmL concentration

Scientific name Microorganisms growth inhibition zones (6 millimeter in diameters)KHW TES Kp Kpr Pa Sa Stp Sp Vd Vd

Adhatoda vasica Nees 28 19 22 18 12 mdash 10 17 16 mdashAegle marmelos (L) Correa ex Roxb 29 17 9 10 mdash 15 mdash mdash mdash mdashAlangium salvifolium (L) f Wangerin 15 10 8 8 mdash 12 9 10 mdash mdashAndrographis echioides L 12 9 mdash mdash mdash 17 8 9 mdash mdashAndrographis paniculataWallich ex Nees 26 21 19 13 mdash 25 8 16 12 mdashAcalypha indica L mdash mdash mdash mdash mdash 18 mdash mdash mdash mdashAcalypha lanceolata L mdash mdash 12 13 mdash 10 mdash mdash 21 mdashAchyranthes aspera L mdash mdash 7 14 mdash 15 12 11 mdash mdashAgeratum conyzoides L 10 8 mdash mdash mdash mdash mdash mdash mdash mdashAsteracantha longifolia L 16 12 mdash mdash mdash 15 mdash mdash mdash mdashAzadirachta indica A Juss 15 17 21 16 14 23 12 14 mdash mdashBorassus flabellifer L 9 10 17 8 mdash mdash 9 10 mdash mdashBoerhavia erecta L mdash mdash 8 9 7 16 mdash mdash mdash mdashCalotropis procera (L) mdash mdash 15 mdash 18 28 9 mdash mdash mdashCalotropis gigantea (L) RBrex Ait 11 9 mdash mdash mdash 20 mdash mdash 9 8Cardiospermum halicacabum L 23 mdash mdash 14 mdash 9 19 mdash mdash mdashCatharanthus roseus (L) GDon 13 7 12 9 mdash 15 11 8 mdash mdashCassia auriculata L 17 13 12 mdash mdash 19 13 mdash mdash mdashCassia occidentalis L 18 mdash mdash mdash mdash mdash mdash mdash mdash mdashCassia tora L mdash mdash mdash mdash mdash mdash mdash mdash mdash mdashCassia fistula L mdash mdash mdash mdash mdash mdash mdash mdash mdash mdashCitrus acida Roxb Hookf 26 22 mdash 12 29 9 13 8 mdashCissus quadrangularis L mdash mdash mdash mdash mdash mdash mdash mdash mdash mdashCinnamomum zeylanicum Garcin exBlume 14 16 7 20 mdash 22 19 7 mdash mdash

Cinnamomum iners Reinw ex Blume 20 16 mdash 15 mdash 16 12 mdash mdash mdashRosmarinus officinalis L mdash 9 mdash 10 mdash 7 mdash 8 mdash mdashCentella asiatica (L) 9 8 mdash mdash mdash 11 mdash mdash mdash mdashClerodendrum inerme (L) Gaertn 13 7 12 mdash mdash 15 11 8 mdash mdashClitoria ternatea L mdash mdash mdash mdash mdash mdash mdash mdash mdash mdashClitoria ternatea L 16 12 mdash 13 mdash 8 10 mdash mdash mdashCleome gynandropsis L mdash mdash 12 19 11 mdash 9 15 12 mdashCleome viscose L mdash mdash 8 10 14 20 mdash mdash mdash mdashCoccinia grandisWampA 9 10 17 8 mdash mdash 9 10 mdash mdashCymbopogon citratus (DC) 16 18 mdash 17 mdash 14 mdash mdash mdash mdashDatura metel L mdash mdash mdash mdash mdash mdash mdash mdash mdash mdashEclipta alba (L) Hassk 20 mdash mdash 9 mdash mdash 10 mdash mdash mdashEuphorbia hirta Linn 11 mdash mdash mdash mdash 16 mdash mdash mdash mdashEucalyptus globulus Labill mdash mdash mdash 7 mdash mdash mdash 7 mdash mdashEugenia caryophyllus Bullock amp Harrison mdash 7 mdash 11 mdash 9 mdash 8 mdash mdashElettaria cardamomumWhite et Mason 21 20 7 14 mdash 22 12 17 mdash mdashHyptis suaveolens (L) Poit 8 10 8 7 mdash mdash mdash mdash mdash mdashIchnocarpus frutescens (L) WJ Aiton mdash mdash mdash mdash mdash mdash mdash mdash mdash mdashJatropha curcas L mdash mdash mdash 8 mdash 11 mdash 12 mdash 7Leucas aspera (Willd) Link 9 10 17 8 mdash 12 9 10 mdash mdashLawsonia inermis L mdash mdash mdash mdash mdash mdash mdash mdash mdash mdashMadhuca longifolia (L) JF Macbr 18 16 14 19 12 mdash 8 7 9 mdash

Evidence-Based Complementary and Alternative Medicine 9

Table 2 Continued

Scientific name Microorganisms growth inhibition zones (6 millimeter in diameters)KHW TES Kp Kpr Pa Sa Stp Sp Vd Vd

Merremia hastate L (Desr) Hallierf mdash mdash mdash mdash mdash mdash mdash mdash mdash mdashMorinda tinctoria Roxb mdash mdash mdash mdash mdash mdash mdash mdash mdash mdashMentha piperita L 23 17 26 12 20 25 19 mdash mdash mdashOcimum sanctum L 12 9 11 7 mdash 15 7 8 mdash mdashOldenlandia umbellata L mdash mdash mdash mdash mdash 17 mdash mdash mdash mdashPiper attenuatum Buch Hamex Miq 13 7 12 21 mdash 17 11 8 mdash mdashPlumbago zeylanica (L) Cav 9 10 17 8 mdash 12 9 10 mdash mdashPlectranthus amboinicus (L) Spreng mdash mdash mdash 8 11 15 mdash mdash mdash mdashPhyllanthus debilis L (Klein ex Willd) 7 8 mdash 9 mdash 9 18 7 mdash mdashPhyllanthus maderaspatensis L 17 mdash mdash mdash mdash mdash mdash mdash mdash mdashPremna tomentosaWilld 13 10 mdash 15 mdash 9 10 mdash mdash mdashGloriosa superba L 17 16 7 8 mdash 15 mdash mdash mdash 8Sebastiania chamaelea (L) Muell Arg 19 12 mdash 17 mdash 13 19 8 mdash mdashSolanum trilobatum L mdash mdash 13 9 mdash 8 mdash mdash mdash mdashSphaeranthus indicus L 20 18 21 7 22 mdash 11 9 mdash 16Swertia chirata (L) Ham mdash mdash mdash mdash mdash mdash mdash mdash mdash mdashTerminalia arjuna (W amp A) mdash mdash mdash mdash mdash 16 mdash mdash mdash mdashTinospora cordifolia (Willd) Miers exHooff ampThoms mdash mdash 23 16 mdash 12 15 mdash mdash mdash

Tridax procumbens L 9 8 7 mdash mdash 14 mdash mdash mdash mdashTragia involucrata L 25 23 20 mdash mdash 31 28 22 19 mdashVitex negundo L mdash mdash mdash mdash mdash 14 mdash mdash mdash mdashVetiveria zizanioides (L) 11 9 7 7 mdash 16 mdash mdash 8Withania somnifera (L) Dunal 12 20 mdash mdash mdash 15 12 mdash mdash mdashZingiber officinale Rosc 14 11 7 15 mdash 7 12 7 mdash mdashZanthoxylum limonella (Dennst) Alston 13 7 12 9 mdash mdash 11 8 mdash mdashChloramphenicol (30120583gdisc) 21 12 15 17 29 16 15 18 13 11Ceftazidime (30 120583gdisc) 33 16 22 19 16 25 21 20 12 15lowastBacteria (+minus) Results obtained in the disc diffusion assay antibacterial activity is expressed as the mean plusmn SD (119899 = 3) of the inhibition by the extract andits diameter around the discs One way analysis of variance was performed (mean plusmn SD 119899 = 3 replicates) Size of inhibition zones were including the sterileblank discs 6 millimeter (mm) in diameters Absence of bacterial inhibition indicates (mdash) antibiotic disc (30 120583gdisc)

Interestingly sixteen plants such as Andrographis echioidesC auriculata C viscose C gigantea T arjuna Oldenlandiaumbellata Boerhavia erecta and E hirta exerted a strongactivity against the Gram-positive S aureus bacteria

37 Phytochemical Screening of Plants The results obtainedfrom the phytochemical screening as shown in Table 1 indi-cate the presence of various types of secondary metabolitessuch as polyphenols tannins saponins alkaloids and gly-cosidespolysaccharides Most of the plant extracts relativelyrich in alkaloids phenols flavonoids polyphenols tanninssterols and terpenoids were found to inhibit the growth oforganisms

38 Antimicrobial Activity of Fractioned Compounds Activecomponents were purified from the most active extractsfor further testing The compound shellsol of T involucrata

and C acida exhibited the most potent action againstthe antibiotic resistant strains of B pseudomallei (KHW)S aureus B pseudomallei (TES) and K pneumoniae Amarmelos was also found to inhibit the growth of B pseudo-mallei (KHW) more effectively than other tested bacteria Avasica showed the broad spectrum growth inhibitory activityon B pseudomallei (KHW) K pneumoniae K pneumoniaeresistant B pseudomallei (TES) S pyogenes and V damselaHowever E cardamomum displayed antimicrobial activityon some of the B pseudomallei (KHW) S pyogenes Bpseudomallei (TES) and S typhi strains Similarly A indicaexerted the growth inhibition on K pneumoniae and Saureus Remarkably Sebastiania chamaelea was more activeagainst K pneumoniae K pneumoniae and S pyogenesThe compound from S indicus inhibited the growth of Kpneumoniae K pneumoniae B pseudomallei (KHW andTES) and S typhi strains as compared to the activity shown

10 Evidence-Based Complementary and Alternative Medicine

Figure 2 Diverse biodiversity richness of medicinal plants in Western and Eastern Ghats (a) Topography of plant covering area in Kollihills (Namakkal district Tamil Nadu) (b) Aerial view of occurrence of medicinal plants in Mundanthurai hills (Tirunelveli district TN) (c)Pachamalai hills (Trichy district) and its natural vegetation inhabitants for Malaiyali tribes (d) Deforestation of natural herbal resources dueto urbanization in Kalrayan hills (Salem district) in the Eastern Ghats of TN (e) Medicinal plants and its various parts used by the natives(traditional healers) for the treatment of diverse human illness with a very high percentage of leaves and whole plants often used for herbaldrug preparation by the local practitioners (f) Various category of plants like shrub herb climbers and tree and the parts used in medicine

Evidence-Based Complementary and Alternative Medicine 11

Figure 3 Medicinal aromatic herbs spices and toxic plants were collected from the tribal areas of the Western and Eastern Ghats region inTamil Nadu India (a) A vasica Nees (leaf) (b) Eclipta alba (L) Hassk (whole plant) (c)Mimosa pudica L (whole plant) (d) P amboinicus(L) Spreng (whole plant) (e) T procumbens (L) (whole plant) (f) Euphorbia hirta Linn (whole plant) (g) A paniculata Wallich ex Nees(H) C roseus (L) GDon (whole plant) used for therapy (i) Cinnamomum iners Reinw ex Blume (leaf) (j) E globulus Labill (leaf and bark)(k) Z officinale Rosc (Rhizome) (l) E caryophyllus (Sprengel) Bullock and Harrison (flower buds) (m) M piperita L (whole plant) (n) Ccitratus (DC) Clitoria ternatea L (whole plant) (o) C zeylanicum Garcin ex Blume (bark) (p) Elettaria cardamomumWhite et Mason (fruitpod) used for medicine and food preparation

by the crude extracts (Figure 4)The antimicrobial efficacy offractions collected from the oil yielding plants was also com-pared with that of the tested compounds C zeylanicum andR officinaliswere themost sensitive in controlling the growthof B pseudomallei (KHW) S aureus K pneumonia and Spneumoniae Fascinatingly all the compounds obtained fromaromatic plants except those from E globules were foundto be very effective against the multidrug resistant humanpathogen B pseudomallei (KHW) that causes melioidosis

On the other hand compounds from C citrates O sanctumE caryophyllus and Z zizanioide showed some promisingeffect only against S aureus (Figure 5) The activity of thecompounds were pronounced more than that of the oilyielding plant fractions

39 Minimum Inhibitory Concentrations (MICs) The antibi-otic potential of the purified fractions was obtained fromthe MIC determination The hydrocarbon ester shellsol

12 Evidence-Based Complementary and Alternative Medicine

(mean plusmn SD n = 3)

0

5

10

15

20

25

30

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn V

d St

Bacteria (+minus)

lowast

lowast

lowast

A vasica

In vitro assay(P

lant

frac

tion)

Inhi

bitio

n zo

nes (

mm

)

(a)

(mean plusmn SD n = 3)

0

5

10

15

20

25

30

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn V

d St

Bacteria (+minus)

lowast

A marmelos

Inhi

bitio

n zo

nes (

mm

)

In vitro assay

(b)

(mean plusmn SD n = 3)

0

5

10

15

20

25

30

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn V

d St

Bacteria (+minus)

lowastlowast

A indica

Inhi

bitio

n zo

nes (

mm

)

Incu

batio

n (37∘C

24 h

)

(c)

(mean plusmn SD n = 3)

0

5

10

15

20

25

30

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn V

d St

Bacteria (+minus)

lowast

lowast lowast

35

C acida

Inhi

bitio

n zo

nes (

mm

)(P

lant

frac

tion)

(d)

(mean plusmn SD n = 3)

0

5

10

15

20

25

30

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn V

d St

Bacteria (+minus)

lowastlowast

lowast

S chamaelea

Inhi

bitio

n zo

nes (

mm

)

(e)

(mean plusmn SD n = 3)

0

5

10

15

20

25

30

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn V

d St

Bacteria (+minus)

lowastlowast

lowastlowast

S indicus

Inhi

bitio

n zo

nes (

mm

)

Incu

batio

n (37∘C

24 h

)

(f)

(mean plusmn SD n = 3)

0

5

10

15

20

25

30

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn V

d St

Bacteria (+minus)

lowast

lowast lowast

E cardamomum

Inhi

bitio

n zo

nes (

mm

)(P

lant

frac

tion)

(g)

(mean plusmn SD n = 3)

0

5

10

15

20

25

30

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn V

d St

Bacteria (+minus)

lowast

lowast

lowast

lowast

Shellsol

35

Inhi

bitio

n zo

nes (

mm

)

Incu

batio

n (37∘C

24 h

)

(h)

Figure 4 In vitro antimicrobial activity of purified fractions from the most active plant extracts tested against bacteria Growth inhibitionzones were measured and analyzed with mean plusmn standard deviation (SD) (119899 = 3) using one way analysis of variance Level of significanceat (lowast119875 gt 001) Most of the fractions exerted a potent inhibitory effect against multidrug resistant Gram-negative bacteria (B pseudomalleistrains KHW and TES) K pneumonia and Gram-positive bacteria S aureus

(T involucrata) andC acida showed an interesting inhibitorypotential against S aureus (MIC of 78 120583gmL) and B pseu-domallei strain of KHW (MIC of 156 120583gmL) A vasicashowed an MIC of 156120583gmL against B pseudomallei(KHW) and an MIC of 3125120583gmL against K pneumo-niae K pneumoniae S pyogenes and V damsela strainsFractions from A marmelos and terpenoid from A indicaexerted bacteriostatic effect with MIC values of 3125120583gmLon some selected bacteria including B pseudomallei ofKHW S aureus and B pseudomallei of TES The MIC of3125 120583gmL was found for E cardamomum against S aureus

K pneumonia and S pyogenes S indicus displayed a verystrong inhibition against MDR K pneumoniae (MIC of156 120583gmL) and against B pseudomallei (KHW and TES)at MIC of 3125 120583gmL When the antimicrobial efficacies ofpurified fractions from aromatic plants were compared theC zeylanicum fraction displayed an important antimicrobialeffect against S aureus (MIC of 78 120583gmL) MDR B pseu-domallei of KHW (MIC of 156 120583gmL) and S pneumoniae(MIC of 3125 120583gmL) The essential oil from M piperateashowed MIC value of 3125120583gmL against K pneumoniae Saureus and B pseudomallei (KHW) respectivelyO sanctum

Evidence-Based Complementary and Alternative Medicine 13

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn

Bacteria (+minus)

0

5

10

15

20

25

30

Inhi

bitio

n zo

nes (

mm

)(P

lant

frac

tion)

lowastlowast

lowast

(mean plusmn SD n = 3)In vitro assay

C citratus

(a)

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn

Bacteria (+minus)

0

5

10

15

20

25

30

Inhi

bitio

n zo

nes (

mm

)

lowast

lowast

lowastlowast

lowast

lowast

(mean plusmn SD n = 3)In vitro assay

35

C zeylanicum

(b)

Bacteria (+minus)

0

5

10

15

20

25

30

Inhi

bitio

n zo

nes (

mm

)

lowast

lowast

lowast

(mean plusmn SD n = 3)

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn V

d St

O sanctum

Incu

batio

n (37∘C

24 h

)

(c)

Bacteria (+minus)

0

5

10

15

20

25

Inhi

bitio

n zo

nes (

mm

)

lowast

lowast

(mean plusmn SD n = 3)

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn V

d St

M piperita

(Pla

nt fr

actio

n)

(d)

Bacteria (+minus)

0

5

10

15

20

25

30

Inhi

bitio

n zo

nes (

mm

)lowast

lowast

lowast

(mean plusmn SD n = 3)

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn V

d St

E caryophyllus

(e)

0

5

10

15

20

25

30

Inhi

bitio

n zo

nes (

mm

) (mean plusmn SD n = 3)

Bacteria (+minus)

KHW

TES

Kp

KprPa Sa

Stp

Spn

E globulus

Incu

batio

n (37∘C

24 h

)

(f)

Bacteria (+minus)

0

5

10

15

20

25

30

Inhi

bitio

n zo

nes (

mm

)(P

lant

frac

tion)

lowast

lowast

lowast

lowast

(mean plusmn SD n = 3)

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn V

d

R officinalis

(g)

0

5

10

15

20

25

30

Inhi

bitio

n zo

nes (

mm

)

lowast

lowast

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn V

d St

Bacteria (+minus)V zizanioides

(mean plusmn SD n = 3)

Incu

batio

n (37∘C

24 h

)

(h)

Figure 5 Comparison of antimicrobial effect of plant compounds obtained from the most popularly used sources of essential oils as assayedby the disc-diffusion method in vitro It displayed a powerful activity against B pseudomallei and S aureus than the other bacteria strainsOther compounds showed only a moderate or weak action against the tested bacteria Values for zone of bacterial growth inhibition werepresented as mean plusmn SD (119899 = 3) with level of significance at (lowast119875 gt 001)

and C citratus fractions also showed antimicrobial activity(MICs of 3125ndash125120583gmL) only at higher concentrationsagainst the tested bacteria In addition to that higher concen-trations (gt250120583gmL) (of Vetiveria fractions) were requiredto inhibit Vibro species and others (including E globulusfractions) failed to show any effect at tested concentrations(78ndash125 120583gmL) However the purified fractions (from mostactive medicinal plants) showed strong bacteriostatic inhibi-tion against the tested organisms (Table 3)

310 Cytotoxic Effects of Plants When the components wereassayed for cytotoxicity against the normal human skinfibroblasts (HEPK) cells the compounds obtained from

E cardamomum T involucrata S indicusC acidaA vasicaA marmelos A indica and A paniculata did not showtoxicity up to 1000120583gmL (see Figures S1 and S2 in Supple-mentaryMaterial available online at httpdxdoiorg1011552013525613) A slight reduction of cell proliferation wasnoted only at higher doses (2000120583gmL) In contrast cellproliferation was markedly reduced after exposure of HEPKcells to O sanctum E globulus V zizanioides C citratusand E globulus compounds There was no gradual reductionin skin cell proliferation seen after exposure to C zeylan-icum R officinalis and M piperita (see Figures S3 and S4)compounds The toxicity was found to be concentration-dependent when the skin fibroblasts (HEPK) cells were

14 Evidence-Based Complementary and Alternative Medicine

Table 3 Minimum inhibitory concentrations (MICs) of purified plant fractions and essential oils against antibiotic resistant bacteria

Botanical name Family Parts used Gram-positive and -negative bacteria (MICs 120583gmL)KHW TES Kp KPr Sa Stp Sp Vd St

A indica Juss Meliaceae Seed (fraction) 3125 625 125 lt250 3125 mdash mdash mdash mdashA marmelos (L) Rutaceae Root-bark (F) 3125 3125 mdash mdash 3125 625 125 250 mdashA paniculata Nees Acanthaceae Leaf (fraction) 250 mdash 125 mdash 625 mdash mdash 125 mdashA vasica Nees Acanthaceae Fraction (Stem) 156 3125 3125 625 mdash lt250 3125 3125 mdashC acida Roxb Rutaceae Leaf (fraction) 156 625 mdash mdash 78 mdash 3125 mdash mdashE cardamomumWhite et Mason Acanthaceae Fraction (WP) 625 mdash 3125 mdash 3125 250 625 mdash mdashS indicus (L) Euphorbiaceae Whole plant (F) 3125 125 156 mdash 625 mdash mdash lt250 mdashT involucrata (L) Euphorbiaceae Shellosol (leaf) 156 3125 mdash mdash 78 625 mdash mdash mdashCinnamomum zeylanicum (L) Lauraceae Bark (fraction) 156 625 mdash mdash 78 3125 mdash mdash 125Cymbopogon citratus (L) Graminae Leaf (fraction) 250 lt250 625 mdash 625 125 mdash mdash mdashEugenia caryophyllus (L) Myrtaceae Flower buds (F) 625 125 mdash mdash 625 mdash mdash mdash mdashEucalyptus globulus (L) Myrtaceae Fraction (leaf) mdash mdash mdash mdash mdash mdash lt250 mdash lt250Mentha piperita (L) Labiatae Fraction (WP) mdash 625 3125 mdash 3125 mdash lt250 mdash mdashOcimum sanctum (L) Labiatae Leaf (fraction) 625 125 mdash 3125 625 mdash 125 lt250 mdashRosmarinus officinalis (L) Labiatae Rosemary oil 3125 mdash mdash mdash 3125 mdash mdash mdash mdashVetiveria zizanioides (L) Graminae Root (fraction) mdash 125 125 mdash mdash mdash gt250 gt250The bacterial growth inhibitory activity was compared for significant differences within the bacterial strains by broth-dilution method at 250 125 625 3125156 and 78 120583gmL F fractions

exposed to various compounds The cell proliferation wasincreased by the influence of the plant components at thelower concentrations Whereas the oil yielding plant com-pounds showed inhibition of cell proliferation and toxicity athigher doses 250ndash1000120583gmL

4 Discussion

The Western Ghats is considered as one of the richestbiodiversity hotspots in the world [41] In this survey wecollected nearly 78 medicinal plants from Western andEastern Ghats that are edible and popularly used for curingvarious ailments including snakebite Traditional remedieshave a long-standing history in many tribal settlements inTN India and they continue to provide useful and applicabletools for treating ailments [42] The ingredients that makeup the ldquoVishakallurdquo stone which is used as an antidote forsnakebite are different herbs and pebbles available from theriver banks Likewise aqueous paste and decoction obtainedfrom the leaves of A paniculata are widely used for snakebitetreatment by indigenous people [43] Previous studies havereported that ethnomedicine plays major roles in conservingthe disappearing knowledge of tribal communities [44ndash47]The traditional beliefs of reliance on a rich diversity ofethnomedicinal plants located at different settlements havealso been confirmed in another study [48 49] Herein weexplored the various types of traditional practices reported bythe primitive tribal communities with a view to gain furtherknowledge from such studies

In the present investigation potentially rich sources oftribal medicine (71 plants) were scientifically evaluated fortheir antibacterial activity against the MDR bacteria and theaccumulated data was disseminated for the first time to the

scientific community Out of the 71medicinal plants screenedfor the antibacterial activity 10 of them (T involucrataC acida A marmelos A vasica C procera A paniculataand M piperita A indica S indicus and E cardamomum)displayed the highest antibacterial activity against the mul-tidrug resistant B pseudomallei (KHW and TES) and Saureus strains The antibacterial activity of those crude plantextracts was as equally effective as that of the standard drugsOur findings corroborated with the previous reports madeon the antistaphylococcal activity of tribal medicinal plants[34 50 51] On the other hand isolated components from themost active extracts of T involucrata shellsol and C acidaexhibited the most potent action against the antibiotic resis-tant B pseudomallei (KHW) K pneumoniae and S aureusstrainsThese results further confirmed our previous findingson the leaves of T involucrata and its compounds hydrocar-bon ester-like shellsol which displayed a high antibacterialeffect against the different bacterial strains especially thatof S aureus [34] Eugenol and caryophyllene are the activeagents contained in the M piperita [52] and O sanctumplants which are believed to be mainly responsible for theantimicrobial properties of these plants [53] Interestinglythe compounds obtained from the aromatic plants such asC zeylanicum and R officinalis were also found to be veryeffective against the multidrug resistant human pathogen Kpneumoniae S aureus S typhi and B pseudomallei (KHW)that causes melioidosis

The inhibitory potential determined for the shellsol(T involucrata) vaseline (A vasica) C acida and C zey-lanicum indicates that the MIC of 78ndash3125120583gmL foundagainst the B pseudomallei of KHW K pneumoniae Kpneumoniae S pyogenes and V damsel and S pneumoniaewas quite low Similarly lower MIC values were found for

Evidence-Based Complementary and Alternative Medicine 15

M piperita (MIC of 113ndash225 mgmL) against the abovebacterial strains [52] MIC was found for the most activealcohol extracts of A salvifolium (MIC 0034ndash0263mgmL)on S aureus [54] and the MIC for S trilobatum aqueousextracts determined against the tested organisms rangedfrom 006 to 05mgmL [55] C zeylanicum was foundto have an effective antibacterial activity (MIC 64 120583gmL)against P aeruginosa E coli B subtilis and S aureus[56] Previously several investigators have demonstrated thatactive agents exert interesting activity against bacteria evenat lower concentrations tested [57 58] The bacteriostaticmechanism involves damage to the cell walls of bacteriafollowed by inhibition of protein synthesis that ultimatelyleads to bacterial death [59] The most active plants arewidely used by various tribes as traditional treatment (iecut wounds skin infection and scabies) thus indicatingthe potential for further development into promising drugsIn order to ascertain the safety and efficacy of the mostactive compounds their effect on human skin fibroblast cellswas evaluated Chemical constituents of E cardamomumT involucrata S indicus C acida A vasica A marmelosA indica and A paniculata plants failed to produce anynoticeable toxicity up to 1000120583gmL Although some ofthe tested compounds exhibited a slight reduction of cellproliferation and some minor morphological changes suchchanges were insignificant at lower doses and became evidentonly at higher doses However certain aromatic compoundsof O sanctum E globulus V zizanioides C citratus and Eglobulus plants showed reduction of cell proliferation againstHEPK cells

Our phytochemical screening also provides evidenceof the presence of several types of compounds that aremainly responsible for the remarkable antibacterial effectof these plants The differences noted for the bactericidalactivity of various plant extracts in this study appears tobe directly related to the diversity of compounds (shown inparentheses) that are accumulated in the following plants(eg A marmelos and O umbellate) [60ndash62] Compoundslike tannins phenol and polyphenols can bind the Gram-negative bacteria to form a heavy soluble complex on the cellsurface which subsequently disturbs the availability of recep-tor on cells and kills the bacteria [63] Several species havingwide spectra of antimicrobial activity mainly due to theactive constituents such as essential oil phenolic compoundslike thymol carvacrol in oregano and thyme eugenol inclove and cinnamon were also identified previously [64 65]Essential oils degrade the cell wall interact with the cell com-ponents and then disrupt the cytoplasmic membrane [66]The antimicrobial effect of phenolic compounds may involvemultiplemodes of action including damage to themembraneprotein interference with membrane integrated enzymes[67] causing leakage of cellular components coagulation ofcytoplasm depletion of the proton motive force alterationof fatty acid and phospholipid constituents impairmentof enzymatic mechanisms for production and metabolismalteration of nutrient uptake and electron transport [68]influencing the synthesis of DNA and RNA and destroyingprotein translation and the function of the mitochondrionin eukaryotes [69] The mode of action of antimicrobial

agents depends on the type of microorganism and is mainlyrelated to their cell wall structure and the outer membranearrangement Most of the plant spices and herbs containcomplex phenolics (ie phenolic acids flavonoids tanninslignans coumarins quinines) In addition the mechanismsof action of each phenolic compound against various bacteriaare also very complicated [26 70] Further investigation istherefore required to understand the relationship betweenthe antimicrobial action and the chemical structure of everyphenolic compound in the tested extracts The informationavailable from previous pharmacological sources combinedwith the findings herein reported on the medicinal plantextracts may serve as essential data for future drug develop-ment to combat diseases caused by theMDRbacterial strains

Abbreviations

MDR Multidrug resistant bacteriaTES Strain of Burkholderia pseudomalleiMRSA Methicillin-resistant Staphylococcus aureusVRE Vancomycin-resistant enterococciMTB Mycobacterium tuberculosisCH3OH Methanol

CFU Colony forming unitHCl Hydrochloric acidTLC Thin layer chromatographyH2SO4 Sulphuric acids

UV Ultraviolet spectrophotometerHEPK Human skin fibroblasts

Acknowledgments

Theauthors are thankful to the ERI Loyola College ChennaiIndia for his support during the survey The authors alsothank the Department of Microbiology NUHS NUS forthe bacterial cultures used in this investigation The authorsare thankful to the tribal people for sharing their healingknowledge on medicinal aromatic plants

References

[1] M N Alekshun and S B Levy ldquoMolecular mechanisms ofantibacterialmultidrug resistancerdquoCell vol 128 no 6 pp 1037ndash1050 2007

[2] A D Russell ldquoAntibiotic and biocide resistance in bacteriaintroductionrdquo Symposium Series Society for Applied Microbiol-ogy vol 31 no 92 pp 1Sndash3S 2002

[3] D R Guay ldquoTreatment of bacterial skin and skin structureinfectionsrdquo Expert Opinion on Pharmacotherapy vol 4 no 8pp 1259ndash1275 2003

[4] A J Brosnahan and P M Schlievert ldquoGram-positive bac-terial superantigen outside-in signaling causes toxic shocksyndromerdquo FEBS Journal vol 278 no 23 pp 4649ndash4667 2011

[5] K Fellermann J Wehkamp K R Herrlinger and E F StangeldquoCrohnrsquos disease a defensin deficiency syndromerdquo EuropeanJournal of Gastroenterology and Hepatology vol 15 no 6 pp627ndash634 2003

16 Evidence-Based Complementary and Alternative Medicine

[6] L Chen S Lin R Agha-Majzoub L Overbergh C Mathieuand L S Chan ldquoCCL27 is a critical factor for the developmentof atopic dermatitis in the keratin-14 IL-4 transgenic mousemodelrdquo International Immunology vol 18 no 8 pp 1233ndash12422006

[7] J H Song V Thamlikitkul and P R Hsueh ldquoClinical andeconomic burden of community-acquired pneumonia amongstadults in the Asia-Pacific regionrdquo International Journal ofAntimicrobial Agents vol 38 no 2 pp 108ndash117 2011

[8] D A B Dance ldquoMelioidosisrdquo Current Opinion in InfectiousDiseases vol 15 no 2 pp 127ndash132 2002

[9] N J White ldquoMelioidosisrdquo The Lancet vol 361 no 9370 pp1715ndash1722 2003

[10] S Yang ldquoMelioidosis research in Chinardquo Acta Tropica vol 77no 2 pp 157ndash165 2000

[11] P R Hsueh L J Teng L N Lee et al ldquoMelioidosis an emerginginfection in Taiwanrdquo Emerging Infectious Diseases vol 7 no 3pp 428ndash433 2001

[12] C Walsh and S Fanning ldquoAntimicrobial resistance in food-borne pathogensmdasha cause for concernrdquo Current Drug Targetsvol 9 no 9 pp 808ndash815 2008

[13] M Vorachit P Chongtrakool S Arkomsean and S BoonsongldquoAntimicrobial resistance in Burkholderia pseudomalleirdquo ActaTropica vol 74 no 2-3 pp 139ndash144 2000

[14] F M Thibault E Hernandez D R Vidal M Girardet and JD Cavallo ldquoAntibiotic susceptibility of 65 isolates of Burkholde-ria pseudomallei and Burkholderia mallei to 35 antimicrobialagentsrdquo Journal of Antimicrobial Chemotherapy vol 54 no 6pp 1134ndash1138 2004

[15] R L S Sikarwar ldquoEthnogynaecological uses of plants new toIndiardquo Ethnobotany vol 12 no 1-2 pp 112ndash115 2002

[16] R Siva ldquoStatus of natural dyes and dye-yielding plants in IndiardquoCurrent Science vol 92 no 7 pp 916ndash925 2007

[17] P Kumar Singh V Kumar R K Tiwari A Sharma ChV Rao and R H Singh ldquoMedico-ethnobotany of lsquochatararsquoblock of district sonebhadra Uttar Pradesh Indiardquo Advances inBiological Research vol 4 no 1 pp 65ndash80 2010

[18] W H Lewis and M P Elwin-Lewis Medical Botany PlantsAffecting Human Health John Wiley amp Sons New York NYUSA 2003

[19] R Perumal Samy P N Pushparaj and P GopalakrishnakoneldquoA compilation of bioactive compounds fromAyurvedardquo Bioin-formation vol 3 no 3 pp 100ndash110 2008

[20] WorldHealthOrganization (WHO)TraditionalMedicine Strat-egy 2002ndash2005 Genera World Health Organization GenevaSwitzerland 2002

[21] P S Haddad G A Azar S Groom and M Boivin ldquoNaturalhealth products modulation of immune function and preven-tion of chronic diseasesrdquo Evidence Based Complementary andAlternative Medicine vol 2 no 4 pp 513ndash520 2005

[22] K Poonam andG S Singh ldquoEthnobotanical study of medicinalplants used by the Taungya community in Terai Arc LandscapeIndiardquo Journal of Ethnopharmacology vol 123 no 1 pp 167ndash1762009

[23] G B Mahady ldquoMedicinal plants for the prevention and treat-ment of bacterial infectionsrdquo Current Pharmaceutical Designvol 11 no 19 pp 2405ndash2427 2005

[24] V P Kumar N S Chauhan H Padh and M Rajani ldquoSearchfor antibacterial and antifungal agents from selected Indianmedicinal plantsrdquo Journal of Ethnopharmacology vol 107 no2 pp 182ndash188 2006

[25] R Nair T Kalariya and S Chanda ldquoAntibacterial activity ofsome plant extracts used in folk medicinerdquo Journal of HerbalPharmacotherapy vol 7 no 3-4 pp 191ndash201 2007

[26] D Kalemba and A Kunicka ldquoAntibacterial and antifungalproperties of essential oilsrdquo Current Medicinal Chemistry vol10 no 10 pp 813ndash829 2003

[27] A H Ashour ldquoAntibacterial antifungal and anticancer activi-ties of volatile oils and extracts from stems leaves and flowers ofEucalyptus sideroxylon and Eucalyptus torquatardquoCancer BiologyandTherapy vol 7 no 3 pp 399ndash403 2008

[28] M J Abad L M Bedoya L Apaza and P Bermejo ldquoTheArtemisia L genus a review of bioactive essential oilsrdquoMolecules vol 17 no 3 pp 2542ndash2566 2012

[29] H N Marzoug M Romdhane A Lebrihi et al ldquoEucalyptusoleosa essential oils chemical composition and antimicrobialand antioxidant activities of the oils from different plant parts(stems leaves flowers and fruits)rdquoMolecules vol 16 no 2 pp1695ndash1709 2011

[30] C Proestos D Sereli and M Komaitis ldquoDetermination ofphenolic compounds in aromatic plants by RP-HPLC and GC-MSrdquo Food Chemistry vol 95 no 1 pp 44ndash52 2006

[31] S Chao G Young C Oberg and K Nakaoka ldquoInhibition ofmethicillin-resistant Staphylococcus aureus (MRSA) by essentialoilsrdquo Flavour and Fragrance Journal vol 23 no 6 pp 444ndash4492008

[32] M Akin D Oguz and H T Saracoglu ldquoAntibacterial effectsof some plant extracts from Labiatae (Lamiaceae) growingnaturally around Sirnak-Silopi Turkeyrdquo International Journal ofPharmaceutical and Applied Sciences vol 1 no 1 pp 4ndash47 2010

[33] K Bayoub T Baibai D Mountassif A Retmane and ASoukri ldquoAntibacterial activities of the crude ethanol extracts ofmedicinal plants against Listeriamonocytogenes and some otherpathogenic strainsrdquo African Journal of Biotechnology vol 9 no27 pp 4251ndash4258 2010

[34] R P Samy P Gopalakrishnakone M Sarumathi P Houghtonand S Ignacimuthu ldquoPurification of antibacterial agents fromTragia involucratamdasha popular tribal medicine for wound heal-ingrdquo Journal of Ethnopharmacology vol 107 no 1 pp 99ndash1062006

[35] K M Matthew The Flora of Tamil Nadu Carnatic Part IndashIIIprinted at the Diocesan Press Madras India 1981ndash1983

[36] AW BauerWMKirby J C Sherris andM Turck ldquoAntibioticsusceptibility testing by a standardized single disk methodrdquoAmerican Journal of Clinical Pathology vol 45 no 4 pp 493ndash496 1966

[37] National Committee for Clinical Laboratory Standards Per-formance Standards For Antimicrobial Susceptibility Testing12th Informational Supplement Approved standard M100-S12National Committee for Clinical Laboratory Standards WaynePa USA 2002

[38] E A Elsinghorst ldquoMeasurement of invasion by gentamicinresistancerdquoMethods in Enzymology vol 236 pp 405ndash420 1994

[39] J B Harborne Phytochemical Methods Chapman and HallNew York NY USA 1976

[40] H Wagner S Bladt and E M Zgairski Plant Drug AnalysisSpringer New York NY USA 1984

[41] N Myers R A Mittermeler C G Mittermeler G A B daFonseca and J Kent ldquoBiodiversity hotspots for conservationprioritiesrdquo Nature vol 403 no 6772 pp 853ndash858 2000

[42] R Perumal Samy and S Ignacimuthu ldquoAntibacterial activity ofsome folkloremedicinal plants used by tribals inWestern Ghats

Evidence-Based Complementary and Alternative Medicine 17

of Indiardquo Journal of Ethnopharmacology vol 69 no 1 pp 63ndash712000

[43] K Nampoothiri A Chrispal A Begum S Jasmine K GGopinath and A Zachariah ldquoA clinical study of renal tubulardysfunction inCleistanthus collinus (Oduvanthalai) poisoningrdquoClinical Toxicology vol 48 no 3 pp 193ndash197 2010

[44] M Strathern UsefulKnowledge The Isiah Berlin Lecture Uni-versity of Manchester Brunswick UK

[45] C N Sunil and A K Pradeep ldquo(2001) Another new species ofTripogon (Poaceae) from Indiardquo Sida vol 19 pp 803ndash806 2005

[46] S Biber-Klemm and T Cottier Right To Plant GeneticResources and Traditional Knowledge Basic Issues and Prospec-tive Oxfordshire Nosworthy Way UK 2006

[47] V S Ramachandran S Shijo Joseph and R Aruna ldquoEth-nobotanical studies from amaravathy range of Indira GandhiWildlife SanctuaryWesternGhats CoimbatoreDistrict South-ern Indiardquo Ethnobotanical Leaflets vol 13 pp 1069ndash1087 2009

[48] A H Rajasab and I Mohamad ldquoDocumentation of folkknowledge on edible wild plants of North Karnatakardquo IndianJournal of Traditional Knowledge vol 3 no 4 pp 419ndash4292004

[49] M Rajadurai V G Vidhya M Ramya and A Bhaskar ldquoEthno-medicinal plants used by the traditional healers of pachamalaihills Tamilnadu Indiardquo Studies on Ethno-Medicine vol 3 no 1pp 39ndash41 2009

[50] S K Adesina O Idowu A O Ogundaini et al ldquoAntimicrobialconstituents of the leaves of Acalypha wilkesiana and Acalyphahispidardquo Phytotherapy Research vol 14 no 5 pp 371ndash374 2000

[51] M T Gutierrez-Lugo M P Singh W M Maiese and BN Timmermann ldquoNew antimicrobial cycloartane triterpenesfrom Acalypha communisrdquo Journal of Natural Products vol 65no 6 pp 872ndash875 2002

[52] A K Tyagi andAMalik ldquoAntimicrobial potential and chemicalcomposition of Mentha piperita oil in liquid and vapour phaseagainst food spoiling microorganismsrdquo Food Control vol 22no 11 pp 1707ndash1714 2011

[53] G Devendran and U Balasubramanian ldquoQualitative phyto-chemical screening and GC-MS analysis of Ocimum sanctumL leavesrdquoAsian Journal of Plant Science and Research vol 1 no4 pp 44ndash48 2011

[54] V C Jain N M Patel D P Shah P K Patel and B HJoshi ldquoAntioxidant and antimicrobial activities of Alangiumsalvifolium (Lf) Wang rootrdquo Global Journal of Pharmacologyvol 4 no 1 pp 13ndash18 2010

[55] P Swapna Latha andK Kannabiran ldquoAntimicrobial activity andphytochemicals of Solanum trilobatum LinnrdquoAfrican Journal ofBiotechnology vol 5 no 23 pp 2402ndash2404 2006

[56] M Usha1 S Ragini and S M A Naqvi ldquoAntibacterial activityof acetone and ethanol extracts of Cinnamon (Cinnamomumzeylanicum) and Ajowan (Trachyspermum ammi) on four foodspoilage bacteriardquo International Research Journal of BiologicalSciences vol 1 no 4 pp 7ndash11 2012

[57] S Prabuseenivasan M Jayakumar and S Ignacimuthu ldquoInvitro antibacterial activity of some plant essential oilsrdquo BMCComplementary and Alternative Medicine vol 6 article 39 pp1ndash8 2006

[58] G A Ayoola F M Lawore T Adelowotan et al ldquoChemi-cal analysis and antimicrobial activity of the essential oil ofSyzigium aromaticum (clove)rdquo African Journal of Biotechnologyvol 7 no 13 pp 2227ndash2231 2008

[59] Y J Fu L Chen Y Zu et al ldquoThe antibacterial activity of cloveessential oil against Propionibacterium acnes and its mechanismof actionrdquo Archives of Dermatology vol 145 no 1 pp 86ndash882009

[60] D Venkatesan C M Karrunakarn S Selva Kumar and P TPalani Swamy ldquoIdentification of phytochemical constituents ofAegle marmelos responsible for antimicrobial activity againstselected pathogenic organismsrdquo Ethnobotanical Leaflets vol 13pp 1362ndash1372 2009

[61] P Arun K G Purushotham J Johnsy jayarani and V KumarildquoIn vitro Antibacterial Activity of Oldenlandia umbellata anIndian medicinal Plantrdquo Journal of Pharmaceutical Science andTechnology vol 2 no 4 pp 198ndash201 2010

[62] A Manjamalai R Sardar Sathyajith Singh C Guruvayoorap-pan and V M Berlin Grace ldquoAnalysis of phytochemicalconstituents and antimicrobial activity of some medicinalplants in Taminadu Indiardquo Global Journal of Biotechnology andBiochemistry vol 5 no 2 pp 120ndash128 2010

[63] E Haslam ldquoNatural polyphenols (vegetable tannins) as drugsPossible modes of actionrdquo Journal of Natural Products vol 59no 2 pp 205ndash215 1996

[64] Y Cai Q Luo M Sun and H Corke ldquoAntioxidant activityand phenolic compounds of 112 traditional Chinese medicinalplants associated with anticancerrdquo Life Sciences vol 74 no 17pp 2157ndash2184 2004

[65] B Shan Y Z Cai M Sun and H Corke ldquoAntioxidant capacityof 26 spice extracts and characterization of their phenolicconstituentsrdquo Journal of Agricultural and Food Chemistry vol53 no 20 pp 7749ndash7759 2005

[66] R J W Lambert P N Skandamis P J Coote and G-J ENychas ldquoA study of the minimum inhibitory concentration andmode of action of oregano essential oil thymol and carvacrolrdquoJournal of AppliedMicrobiology vol 91 no 3 pp 453ndash462 2001

[67] M Raccach ldquoThe antimicrobial activity of phenolic antioxi-dants in food a reviewrdquo Journal of Food Safety vol 6 no 3 pp141ndash170 1984

[68] M Taniguchi Y Yano E Tada et al ldquoMode of action of polygo-dial an antifungal sesquiterpene dialdehyderdquo Agricultural andBiological Chemistry vol 52 no 6 pp 1409ndash1414 1988

[69] G J E Nychas ldquoNatural antimicrobials from plantsrdquo in NewMethods of Food Preservation G W Gould Ed pp 58ndash89Blackie Academic London UK 1995

[70] S Burt ldquoEssential oils their antibacterial properties and poten-tial applications in foodsmdasha reviewrdquo International Journal ofFood Microbiology vol 94 no 3 pp 223ndash253 2004

Submit your manuscripts athttpwwwhindawicom

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Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

MEDIATORSINFLAMMATION

of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Behavioural Neurology

EndocrinologyInternational Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Disease Markers

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

BioMed Research International

OncologyJournal of

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Oxidative Medicine and Cellular Longevity

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PPAR Research

The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014

Immunology ResearchHindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Journal of

ObesityJournal of

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Computational and Mathematical Methods in Medicine

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Research and TreatmentAIDS

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Gastroenterology Research and Practice

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Parkinsonrsquos Disease

Evidence-Based Complementary and Alternative Medicine

Volume 2014Hindawi Publishing Corporationhttpwwwhindawicom

8 Evidence-Based Complementary and Alternative Medicine

Table 2 Antimicrobial activity of methanol extract of aromatic medicinal plants and essential oils evaluated against multidrug resistant(MDR) human pathogens at 100120583gmL concentration

Scientific name Microorganisms growth inhibition zones (6 millimeter in diameters)KHW TES Kp Kpr Pa Sa Stp Sp Vd Vd

Adhatoda vasica Nees 28 19 22 18 12 mdash 10 17 16 mdashAegle marmelos (L) Correa ex Roxb 29 17 9 10 mdash 15 mdash mdash mdash mdashAlangium salvifolium (L) f Wangerin 15 10 8 8 mdash 12 9 10 mdash mdashAndrographis echioides L 12 9 mdash mdash mdash 17 8 9 mdash mdashAndrographis paniculataWallich ex Nees 26 21 19 13 mdash 25 8 16 12 mdashAcalypha indica L mdash mdash mdash mdash mdash 18 mdash mdash mdash mdashAcalypha lanceolata L mdash mdash 12 13 mdash 10 mdash mdash 21 mdashAchyranthes aspera L mdash mdash 7 14 mdash 15 12 11 mdash mdashAgeratum conyzoides L 10 8 mdash mdash mdash mdash mdash mdash mdash mdashAsteracantha longifolia L 16 12 mdash mdash mdash 15 mdash mdash mdash mdashAzadirachta indica A Juss 15 17 21 16 14 23 12 14 mdash mdashBorassus flabellifer L 9 10 17 8 mdash mdash 9 10 mdash mdashBoerhavia erecta L mdash mdash 8 9 7 16 mdash mdash mdash mdashCalotropis procera (L) mdash mdash 15 mdash 18 28 9 mdash mdash mdashCalotropis gigantea (L) RBrex Ait 11 9 mdash mdash mdash 20 mdash mdash 9 8Cardiospermum halicacabum L 23 mdash mdash 14 mdash 9 19 mdash mdash mdashCatharanthus roseus (L) GDon 13 7 12 9 mdash 15 11 8 mdash mdashCassia auriculata L 17 13 12 mdash mdash 19 13 mdash mdash mdashCassia occidentalis L 18 mdash mdash mdash mdash mdash mdash mdash mdash mdashCassia tora L mdash mdash mdash mdash mdash mdash mdash mdash mdash mdashCassia fistula L mdash mdash mdash mdash mdash mdash mdash mdash mdash mdashCitrus acida Roxb Hookf 26 22 mdash 12 29 9 13 8 mdashCissus quadrangularis L mdash mdash mdash mdash mdash mdash mdash mdash mdash mdashCinnamomum zeylanicum Garcin exBlume 14 16 7 20 mdash 22 19 7 mdash mdash

Cinnamomum iners Reinw ex Blume 20 16 mdash 15 mdash 16 12 mdash mdash mdashRosmarinus officinalis L mdash 9 mdash 10 mdash 7 mdash 8 mdash mdashCentella asiatica (L) 9 8 mdash mdash mdash 11 mdash mdash mdash mdashClerodendrum inerme (L) Gaertn 13 7 12 mdash mdash 15 11 8 mdash mdashClitoria ternatea L mdash mdash mdash mdash mdash mdash mdash mdash mdash mdashClitoria ternatea L 16 12 mdash 13 mdash 8 10 mdash mdash mdashCleome gynandropsis L mdash mdash 12 19 11 mdash 9 15 12 mdashCleome viscose L mdash mdash 8 10 14 20 mdash mdash mdash mdashCoccinia grandisWampA 9 10 17 8 mdash mdash 9 10 mdash mdashCymbopogon citratus (DC) 16 18 mdash 17 mdash 14 mdash mdash mdash mdashDatura metel L mdash mdash mdash mdash mdash mdash mdash mdash mdash mdashEclipta alba (L) Hassk 20 mdash mdash 9 mdash mdash 10 mdash mdash mdashEuphorbia hirta Linn 11 mdash mdash mdash mdash 16 mdash mdash mdash mdashEucalyptus globulus Labill mdash mdash mdash 7 mdash mdash mdash 7 mdash mdashEugenia caryophyllus Bullock amp Harrison mdash 7 mdash 11 mdash 9 mdash 8 mdash mdashElettaria cardamomumWhite et Mason 21 20 7 14 mdash 22 12 17 mdash mdashHyptis suaveolens (L) Poit 8 10 8 7 mdash mdash mdash mdash mdash mdashIchnocarpus frutescens (L) WJ Aiton mdash mdash mdash mdash mdash mdash mdash mdash mdash mdashJatropha curcas L mdash mdash mdash 8 mdash 11 mdash 12 mdash 7Leucas aspera (Willd) Link 9 10 17 8 mdash 12 9 10 mdash mdashLawsonia inermis L mdash mdash mdash mdash mdash mdash mdash mdash mdash mdashMadhuca longifolia (L) JF Macbr 18 16 14 19 12 mdash 8 7 9 mdash

Evidence-Based Complementary and Alternative Medicine 9

Table 2 Continued

Scientific name Microorganisms growth inhibition zones (6 millimeter in diameters)KHW TES Kp Kpr Pa Sa Stp Sp Vd Vd

Merremia hastate L (Desr) Hallierf mdash mdash mdash mdash mdash mdash mdash mdash mdash mdashMorinda tinctoria Roxb mdash mdash mdash mdash mdash mdash mdash mdash mdash mdashMentha piperita L 23 17 26 12 20 25 19 mdash mdash mdashOcimum sanctum L 12 9 11 7 mdash 15 7 8 mdash mdashOldenlandia umbellata L mdash mdash mdash mdash mdash 17 mdash mdash mdash mdashPiper attenuatum Buch Hamex Miq 13 7 12 21 mdash 17 11 8 mdash mdashPlumbago zeylanica (L) Cav 9 10 17 8 mdash 12 9 10 mdash mdashPlectranthus amboinicus (L) Spreng mdash mdash mdash 8 11 15 mdash mdash mdash mdashPhyllanthus debilis L (Klein ex Willd) 7 8 mdash 9 mdash 9 18 7 mdash mdashPhyllanthus maderaspatensis L 17 mdash mdash mdash mdash mdash mdash mdash mdash mdashPremna tomentosaWilld 13 10 mdash 15 mdash 9 10 mdash mdash mdashGloriosa superba L 17 16 7 8 mdash 15 mdash mdash mdash 8Sebastiania chamaelea (L) Muell Arg 19 12 mdash 17 mdash 13 19 8 mdash mdashSolanum trilobatum L mdash mdash 13 9 mdash 8 mdash mdash mdash mdashSphaeranthus indicus L 20 18 21 7 22 mdash 11 9 mdash 16Swertia chirata (L) Ham mdash mdash mdash mdash mdash mdash mdash mdash mdash mdashTerminalia arjuna (W amp A) mdash mdash mdash mdash mdash 16 mdash mdash mdash mdashTinospora cordifolia (Willd) Miers exHooff ampThoms mdash mdash 23 16 mdash 12 15 mdash mdash mdash

Tridax procumbens L 9 8 7 mdash mdash 14 mdash mdash mdash mdashTragia involucrata L 25 23 20 mdash mdash 31 28 22 19 mdashVitex negundo L mdash mdash mdash mdash mdash 14 mdash mdash mdash mdashVetiveria zizanioides (L) 11 9 7 7 mdash 16 mdash mdash 8Withania somnifera (L) Dunal 12 20 mdash mdash mdash 15 12 mdash mdash mdashZingiber officinale Rosc 14 11 7 15 mdash 7 12 7 mdash mdashZanthoxylum limonella (Dennst) Alston 13 7 12 9 mdash mdash 11 8 mdash mdashChloramphenicol (30120583gdisc) 21 12 15 17 29 16 15 18 13 11Ceftazidime (30 120583gdisc) 33 16 22 19 16 25 21 20 12 15lowastBacteria (+minus) Results obtained in the disc diffusion assay antibacterial activity is expressed as the mean plusmn SD (119899 = 3) of the inhibition by the extract andits diameter around the discs One way analysis of variance was performed (mean plusmn SD 119899 = 3 replicates) Size of inhibition zones were including the sterileblank discs 6 millimeter (mm) in diameters Absence of bacterial inhibition indicates (mdash) antibiotic disc (30 120583gdisc)

Interestingly sixteen plants such as Andrographis echioidesC auriculata C viscose C gigantea T arjuna Oldenlandiaumbellata Boerhavia erecta and E hirta exerted a strongactivity against the Gram-positive S aureus bacteria

37 Phytochemical Screening of Plants The results obtainedfrom the phytochemical screening as shown in Table 1 indi-cate the presence of various types of secondary metabolitessuch as polyphenols tannins saponins alkaloids and gly-cosidespolysaccharides Most of the plant extracts relativelyrich in alkaloids phenols flavonoids polyphenols tanninssterols and terpenoids were found to inhibit the growth oforganisms

38 Antimicrobial Activity of Fractioned Compounds Activecomponents were purified from the most active extractsfor further testing The compound shellsol of T involucrata

and C acida exhibited the most potent action againstthe antibiotic resistant strains of B pseudomallei (KHW)S aureus B pseudomallei (TES) and K pneumoniae Amarmelos was also found to inhibit the growth of B pseudo-mallei (KHW) more effectively than other tested bacteria Avasica showed the broad spectrum growth inhibitory activityon B pseudomallei (KHW) K pneumoniae K pneumoniaeresistant B pseudomallei (TES) S pyogenes and V damselaHowever E cardamomum displayed antimicrobial activityon some of the B pseudomallei (KHW) S pyogenes Bpseudomallei (TES) and S typhi strains Similarly A indicaexerted the growth inhibition on K pneumoniae and Saureus Remarkably Sebastiania chamaelea was more activeagainst K pneumoniae K pneumoniae and S pyogenesThe compound from S indicus inhibited the growth of Kpneumoniae K pneumoniae B pseudomallei (KHW andTES) and S typhi strains as compared to the activity shown

10 Evidence-Based Complementary and Alternative Medicine

Figure 2 Diverse biodiversity richness of medicinal plants in Western and Eastern Ghats (a) Topography of plant covering area in Kollihills (Namakkal district Tamil Nadu) (b) Aerial view of occurrence of medicinal plants in Mundanthurai hills (Tirunelveli district TN) (c)Pachamalai hills (Trichy district) and its natural vegetation inhabitants for Malaiyali tribes (d) Deforestation of natural herbal resources dueto urbanization in Kalrayan hills (Salem district) in the Eastern Ghats of TN (e) Medicinal plants and its various parts used by the natives(traditional healers) for the treatment of diverse human illness with a very high percentage of leaves and whole plants often used for herbaldrug preparation by the local practitioners (f) Various category of plants like shrub herb climbers and tree and the parts used in medicine

Evidence-Based Complementary and Alternative Medicine 11

Figure 3 Medicinal aromatic herbs spices and toxic plants were collected from the tribal areas of the Western and Eastern Ghats region inTamil Nadu India (a) A vasica Nees (leaf) (b) Eclipta alba (L) Hassk (whole plant) (c)Mimosa pudica L (whole plant) (d) P amboinicus(L) Spreng (whole plant) (e) T procumbens (L) (whole plant) (f) Euphorbia hirta Linn (whole plant) (g) A paniculata Wallich ex Nees(H) C roseus (L) GDon (whole plant) used for therapy (i) Cinnamomum iners Reinw ex Blume (leaf) (j) E globulus Labill (leaf and bark)(k) Z officinale Rosc (Rhizome) (l) E caryophyllus (Sprengel) Bullock and Harrison (flower buds) (m) M piperita L (whole plant) (n) Ccitratus (DC) Clitoria ternatea L (whole plant) (o) C zeylanicum Garcin ex Blume (bark) (p) Elettaria cardamomumWhite et Mason (fruitpod) used for medicine and food preparation

by the crude extracts (Figure 4)The antimicrobial efficacy offractions collected from the oil yielding plants was also com-pared with that of the tested compounds C zeylanicum andR officinaliswere themost sensitive in controlling the growthof B pseudomallei (KHW) S aureus K pneumonia and Spneumoniae Fascinatingly all the compounds obtained fromaromatic plants except those from E globules were foundto be very effective against the multidrug resistant humanpathogen B pseudomallei (KHW) that causes melioidosis

On the other hand compounds from C citrates O sanctumE caryophyllus and Z zizanioide showed some promisingeffect only against S aureus (Figure 5) The activity of thecompounds were pronounced more than that of the oilyielding plant fractions

39 Minimum Inhibitory Concentrations (MICs) The antibi-otic potential of the purified fractions was obtained fromthe MIC determination The hydrocarbon ester shellsol

12 Evidence-Based Complementary and Alternative Medicine

(mean plusmn SD n = 3)

0

5

10

15

20

25

30

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn V

d St

Bacteria (+minus)

lowast

lowast

lowast

A vasica

In vitro assay(P

lant

frac

tion)

Inhi

bitio

n zo

nes (

mm

)

(a)

(mean plusmn SD n = 3)

0

5

10

15

20

25

30

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn V

d St

Bacteria (+minus)

lowast

A marmelos

Inhi

bitio

n zo

nes (

mm

)

In vitro assay

(b)

(mean plusmn SD n = 3)

0

5

10

15

20

25

30

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn V

d St

Bacteria (+minus)

lowastlowast

A indica

Inhi

bitio

n zo

nes (

mm

)

Incu

batio

n (37∘C

24 h

)

(c)

(mean plusmn SD n = 3)

0

5

10

15

20

25

30

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn V

d St

Bacteria (+minus)

lowast

lowast lowast

35

C acida

Inhi

bitio

n zo

nes (

mm

)(P

lant

frac

tion)

(d)

(mean plusmn SD n = 3)

0

5

10

15

20

25

30

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn V

d St

Bacteria (+minus)

lowastlowast

lowast

S chamaelea

Inhi

bitio

n zo

nes (

mm

)

(e)

(mean plusmn SD n = 3)

0

5

10

15

20

25

30

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn V

d St

Bacteria (+minus)

lowastlowast

lowastlowast

S indicus

Inhi

bitio

n zo

nes (

mm

)

Incu

batio

n (37∘C

24 h

)

(f)

(mean plusmn SD n = 3)

0

5

10

15

20

25

30

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn V

d St

Bacteria (+minus)

lowast

lowast lowast

E cardamomum

Inhi

bitio

n zo

nes (

mm

)(P

lant

frac

tion)

(g)

(mean plusmn SD n = 3)

0

5

10

15

20

25

30

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn V

d St

Bacteria (+minus)

lowast

lowast

lowast

lowast

Shellsol

35

Inhi

bitio

n zo

nes (

mm

)

Incu

batio

n (37∘C

24 h

)

(h)

Figure 4 In vitro antimicrobial activity of purified fractions from the most active plant extracts tested against bacteria Growth inhibitionzones were measured and analyzed with mean plusmn standard deviation (SD) (119899 = 3) using one way analysis of variance Level of significanceat (lowast119875 gt 001) Most of the fractions exerted a potent inhibitory effect against multidrug resistant Gram-negative bacteria (B pseudomalleistrains KHW and TES) K pneumonia and Gram-positive bacteria S aureus

(T involucrata) andC acida showed an interesting inhibitorypotential against S aureus (MIC of 78 120583gmL) and B pseu-domallei strain of KHW (MIC of 156 120583gmL) A vasicashowed an MIC of 156120583gmL against B pseudomallei(KHW) and an MIC of 3125120583gmL against K pneumo-niae K pneumoniae S pyogenes and V damsela strainsFractions from A marmelos and terpenoid from A indicaexerted bacteriostatic effect with MIC values of 3125120583gmLon some selected bacteria including B pseudomallei ofKHW S aureus and B pseudomallei of TES The MIC of3125 120583gmL was found for E cardamomum against S aureus

K pneumonia and S pyogenes S indicus displayed a verystrong inhibition against MDR K pneumoniae (MIC of156 120583gmL) and against B pseudomallei (KHW and TES)at MIC of 3125 120583gmL When the antimicrobial efficacies ofpurified fractions from aromatic plants were compared theC zeylanicum fraction displayed an important antimicrobialeffect against S aureus (MIC of 78 120583gmL) MDR B pseu-domallei of KHW (MIC of 156 120583gmL) and S pneumoniae(MIC of 3125 120583gmL) The essential oil from M piperateashowed MIC value of 3125120583gmL against K pneumoniae Saureus and B pseudomallei (KHW) respectivelyO sanctum

Evidence-Based Complementary and Alternative Medicine 13

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn

Bacteria (+minus)

0

5

10

15

20

25

30

Inhi

bitio

n zo

nes (

mm

)(P

lant

frac

tion)

lowastlowast

lowast

(mean plusmn SD n = 3)In vitro assay

C citratus

(a)

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn

Bacteria (+minus)

0

5

10

15

20

25

30

Inhi

bitio

n zo

nes (

mm

)

lowast

lowast

lowastlowast

lowast

lowast

(mean plusmn SD n = 3)In vitro assay

35

C zeylanicum

(b)

Bacteria (+minus)

0

5

10

15

20

25

30

Inhi

bitio

n zo

nes (

mm

)

lowast

lowast

lowast

(mean plusmn SD n = 3)

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn V

d St

O sanctum

Incu

batio

n (37∘C

24 h

)

(c)

Bacteria (+minus)

0

5

10

15

20

25

Inhi

bitio

n zo

nes (

mm

)

lowast

lowast

(mean plusmn SD n = 3)

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn V

d St

M piperita

(Pla

nt fr

actio

n)

(d)

Bacteria (+minus)

0

5

10

15

20

25

30

Inhi

bitio

n zo

nes (

mm

)lowast

lowast

lowast

(mean plusmn SD n = 3)

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn V

d St

E caryophyllus

(e)

0

5

10

15

20

25

30

Inhi

bitio

n zo

nes (

mm

) (mean plusmn SD n = 3)

Bacteria (+minus)

KHW

TES

Kp

KprPa Sa

Stp

Spn

E globulus

Incu

batio

n (37∘C

24 h

)

(f)

Bacteria (+minus)

0

5

10

15

20

25

30

Inhi

bitio

n zo

nes (

mm

)(P

lant

frac

tion)

lowast

lowast

lowast

lowast

(mean plusmn SD n = 3)

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn V

d

R officinalis

(g)

0

5

10

15

20

25

30

Inhi

bitio

n zo

nes (

mm

)

lowast

lowast

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn V

d St

Bacteria (+minus)V zizanioides

(mean plusmn SD n = 3)

Incu

batio

n (37∘C

24 h

)

(h)

Figure 5 Comparison of antimicrobial effect of plant compounds obtained from the most popularly used sources of essential oils as assayedby the disc-diffusion method in vitro It displayed a powerful activity against B pseudomallei and S aureus than the other bacteria strainsOther compounds showed only a moderate or weak action against the tested bacteria Values for zone of bacterial growth inhibition werepresented as mean plusmn SD (119899 = 3) with level of significance at (lowast119875 gt 001)

and C citratus fractions also showed antimicrobial activity(MICs of 3125ndash125120583gmL) only at higher concentrationsagainst the tested bacteria In addition to that higher concen-trations (gt250120583gmL) (of Vetiveria fractions) were requiredto inhibit Vibro species and others (including E globulusfractions) failed to show any effect at tested concentrations(78ndash125 120583gmL) However the purified fractions (from mostactive medicinal plants) showed strong bacteriostatic inhibi-tion against the tested organisms (Table 3)

310 Cytotoxic Effects of Plants When the components wereassayed for cytotoxicity against the normal human skinfibroblasts (HEPK) cells the compounds obtained from

E cardamomum T involucrata S indicusC acidaA vasicaA marmelos A indica and A paniculata did not showtoxicity up to 1000120583gmL (see Figures S1 and S2 in Supple-mentaryMaterial available online at httpdxdoiorg1011552013525613) A slight reduction of cell proliferation wasnoted only at higher doses (2000120583gmL) In contrast cellproliferation was markedly reduced after exposure of HEPKcells to O sanctum E globulus V zizanioides C citratusand E globulus compounds There was no gradual reductionin skin cell proliferation seen after exposure to C zeylan-icum R officinalis and M piperita (see Figures S3 and S4)compounds The toxicity was found to be concentration-dependent when the skin fibroblasts (HEPK) cells were

14 Evidence-Based Complementary and Alternative Medicine

Table 3 Minimum inhibitory concentrations (MICs) of purified plant fractions and essential oils against antibiotic resistant bacteria

Botanical name Family Parts used Gram-positive and -negative bacteria (MICs 120583gmL)KHW TES Kp KPr Sa Stp Sp Vd St

A indica Juss Meliaceae Seed (fraction) 3125 625 125 lt250 3125 mdash mdash mdash mdashA marmelos (L) Rutaceae Root-bark (F) 3125 3125 mdash mdash 3125 625 125 250 mdashA paniculata Nees Acanthaceae Leaf (fraction) 250 mdash 125 mdash 625 mdash mdash 125 mdashA vasica Nees Acanthaceae Fraction (Stem) 156 3125 3125 625 mdash lt250 3125 3125 mdashC acida Roxb Rutaceae Leaf (fraction) 156 625 mdash mdash 78 mdash 3125 mdash mdashE cardamomumWhite et Mason Acanthaceae Fraction (WP) 625 mdash 3125 mdash 3125 250 625 mdash mdashS indicus (L) Euphorbiaceae Whole plant (F) 3125 125 156 mdash 625 mdash mdash lt250 mdashT involucrata (L) Euphorbiaceae Shellosol (leaf) 156 3125 mdash mdash 78 625 mdash mdash mdashCinnamomum zeylanicum (L) Lauraceae Bark (fraction) 156 625 mdash mdash 78 3125 mdash mdash 125Cymbopogon citratus (L) Graminae Leaf (fraction) 250 lt250 625 mdash 625 125 mdash mdash mdashEugenia caryophyllus (L) Myrtaceae Flower buds (F) 625 125 mdash mdash 625 mdash mdash mdash mdashEucalyptus globulus (L) Myrtaceae Fraction (leaf) mdash mdash mdash mdash mdash mdash lt250 mdash lt250Mentha piperita (L) Labiatae Fraction (WP) mdash 625 3125 mdash 3125 mdash lt250 mdash mdashOcimum sanctum (L) Labiatae Leaf (fraction) 625 125 mdash 3125 625 mdash 125 lt250 mdashRosmarinus officinalis (L) Labiatae Rosemary oil 3125 mdash mdash mdash 3125 mdash mdash mdash mdashVetiveria zizanioides (L) Graminae Root (fraction) mdash 125 125 mdash mdash mdash gt250 gt250The bacterial growth inhibitory activity was compared for significant differences within the bacterial strains by broth-dilution method at 250 125 625 3125156 and 78 120583gmL F fractions

exposed to various compounds The cell proliferation wasincreased by the influence of the plant components at thelower concentrations Whereas the oil yielding plant com-pounds showed inhibition of cell proliferation and toxicity athigher doses 250ndash1000120583gmL

4 Discussion

The Western Ghats is considered as one of the richestbiodiversity hotspots in the world [41] In this survey wecollected nearly 78 medicinal plants from Western andEastern Ghats that are edible and popularly used for curingvarious ailments including snakebite Traditional remedieshave a long-standing history in many tribal settlements inTN India and they continue to provide useful and applicabletools for treating ailments [42] The ingredients that makeup the ldquoVishakallurdquo stone which is used as an antidote forsnakebite are different herbs and pebbles available from theriver banks Likewise aqueous paste and decoction obtainedfrom the leaves of A paniculata are widely used for snakebitetreatment by indigenous people [43] Previous studies havereported that ethnomedicine plays major roles in conservingthe disappearing knowledge of tribal communities [44ndash47]The traditional beliefs of reliance on a rich diversity ofethnomedicinal plants located at different settlements havealso been confirmed in another study [48 49] Herein weexplored the various types of traditional practices reported bythe primitive tribal communities with a view to gain furtherknowledge from such studies

In the present investigation potentially rich sources oftribal medicine (71 plants) were scientifically evaluated fortheir antibacterial activity against the MDR bacteria and theaccumulated data was disseminated for the first time to the

scientific community Out of the 71medicinal plants screenedfor the antibacterial activity 10 of them (T involucrataC acida A marmelos A vasica C procera A paniculataand M piperita A indica S indicus and E cardamomum)displayed the highest antibacterial activity against the mul-tidrug resistant B pseudomallei (KHW and TES) and Saureus strains The antibacterial activity of those crude plantextracts was as equally effective as that of the standard drugsOur findings corroborated with the previous reports madeon the antistaphylococcal activity of tribal medicinal plants[34 50 51] On the other hand isolated components from themost active extracts of T involucrata shellsol and C acidaexhibited the most potent action against the antibiotic resis-tant B pseudomallei (KHW) K pneumoniae and S aureusstrainsThese results further confirmed our previous findingson the leaves of T involucrata and its compounds hydrocar-bon ester-like shellsol which displayed a high antibacterialeffect against the different bacterial strains especially thatof S aureus [34] Eugenol and caryophyllene are the activeagents contained in the M piperita [52] and O sanctumplants which are believed to be mainly responsible for theantimicrobial properties of these plants [53] Interestinglythe compounds obtained from the aromatic plants such asC zeylanicum and R officinalis were also found to be veryeffective against the multidrug resistant human pathogen Kpneumoniae S aureus S typhi and B pseudomallei (KHW)that causes melioidosis

The inhibitory potential determined for the shellsol(T involucrata) vaseline (A vasica) C acida and C zey-lanicum indicates that the MIC of 78ndash3125120583gmL foundagainst the B pseudomallei of KHW K pneumoniae Kpneumoniae S pyogenes and V damsel and S pneumoniaewas quite low Similarly lower MIC values were found for

Evidence-Based Complementary and Alternative Medicine 15

M piperita (MIC of 113ndash225 mgmL) against the abovebacterial strains [52] MIC was found for the most activealcohol extracts of A salvifolium (MIC 0034ndash0263mgmL)on S aureus [54] and the MIC for S trilobatum aqueousextracts determined against the tested organisms rangedfrom 006 to 05mgmL [55] C zeylanicum was foundto have an effective antibacterial activity (MIC 64 120583gmL)against P aeruginosa E coli B subtilis and S aureus[56] Previously several investigators have demonstrated thatactive agents exert interesting activity against bacteria evenat lower concentrations tested [57 58] The bacteriostaticmechanism involves damage to the cell walls of bacteriafollowed by inhibition of protein synthesis that ultimatelyleads to bacterial death [59] The most active plants arewidely used by various tribes as traditional treatment (iecut wounds skin infection and scabies) thus indicatingthe potential for further development into promising drugsIn order to ascertain the safety and efficacy of the mostactive compounds their effect on human skin fibroblast cellswas evaluated Chemical constituents of E cardamomumT involucrata S indicus C acida A vasica A marmelosA indica and A paniculata plants failed to produce anynoticeable toxicity up to 1000120583gmL Although some ofthe tested compounds exhibited a slight reduction of cellproliferation and some minor morphological changes suchchanges were insignificant at lower doses and became evidentonly at higher doses However certain aromatic compoundsof O sanctum E globulus V zizanioides C citratus and Eglobulus plants showed reduction of cell proliferation againstHEPK cells

Our phytochemical screening also provides evidenceof the presence of several types of compounds that aremainly responsible for the remarkable antibacterial effectof these plants The differences noted for the bactericidalactivity of various plant extracts in this study appears tobe directly related to the diversity of compounds (shown inparentheses) that are accumulated in the following plants(eg A marmelos and O umbellate) [60ndash62] Compoundslike tannins phenol and polyphenols can bind the Gram-negative bacteria to form a heavy soluble complex on the cellsurface which subsequently disturbs the availability of recep-tor on cells and kills the bacteria [63] Several species havingwide spectra of antimicrobial activity mainly due to theactive constituents such as essential oil phenolic compoundslike thymol carvacrol in oregano and thyme eugenol inclove and cinnamon were also identified previously [64 65]Essential oils degrade the cell wall interact with the cell com-ponents and then disrupt the cytoplasmic membrane [66]The antimicrobial effect of phenolic compounds may involvemultiplemodes of action including damage to themembraneprotein interference with membrane integrated enzymes[67] causing leakage of cellular components coagulation ofcytoplasm depletion of the proton motive force alterationof fatty acid and phospholipid constituents impairmentof enzymatic mechanisms for production and metabolismalteration of nutrient uptake and electron transport [68]influencing the synthesis of DNA and RNA and destroyingprotein translation and the function of the mitochondrionin eukaryotes [69] The mode of action of antimicrobial

agents depends on the type of microorganism and is mainlyrelated to their cell wall structure and the outer membranearrangement Most of the plant spices and herbs containcomplex phenolics (ie phenolic acids flavonoids tanninslignans coumarins quinines) In addition the mechanismsof action of each phenolic compound against various bacteriaare also very complicated [26 70] Further investigation istherefore required to understand the relationship betweenthe antimicrobial action and the chemical structure of everyphenolic compound in the tested extracts The informationavailable from previous pharmacological sources combinedwith the findings herein reported on the medicinal plantextracts may serve as essential data for future drug develop-ment to combat diseases caused by theMDRbacterial strains

Abbreviations

MDR Multidrug resistant bacteriaTES Strain of Burkholderia pseudomalleiMRSA Methicillin-resistant Staphylococcus aureusVRE Vancomycin-resistant enterococciMTB Mycobacterium tuberculosisCH3OH Methanol

CFU Colony forming unitHCl Hydrochloric acidTLC Thin layer chromatographyH2SO4 Sulphuric acids

UV Ultraviolet spectrophotometerHEPK Human skin fibroblasts

Acknowledgments

Theauthors are thankful to the ERI Loyola College ChennaiIndia for his support during the survey The authors alsothank the Department of Microbiology NUHS NUS forthe bacterial cultures used in this investigation The authorsare thankful to the tribal people for sharing their healingknowledge on medicinal aromatic plants

References

[1] M N Alekshun and S B Levy ldquoMolecular mechanisms ofantibacterialmultidrug resistancerdquoCell vol 128 no 6 pp 1037ndash1050 2007

[2] A D Russell ldquoAntibiotic and biocide resistance in bacteriaintroductionrdquo Symposium Series Society for Applied Microbiol-ogy vol 31 no 92 pp 1Sndash3S 2002

[3] D R Guay ldquoTreatment of bacterial skin and skin structureinfectionsrdquo Expert Opinion on Pharmacotherapy vol 4 no 8pp 1259ndash1275 2003

[4] A J Brosnahan and P M Schlievert ldquoGram-positive bac-terial superantigen outside-in signaling causes toxic shocksyndromerdquo FEBS Journal vol 278 no 23 pp 4649ndash4667 2011

[5] K Fellermann J Wehkamp K R Herrlinger and E F StangeldquoCrohnrsquos disease a defensin deficiency syndromerdquo EuropeanJournal of Gastroenterology and Hepatology vol 15 no 6 pp627ndash634 2003

16 Evidence-Based Complementary and Alternative Medicine

[6] L Chen S Lin R Agha-Majzoub L Overbergh C Mathieuand L S Chan ldquoCCL27 is a critical factor for the developmentof atopic dermatitis in the keratin-14 IL-4 transgenic mousemodelrdquo International Immunology vol 18 no 8 pp 1233ndash12422006

[7] J H Song V Thamlikitkul and P R Hsueh ldquoClinical andeconomic burden of community-acquired pneumonia amongstadults in the Asia-Pacific regionrdquo International Journal ofAntimicrobial Agents vol 38 no 2 pp 108ndash117 2011

[8] D A B Dance ldquoMelioidosisrdquo Current Opinion in InfectiousDiseases vol 15 no 2 pp 127ndash132 2002

[9] N J White ldquoMelioidosisrdquo The Lancet vol 361 no 9370 pp1715ndash1722 2003

[10] S Yang ldquoMelioidosis research in Chinardquo Acta Tropica vol 77no 2 pp 157ndash165 2000

[11] P R Hsueh L J Teng L N Lee et al ldquoMelioidosis an emerginginfection in Taiwanrdquo Emerging Infectious Diseases vol 7 no 3pp 428ndash433 2001

[12] C Walsh and S Fanning ldquoAntimicrobial resistance in food-borne pathogensmdasha cause for concernrdquo Current Drug Targetsvol 9 no 9 pp 808ndash815 2008

[13] M Vorachit P Chongtrakool S Arkomsean and S BoonsongldquoAntimicrobial resistance in Burkholderia pseudomalleirdquo ActaTropica vol 74 no 2-3 pp 139ndash144 2000

[14] F M Thibault E Hernandez D R Vidal M Girardet and JD Cavallo ldquoAntibiotic susceptibility of 65 isolates of Burkholde-ria pseudomallei and Burkholderia mallei to 35 antimicrobialagentsrdquo Journal of Antimicrobial Chemotherapy vol 54 no 6pp 1134ndash1138 2004

[15] R L S Sikarwar ldquoEthnogynaecological uses of plants new toIndiardquo Ethnobotany vol 12 no 1-2 pp 112ndash115 2002

[16] R Siva ldquoStatus of natural dyes and dye-yielding plants in IndiardquoCurrent Science vol 92 no 7 pp 916ndash925 2007

[17] P Kumar Singh V Kumar R K Tiwari A Sharma ChV Rao and R H Singh ldquoMedico-ethnobotany of lsquochatararsquoblock of district sonebhadra Uttar Pradesh Indiardquo Advances inBiological Research vol 4 no 1 pp 65ndash80 2010

[18] W H Lewis and M P Elwin-Lewis Medical Botany PlantsAffecting Human Health John Wiley amp Sons New York NYUSA 2003

[19] R Perumal Samy P N Pushparaj and P GopalakrishnakoneldquoA compilation of bioactive compounds fromAyurvedardquo Bioin-formation vol 3 no 3 pp 100ndash110 2008

[20] WorldHealthOrganization (WHO)TraditionalMedicine Strat-egy 2002ndash2005 Genera World Health Organization GenevaSwitzerland 2002

[21] P S Haddad G A Azar S Groom and M Boivin ldquoNaturalhealth products modulation of immune function and preven-tion of chronic diseasesrdquo Evidence Based Complementary andAlternative Medicine vol 2 no 4 pp 513ndash520 2005

[22] K Poonam andG S Singh ldquoEthnobotanical study of medicinalplants used by the Taungya community in Terai Arc LandscapeIndiardquo Journal of Ethnopharmacology vol 123 no 1 pp 167ndash1762009

[23] G B Mahady ldquoMedicinal plants for the prevention and treat-ment of bacterial infectionsrdquo Current Pharmaceutical Designvol 11 no 19 pp 2405ndash2427 2005

[24] V P Kumar N S Chauhan H Padh and M Rajani ldquoSearchfor antibacterial and antifungal agents from selected Indianmedicinal plantsrdquo Journal of Ethnopharmacology vol 107 no2 pp 182ndash188 2006

[25] R Nair T Kalariya and S Chanda ldquoAntibacterial activity ofsome plant extracts used in folk medicinerdquo Journal of HerbalPharmacotherapy vol 7 no 3-4 pp 191ndash201 2007

[26] D Kalemba and A Kunicka ldquoAntibacterial and antifungalproperties of essential oilsrdquo Current Medicinal Chemistry vol10 no 10 pp 813ndash829 2003

[27] A H Ashour ldquoAntibacterial antifungal and anticancer activi-ties of volatile oils and extracts from stems leaves and flowers ofEucalyptus sideroxylon and Eucalyptus torquatardquoCancer BiologyandTherapy vol 7 no 3 pp 399ndash403 2008

[28] M J Abad L M Bedoya L Apaza and P Bermejo ldquoTheArtemisia L genus a review of bioactive essential oilsrdquoMolecules vol 17 no 3 pp 2542ndash2566 2012

[29] H N Marzoug M Romdhane A Lebrihi et al ldquoEucalyptusoleosa essential oils chemical composition and antimicrobialand antioxidant activities of the oils from different plant parts(stems leaves flowers and fruits)rdquoMolecules vol 16 no 2 pp1695ndash1709 2011

[30] C Proestos D Sereli and M Komaitis ldquoDetermination ofphenolic compounds in aromatic plants by RP-HPLC and GC-MSrdquo Food Chemistry vol 95 no 1 pp 44ndash52 2006

[31] S Chao G Young C Oberg and K Nakaoka ldquoInhibition ofmethicillin-resistant Staphylococcus aureus (MRSA) by essentialoilsrdquo Flavour and Fragrance Journal vol 23 no 6 pp 444ndash4492008

[32] M Akin D Oguz and H T Saracoglu ldquoAntibacterial effectsof some plant extracts from Labiatae (Lamiaceae) growingnaturally around Sirnak-Silopi Turkeyrdquo International Journal ofPharmaceutical and Applied Sciences vol 1 no 1 pp 4ndash47 2010

[33] K Bayoub T Baibai D Mountassif A Retmane and ASoukri ldquoAntibacterial activities of the crude ethanol extracts ofmedicinal plants against Listeriamonocytogenes and some otherpathogenic strainsrdquo African Journal of Biotechnology vol 9 no27 pp 4251ndash4258 2010

[34] R P Samy P Gopalakrishnakone M Sarumathi P Houghtonand S Ignacimuthu ldquoPurification of antibacterial agents fromTragia involucratamdasha popular tribal medicine for wound heal-ingrdquo Journal of Ethnopharmacology vol 107 no 1 pp 99ndash1062006

[35] K M Matthew The Flora of Tamil Nadu Carnatic Part IndashIIIprinted at the Diocesan Press Madras India 1981ndash1983

[36] AW BauerWMKirby J C Sherris andM Turck ldquoAntibioticsusceptibility testing by a standardized single disk methodrdquoAmerican Journal of Clinical Pathology vol 45 no 4 pp 493ndash496 1966

[37] National Committee for Clinical Laboratory Standards Per-formance Standards For Antimicrobial Susceptibility Testing12th Informational Supplement Approved standard M100-S12National Committee for Clinical Laboratory Standards WaynePa USA 2002

[38] E A Elsinghorst ldquoMeasurement of invasion by gentamicinresistancerdquoMethods in Enzymology vol 236 pp 405ndash420 1994

[39] J B Harborne Phytochemical Methods Chapman and HallNew York NY USA 1976

[40] H Wagner S Bladt and E M Zgairski Plant Drug AnalysisSpringer New York NY USA 1984

[41] N Myers R A Mittermeler C G Mittermeler G A B daFonseca and J Kent ldquoBiodiversity hotspots for conservationprioritiesrdquo Nature vol 403 no 6772 pp 853ndash858 2000

[42] R Perumal Samy and S Ignacimuthu ldquoAntibacterial activity ofsome folkloremedicinal plants used by tribals inWestern Ghats

Evidence-Based Complementary and Alternative Medicine 17

of Indiardquo Journal of Ethnopharmacology vol 69 no 1 pp 63ndash712000

[43] K Nampoothiri A Chrispal A Begum S Jasmine K GGopinath and A Zachariah ldquoA clinical study of renal tubulardysfunction inCleistanthus collinus (Oduvanthalai) poisoningrdquoClinical Toxicology vol 48 no 3 pp 193ndash197 2010

[44] M Strathern UsefulKnowledge The Isiah Berlin Lecture Uni-versity of Manchester Brunswick UK

[45] C N Sunil and A K Pradeep ldquo(2001) Another new species ofTripogon (Poaceae) from Indiardquo Sida vol 19 pp 803ndash806 2005

[46] S Biber-Klemm and T Cottier Right To Plant GeneticResources and Traditional Knowledge Basic Issues and Prospec-tive Oxfordshire Nosworthy Way UK 2006

[47] V S Ramachandran S Shijo Joseph and R Aruna ldquoEth-nobotanical studies from amaravathy range of Indira GandhiWildlife SanctuaryWesternGhats CoimbatoreDistrict South-ern Indiardquo Ethnobotanical Leaflets vol 13 pp 1069ndash1087 2009

[48] A H Rajasab and I Mohamad ldquoDocumentation of folkknowledge on edible wild plants of North Karnatakardquo IndianJournal of Traditional Knowledge vol 3 no 4 pp 419ndash4292004

[49] M Rajadurai V G Vidhya M Ramya and A Bhaskar ldquoEthno-medicinal plants used by the traditional healers of pachamalaihills Tamilnadu Indiardquo Studies on Ethno-Medicine vol 3 no 1pp 39ndash41 2009

[50] S K Adesina O Idowu A O Ogundaini et al ldquoAntimicrobialconstituents of the leaves of Acalypha wilkesiana and Acalyphahispidardquo Phytotherapy Research vol 14 no 5 pp 371ndash374 2000

[51] M T Gutierrez-Lugo M P Singh W M Maiese and BN Timmermann ldquoNew antimicrobial cycloartane triterpenesfrom Acalypha communisrdquo Journal of Natural Products vol 65no 6 pp 872ndash875 2002

[52] A K Tyagi andAMalik ldquoAntimicrobial potential and chemicalcomposition of Mentha piperita oil in liquid and vapour phaseagainst food spoiling microorganismsrdquo Food Control vol 22no 11 pp 1707ndash1714 2011

[53] G Devendran and U Balasubramanian ldquoQualitative phyto-chemical screening and GC-MS analysis of Ocimum sanctumL leavesrdquoAsian Journal of Plant Science and Research vol 1 no4 pp 44ndash48 2011

[54] V C Jain N M Patel D P Shah P K Patel and B HJoshi ldquoAntioxidant and antimicrobial activities of Alangiumsalvifolium (Lf) Wang rootrdquo Global Journal of Pharmacologyvol 4 no 1 pp 13ndash18 2010

[55] P Swapna Latha andK Kannabiran ldquoAntimicrobial activity andphytochemicals of Solanum trilobatum LinnrdquoAfrican Journal ofBiotechnology vol 5 no 23 pp 2402ndash2404 2006

[56] M Usha1 S Ragini and S M A Naqvi ldquoAntibacterial activityof acetone and ethanol extracts of Cinnamon (Cinnamomumzeylanicum) and Ajowan (Trachyspermum ammi) on four foodspoilage bacteriardquo International Research Journal of BiologicalSciences vol 1 no 4 pp 7ndash11 2012

[57] S Prabuseenivasan M Jayakumar and S Ignacimuthu ldquoInvitro antibacterial activity of some plant essential oilsrdquo BMCComplementary and Alternative Medicine vol 6 article 39 pp1ndash8 2006

[58] G A Ayoola F M Lawore T Adelowotan et al ldquoChemi-cal analysis and antimicrobial activity of the essential oil ofSyzigium aromaticum (clove)rdquo African Journal of Biotechnologyvol 7 no 13 pp 2227ndash2231 2008

[59] Y J Fu L Chen Y Zu et al ldquoThe antibacterial activity of cloveessential oil against Propionibacterium acnes and its mechanismof actionrdquo Archives of Dermatology vol 145 no 1 pp 86ndash882009

[60] D Venkatesan C M Karrunakarn S Selva Kumar and P TPalani Swamy ldquoIdentification of phytochemical constituents ofAegle marmelos responsible for antimicrobial activity againstselected pathogenic organismsrdquo Ethnobotanical Leaflets vol 13pp 1362ndash1372 2009

[61] P Arun K G Purushotham J Johnsy jayarani and V KumarildquoIn vitro Antibacterial Activity of Oldenlandia umbellata anIndian medicinal Plantrdquo Journal of Pharmaceutical Science andTechnology vol 2 no 4 pp 198ndash201 2010

[62] A Manjamalai R Sardar Sathyajith Singh C Guruvayoorap-pan and V M Berlin Grace ldquoAnalysis of phytochemicalconstituents and antimicrobial activity of some medicinalplants in Taminadu Indiardquo Global Journal of Biotechnology andBiochemistry vol 5 no 2 pp 120ndash128 2010

[63] E Haslam ldquoNatural polyphenols (vegetable tannins) as drugsPossible modes of actionrdquo Journal of Natural Products vol 59no 2 pp 205ndash215 1996

[64] Y Cai Q Luo M Sun and H Corke ldquoAntioxidant activityand phenolic compounds of 112 traditional Chinese medicinalplants associated with anticancerrdquo Life Sciences vol 74 no 17pp 2157ndash2184 2004

[65] B Shan Y Z Cai M Sun and H Corke ldquoAntioxidant capacityof 26 spice extracts and characterization of their phenolicconstituentsrdquo Journal of Agricultural and Food Chemistry vol53 no 20 pp 7749ndash7759 2005

[66] R J W Lambert P N Skandamis P J Coote and G-J ENychas ldquoA study of the minimum inhibitory concentration andmode of action of oregano essential oil thymol and carvacrolrdquoJournal of AppliedMicrobiology vol 91 no 3 pp 453ndash462 2001

[67] M Raccach ldquoThe antimicrobial activity of phenolic antioxi-dants in food a reviewrdquo Journal of Food Safety vol 6 no 3 pp141ndash170 1984

[68] M Taniguchi Y Yano E Tada et al ldquoMode of action of polygo-dial an antifungal sesquiterpene dialdehyderdquo Agricultural andBiological Chemistry vol 52 no 6 pp 1409ndash1414 1988

[69] G J E Nychas ldquoNatural antimicrobials from plantsrdquo in NewMethods of Food Preservation G W Gould Ed pp 58ndash89Blackie Academic London UK 1995

[70] S Burt ldquoEssential oils their antibacterial properties and poten-tial applications in foodsmdasha reviewrdquo International Journal ofFood Microbiology vol 94 no 3 pp 223ndash253 2004

Submit your manuscripts athttpwwwhindawicom

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Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

MEDIATORSINFLAMMATION

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Behavioural Neurology

EndocrinologyInternational Journal of

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Disease Markers

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OncologyJournal of

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Oxidative Medicine and Cellular Longevity

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The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014

Immunology ResearchHindawi Publishing Corporationhttpwwwhindawicom Volume 2014

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Research and TreatmentAIDS

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Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Parkinsonrsquos Disease

Evidence-Based Complementary and Alternative Medicine

Volume 2014Hindawi Publishing Corporationhttpwwwhindawicom

Evidence-Based Complementary and Alternative Medicine 9

Table 2 Continued

Scientific name Microorganisms growth inhibition zones (6 millimeter in diameters)KHW TES Kp Kpr Pa Sa Stp Sp Vd Vd

Merremia hastate L (Desr) Hallierf mdash mdash mdash mdash mdash mdash mdash mdash mdash mdashMorinda tinctoria Roxb mdash mdash mdash mdash mdash mdash mdash mdash mdash mdashMentha piperita L 23 17 26 12 20 25 19 mdash mdash mdashOcimum sanctum L 12 9 11 7 mdash 15 7 8 mdash mdashOldenlandia umbellata L mdash mdash mdash mdash mdash 17 mdash mdash mdash mdashPiper attenuatum Buch Hamex Miq 13 7 12 21 mdash 17 11 8 mdash mdashPlumbago zeylanica (L) Cav 9 10 17 8 mdash 12 9 10 mdash mdashPlectranthus amboinicus (L) Spreng mdash mdash mdash 8 11 15 mdash mdash mdash mdashPhyllanthus debilis L (Klein ex Willd) 7 8 mdash 9 mdash 9 18 7 mdash mdashPhyllanthus maderaspatensis L 17 mdash mdash mdash mdash mdash mdash mdash mdash mdashPremna tomentosaWilld 13 10 mdash 15 mdash 9 10 mdash mdash mdashGloriosa superba L 17 16 7 8 mdash 15 mdash mdash mdash 8Sebastiania chamaelea (L) Muell Arg 19 12 mdash 17 mdash 13 19 8 mdash mdashSolanum trilobatum L mdash mdash 13 9 mdash 8 mdash mdash mdash mdashSphaeranthus indicus L 20 18 21 7 22 mdash 11 9 mdash 16Swertia chirata (L) Ham mdash mdash mdash mdash mdash mdash mdash mdash mdash mdashTerminalia arjuna (W amp A) mdash mdash mdash mdash mdash 16 mdash mdash mdash mdashTinospora cordifolia (Willd) Miers exHooff ampThoms mdash mdash 23 16 mdash 12 15 mdash mdash mdash

Tridax procumbens L 9 8 7 mdash mdash 14 mdash mdash mdash mdashTragia involucrata L 25 23 20 mdash mdash 31 28 22 19 mdashVitex negundo L mdash mdash mdash mdash mdash 14 mdash mdash mdash mdashVetiveria zizanioides (L) 11 9 7 7 mdash 16 mdash mdash 8Withania somnifera (L) Dunal 12 20 mdash mdash mdash 15 12 mdash mdash mdashZingiber officinale Rosc 14 11 7 15 mdash 7 12 7 mdash mdashZanthoxylum limonella (Dennst) Alston 13 7 12 9 mdash mdash 11 8 mdash mdashChloramphenicol (30120583gdisc) 21 12 15 17 29 16 15 18 13 11Ceftazidime (30 120583gdisc) 33 16 22 19 16 25 21 20 12 15lowastBacteria (+minus) Results obtained in the disc diffusion assay antibacterial activity is expressed as the mean plusmn SD (119899 = 3) of the inhibition by the extract andits diameter around the discs One way analysis of variance was performed (mean plusmn SD 119899 = 3 replicates) Size of inhibition zones were including the sterileblank discs 6 millimeter (mm) in diameters Absence of bacterial inhibition indicates (mdash) antibiotic disc (30 120583gdisc)

Interestingly sixteen plants such as Andrographis echioidesC auriculata C viscose C gigantea T arjuna Oldenlandiaumbellata Boerhavia erecta and E hirta exerted a strongactivity against the Gram-positive S aureus bacteria

37 Phytochemical Screening of Plants The results obtainedfrom the phytochemical screening as shown in Table 1 indi-cate the presence of various types of secondary metabolitessuch as polyphenols tannins saponins alkaloids and gly-cosidespolysaccharides Most of the plant extracts relativelyrich in alkaloids phenols flavonoids polyphenols tanninssterols and terpenoids were found to inhibit the growth oforganisms

38 Antimicrobial Activity of Fractioned Compounds Activecomponents were purified from the most active extractsfor further testing The compound shellsol of T involucrata

and C acida exhibited the most potent action againstthe antibiotic resistant strains of B pseudomallei (KHW)S aureus B pseudomallei (TES) and K pneumoniae Amarmelos was also found to inhibit the growth of B pseudo-mallei (KHW) more effectively than other tested bacteria Avasica showed the broad spectrum growth inhibitory activityon B pseudomallei (KHW) K pneumoniae K pneumoniaeresistant B pseudomallei (TES) S pyogenes and V damselaHowever E cardamomum displayed antimicrobial activityon some of the B pseudomallei (KHW) S pyogenes Bpseudomallei (TES) and S typhi strains Similarly A indicaexerted the growth inhibition on K pneumoniae and Saureus Remarkably Sebastiania chamaelea was more activeagainst K pneumoniae K pneumoniae and S pyogenesThe compound from S indicus inhibited the growth of Kpneumoniae K pneumoniae B pseudomallei (KHW andTES) and S typhi strains as compared to the activity shown

10 Evidence-Based Complementary and Alternative Medicine

Figure 2 Diverse biodiversity richness of medicinal plants in Western and Eastern Ghats (a) Topography of plant covering area in Kollihills (Namakkal district Tamil Nadu) (b) Aerial view of occurrence of medicinal plants in Mundanthurai hills (Tirunelveli district TN) (c)Pachamalai hills (Trichy district) and its natural vegetation inhabitants for Malaiyali tribes (d) Deforestation of natural herbal resources dueto urbanization in Kalrayan hills (Salem district) in the Eastern Ghats of TN (e) Medicinal plants and its various parts used by the natives(traditional healers) for the treatment of diverse human illness with a very high percentage of leaves and whole plants often used for herbaldrug preparation by the local practitioners (f) Various category of plants like shrub herb climbers and tree and the parts used in medicine

Evidence-Based Complementary and Alternative Medicine 11

Figure 3 Medicinal aromatic herbs spices and toxic plants were collected from the tribal areas of the Western and Eastern Ghats region inTamil Nadu India (a) A vasica Nees (leaf) (b) Eclipta alba (L) Hassk (whole plant) (c)Mimosa pudica L (whole plant) (d) P amboinicus(L) Spreng (whole plant) (e) T procumbens (L) (whole plant) (f) Euphorbia hirta Linn (whole plant) (g) A paniculata Wallich ex Nees(H) C roseus (L) GDon (whole plant) used for therapy (i) Cinnamomum iners Reinw ex Blume (leaf) (j) E globulus Labill (leaf and bark)(k) Z officinale Rosc (Rhizome) (l) E caryophyllus (Sprengel) Bullock and Harrison (flower buds) (m) M piperita L (whole plant) (n) Ccitratus (DC) Clitoria ternatea L (whole plant) (o) C zeylanicum Garcin ex Blume (bark) (p) Elettaria cardamomumWhite et Mason (fruitpod) used for medicine and food preparation

by the crude extracts (Figure 4)The antimicrobial efficacy offractions collected from the oil yielding plants was also com-pared with that of the tested compounds C zeylanicum andR officinaliswere themost sensitive in controlling the growthof B pseudomallei (KHW) S aureus K pneumonia and Spneumoniae Fascinatingly all the compounds obtained fromaromatic plants except those from E globules were foundto be very effective against the multidrug resistant humanpathogen B pseudomallei (KHW) that causes melioidosis

On the other hand compounds from C citrates O sanctumE caryophyllus and Z zizanioide showed some promisingeffect only against S aureus (Figure 5) The activity of thecompounds were pronounced more than that of the oilyielding plant fractions

39 Minimum Inhibitory Concentrations (MICs) The antibi-otic potential of the purified fractions was obtained fromthe MIC determination The hydrocarbon ester shellsol

12 Evidence-Based Complementary and Alternative Medicine

(mean plusmn SD n = 3)

0

5

10

15

20

25

30

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn V

d St

Bacteria (+minus)

lowast

lowast

lowast

A vasica

In vitro assay(P

lant

frac

tion)

Inhi

bitio

n zo

nes (

mm

)

(a)

(mean plusmn SD n = 3)

0

5

10

15

20

25

30

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn V

d St

Bacteria (+minus)

lowast

A marmelos

Inhi

bitio

n zo

nes (

mm

)

In vitro assay

(b)

(mean plusmn SD n = 3)

0

5

10

15

20

25

30

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn V

d St

Bacteria (+minus)

lowastlowast

A indica

Inhi

bitio

n zo

nes (

mm

)

Incu

batio

n (37∘C

24 h

)

(c)

(mean plusmn SD n = 3)

0

5

10

15

20

25

30

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn V

d St

Bacteria (+minus)

lowast

lowast lowast

35

C acida

Inhi

bitio

n zo

nes (

mm

)(P

lant

frac

tion)

(d)

(mean plusmn SD n = 3)

0

5

10

15

20

25

30

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn V

d St

Bacteria (+minus)

lowastlowast

lowast

S chamaelea

Inhi

bitio

n zo

nes (

mm

)

(e)

(mean plusmn SD n = 3)

0

5

10

15

20

25

30

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn V

d St

Bacteria (+minus)

lowastlowast

lowastlowast

S indicus

Inhi

bitio

n zo

nes (

mm

)

Incu

batio

n (37∘C

24 h

)

(f)

(mean plusmn SD n = 3)

0

5

10

15

20

25

30

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn V

d St

Bacteria (+minus)

lowast

lowast lowast

E cardamomum

Inhi

bitio

n zo

nes (

mm

)(P

lant

frac

tion)

(g)

(mean plusmn SD n = 3)

0

5

10

15

20

25

30

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn V

d St

Bacteria (+minus)

lowast

lowast

lowast

lowast

Shellsol

35

Inhi

bitio

n zo

nes (

mm

)

Incu

batio

n (37∘C

24 h

)

(h)

Figure 4 In vitro antimicrobial activity of purified fractions from the most active plant extracts tested against bacteria Growth inhibitionzones were measured and analyzed with mean plusmn standard deviation (SD) (119899 = 3) using one way analysis of variance Level of significanceat (lowast119875 gt 001) Most of the fractions exerted a potent inhibitory effect against multidrug resistant Gram-negative bacteria (B pseudomalleistrains KHW and TES) K pneumonia and Gram-positive bacteria S aureus

(T involucrata) andC acida showed an interesting inhibitorypotential against S aureus (MIC of 78 120583gmL) and B pseu-domallei strain of KHW (MIC of 156 120583gmL) A vasicashowed an MIC of 156120583gmL against B pseudomallei(KHW) and an MIC of 3125120583gmL against K pneumo-niae K pneumoniae S pyogenes and V damsela strainsFractions from A marmelos and terpenoid from A indicaexerted bacteriostatic effect with MIC values of 3125120583gmLon some selected bacteria including B pseudomallei ofKHW S aureus and B pseudomallei of TES The MIC of3125 120583gmL was found for E cardamomum against S aureus

K pneumonia and S pyogenes S indicus displayed a verystrong inhibition against MDR K pneumoniae (MIC of156 120583gmL) and against B pseudomallei (KHW and TES)at MIC of 3125 120583gmL When the antimicrobial efficacies ofpurified fractions from aromatic plants were compared theC zeylanicum fraction displayed an important antimicrobialeffect against S aureus (MIC of 78 120583gmL) MDR B pseu-domallei of KHW (MIC of 156 120583gmL) and S pneumoniae(MIC of 3125 120583gmL) The essential oil from M piperateashowed MIC value of 3125120583gmL against K pneumoniae Saureus and B pseudomallei (KHW) respectivelyO sanctum

Evidence-Based Complementary and Alternative Medicine 13

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn

Bacteria (+minus)

0

5

10

15

20

25

30

Inhi

bitio

n zo

nes (

mm

)(P

lant

frac

tion)

lowastlowast

lowast

(mean plusmn SD n = 3)In vitro assay

C citratus

(a)

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn

Bacteria (+minus)

0

5

10

15

20

25

30

Inhi

bitio

n zo

nes (

mm

)

lowast

lowast

lowastlowast

lowast

lowast

(mean plusmn SD n = 3)In vitro assay

35

C zeylanicum

(b)

Bacteria (+minus)

0

5

10

15

20

25

30

Inhi

bitio

n zo

nes (

mm

)

lowast

lowast

lowast

(mean plusmn SD n = 3)

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn V

d St

O sanctum

Incu

batio

n (37∘C

24 h

)

(c)

Bacteria (+minus)

0

5

10

15

20

25

Inhi

bitio

n zo

nes (

mm

)

lowast

lowast

(mean plusmn SD n = 3)

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn V

d St

M piperita

(Pla

nt fr

actio

n)

(d)

Bacteria (+minus)

0

5

10

15

20

25

30

Inhi

bitio

n zo

nes (

mm

)lowast

lowast

lowast

(mean plusmn SD n = 3)

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn V

d St

E caryophyllus

(e)

0

5

10

15

20

25

30

Inhi

bitio

n zo

nes (

mm

) (mean plusmn SD n = 3)

Bacteria (+minus)

KHW

TES

Kp

KprPa Sa

Stp

Spn

E globulus

Incu

batio

n (37∘C

24 h

)

(f)

Bacteria (+minus)

0

5

10

15

20

25

30

Inhi

bitio

n zo

nes (

mm

)(P

lant

frac

tion)

lowast

lowast

lowast

lowast

(mean plusmn SD n = 3)

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn V

d

R officinalis

(g)

0

5

10

15

20

25

30

Inhi

bitio

n zo

nes (

mm

)

lowast

lowast

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn V

d St

Bacteria (+minus)V zizanioides

(mean plusmn SD n = 3)

Incu

batio

n (37∘C

24 h

)

(h)

Figure 5 Comparison of antimicrobial effect of plant compounds obtained from the most popularly used sources of essential oils as assayedby the disc-diffusion method in vitro It displayed a powerful activity against B pseudomallei and S aureus than the other bacteria strainsOther compounds showed only a moderate or weak action against the tested bacteria Values for zone of bacterial growth inhibition werepresented as mean plusmn SD (119899 = 3) with level of significance at (lowast119875 gt 001)

and C citratus fractions also showed antimicrobial activity(MICs of 3125ndash125120583gmL) only at higher concentrationsagainst the tested bacteria In addition to that higher concen-trations (gt250120583gmL) (of Vetiveria fractions) were requiredto inhibit Vibro species and others (including E globulusfractions) failed to show any effect at tested concentrations(78ndash125 120583gmL) However the purified fractions (from mostactive medicinal plants) showed strong bacteriostatic inhibi-tion against the tested organisms (Table 3)

310 Cytotoxic Effects of Plants When the components wereassayed for cytotoxicity against the normal human skinfibroblasts (HEPK) cells the compounds obtained from

E cardamomum T involucrata S indicusC acidaA vasicaA marmelos A indica and A paniculata did not showtoxicity up to 1000120583gmL (see Figures S1 and S2 in Supple-mentaryMaterial available online at httpdxdoiorg1011552013525613) A slight reduction of cell proliferation wasnoted only at higher doses (2000120583gmL) In contrast cellproliferation was markedly reduced after exposure of HEPKcells to O sanctum E globulus V zizanioides C citratusand E globulus compounds There was no gradual reductionin skin cell proliferation seen after exposure to C zeylan-icum R officinalis and M piperita (see Figures S3 and S4)compounds The toxicity was found to be concentration-dependent when the skin fibroblasts (HEPK) cells were

14 Evidence-Based Complementary and Alternative Medicine

Table 3 Minimum inhibitory concentrations (MICs) of purified plant fractions and essential oils against antibiotic resistant bacteria

Botanical name Family Parts used Gram-positive and -negative bacteria (MICs 120583gmL)KHW TES Kp KPr Sa Stp Sp Vd St

A indica Juss Meliaceae Seed (fraction) 3125 625 125 lt250 3125 mdash mdash mdash mdashA marmelos (L) Rutaceae Root-bark (F) 3125 3125 mdash mdash 3125 625 125 250 mdashA paniculata Nees Acanthaceae Leaf (fraction) 250 mdash 125 mdash 625 mdash mdash 125 mdashA vasica Nees Acanthaceae Fraction (Stem) 156 3125 3125 625 mdash lt250 3125 3125 mdashC acida Roxb Rutaceae Leaf (fraction) 156 625 mdash mdash 78 mdash 3125 mdash mdashE cardamomumWhite et Mason Acanthaceae Fraction (WP) 625 mdash 3125 mdash 3125 250 625 mdash mdashS indicus (L) Euphorbiaceae Whole plant (F) 3125 125 156 mdash 625 mdash mdash lt250 mdashT involucrata (L) Euphorbiaceae Shellosol (leaf) 156 3125 mdash mdash 78 625 mdash mdash mdashCinnamomum zeylanicum (L) Lauraceae Bark (fraction) 156 625 mdash mdash 78 3125 mdash mdash 125Cymbopogon citratus (L) Graminae Leaf (fraction) 250 lt250 625 mdash 625 125 mdash mdash mdashEugenia caryophyllus (L) Myrtaceae Flower buds (F) 625 125 mdash mdash 625 mdash mdash mdash mdashEucalyptus globulus (L) Myrtaceae Fraction (leaf) mdash mdash mdash mdash mdash mdash lt250 mdash lt250Mentha piperita (L) Labiatae Fraction (WP) mdash 625 3125 mdash 3125 mdash lt250 mdash mdashOcimum sanctum (L) Labiatae Leaf (fraction) 625 125 mdash 3125 625 mdash 125 lt250 mdashRosmarinus officinalis (L) Labiatae Rosemary oil 3125 mdash mdash mdash 3125 mdash mdash mdash mdashVetiveria zizanioides (L) Graminae Root (fraction) mdash 125 125 mdash mdash mdash gt250 gt250The bacterial growth inhibitory activity was compared for significant differences within the bacterial strains by broth-dilution method at 250 125 625 3125156 and 78 120583gmL F fractions

exposed to various compounds The cell proliferation wasincreased by the influence of the plant components at thelower concentrations Whereas the oil yielding plant com-pounds showed inhibition of cell proliferation and toxicity athigher doses 250ndash1000120583gmL

4 Discussion

The Western Ghats is considered as one of the richestbiodiversity hotspots in the world [41] In this survey wecollected nearly 78 medicinal plants from Western andEastern Ghats that are edible and popularly used for curingvarious ailments including snakebite Traditional remedieshave a long-standing history in many tribal settlements inTN India and they continue to provide useful and applicabletools for treating ailments [42] The ingredients that makeup the ldquoVishakallurdquo stone which is used as an antidote forsnakebite are different herbs and pebbles available from theriver banks Likewise aqueous paste and decoction obtainedfrom the leaves of A paniculata are widely used for snakebitetreatment by indigenous people [43] Previous studies havereported that ethnomedicine plays major roles in conservingthe disappearing knowledge of tribal communities [44ndash47]The traditional beliefs of reliance on a rich diversity ofethnomedicinal plants located at different settlements havealso been confirmed in another study [48 49] Herein weexplored the various types of traditional practices reported bythe primitive tribal communities with a view to gain furtherknowledge from such studies

In the present investigation potentially rich sources oftribal medicine (71 plants) were scientifically evaluated fortheir antibacterial activity against the MDR bacteria and theaccumulated data was disseminated for the first time to the

scientific community Out of the 71medicinal plants screenedfor the antibacterial activity 10 of them (T involucrataC acida A marmelos A vasica C procera A paniculataand M piperita A indica S indicus and E cardamomum)displayed the highest antibacterial activity against the mul-tidrug resistant B pseudomallei (KHW and TES) and Saureus strains The antibacterial activity of those crude plantextracts was as equally effective as that of the standard drugsOur findings corroborated with the previous reports madeon the antistaphylococcal activity of tribal medicinal plants[34 50 51] On the other hand isolated components from themost active extracts of T involucrata shellsol and C acidaexhibited the most potent action against the antibiotic resis-tant B pseudomallei (KHW) K pneumoniae and S aureusstrainsThese results further confirmed our previous findingson the leaves of T involucrata and its compounds hydrocar-bon ester-like shellsol which displayed a high antibacterialeffect against the different bacterial strains especially thatof S aureus [34] Eugenol and caryophyllene are the activeagents contained in the M piperita [52] and O sanctumplants which are believed to be mainly responsible for theantimicrobial properties of these plants [53] Interestinglythe compounds obtained from the aromatic plants such asC zeylanicum and R officinalis were also found to be veryeffective against the multidrug resistant human pathogen Kpneumoniae S aureus S typhi and B pseudomallei (KHW)that causes melioidosis

The inhibitory potential determined for the shellsol(T involucrata) vaseline (A vasica) C acida and C zey-lanicum indicates that the MIC of 78ndash3125120583gmL foundagainst the B pseudomallei of KHW K pneumoniae Kpneumoniae S pyogenes and V damsel and S pneumoniaewas quite low Similarly lower MIC values were found for

Evidence-Based Complementary and Alternative Medicine 15

M piperita (MIC of 113ndash225 mgmL) against the abovebacterial strains [52] MIC was found for the most activealcohol extracts of A salvifolium (MIC 0034ndash0263mgmL)on S aureus [54] and the MIC for S trilobatum aqueousextracts determined against the tested organisms rangedfrom 006 to 05mgmL [55] C zeylanicum was foundto have an effective antibacterial activity (MIC 64 120583gmL)against P aeruginosa E coli B subtilis and S aureus[56] Previously several investigators have demonstrated thatactive agents exert interesting activity against bacteria evenat lower concentrations tested [57 58] The bacteriostaticmechanism involves damage to the cell walls of bacteriafollowed by inhibition of protein synthesis that ultimatelyleads to bacterial death [59] The most active plants arewidely used by various tribes as traditional treatment (iecut wounds skin infection and scabies) thus indicatingthe potential for further development into promising drugsIn order to ascertain the safety and efficacy of the mostactive compounds their effect on human skin fibroblast cellswas evaluated Chemical constituents of E cardamomumT involucrata S indicus C acida A vasica A marmelosA indica and A paniculata plants failed to produce anynoticeable toxicity up to 1000120583gmL Although some ofthe tested compounds exhibited a slight reduction of cellproliferation and some minor morphological changes suchchanges were insignificant at lower doses and became evidentonly at higher doses However certain aromatic compoundsof O sanctum E globulus V zizanioides C citratus and Eglobulus plants showed reduction of cell proliferation againstHEPK cells

Our phytochemical screening also provides evidenceof the presence of several types of compounds that aremainly responsible for the remarkable antibacterial effectof these plants The differences noted for the bactericidalactivity of various plant extracts in this study appears tobe directly related to the diversity of compounds (shown inparentheses) that are accumulated in the following plants(eg A marmelos and O umbellate) [60ndash62] Compoundslike tannins phenol and polyphenols can bind the Gram-negative bacteria to form a heavy soluble complex on the cellsurface which subsequently disturbs the availability of recep-tor on cells and kills the bacteria [63] Several species havingwide spectra of antimicrobial activity mainly due to theactive constituents such as essential oil phenolic compoundslike thymol carvacrol in oregano and thyme eugenol inclove and cinnamon were also identified previously [64 65]Essential oils degrade the cell wall interact with the cell com-ponents and then disrupt the cytoplasmic membrane [66]The antimicrobial effect of phenolic compounds may involvemultiplemodes of action including damage to themembraneprotein interference with membrane integrated enzymes[67] causing leakage of cellular components coagulation ofcytoplasm depletion of the proton motive force alterationof fatty acid and phospholipid constituents impairmentof enzymatic mechanisms for production and metabolismalteration of nutrient uptake and electron transport [68]influencing the synthesis of DNA and RNA and destroyingprotein translation and the function of the mitochondrionin eukaryotes [69] The mode of action of antimicrobial

agents depends on the type of microorganism and is mainlyrelated to their cell wall structure and the outer membranearrangement Most of the plant spices and herbs containcomplex phenolics (ie phenolic acids flavonoids tanninslignans coumarins quinines) In addition the mechanismsof action of each phenolic compound against various bacteriaare also very complicated [26 70] Further investigation istherefore required to understand the relationship betweenthe antimicrobial action and the chemical structure of everyphenolic compound in the tested extracts The informationavailable from previous pharmacological sources combinedwith the findings herein reported on the medicinal plantextracts may serve as essential data for future drug develop-ment to combat diseases caused by theMDRbacterial strains

Abbreviations

MDR Multidrug resistant bacteriaTES Strain of Burkholderia pseudomalleiMRSA Methicillin-resistant Staphylococcus aureusVRE Vancomycin-resistant enterococciMTB Mycobacterium tuberculosisCH3OH Methanol

CFU Colony forming unitHCl Hydrochloric acidTLC Thin layer chromatographyH2SO4 Sulphuric acids

UV Ultraviolet spectrophotometerHEPK Human skin fibroblasts

Acknowledgments

Theauthors are thankful to the ERI Loyola College ChennaiIndia for his support during the survey The authors alsothank the Department of Microbiology NUHS NUS forthe bacterial cultures used in this investigation The authorsare thankful to the tribal people for sharing their healingknowledge on medicinal aromatic plants

References

[1] M N Alekshun and S B Levy ldquoMolecular mechanisms ofantibacterialmultidrug resistancerdquoCell vol 128 no 6 pp 1037ndash1050 2007

[2] A D Russell ldquoAntibiotic and biocide resistance in bacteriaintroductionrdquo Symposium Series Society for Applied Microbiol-ogy vol 31 no 92 pp 1Sndash3S 2002

[3] D R Guay ldquoTreatment of bacterial skin and skin structureinfectionsrdquo Expert Opinion on Pharmacotherapy vol 4 no 8pp 1259ndash1275 2003

[4] A J Brosnahan and P M Schlievert ldquoGram-positive bac-terial superantigen outside-in signaling causes toxic shocksyndromerdquo FEBS Journal vol 278 no 23 pp 4649ndash4667 2011

[5] K Fellermann J Wehkamp K R Herrlinger and E F StangeldquoCrohnrsquos disease a defensin deficiency syndromerdquo EuropeanJournal of Gastroenterology and Hepatology vol 15 no 6 pp627ndash634 2003

16 Evidence-Based Complementary and Alternative Medicine

[6] L Chen S Lin R Agha-Majzoub L Overbergh C Mathieuand L S Chan ldquoCCL27 is a critical factor for the developmentof atopic dermatitis in the keratin-14 IL-4 transgenic mousemodelrdquo International Immunology vol 18 no 8 pp 1233ndash12422006

[7] J H Song V Thamlikitkul and P R Hsueh ldquoClinical andeconomic burden of community-acquired pneumonia amongstadults in the Asia-Pacific regionrdquo International Journal ofAntimicrobial Agents vol 38 no 2 pp 108ndash117 2011

[8] D A B Dance ldquoMelioidosisrdquo Current Opinion in InfectiousDiseases vol 15 no 2 pp 127ndash132 2002

[9] N J White ldquoMelioidosisrdquo The Lancet vol 361 no 9370 pp1715ndash1722 2003

[10] S Yang ldquoMelioidosis research in Chinardquo Acta Tropica vol 77no 2 pp 157ndash165 2000

[11] P R Hsueh L J Teng L N Lee et al ldquoMelioidosis an emerginginfection in Taiwanrdquo Emerging Infectious Diseases vol 7 no 3pp 428ndash433 2001

[12] C Walsh and S Fanning ldquoAntimicrobial resistance in food-borne pathogensmdasha cause for concernrdquo Current Drug Targetsvol 9 no 9 pp 808ndash815 2008

[13] M Vorachit P Chongtrakool S Arkomsean and S BoonsongldquoAntimicrobial resistance in Burkholderia pseudomalleirdquo ActaTropica vol 74 no 2-3 pp 139ndash144 2000

[14] F M Thibault E Hernandez D R Vidal M Girardet and JD Cavallo ldquoAntibiotic susceptibility of 65 isolates of Burkholde-ria pseudomallei and Burkholderia mallei to 35 antimicrobialagentsrdquo Journal of Antimicrobial Chemotherapy vol 54 no 6pp 1134ndash1138 2004

[15] R L S Sikarwar ldquoEthnogynaecological uses of plants new toIndiardquo Ethnobotany vol 12 no 1-2 pp 112ndash115 2002

[16] R Siva ldquoStatus of natural dyes and dye-yielding plants in IndiardquoCurrent Science vol 92 no 7 pp 916ndash925 2007

[17] P Kumar Singh V Kumar R K Tiwari A Sharma ChV Rao and R H Singh ldquoMedico-ethnobotany of lsquochatararsquoblock of district sonebhadra Uttar Pradesh Indiardquo Advances inBiological Research vol 4 no 1 pp 65ndash80 2010

[18] W H Lewis and M P Elwin-Lewis Medical Botany PlantsAffecting Human Health John Wiley amp Sons New York NYUSA 2003

[19] R Perumal Samy P N Pushparaj and P GopalakrishnakoneldquoA compilation of bioactive compounds fromAyurvedardquo Bioin-formation vol 3 no 3 pp 100ndash110 2008

[20] WorldHealthOrganization (WHO)TraditionalMedicine Strat-egy 2002ndash2005 Genera World Health Organization GenevaSwitzerland 2002

[21] P S Haddad G A Azar S Groom and M Boivin ldquoNaturalhealth products modulation of immune function and preven-tion of chronic diseasesrdquo Evidence Based Complementary andAlternative Medicine vol 2 no 4 pp 513ndash520 2005

[22] K Poonam andG S Singh ldquoEthnobotanical study of medicinalplants used by the Taungya community in Terai Arc LandscapeIndiardquo Journal of Ethnopharmacology vol 123 no 1 pp 167ndash1762009

[23] G B Mahady ldquoMedicinal plants for the prevention and treat-ment of bacterial infectionsrdquo Current Pharmaceutical Designvol 11 no 19 pp 2405ndash2427 2005

[24] V P Kumar N S Chauhan H Padh and M Rajani ldquoSearchfor antibacterial and antifungal agents from selected Indianmedicinal plantsrdquo Journal of Ethnopharmacology vol 107 no2 pp 182ndash188 2006

[25] R Nair T Kalariya and S Chanda ldquoAntibacterial activity ofsome plant extracts used in folk medicinerdquo Journal of HerbalPharmacotherapy vol 7 no 3-4 pp 191ndash201 2007

[26] D Kalemba and A Kunicka ldquoAntibacterial and antifungalproperties of essential oilsrdquo Current Medicinal Chemistry vol10 no 10 pp 813ndash829 2003

[27] A H Ashour ldquoAntibacterial antifungal and anticancer activi-ties of volatile oils and extracts from stems leaves and flowers ofEucalyptus sideroxylon and Eucalyptus torquatardquoCancer BiologyandTherapy vol 7 no 3 pp 399ndash403 2008

[28] M J Abad L M Bedoya L Apaza and P Bermejo ldquoTheArtemisia L genus a review of bioactive essential oilsrdquoMolecules vol 17 no 3 pp 2542ndash2566 2012

[29] H N Marzoug M Romdhane A Lebrihi et al ldquoEucalyptusoleosa essential oils chemical composition and antimicrobialand antioxidant activities of the oils from different plant parts(stems leaves flowers and fruits)rdquoMolecules vol 16 no 2 pp1695ndash1709 2011

[30] C Proestos D Sereli and M Komaitis ldquoDetermination ofphenolic compounds in aromatic plants by RP-HPLC and GC-MSrdquo Food Chemistry vol 95 no 1 pp 44ndash52 2006

[31] S Chao G Young C Oberg and K Nakaoka ldquoInhibition ofmethicillin-resistant Staphylococcus aureus (MRSA) by essentialoilsrdquo Flavour and Fragrance Journal vol 23 no 6 pp 444ndash4492008

[32] M Akin D Oguz and H T Saracoglu ldquoAntibacterial effectsof some plant extracts from Labiatae (Lamiaceae) growingnaturally around Sirnak-Silopi Turkeyrdquo International Journal ofPharmaceutical and Applied Sciences vol 1 no 1 pp 4ndash47 2010

[33] K Bayoub T Baibai D Mountassif A Retmane and ASoukri ldquoAntibacterial activities of the crude ethanol extracts ofmedicinal plants against Listeriamonocytogenes and some otherpathogenic strainsrdquo African Journal of Biotechnology vol 9 no27 pp 4251ndash4258 2010

[34] R P Samy P Gopalakrishnakone M Sarumathi P Houghtonand S Ignacimuthu ldquoPurification of antibacterial agents fromTragia involucratamdasha popular tribal medicine for wound heal-ingrdquo Journal of Ethnopharmacology vol 107 no 1 pp 99ndash1062006

[35] K M Matthew The Flora of Tamil Nadu Carnatic Part IndashIIIprinted at the Diocesan Press Madras India 1981ndash1983

[36] AW BauerWMKirby J C Sherris andM Turck ldquoAntibioticsusceptibility testing by a standardized single disk methodrdquoAmerican Journal of Clinical Pathology vol 45 no 4 pp 493ndash496 1966

[37] National Committee for Clinical Laboratory Standards Per-formance Standards For Antimicrobial Susceptibility Testing12th Informational Supplement Approved standard M100-S12National Committee for Clinical Laboratory Standards WaynePa USA 2002

[38] E A Elsinghorst ldquoMeasurement of invasion by gentamicinresistancerdquoMethods in Enzymology vol 236 pp 405ndash420 1994

[39] J B Harborne Phytochemical Methods Chapman and HallNew York NY USA 1976

[40] H Wagner S Bladt and E M Zgairski Plant Drug AnalysisSpringer New York NY USA 1984

[41] N Myers R A Mittermeler C G Mittermeler G A B daFonseca and J Kent ldquoBiodiversity hotspots for conservationprioritiesrdquo Nature vol 403 no 6772 pp 853ndash858 2000

[42] R Perumal Samy and S Ignacimuthu ldquoAntibacterial activity ofsome folkloremedicinal plants used by tribals inWestern Ghats

Evidence-Based Complementary and Alternative Medicine 17

of Indiardquo Journal of Ethnopharmacology vol 69 no 1 pp 63ndash712000

[43] K Nampoothiri A Chrispal A Begum S Jasmine K GGopinath and A Zachariah ldquoA clinical study of renal tubulardysfunction inCleistanthus collinus (Oduvanthalai) poisoningrdquoClinical Toxicology vol 48 no 3 pp 193ndash197 2010

[44] M Strathern UsefulKnowledge The Isiah Berlin Lecture Uni-versity of Manchester Brunswick UK

[45] C N Sunil and A K Pradeep ldquo(2001) Another new species ofTripogon (Poaceae) from Indiardquo Sida vol 19 pp 803ndash806 2005

[46] S Biber-Klemm and T Cottier Right To Plant GeneticResources and Traditional Knowledge Basic Issues and Prospec-tive Oxfordshire Nosworthy Way UK 2006

[47] V S Ramachandran S Shijo Joseph and R Aruna ldquoEth-nobotanical studies from amaravathy range of Indira GandhiWildlife SanctuaryWesternGhats CoimbatoreDistrict South-ern Indiardquo Ethnobotanical Leaflets vol 13 pp 1069ndash1087 2009

[48] A H Rajasab and I Mohamad ldquoDocumentation of folkknowledge on edible wild plants of North Karnatakardquo IndianJournal of Traditional Knowledge vol 3 no 4 pp 419ndash4292004

[49] M Rajadurai V G Vidhya M Ramya and A Bhaskar ldquoEthno-medicinal plants used by the traditional healers of pachamalaihills Tamilnadu Indiardquo Studies on Ethno-Medicine vol 3 no 1pp 39ndash41 2009

[50] S K Adesina O Idowu A O Ogundaini et al ldquoAntimicrobialconstituents of the leaves of Acalypha wilkesiana and Acalyphahispidardquo Phytotherapy Research vol 14 no 5 pp 371ndash374 2000

[51] M T Gutierrez-Lugo M P Singh W M Maiese and BN Timmermann ldquoNew antimicrobial cycloartane triterpenesfrom Acalypha communisrdquo Journal of Natural Products vol 65no 6 pp 872ndash875 2002

[52] A K Tyagi andAMalik ldquoAntimicrobial potential and chemicalcomposition of Mentha piperita oil in liquid and vapour phaseagainst food spoiling microorganismsrdquo Food Control vol 22no 11 pp 1707ndash1714 2011

[53] G Devendran and U Balasubramanian ldquoQualitative phyto-chemical screening and GC-MS analysis of Ocimum sanctumL leavesrdquoAsian Journal of Plant Science and Research vol 1 no4 pp 44ndash48 2011

[54] V C Jain N M Patel D P Shah P K Patel and B HJoshi ldquoAntioxidant and antimicrobial activities of Alangiumsalvifolium (Lf) Wang rootrdquo Global Journal of Pharmacologyvol 4 no 1 pp 13ndash18 2010

[55] P Swapna Latha andK Kannabiran ldquoAntimicrobial activity andphytochemicals of Solanum trilobatum LinnrdquoAfrican Journal ofBiotechnology vol 5 no 23 pp 2402ndash2404 2006

[56] M Usha1 S Ragini and S M A Naqvi ldquoAntibacterial activityof acetone and ethanol extracts of Cinnamon (Cinnamomumzeylanicum) and Ajowan (Trachyspermum ammi) on four foodspoilage bacteriardquo International Research Journal of BiologicalSciences vol 1 no 4 pp 7ndash11 2012

[57] S Prabuseenivasan M Jayakumar and S Ignacimuthu ldquoInvitro antibacterial activity of some plant essential oilsrdquo BMCComplementary and Alternative Medicine vol 6 article 39 pp1ndash8 2006

[58] G A Ayoola F M Lawore T Adelowotan et al ldquoChemi-cal analysis and antimicrobial activity of the essential oil ofSyzigium aromaticum (clove)rdquo African Journal of Biotechnologyvol 7 no 13 pp 2227ndash2231 2008

[59] Y J Fu L Chen Y Zu et al ldquoThe antibacterial activity of cloveessential oil against Propionibacterium acnes and its mechanismof actionrdquo Archives of Dermatology vol 145 no 1 pp 86ndash882009

[60] D Venkatesan C M Karrunakarn S Selva Kumar and P TPalani Swamy ldquoIdentification of phytochemical constituents ofAegle marmelos responsible for antimicrobial activity againstselected pathogenic organismsrdquo Ethnobotanical Leaflets vol 13pp 1362ndash1372 2009

[61] P Arun K G Purushotham J Johnsy jayarani and V KumarildquoIn vitro Antibacterial Activity of Oldenlandia umbellata anIndian medicinal Plantrdquo Journal of Pharmaceutical Science andTechnology vol 2 no 4 pp 198ndash201 2010

[62] A Manjamalai R Sardar Sathyajith Singh C Guruvayoorap-pan and V M Berlin Grace ldquoAnalysis of phytochemicalconstituents and antimicrobial activity of some medicinalplants in Taminadu Indiardquo Global Journal of Biotechnology andBiochemistry vol 5 no 2 pp 120ndash128 2010

[63] E Haslam ldquoNatural polyphenols (vegetable tannins) as drugsPossible modes of actionrdquo Journal of Natural Products vol 59no 2 pp 205ndash215 1996

[64] Y Cai Q Luo M Sun and H Corke ldquoAntioxidant activityand phenolic compounds of 112 traditional Chinese medicinalplants associated with anticancerrdquo Life Sciences vol 74 no 17pp 2157ndash2184 2004

[65] B Shan Y Z Cai M Sun and H Corke ldquoAntioxidant capacityof 26 spice extracts and characterization of their phenolicconstituentsrdquo Journal of Agricultural and Food Chemistry vol53 no 20 pp 7749ndash7759 2005

[66] R J W Lambert P N Skandamis P J Coote and G-J ENychas ldquoA study of the minimum inhibitory concentration andmode of action of oregano essential oil thymol and carvacrolrdquoJournal of AppliedMicrobiology vol 91 no 3 pp 453ndash462 2001

[67] M Raccach ldquoThe antimicrobial activity of phenolic antioxi-dants in food a reviewrdquo Journal of Food Safety vol 6 no 3 pp141ndash170 1984

[68] M Taniguchi Y Yano E Tada et al ldquoMode of action of polygo-dial an antifungal sesquiterpene dialdehyderdquo Agricultural andBiological Chemistry vol 52 no 6 pp 1409ndash1414 1988

[69] G J E Nychas ldquoNatural antimicrobials from plantsrdquo in NewMethods of Food Preservation G W Gould Ed pp 58ndash89Blackie Academic London UK 1995

[70] S Burt ldquoEssential oils their antibacterial properties and poten-tial applications in foodsmdasha reviewrdquo International Journal ofFood Microbiology vol 94 no 3 pp 223ndash253 2004

Submit your manuscripts athttpwwwhindawicom

Stem CellsInternational

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

MEDIATORSINFLAMMATION

of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Behavioural Neurology

EndocrinologyInternational Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Disease Markers

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

BioMed Research International

OncologyJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Oxidative Medicine and Cellular Longevity

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

PPAR Research

The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014

Immunology ResearchHindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Journal of

ObesityJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Computational and Mathematical Methods in Medicine

OphthalmologyJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Diabetes ResearchJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Research and TreatmentAIDS

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Gastroenterology Research and Practice

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Parkinsonrsquos Disease

Evidence-Based Complementary and Alternative Medicine

Volume 2014Hindawi Publishing Corporationhttpwwwhindawicom

10 Evidence-Based Complementary and Alternative Medicine

Figure 2 Diverse biodiversity richness of medicinal plants in Western and Eastern Ghats (a) Topography of plant covering area in Kollihills (Namakkal district Tamil Nadu) (b) Aerial view of occurrence of medicinal plants in Mundanthurai hills (Tirunelveli district TN) (c)Pachamalai hills (Trichy district) and its natural vegetation inhabitants for Malaiyali tribes (d) Deforestation of natural herbal resources dueto urbanization in Kalrayan hills (Salem district) in the Eastern Ghats of TN (e) Medicinal plants and its various parts used by the natives(traditional healers) for the treatment of diverse human illness with a very high percentage of leaves and whole plants often used for herbaldrug preparation by the local practitioners (f) Various category of plants like shrub herb climbers and tree and the parts used in medicine

Evidence-Based Complementary and Alternative Medicine 11

Figure 3 Medicinal aromatic herbs spices and toxic plants were collected from the tribal areas of the Western and Eastern Ghats region inTamil Nadu India (a) A vasica Nees (leaf) (b) Eclipta alba (L) Hassk (whole plant) (c)Mimosa pudica L (whole plant) (d) P amboinicus(L) Spreng (whole plant) (e) T procumbens (L) (whole plant) (f) Euphorbia hirta Linn (whole plant) (g) A paniculata Wallich ex Nees(H) C roseus (L) GDon (whole plant) used for therapy (i) Cinnamomum iners Reinw ex Blume (leaf) (j) E globulus Labill (leaf and bark)(k) Z officinale Rosc (Rhizome) (l) E caryophyllus (Sprengel) Bullock and Harrison (flower buds) (m) M piperita L (whole plant) (n) Ccitratus (DC) Clitoria ternatea L (whole plant) (o) C zeylanicum Garcin ex Blume (bark) (p) Elettaria cardamomumWhite et Mason (fruitpod) used for medicine and food preparation

by the crude extracts (Figure 4)The antimicrobial efficacy offractions collected from the oil yielding plants was also com-pared with that of the tested compounds C zeylanicum andR officinaliswere themost sensitive in controlling the growthof B pseudomallei (KHW) S aureus K pneumonia and Spneumoniae Fascinatingly all the compounds obtained fromaromatic plants except those from E globules were foundto be very effective against the multidrug resistant humanpathogen B pseudomallei (KHW) that causes melioidosis

On the other hand compounds from C citrates O sanctumE caryophyllus and Z zizanioide showed some promisingeffect only against S aureus (Figure 5) The activity of thecompounds were pronounced more than that of the oilyielding plant fractions

39 Minimum Inhibitory Concentrations (MICs) The antibi-otic potential of the purified fractions was obtained fromthe MIC determination The hydrocarbon ester shellsol

12 Evidence-Based Complementary and Alternative Medicine

(mean plusmn SD n = 3)

0

5

10

15

20

25

30

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn V

d St

Bacteria (+minus)

lowast

lowast

lowast

A vasica

In vitro assay(P

lant

frac

tion)

Inhi

bitio

n zo

nes (

mm

)

(a)

(mean plusmn SD n = 3)

0

5

10

15

20

25

30

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn V

d St

Bacteria (+minus)

lowast

A marmelos

Inhi

bitio

n zo

nes (

mm

)

In vitro assay

(b)

(mean plusmn SD n = 3)

0

5

10

15

20

25

30

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn V

d St

Bacteria (+minus)

lowastlowast

A indica

Inhi

bitio

n zo

nes (

mm

)

Incu

batio

n (37∘C

24 h

)

(c)

(mean plusmn SD n = 3)

0

5

10

15

20

25

30

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn V

d St

Bacteria (+minus)

lowast

lowast lowast

35

C acida

Inhi

bitio

n zo

nes (

mm

)(P

lant

frac

tion)

(d)

(mean plusmn SD n = 3)

0

5

10

15

20

25

30

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn V

d St

Bacteria (+minus)

lowastlowast

lowast

S chamaelea

Inhi

bitio

n zo

nes (

mm

)

(e)

(mean plusmn SD n = 3)

0

5

10

15

20

25

30

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn V

d St

Bacteria (+minus)

lowastlowast

lowastlowast

S indicus

Inhi

bitio

n zo

nes (

mm

)

Incu

batio

n (37∘C

24 h

)

(f)

(mean plusmn SD n = 3)

0

5

10

15

20

25

30

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn V

d St

Bacteria (+minus)

lowast

lowast lowast

E cardamomum

Inhi

bitio

n zo

nes (

mm

)(P

lant

frac

tion)

(g)

(mean plusmn SD n = 3)

0

5

10

15

20

25

30

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn V

d St

Bacteria (+minus)

lowast

lowast

lowast

lowast

Shellsol

35

Inhi

bitio

n zo

nes (

mm

)

Incu

batio

n (37∘C

24 h

)

(h)

Figure 4 In vitro antimicrobial activity of purified fractions from the most active plant extracts tested against bacteria Growth inhibitionzones were measured and analyzed with mean plusmn standard deviation (SD) (119899 = 3) using one way analysis of variance Level of significanceat (lowast119875 gt 001) Most of the fractions exerted a potent inhibitory effect against multidrug resistant Gram-negative bacteria (B pseudomalleistrains KHW and TES) K pneumonia and Gram-positive bacteria S aureus

(T involucrata) andC acida showed an interesting inhibitorypotential against S aureus (MIC of 78 120583gmL) and B pseu-domallei strain of KHW (MIC of 156 120583gmL) A vasicashowed an MIC of 156120583gmL against B pseudomallei(KHW) and an MIC of 3125120583gmL against K pneumo-niae K pneumoniae S pyogenes and V damsela strainsFractions from A marmelos and terpenoid from A indicaexerted bacteriostatic effect with MIC values of 3125120583gmLon some selected bacteria including B pseudomallei ofKHW S aureus and B pseudomallei of TES The MIC of3125 120583gmL was found for E cardamomum against S aureus

K pneumonia and S pyogenes S indicus displayed a verystrong inhibition against MDR K pneumoniae (MIC of156 120583gmL) and against B pseudomallei (KHW and TES)at MIC of 3125 120583gmL When the antimicrobial efficacies ofpurified fractions from aromatic plants were compared theC zeylanicum fraction displayed an important antimicrobialeffect against S aureus (MIC of 78 120583gmL) MDR B pseu-domallei of KHW (MIC of 156 120583gmL) and S pneumoniae(MIC of 3125 120583gmL) The essential oil from M piperateashowed MIC value of 3125120583gmL against K pneumoniae Saureus and B pseudomallei (KHW) respectivelyO sanctum

Evidence-Based Complementary and Alternative Medicine 13

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn

Bacteria (+minus)

0

5

10

15

20

25

30

Inhi

bitio

n zo

nes (

mm

)(P

lant

frac

tion)

lowastlowast

lowast

(mean plusmn SD n = 3)In vitro assay

C citratus

(a)

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn

Bacteria (+minus)

0

5

10

15

20

25

30

Inhi

bitio

n zo

nes (

mm

)

lowast

lowast

lowastlowast

lowast

lowast

(mean plusmn SD n = 3)In vitro assay

35

C zeylanicum

(b)

Bacteria (+minus)

0

5

10

15

20

25

30

Inhi

bitio

n zo

nes (

mm

)

lowast

lowast

lowast

(mean plusmn SD n = 3)

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn V

d St

O sanctum

Incu

batio

n (37∘C

24 h

)

(c)

Bacteria (+minus)

0

5

10

15

20

25

Inhi

bitio

n zo

nes (

mm

)

lowast

lowast

(mean plusmn SD n = 3)

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn V

d St

M piperita

(Pla

nt fr

actio

n)

(d)

Bacteria (+minus)

0

5

10

15

20

25

30

Inhi

bitio

n zo

nes (

mm

)lowast

lowast

lowast

(mean plusmn SD n = 3)

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn V

d St

E caryophyllus

(e)

0

5

10

15

20

25

30

Inhi

bitio

n zo

nes (

mm

) (mean plusmn SD n = 3)

Bacteria (+minus)

KHW

TES

Kp

KprPa Sa

Stp

Spn

E globulus

Incu

batio

n (37∘C

24 h

)

(f)

Bacteria (+minus)

0

5

10

15

20

25

30

Inhi

bitio

n zo

nes (

mm

)(P

lant

frac

tion)

lowast

lowast

lowast

lowast

(mean plusmn SD n = 3)

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn V

d

R officinalis

(g)

0

5

10

15

20

25

30

Inhi

bitio

n zo

nes (

mm

)

lowast

lowast

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn V

d St

Bacteria (+minus)V zizanioides

(mean plusmn SD n = 3)

Incu

batio

n (37∘C

24 h

)

(h)

Figure 5 Comparison of antimicrobial effect of plant compounds obtained from the most popularly used sources of essential oils as assayedby the disc-diffusion method in vitro It displayed a powerful activity against B pseudomallei and S aureus than the other bacteria strainsOther compounds showed only a moderate or weak action against the tested bacteria Values for zone of bacterial growth inhibition werepresented as mean plusmn SD (119899 = 3) with level of significance at (lowast119875 gt 001)

and C citratus fractions also showed antimicrobial activity(MICs of 3125ndash125120583gmL) only at higher concentrationsagainst the tested bacteria In addition to that higher concen-trations (gt250120583gmL) (of Vetiveria fractions) were requiredto inhibit Vibro species and others (including E globulusfractions) failed to show any effect at tested concentrations(78ndash125 120583gmL) However the purified fractions (from mostactive medicinal plants) showed strong bacteriostatic inhibi-tion against the tested organisms (Table 3)

310 Cytotoxic Effects of Plants When the components wereassayed for cytotoxicity against the normal human skinfibroblasts (HEPK) cells the compounds obtained from

E cardamomum T involucrata S indicusC acidaA vasicaA marmelos A indica and A paniculata did not showtoxicity up to 1000120583gmL (see Figures S1 and S2 in Supple-mentaryMaterial available online at httpdxdoiorg1011552013525613) A slight reduction of cell proliferation wasnoted only at higher doses (2000120583gmL) In contrast cellproliferation was markedly reduced after exposure of HEPKcells to O sanctum E globulus V zizanioides C citratusand E globulus compounds There was no gradual reductionin skin cell proliferation seen after exposure to C zeylan-icum R officinalis and M piperita (see Figures S3 and S4)compounds The toxicity was found to be concentration-dependent when the skin fibroblasts (HEPK) cells were

14 Evidence-Based Complementary and Alternative Medicine

Table 3 Minimum inhibitory concentrations (MICs) of purified plant fractions and essential oils against antibiotic resistant bacteria

Botanical name Family Parts used Gram-positive and -negative bacteria (MICs 120583gmL)KHW TES Kp KPr Sa Stp Sp Vd St

A indica Juss Meliaceae Seed (fraction) 3125 625 125 lt250 3125 mdash mdash mdash mdashA marmelos (L) Rutaceae Root-bark (F) 3125 3125 mdash mdash 3125 625 125 250 mdashA paniculata Nees Acanthaceae Leaf (fraction) 250 mdash 125 mdash 625 mdash mdash 125 mdashA vasica Nees Acanthaceae Fraction (Stem) 156 3125 3125 625 mdash lt250 3125 3125 mdashC acida Roxb Rutaceae Leaf (fraction) 156 625 mdash mdash 78 mdash 3125 mdash mdashE cardamomumWhite et Mason Acanthaceae Fraction (WP) 625 mdash 3125 mdash 3125 250 625 mdash mdashS indicus (L) Euphorbiaceae Whole plant (F) 3125 125 156 mdash 625 mdash mdash lt250 mdashT involucrata (L) Euphorbiaceae Shellosol (leaf) 156 3125 mdash mdash 78 625 mdash mdash mdashCinnamomum zeylanicum (L) Lauraceae Bark (fraction) 156 625 mdash mdash 78 3125 mdash mdash 125Cymbopogon citratus (L) Graminae Leaf (fraction) 250 lt250 625 mdash 625 125 mdash mdash mdashEugenia caryophyllus (L) Myrtaceae Flower buds (F) 625 125 mdash mdash 625 mdash mdash mdash mdashEucalyptus globulus (L) Myrtaceae Fraction (leaf) mdash mdash mdash mdash mdash mdash lt250 mdash lt250Mentha piperita (L) Labiatae Fraction (WP) mdash 625 3125 mdash 3125 mdash lt250 mdash mdashOcimum sanctum (L) Labiatae Leaf (fraction) 625 125 mdash 3125 625 mdash 125 lt250 mdashRosmarinus officinalis (L) Labiatae Rosemary oil 3125 mdash mdash mdash 3125 mdash mdash mdash mdashVetiveria zizanioides (L) Graminae Root (fraction) mdash 125 125 mdash mdash mdash gt250 gt250The bacterial growth inhibitory activity was compared for significant differences within the bacterial strains by broth-dilution method at 250 125 625 3125156 and 78 120583gmL F fractions

exposed to various compounds The cell proliferation wasincreased by the influence of the plant components at thelower concentrations Whereas the oil yielding plant com-pounds showed inhibition of cell proliferation and toxicity athigher doses 250ndash1000120583gmL

4 Discussion

The Western Ghats is considered as one of the richestbiodiversity hotspots in the world [41] In this survey wecollected nearly 78 medicinal plants from Western andEastern Ghats that are edible and popularly used for curingvarious ailments including snakebite Traditional remedieshave a long-standing history in many tribal settlements inTN India and they continue to provide useful and applicabletools for treating ailments [42] The ingredients that makeup the ldquoVishakallurdquo stone which is used as an antidote forsnakebite are different herbs and pebbles available from theriver banks Likewise aqueous paste and decoction obtainedfrom the leaves of A paniculata are widely used for snakebitetreatment by indigenous people [43] Previous studies havereported that ethnomedicine plays major roles in conservingthe disappearing knowledge of tribal communities [44ndash47]The traditional beliefs of reliance on a rich diversity ofethnomedicinal plants located at different settlements havealso been confirmed in another study [48 49] Herein weexplored the various types of traditional practices reported bythe primitive tribal communities with a view to gain furtherknowledge from such studies

In the present investigation potentially rich sources oftribal medicine (71 plants) were scientifically evaluated fortheir antibacterial activity against the MDR bacteria and theaccumulated data was disseminated for the first time to the

scientific community Out of the 71medicinal plants screenedfor the antibacterial activity 10 of them (T involucrataC acida A marmelos A vasica C procera A paniculataand M piperita A indica S indicus and E cardamomum)displayed the highest antibacterial activity against the mul-tidrug resistant B pseudomallei (KHW and TES) and Saureus strains The antibacterial activity of those crude plantextracts was as equally effective as that of the standard drugsOur findings corroborated with the previous reports madeon the antistaphylococcal activity of tribal medicinal plants[34 50 51] On the other hand isolated components from themost active extracts of T involucrata shellsol and C acidaexhibited the most potent action against the antibiotic resis-tant B pseudomallei (KHW) K pneumoniae and S aureusstrainsThese results further confirmed our previous findingson the leaves of T involucrata and its compounds hydrocar-bon ester-like shellsol which displayed a high antibacterialeffect against the different bacterial strains especially thatof S aureus [34] Eugenol and caryophyllene are the activeagents contained in the M piperita [52] and O sanctumplants which are believed to be mainly responsible for theantimicrobial properties of these plants [53] Interestinglythe compounds obtained from the aromatic plants such asC zeylanicum and R officinalis were also found to be veryeffective against the multidrug resistant human pathogen Kpneumoniae S aureus S typhi and B pseudomallei (KHW)that causes melioidosis

The inhibitory potential determined for the shellsol(T involucrata) vaseline (A vasica) C acida and C zey-lanicum indicates that the MIC of 78ndash3125120583gmL foundagainst the B pseudomallei of KHW K pneumoniae Kpneumoniae S pyogenes and V damsel and S pneumoniaewas quite low Similarly lower MIC values were found for

Evidence-Based Complementary and Alternative Medicine 15

M piperita (MIC of 113ndash225 mgmL) against the abovebacterial strains [52] MIC was found for the most activealcohol extracts of A salvifolium (MIC 0034ndash0263mgmL)on S aureus [54] and the MIC for S trilobatum aqueousextracts determined against the tested organisms rangedfrom 006 to 05mgmL [55] C zeylanicum was foundto have an effective antibacterial activity (MIC 64 120583gmL)against P aeruginosa E coli B subtilis and S aureus[56] Previously several investigators have demonstrated thatactive agents exert interesting activity against bacteria evenat lower concentrations tested [57 58] The bacteriostaticmechanism involves damage to the cell walls of bacteriafollowed by inhibition of protein synthesis that ultimatelyleads to bacterial death [59] The most active plants arewidely used by various tribes as traditional treatment (iecut wounds skin infection and scabies) thus indicatingthe potential for further development into promising drugsIn order to ascertain the safety and efficacy of the mostactive compounds their effect on human skin fibroblast cellswas evaluated Chemical constituents of E cardamomumT involucrata S indicus C acida A vasica A marmelosA indica and A paniculata plants failed to produce anynoticeable toxicity up to 1000120583gmL Although some ofthe tested compounds exhibited a slight reduction of cellproliferation and some minor morphological changes suchchanges were insignificant at lower doses and became evidentonly at higher doses However certain aromatic compoundsof O sanctum E globulus V zizanioides C citratus and Eglobulus plants showed reduction of cell proliferation againstHEPK cells

Our phytochemical screening also provides evidenceof the presence of several types of compounds that aremainly responsible for the remarkable antibacterial effectof these plants The differences noted for the bactericidalactivity of various plant extracts in this study appears tobe directly related to the diversity of compounds (shown inparentheses) that are accumulated in the following plants(eg A marmelos and O umbellate) [60ndash62] Compoundslike tannins phenol and polyphenols can bind the Gram-negative bacteria to form a heavy soluble complex on the cellsurface which subsequently disturbs the availability of recep-tor on cells and kills the bacteria [63] Several species havingwide spectra of antimicrobial activity mainly due to theactive constituents such as essential oil phenolic compoundslike thymol carvacrol in oregano and thyme eugenol inclove and cinnamon were also identified previously [64 65]Essential oils degrade the cell wall interact with the cell com-ponents and then disrupt the cytoplasmic membrane [66]The antimicrobial effect of phenolic compounds may involvemultiplemodes of action including damage to themembraneprotein interference with membrane integrated enzymes[67] causing leakage of cellular components coagulation ofcytoplasm depletion of the proton motive force alterationof fatty acid and phospholipid constituents impairmentof enzymatic mechanisms for production and metabolismalteration of nutrient uptake and electron transport [68]influencing the synthesis of DNA and RNA and destroyingprotein translation and the function of the mitochondrionin eukaryotes [69] The mode of action of antimicrobial

agents depends on the type of microorganism and is mainlyrelated to their cell wall structure and the outer membranearrangement Most of the plant spices and herbs containcomplex phenolics (ie phenolic acids flavonoids tanninslignans coumarins quinines) In addition the mechanismsof action of each phenolic compound against various bacteriaare also very complicated [26 70] Further investigation istherefore required to understand the relationship betweenthe antimicrobial action and the chemical structure of everyphenolic compound in the tested extracts The informationavailable from previous pharmacological sources combinedwith the findings herein reported on the medicinal plantextracts may serve as essential data for future drug develop-ment to combat diseases caused by theMDRbacterial strains

Abbreviations

MDR Multidrug resistant bacteriaTES Strain of Burkholderia pseudomalleiMRSA Methicillin-resistant Staphylococcus aureusVRE Vancomycin-resistant enterococciMTB Mycobacterium tuberculosisCH3OH Methanol

CFU Colony forming unitHCl Hydrochloric acidTLC Thin layer chromatographyH2SO4 Sulphuric acids

UV Ultraviolet spectrophotometerHEPK Human skin fibroblasts

Acknowledgments

Theauthors are thankful to the ERI Loyola College ChennaiIndia for his support during the survey The authors alsothank the Department of Microbiology NUHS NUS forthe bacterial cultures used in this investigation The authorsare thankful to the tribal people for sharing their healingknowledge on medicinal aromatic plants

References

[1] M N Alekshun and S B Levy ldquoMolecular mechanisms ofantibacterialmultidrug resistancerdquoCell vol 128 no 6 pp 1037ndash1050 2007

[2] A D Russell ldquoAntibiotic and biocide resistance in bacteriaintroductionrdquo Symposium Series Society for Applied Microbiol-ogy vol 31 no 92 pp 1Sndash3S 2002

[3] D R Guay ldquoTreatment of bacterial skin and skin structureinfectionsrdquo Expert Opinion on Pharmacotherapy vol 4 no 8pp 1259ndash1275 2003

[4] A J Brosnahan and P M Schlievert ldquoGram-positive bac-terial superantigen outside-in signaling causes toxic shocksyndromerdquo FEBS Journal vol 278 no 23 pp 4649ndash4667 2011

[5] K Fellermann J Wehkamp K R Herrlinger and E F StangeldquoCrohnrsquos disease a defensin deficiency syndromerdquo EuropeanJournal of Gastroenterology and Hepatology vol 15 no 6 pp627ndash634 2003

16 Evidence-Based Complementary and Alternative Medicine

[6] L Chen S Lin R Agha-Majzoub L Overbergh C Mathieuand L S Chan ldquoCCL27 is a critical factor for the developmentof atopic dermatitis in the keratin-14 IL-4 transgenic mousemodelrdquo International Immunology vol 18 no 8 pp 1233ndash12422006

[7] J H Song V Thamlikitkul and P R Hsueh ldquoClinical andeconomic burden of community-acquired pneumonia amongstadults in the Asia-Pacific regionrdquo International Journal ofAntimicrobial Agents vol 38 no 2 pp 108ndash117 2011

[8] D A B Dance ldquoMelioidosisrdquo Current Opinion in InfectiousDiseases vol 15 no 2 pp 127ndash132 2002

[9] N J White ldquoMelioidosisrdquo The Lancet vol 361 no 9370 pp1715ndash1722 2003

[10] S Yang ldquoMelioidosis research in Chinardquo Acta Tropica vol 77no 2 pp 157ndash165 2000

[11] P R Hsueh L J Teng L N Lee et al ldquoMelioidosis an emerginginfection in Taiwanrdquo Emerging Infectious Diseases vol 7 no 3pp 428ndash433 2001

[12] C Walsh and S Fanning ldquoAntimicrobial resistance in food-borne pathogensmdasha cause for concernrdquo Current Drug Targetsvol 9 no 9 pp 808ndash815 2008

[13] M Vorachit P Chongtrakool S Arkomsean and S BoonsongldquoAntimicrobial resistance in Burkholderia pseudomalleirdquo ActaTropica vol 74 no 2-3 pp 139ndash144 2000

[14] F M Thibault E Hernandez D R Vidal M Girardet and JD Cavallo ldquoAntibiotic susceptibility of 65 isolates of Burkholde-ria pseudomallei and Burkholderia mallei to 35 antimicrobialagentsrdquo Journal of Antimicrobial Chemotherapy vol 54 no 6pp 1134ndash1138 2004

[15] R L S Sikarwar ldquoEthnogynaecological uses of plants new toIndiardquo Ethnobotany vol 12 no 1-2 pp 112ndash115 2002

[16] R Siva ldquoStatus of natural dyes and dye-yielding plants in IndiardquoCurrent Science vol 92 no 7 pp 916ndash925 2007

[17] P Kumar Singh V Kumar R K Tiwari A Sharma ChV Rao and R H Singh ldquoMedico-ethnobotany of lsquochatararsquoblock of district sonebhadra Uttar Pradesh Indiardquo Advances inBiological Research vol 4 no 1 pp 65ndash80 2010

[18] W H Lewis and M P Elwin-Lewis Medical Botany PlantsAffecting Human Health John Wiley amp Sons New York NYUSA 2003

[19] R Perumal Samy P N Pushparaj and P GopalakrishnakoneldquoA compilation of bioactive compounds fromAyurvedardquo Bioin-formation vol 3 no 3 pp 100ndash110 2008

[20] WorldHealthOrganization (WHO)TraditionalMedicine Strat-egy 2002ndash2005 Genera World Health Organization GenevaSwitzerland 2002

[21] P S Haddad G A Azar S Groom and M Boivin ldquoNaturalhealth products modulation of immune function and preven-tion of chronic diseasesrdquo Evidence Based Complementary andAlternative Medicine vol 2 no 4 pp 513ndash520 2005

[22] K Poonam andG S Singh ldquoEthnobotanical study of medicinalplants used by the Taungya community in Terai Arc LandscapeIndiardquo Journal of Ethnopharmacology vol 123 no 1 pp 167ndash1762009

[23] G B Mahady ldquoMedicinal plants for the prevention and treat-ment of bacterial infectionsrdquo Current Pharmaceutical Designvol 11 no 19 pp 2405ndash2427 2005

[24] V P Kumar N S Chauhan H Padh and M Rajani ldquoSearchfor antibacterial and antifungal agents from selected Indianmedicinal plantsrdquo Journal of Ethnopharmacology vol 107 no2 pp 182ndash188 2006

[25] R Nair T Kalariya and S Chanda ldquoAntibacterial activity ofsome plant extracts used in folk medicinerdquo Journal of HerbalPharmacotherapy vol 7 no 3-4 pp 191ndash201 2007

[26] D Kalemba and A Kunicka ldquoAntibacterial and antifungalproperties of essential oilsrdquo Current Medicinal Chemistry vol10 no 10 pp 813ndash829 2003

[27] A H Ashour ldquoAntibacterial antifungal and anticancer activi-ties of volatile oils and extracts from stems leaves and flowers ofEucalyptus sideroxylon and Eucalyptus torquatardquoCancer BiologyandTherapy vol 7 no 3 pp 399ndash403 2008

[28] M J Abad L M Bedoya L Apaza and P Bermejo ldquoTheArtemisia L genus a review of bioactive essential oilsrdquoMolecules vol 17 no 3 pp 2542ndash2566 2012

[29] H N Marzoug M Romdhane A Lebrihi et al ldquoEucalyptusoleosa essential oils chemical composition and antimicrobialand antioxidant activities of the oils from different plant parts(stems leaves flowers and fruits)rdquoMolecules vol 16 no 2 pp1695ndash1709 2011

[30] C Proestos D Sereli and M Komaitis ldquoDetermination ofphenolic compounds in aromatic plants by RP-HPLC and GC-MSrdquo Food Chemistry vol 95 no 1 pp 44ndash52 2006

[31] S Chao G Young C Oberg and K Nakaoka ldquoInhibition ofmethicillin-resistant Staphylococcus aureus (MRSA) by essentialoilsrdquo Flavour and Fragrance Journal vol 23 no 6 pp 444ndash4492008

[32] M Akin D Oguz and H T Saracoglu ldquoAntibacterial effectsof some plant extracts from Labiatae (Lamiaceae) growingnaturally around Sirnak-Silopi Turkeyrdquo International Journal ofPharmaceutical and Applied Sciences vol 1 no 1 pp 4ndash47 2010

[33] K Bayoub T Baibai D Mountassif A Retmane and ASoukri ldquoAntibacterial activities of the crude ethanol extracts ofmedicinal plants against Listeriamonocytogenes and some otherpathogenic strainsrdquo African Journal of Biotechnology vol 9 no27 pp 4251ndash4258 2010

[34] R P Samy P Gopalakrishnakone M Sarumathi P Houghtonand S Ignacimuthu ldquoPurification of antibacterial agents fromTragia involucratamdasha popular tribal medicine for wound heal-ingrdquo Journal of Ethnopharmacology vol 107 no 1 pp 99ndash1062006

[35] K M Matthew The Flora of Tamil Nadu Carnatic Part IndashIIIprinted at the Diocesan Press Madras India 1981ndash1983

[36] AW BauerWMKirby J C Sherris andM Turck ldquoAntibioticsusceptibility testing by a standardized single disk methodrdquoAmerican Journal of Clinical Pathology vol 45 no 4 pp 493ndash496 1966

[37] National Committee for Clinical Laboratory Standards Per-formance Standards For Antimicrobial Susceptibility Testing12th Informational Supplement Approved standard M100-S12National Committee for Clinical Laboratory Standards WaynePa USA 2002

[38] E A Elsinghorst ldquoMeasurement of invasion by gentamicinresistancerdquoMethods in Enzymology vol 236 pp 405ndash420 1994

[39] J B Harborne Phytochemical Methods Chapman and HallNew York NY USA 1976

[40] H Wagner S Bladt and E M Zgairski Plant Drug AnalysisSpringer New York NY USA 1984

[41] N Myers R A Mittermeler C G Mittermeler G A B daFonseca and J Kent ldquoBiodiversity hotspots for conservationprioritiesrdquo Nature vol 403 no 6772 pp 853ndash858 2000

[42] R Perumal Samy and S Ignacimuthu ldquoAntibacterial activity ofsome folkloremedicinal plants used by tribals inWestern Ghats

Evidence-Based Complementary and Alternative Medicine 17

of Indiardquo Journal of Ethnopharmacology vol 69 no 1 pp 63ndash712000

[43] K Nampoothiri A Chrispal A Begum S Jasmine K GGopinath and A Zachariah ldquoA clinical study of renal tubulardysfunction inCleistanthus collinus (Oduvanthalai) poisoningrdquoClinical Toxicology vol 48 no 3 pp 193ndash197 2010

[44] M Strathern UsefulKnowledge The Isiah Berlin Lecture Uni-versity of Manchester Brunswick UK

[45] C N Sunil and A K Pradeep ldquo(2001) Another new species ofTripogon (Poaceae) from Indiardquo Sida vol 19 pp 803ndash806 2005

[46] S Biber-Klemm and T Cottier Right To Plant GeneticResources and Traditional Knowledge Basic Issues and Prospec-tive Oxfordshire Nosworthy Way UK 2006

[47] V S Ramachandran S Shijo Joseph and R Aruna ldquoEth-nobotanical studies from amaravathy range of Indira GandhiWildlife SanctuaryWesternGhats CoimbatoreDistrict South-ern Indiardquo Ethnobotanical Leaflets vol 13 pp 1069ndash1087 2009

[48] A H Rajasab and I Mohamad ldquoDocumentation of folkknowledge on edible wild plants of North Karnatakardquo IndianJournal of Traditional Knowledge vol 3 no 4 pp 419ndash4292004

[49] M Rajadurai V G Vidhya M Ramya and A Bhaskar ldquoEthno-medicinal plants used by the traditional healers of pachamalaihills Tamilnadu Indiardquo Studies on Ethno-Medicine vol 3 no 1pp 39ndash41 2009

[50] S K Adesina O Idowu A O Ogundaini et al ldquoAntimicrobialconstituents of the leaves of Acalypha wilkesiana and Acalyphahispidardquo Phytotherapy Research vol 14 no 5 pp 371ndash374 2000

[51] M T Gutierrez-Lugo M P Singh W M Maiese and BN Timmermann ldquoNew antimicrobial cycloartane triterpenesfrom Acalypha communisrdquo Journal of Natural Products vol 65no 6 pp 872ndash875 2002

[52] A K Tyagi andAMalik ldquoAntimicrobial potential and chemicalcomposition of Mentha piperita oil in liquid and vapour phaseagainst food spoiling microorganismsrdquo Food Control vol 22no 11 pp 1707ndash1714 2011

[53] G Devendran and U Balasubramanian ldquoQualitative phyto-chemical screening and GC-MS analysis of Ocimum sanctumL leavesrdquoAsian Journal of Plant Science and Research vol 1 no4 pp 44ndash48 2011

[54] V C Jain N M Patel D P Shah P K Patel and B HJoshi ldquoAntioxidant and antimicrobial activities of Alangiumsalvifolium (Lf) Wang rootrdquo Global Journal of Pharmacologyvol 4 no 1 pp 13ndash18 2010

[55] P Swapna Latha andK Kannabiran ldquoAntimicrobial activity andphytochemicals of Solanum trilobatum LinnrdquoAfrican Journal ofBiotechnology vol 5 no 23 pp 2402ndash2404 2006

[56] M Usha1 S Ragini and S M A Naqvi ldquoAntibacterial activityof acetone and ethanol extracts of Cinnamon (Cinnamomumzeylanicum) and Ajowan (Trachyspermum ammi) on four foodspoilage bacteriardquo International Research Journal of BiologicalSciences vol 1 no 4 pp 7ndash11 2012

[57] S Prabuseenivasan M Jayakumar and S Ignacimuthu ldquoInvitro antibacterial activity of some plant essential oilsrdquo BMCComplementary and Alternative Medicine vol 6 article 39 pp1ndash8 2006

[58] G A Ayoola F M Lawore T Adelowotan et al ldquoChemi-cal analysis and antimicrobial activity of the essential oil ofSyzigium aromaticum (clove)rdquo African Journal of Biotechnologyvol 7 no 13 pp 2227ndash2231 2008

[59] Y J Fu L Chen Y Zu et al ldquoThe antibacterial activity of cloveessential oil against Propionibacterium acnes and its mechanismof actionrdquo Archives of Dermatology vol 145 no 1 pp 86ndash882009

[60] D Venkatesan C M Karrunakarn S Selva Kumar and P TPalani Swamy ldquoIdentification of phytochemical constituents ofAegle marmelos responsible for antimicrobial activity againstselected pathogenic organismsrdquo Ethnobotanical Leaflets vol 13pp 1362ndash1372 2009

[61] P Arun K G Purushotham J Johnsy jayarani and V KumarildquoIn vitro Antibacterial Activity of Oldenlandia umbellata anIndian medicinal Plantrdquo Journal of Pharmaceutical Science andTechnology vol 2 no 4 pp 198ndash201 2010

[62] A Manjamalai R Sardar Sathyajith Singh C Guruvayoorap-pan and V M Berlin Grace ldquoAnalysis of phytochemicalconstituents and antimicrobial activity of some medicinalplants in Taminadu Indiardquo Global Journal of Biotechnology andBiochemistry vol 5 no 2 pp 120ndash128 2010

[63] E Haslam ldquoNatural polyphenols (vegetable tannins) as drugsPossible modes of actionrdquo Journal of Natural Products vol 59no 2 pp 205ndash215 1996

[64] Y Cai Q Luo M Sun and H Corke ldquoAntioxidant activityand phenolic compounds of 112 traditional Chinese medicinalplants associated with anticancerrdquo Life Sciences vol 74 no 17pp 2157ndash2184 2004

[65] B Shan Y Z Cai M Sun and H Corke ldquoAntioxidant capacityof 26 spice extracts and characterization of their phenolicconstituentsrdquo Journal of Agricultural and Food Chemistry vol53 no 20 pp 7749ndash7759 2005

[66] R J W Lambert P N Skandamis P J Coote and G-J ENychas ldquoA study of the minimum inhibitory concentration andmode of action of oregano essential oil thymol and carvacrolrdquoJournal of AppliedMicrobiology vol 91 no 3 pp 453ndash462 2001

[67] M Raccach ldquoThe antimicrobial activity of phenolic antioxi-dants in food a reviewrdquo Journal of Food Safety vol 6 no 3 pp141ndash170 1984

[68] M Taniguchi Y Yano E Tada et al ldquoMode of action of polygo-dial an antifungal sesquiterpene dialdehyderdquo Agricultural andBiological Chemistry vol 52 no 6 pp 1409ndash1414 1988

[69] G J E Nychas ldquoNatural antimicrobials from plantsrdquo in NewMethods of Food Preservation G W Gould Ed pp 58ndash89Blackie Academic London UK 1995

[70] S Burt ldquoEssential oils their antibacterial properties and poten-tial applications in foodsmdasha reviewrdquo International Journal ofFood Microbiology vol 94 no 3 pp 223ndash253 2004

Submit your manuscripts athttpwwwhindawicom

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Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

MEDIATORSINFLAMMATION

of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Behavioural Neurology

EndocrinologyInternational Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Disease Markers

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

BioMed Research International

OncologyJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Oxidative Medicine and Cellular Longevity

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

PPAR Research

The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014

Immunology ResearchHindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Journal of

ObesityJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Computational and Mathematical Methods in Medicine

OphthalmologyJournal of

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Diabetes ResearchJournal of

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Research and TreatmentAIDS

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Gastroenterology Research and Practice

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Parkinsonrsquos Disease

Evidence-Based Complementary and Alternative Medicine

Volume 2014Hindawi Publishing Corporationhttpwwwhindawicom

Evidence-Based Complementary and Alternative Medicine 11

Figure 3 Medicinal aromatic herbs spices and toxic plants were collected from the tribal areas of the Western and Eastern Ghats region inTamil Nadu India (a) A vasica Nees (leaf) (b) Eclipta alba (L) Hassk (whole plant) (c)Mimosa pudica L (whole plant) (d) P amboinicus(L) Spreng (whole plant) (e) T procumbens (L) (whole plant) (f) Euphorbia hirta Linn (whole plant) (g) A paniculata Wallich ex Nees(H) C roseus (L) GDon (whole plant) used for therapy (i) Cinnamomum iners Reinw ex Blume (leaf) (j) E globulus Labill (leaf and bark)(k) Z officinale Rosc (Rhizome) (l) E caryophyllus (Sprengel) Bullock and Harrison (flower buds) (m) M piperita L (whole plant) (n) Ccitratus (DC) Clitoria ternatea L (whole plant) (o) C zeylanicum Garcin ex Blume (bark) (p) Elettaria cardamomumWhite et Mason (fruitpod) used for medicine and food preparation

by the crude extracts (Figure 4)The antimicrobial efficacy offractions collected from the oil yielding plants was also com-pared with that of the tested compounds C zeylanicum andR officinaliswere themost sensitive in controlling the growthof B pseudomallei (KHW) S aureus K pneumonia and Spneumoniae Fascinatingly all the compounds obtained fromaromatic plants except those from E globules were foundto be very effective against the multidrug resistant humanpathogen B pseudomallei (KHW) that causes melioidosis

On the other hand compounds from C citrates O sanctumE caryophyllus and Z zizanioide showed some promisingeffect only against S aureus (Figure 5) The activity of thecompounds were pronounced more than that of the oilyielding plant fractions

39 Minimum Inhibitory Concentrations (MICs) The antibi-otic potential of the purified fractions was obtained fromthe MIC determination The hydrocarbon ester shellsol

12 Evidence-Based Complementary and Alternative Medicine

(mean plusmn SD n = 3)

0

5

10

15

20

25

30

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn V

d St

Bacteria (+minus)

lowast

lowast

lowast

A vasica

In vitro assay(P

lant

frac

tion)

Inhi

bitio

n zo

nes (

mm

)

(a)

(mean plusmn SD n = 3)

0

5

10

15

20

25

30

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn V

d St

Bacteria (+minus)

lowast

A marmelos

Inhi

bitio

n zo

nes (

mm

)

In vitro assay

(b)

(mean plusmn SD n = 3)

0

5

10

15

20

25

30

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn V

d St

Bacteria (+minus)

lowastlowast

A indica

Inhi

bitio

n zo

nes (

mm

)

Incu

batio

n (37∘C

24 h

)

(c)

(mean plusmn SD n = 3)

0

5

10

15

20

25

30

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn V

d St

Bacteria (+minus)

lowast

lowast lowast

35

C acida

Inhi

bitio

n zo

nes (

mm

)(P

lant

frac

tion)

(d)

(mean plusmn SD n = 3)

0

5

10

15

20

25

30

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn V

d St

Bacteria (+minus)

lowastlowast

lowast

S chamaelea

Inhi

bitio

n zo

nes (

mm

)

(e)

(mean plusmn SD n = 3)

0

5

10

15

20

25

30

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn V

d St

Bacteria (+minus)

lowastlowast

lowastlowast

S indicus

Inhi

bitio

n zo

nes (

mm

)

Incu

batio

n (37∘C

24 h

)

(f)

(mean plusmn SD n = 3)

0

5

10

15

20

25

30

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn V

d St

Bacteria (+minus)

lowast

lowast lowast

E cardamomum

Inhi

bitio

n zo

nes (

mm

)(P

lant

frac

tion)

(g)

(mean plusmn SD n = 3)

0

5

10

15

20

25

30

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn V

d St

Bacteria (+minus)

lowast

lowast

lowast

lowast

Shellsol

35

Inhi

bitio

n zo

nes (

mm

)

Incu

batio

n (37∘C

24 h

)

(h)

Figure 4 In vitro antimicrobial activity of purified fractions from the most active plant extracts tested against bacteria Growth inhibitionzones were measured and analyzed with mean plusmn standard deviation (SD) (119899 = 3) using one way analysis of variance Level of significanceat (lowast119875 gt 001) Most of the fractions exerted a potent inhibitory effect against multidrug resistant Gram-negative bacteria (B pseudomalleistrains KHW and TES) K pneumonia and Gram-positive bacteria S aureus

(T involucrata) andC acida showed an interesting inhibitorypotential against S aureus (MIC of 78 120583gmL) and B pseu-domallei strain of KHW (MIC of 156 120583gmL) A vasicashowed an MIC of 156120583gmL against B pseudomallei(KHW) and an MIC of 3125120583gmL against K pneumo-niae K pneumoniae S pyogenes and V damsela strainsFractions from A marmelos and terpenoid from A indicaexerted bacteriostatic effect with MIC values of 3125120583gmLon some selected bacteria including B pseudomallei ofKHW S aureus and B pseudomallei of TES The MIC of3125 120583gmL was found for E cardamomum against S aureus

K pneumonia and S pyogenes S indicus displayed a verystrong inhibition against MDR K pneumoniae (MIC of156 120583gmL) and against B pseudomallei (KHW and TES)at MIC of 3125 120583gmL When the antimicrobial efficacies ofpurified fractions from aromatic plants were compared theC zeylanicum fraction displayed an important antimicrobialeffect against S aureus (MIC of 78 120583gmL) MDR B pseu-domallei of KHW (MIC of 156 120583gmL) and S pneumoniae(MIC of 3125 120583gmL) The essential oil from M piperateashowed MIC value of 3125120583gmL against K pneumoniae Saureus and B pseudomallei (KHW) respectivelyO sanctum

Evidence-Based Complementary and Alternative Medicine 13

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn

Bacteria (+minus)

0

5

10

15

20

25

30

Inhi

bitio

n zo

nes (

mm

)(P

lant

frac

tion)

lowastlowast

lowast

(mean plusmn SD n = 3)In vitro assay

C citratus

(a)

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn

Bacteria (+minus)

0

5

10

15

20

25

30

Inhi

bitio

n zo

nes (

mm

)

lowast

lowast

lowastlowast

lowast

lowast

(mean plusmn SD n = 3)In vitro assay

35

C zeylanicum

(b)

Bacteria (+minus)

0

5

10

15

20

25

30

Inhi

bitio

n zo

nes (

mm

)

lowast

lowast

lowast

(mean plusmn SD n = 3)

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn V

d St

O sanctum

Incu

batio

n (37∘C

24 h

)

(c)

Bacteria (+minus)

0

5

10

15

20

25

Inhi

bitio

n zo

nes (

mm

)

lowast

lowast

(mean plusmn SD n = 3)

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn V

d St

M piperita

(Pla

nt fr

actio

n)

(d)

Bacteria (+minus)

0

5

10

15

20

25

30

Inhi

bitio

n zo

nes (

mm

)lowast

lowast

lowast

(mean plusmn SD n = 3)

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn V

d St

E caryophyllus

(e)

0

5

10

15

20

25

30

Inhi

bitio

n zo

nes (

mm

) (mean plusmn SD n = 3)

Bacteria (+minus)

KHW

TES

Kp

KprPa Sa

Stp

Spn

E globulus

Incu

batio

n (37∘C

24 h

)

(f)

Bacteria (+minus)

0

5

10

15

20

25

30

Inhi

bitio

n zo

nes (

mm

)(P

lant

frac

tion)

lowast

lowast

lowast

lowast

(mean plusmn SD n = 3)

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn V

d

R officinalis

(g)

0

5

10

15

20

25

30

Inhi

bitio

n zo

nes (

mm

)

lowast

lowast

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn V

d St

Bacteria (+minus)V zizanioides

(mean plusmn SD n = 3)

Incu

batio

n (37∘C

24 h

)

(h)

Figure 5 Comparison of antimicrobial effect of plant compounds obtained from the most popularly used sources of essential oils as assayedby the disc-diffusion method in vitro It displayed a powerful activity against B pseudomallei and S aureus than the other bacteria strainsOther compounds showed only a moderate or weak action against the tested bacteria Values for zone of bacterial growth inhibition werepresented as mean plusmn SD (119899 = 3) with level of significance at (lowast119875 gt 001)

and C citratus fractions also showed antimicrobial activity(MICs of 3125ndash125120583gmL) only at higher concentrationsagainst the tested bacteria In addition to that higher concen-trations (gt250120583gmL) (of Vetiveria fractions) were requiredto inhibit Vibro species and others (including E globulusfractions) failed to show any effect at tested concentrations(78ndash125 120583gmL) However the purified fractions (from mostactive medicinal plants) showed strong bacteriostatic inhibi-tion against the tested organisms (Table 3)

310 Cytotoxic Effects of Plants When the components wereassayed for cytotoxicity against the normal human skinfibroblasts (HEPK) cells the compounds obtained from

E cardamomum T involucrata S indicusC acidaA vasicaA marmelos A indica and A paniculata did not showtoxicity up to 1000120583gmL (see Figures S1 and S2 in Supple-mentaryMaterial available online at httpdxdoiorg1011552013525613) A slight reduction of cell proliferation wasnoted only at higher doses (2000120583gmL) In contrast cellproliferation was markedly reduced after exposure of HEPKcells to O sanctum E globulus V zizanioides C citratusand E globulus compounds There was no gradual reductionin skin cell proliferation seen after exposure to C zeylan-icum R officinalis and M piperita (see Figures S3 and S4)compounds The toxicity was found to be concentration-dependent when the skin fibroblasts (HEPK) cells were

14 Evidence-Based Complementary and Alternative Medicine

Table 3 Minimum inhibitory concentrations (MICs) of purified plant fractions and essential oils against antibiotic resistant bacteria

Botanical name Family Parts used Gram-positive and -negative bacteria (MICs 120583gmL)KHW TES Kp KPr Sa Stp Sp Vd St

A indica Juss Meliaceae Seed (fraction) 3125 625 125 lt250 3125 mdash mdash mdash mdashA marmelos (L) Rutaceae Root-bark (F) 3125 3125 mdash mdash 3125 625 125 250 mdashA paniculata Nees Acanthaceae Leaf (fraction) 250 mdash 125 mdash 625 mdash mdash 125 mdashA vasica Nees Acanthaceae Fraction (Stem) 156 3125 3125 625 mdash lt250 3125 3125 mdashC acida Roxb Rutaceae Leaf (fraction) 156 625 mdash mdash 78 mdash 3125 mdash mdashE cardamomumWhite et Mason Acanthaceae Fraction (WP) 625 mdash 3125 mdash 3125 250 625 mdash mdashS indicus (L) Euphorbiaceae Whole plant (F) 3125 125 156 mdash 625 mdash mdash lt250 mdashT involucrata (L) Euphorbiaceae Shellosol (leaf) 156 3125 mdash mdash 78 625 mdash mdash mdashCinnamomum zeylanicum (L) Lauraceae Bark (fraction) 156 625 mdash mdash 78 3125 mdash mdash 125Cymbopogon citratus (L) Graminae Leaf (fraction) 250 lt250 625 mdash 625 125 mdash mdash mdashEugenia caryophyllus (L) Myrtaceae Flower buds (F) 625 125 mdash mdash 625 mdash mdash mdash mdashEucalyptus globulus (L) Myrtaceae Fraction (leaf) mdash mdash mdash mdash mdash mdash lt250 mdash lt250Mentha piperita (L) Labiatae Fraction (WP) mdash 625 3125 mdash 3125 mdash lt250 mdash mdashOcimum sanctum (L) Labiatae Leaf (fraction) 625 125 mdash 3125 625 mdash 125 lt250 mdashRosmarinus officinalis (L) Labiatae Rosemary oil 3125 mdash mdash mdash 3125 mdash mdash mdash mdashVetiveria zizanioides (L) Graminae Root (fraction) mdash 125 125 mdash mdash mdash gt250 gt250The bacterial growth inhibitory activity was compared for significant differences within the bacterial strains by broth-dilution method at 250 125 625 3125156 and 78 120583gmL F fractions

exposed to various compounds The cell proliferation wasincreased by the influence of the plant components at thelower concentrations Whereas the oil yielding plant com-pounds showed inhibition of cell proliferation and toxicity athigher doses 250ndash1000120583gmL

4 Discussion

The Western Ghats is considered as one of the richestbiodiversity hotspots in the world [41] In this survey wecollected nearly 78 medicinal plants from Western andEastern Ghats that are edible and popularly used for curingvarious ailments including snakebite Traditional remedieshave a long-standing history in many tribal settlements inTN India and they continue to provide useful and applicabletools for treating ailments [42] The ingredients that makeup the ldquoVishakallurdquo stone which is used as an antidote forsnakebite are different herbs and pebbles available from theriver banks Likewise aqueous paste and decoction obtainedfrom the leaves of A paniculata are widely used for snakebitetreatment by indigenous people [43] Previous studies havereported that ethnomedicine plays major roles in conservingthe disappearing knowledge of tribal communities [44ndash47]The traditional beliefs of reliance on a rich diversity ofethnomedicinal plants located at different settlements havealso been confirmed in another study [48 49] Herein weexplored the various types of traditional practices reported bythe primitive tribal communities with a view to gain furtherknowledge from such studies

In the present investigation potentially rich sources oftribal medicine (71 plants) were scientifically evaluated fortheir antibacterial activity against the MDR bacteria and theaccumulated data was disseminated for the first time to the

scientific community Out of the 71medicinal plants screenedfor the antibacterial activity 10 of them (T involucrataC acida A marmelos A vasica C procera A paniculataand M piperita A indica S indicus and E cardamomum)displayed the highest antibacterial activity against the mul-tidrug resistant B pseudomallei (KHW and TES) and Saureus strains The antibacterial activity of those crude plantextracts was as equally effective as that of the standard drugsOur findings corroborated with the previous reports madeon the antistaphylococcal activity of tribal medicinal plants[34 50 51] On the other hand isolated components from themost active extracts of T involucrata shellsol and C acidaexhibited the most potent action against the antibiotic resis-tant B pseudomallei (KHW) K pneumoniae and S aureusstrainsThese results further confirmed our previous findingson the leaves of T involucrata and its compounds hydrocar-bon ester-like shellsol which displayed a high antibacterialeffect against the different bacterial strains especially thatof S aureus [34] Eugenol and caryophyllene are the activeagents contained in the M piperita [52] and O sanctumplants which are believed to be mainly responsible for theantimicrobial properties of these plants [53] Interestinglythe compounds obtained from the aromatic plants such asC zeylanicum and R officinalis were also found to be veryeffective against the multidrug resistant human pathogen Kpneumoniae S aureus S typhi and B pseudomallei (KHW)that causes melioidosis

The inhibitory potential determined for the shellsol(T involucrata) vaseline (A vasica) C acida and C zey-lanicum indicates that the MIC of 78ndash3125120583gmL foundagainst the B pseudomallei of KHW K pneumoniae Kpneumoniae S pyogenes and V damsel and S pneumoniaewas quite low Similarly lower MIC values were found for

Evidence-Based Complementary and Alternative Medicine 15

M piperita (MIC of 113ndash225 mgmL) against the abovebacterial strains [52] MIC was found for the most activealcohol extracts of A salvifolium (MIC 0034ndash0263mgmL)on S aureus [54] and the MIC for S trilobatum aqueousextracts determined against the tested organisms rangedfrom 006 to 05mgmL [55] C zeylanicum was foundto have an effective antibacterial activity (MIC 64 120583gmL)against P aeruginosa E coli B subtilis and S aureus[56] Previously several investigators have demonstrated thatactive agents exert interesting activity against bacteria evenat lower concentrations tested [57 58] The bacteriostaticmechanism involves damage to the cell walls of bacteriafollowed by inhibition of protein synthesis that ultimatelyleads to bacterial death [59] The most active plants arewidely used by various tribes as traditional treatment (iecut wounds skin infection and scabies) thus indicatingthe potential for further development into promising drugsIn order to ascertain the safety and efficacy of the mostactive compounds their effect on human skin fibroblast cellswas evaluated Chemical constituents of E cardamomumT involucrata S indicus C acida A vasica A marmelosA indica and A paniculata plants failed to produce anynoticeable toxicity up to 1000120583gmL Although some ofthe tested compounds exhibited a slight reduction of cellproliferation and some minor morphological changes suchchanges were insignificant at lower doses and became evidentonly at higher doses However certain aromatic compoundsof O sanctum E globulus V zizanioides C citratus and Eglobulus plants showed reduction of cell proliferation againstHEPK cells

Our phytochemical screening also provides evidenceof the presence of several types of compounds that aremainly responsible for the remarkable antibacterial effectof these plants The differences noted for the bactericidalactivity of various plant extracts in this study appears tobe directly related to the diversity of compounds (shown inparentheses) that are accumulated in the following plants(eg A marmelos and O umbellate) [60ndash62] Compoundslike tannins phenol and polyphenols can bind the Gram-negative bacteria to form a heavy soluble complex on the cellsurface which subsequently disturbs the availability of recep-tor on cells and kills the bacteria [63] Several species havingwide spectra of antimicrobial activity mainly due to theactive constituents such as essential oil phenolic compoundslike thymol carvacrol in oregano and thyme eugenol inclove and cinnamon were also identified previously [64 65]Essential oils degrade the cell wall interact with the cell com-ponents and then disrupt the cytoplasmic membrane [66]The antimicrobial effect of phenolic compounds may involvemultiplemodes of action including damage to themembraneprotein interference with membrane integrated enzymes[67] causing leakage of cellular components coagulation ofcytoplasm depletion of the proton motive force alterationof fatty acid and phospholipid constituents impairmentof enzymatic mechanisms for production and metabolismalteration of nutrient uptake and electron transport [68]influencing the synthesis of DNA and RNA and destroyingprotein translation and the function of the mitochondrionin eukaryotes [69] The mode of action of antimicrobial

agents depends on the type of microorganism and is mainlyrelated to their cell wall structure and the outer membranearrangement Most of the plant spices and herbs containcomplex phenolics (ie phenolic acids flavonoids tanninslignans coumarins quinines) In addition the mechanismsof action of each phenolic compound against various bacteriaare also very complicated [26 70] Further investigation istherefore required to understand the relationship betweenthe antimicrobial action and the chemical structure of everyphenolic compound in the tested extracts The informationavailable from previous pharmacological sources combinedwith the findings herein reported on the medicinal plantextracts may serve as essential data for future drug develop-ment to combat diseases caused by theMDRbacterial strains

Abbreviations

MDR Multidrug resistant bacteriaTES Strain of Burkholderia pseudomalleiMRSA Methicillin-resistant Staphylococcus aureusVRE Vancomycin-resistant enterococciMTB Mycobacterium tuberculosisCH3OH Methanol

CFU Colony forming unitHCl Hydrochloric acidTLC Thin layer chromatographyH2SO4 Sulphuric acids

UV Ultraviolet spectrophotometerHEPK Human skin fibroblasts

Acknowledgments

Theauthors are thankful to the ERI Loyola College ChennaiIndia for his support during the survey The authors alsothank the Department of Microbiology NUHS NUS forthe bacterial cultures used in this investigation The authorsare thankful to the tribal people for sharing their healingknowledge on medicinal aromatic plants

References

[1] M N Alekshun and S B Levy ldquoMolecular mechanisms ofantibacterialmultidrug resistancerdquoCell vol 128 no 6 pp 1037ndash1050 2007

[2] A D Russell ldquoAntibiotic and biocide resistance in bacteriaintroductionrdquo Symposium Series Society for Applied Microbiol-ogy vol 31 no 92 pp 1Sndash3S 2002

[3] D R Guay ldquoTreatment of bacterial skin and skin structureinfectionsrdquo Expert Opinion on Pharmacotherapy vol 4 no 8pp 1259ndash1275 2003

[4] A J Brosnahan and P M Schlievert ldquoGram-positive bac-terial superantigen outside-in signaling causes toxic shocksyndromerdquo FEBS Journal vol 278 no 23 pp 4649ndash4667 2011

[5] K Fellermann J Wehkamp K R Herrlinger and E F StangeldquoCrohnrsquos disease a defensin deficiency syndromerdquo EuropeanJournal of Gastroenterology and Hepatology vol 15 no 6 pp627ndash634 2003

16 Evidence-Based Complementary and Alternative Medicine

[6] L Chen S Lin R Agha-Majzoub L Overbergh C Mathieuand L S Chan ldquoCCL27 is a critical factor for the developmentof atopic dermatitis in the keratin-14 IL-4 transgenic mousemodelrdquo International Immunology vol 18 no 8 pp 1233ndash12422006

[7] J H Song V Thamlikitkul and P R Hsueh ldquoClinical andeconomic burden of community-acquired pneumonia amongstadults in the Asia-Pacific regionrdquo International Journal ofAntimicrobial Agents vol 38 no 2 pp 108ndash117 2011

[8] D A B Dance ldquoMelioidosisrdquo Current Opinion in InfectiousDiseases vol 15 no 2 pp 127ndash132 2002

[9] N J White ldquoMelioidosisrdquo The Lancet vol 361 no 9370 pp1715ndash1722 2003

[10] S Yang ldquoMelioidosis research in Chinardquo Acta Tropica vol 77no 2 pp 157ndash165 2000

[11] P R Hsueh L J Teng L N Lee et al ldquoMelioidosis an emerginginfection in Taiwanrdquo Emerging Infectious Diseases vol 7 no 3pp 428ndash433 2001

[12] C Walsh and S Fanning ldquoAntimicrobial resistance in food-borne pathogensmdasha cause for concernrdquo Current Drug Targetsvol 9 no 9 pp 808ndash815 2008

[13] M Vorachit P Chongtrakool S Arkomsean and S BoonsongldquoAntimicrobial resistance in Burkholderia pseudomalleirdquo ActaTropica vol 74 no 2-3 pp 139ndash144 2000

[14] F M Thibault E Hernandez D R Vidal M Girardet and JD Cavallo ldquoAntibiotic susceptibility of 65 isolates of Burkholde-ria pseudomallei and Burkholderia mallei to 35 antimicrobialagentsrdquo Journal of Antimicrobial Chemotherapy vol 54 no 6pp 1134ndash1138 2004

[15] R L S Sikarwar ldquoEthnogynaecological uses of plants new toIndiardquo Ethnobotany vol 12 no 1-2 pp 112ndash115 2002

[16] R Siva ldquoStatus of natural dyes and dye-yielding plants in IndiardquoCurrent Science vol 92 no 7 pp 916ndash925 2007

[17] P Kumar Singh V Kumar R K Tiwari A Sharma ChV Rao and R H Singh ldquoMedico-ethnobotany of lsquochatararsquoblock of district sonebhadra Uttar Pradesh Indiardquo Advances inBiological Research vol 4 no 1 pp 65ndash80 2010

[18] W H Lewis and M P Elwin-Lewis Medical Botany PlantsAffecting Human Health John Wiley amp Sons New York NYUSA 2003

[19] R Perumal Samy P N Pushparaj and P GopalakrishnakoneldquoA compilation of bioactive compounds fromAyurvedardquo Bioin-formation vol 3 no 3 pp 100ndash110 2008

[20] WorldHealthOrganization (WHO)TraditionalMedicine Strat-egy 2002ndash2005 Genera World Health Organization GenevaSwitzerland 2002

[21] P S Haddad G A Azar S Groom and M Boivin ldquoNaturalhealth products modulation of immune function and preven-tion of chronic diseasesrdquo Evidence Based Complementary andAlternative Medicine vol 2 no 4 pp 513ndash520 2005

[22] K Poonam andG S Singh ldquoEthnobotanical study of medicinalplants used by the Taungya community in Terai Arc LandscapeIndiardquo Journal of Ethnopharmacology vol 123 no 1 pp 167ndash1762009

[23] G B Mahady ldquoMedicinal plants for the prevention and treat-ment of bacterial infectionsrdquo Current Pharmaceutical Designvol 11 no 19 pp 2405ndash2427 2005

[24] V P Kumar N S Chauhan H Padh and M Rajani ldquoSearchfor antibacterial and antifungal agents from selected Indianmedicinal plantsrdquo Journal of Ethnopharmacology vol 107 no2 pp 182ndash188 2006

[25] R Nair T Kalariya and S Chanda ldquoAntibacterial activity ofsome plant extracts used in folk medicinerdquo Journal of HerbalPharmacotherapy vol 7 no 3-4 pp 191ndash201 2007

[26] D Kalemba and A Kunicka ldquoAntibacterial and antifungalproperties of essential oilsrdquo Current Medicinal Chemistry vol10 no 10 pp 813ndash829 2003

[27] A H Ashour ldquoAntibacterial antifungal and anticancer activi-ties of volatile oils and extracts from stems leaves and flowers ofEucalyptus sideroxylon and Eucalyptus torquatardquoCancer BiologyandTherapy vol 7 no 3 pp 399ndash403 2008

[28] M J Abad L M Bedoya L Apaza and P Bermejo ldquoTheArtemisia L genus a review of bioactive essential oilsrdquoMolecules vol 17 no 3 pp 2542ndash2566 2012

[29] H N Marzoug M Romdhane A Lebrihi et al ldquoEucalyptusoleosa essential oils chemical composition and antimicrobialand antioxidant activities of the oils from different plant parts(stems leaves flowers and fruits)rdquoMolecules vol 16 no 2 pp1695ndash1709 2011

[30] C Proestos D Sereli and M Komaitis ldquoDetermination ofphenolic compounds in aromatic plants by RP-HPLC and GC-MSrdquo Food Chemistry vol 95 no 1 pp 44ndash52 2006

[31] S Chao G Young C Oberg and K Nakaoka ldquoInhibition ofmethicillin-resistant Staphylococcus aureus (MRSA) by essentialoilsrdquo Flavour and Fragrance Journal vol 23 no 6 pp 444ndash4492008

[32] M Akin D Oguz and H T Saracoglu ldquoAntibacterial effectsof some plant extracts from Labiatae (Lamiaceae) growingnaturally around Sirnak-Silopi Turkeyrdquo International Journal ofPharmaceutical and Applied Sciences vol 1 no 1 pp 4ndash47 2010

[33] K Bayoub T Baibai D Mountassif A Retmane and ASoukri ldquoAntibacterial activities of the crude ethanol extracts ofmedicinal plants against Listeriamonocytogenes and some otherpathogenic strainsrdquo African Journal of Biotechnology vol 9 no27 pp 4251ndash4258 2010

[34] R P Samy P Gopalakrishnakone M Sarumathi P Houghtonand S Ignacimuthu ldquoPurification of antibacterial agents fromTragia involucratamdasha popular tribal medicine for wound heal-ingrdquo Journal of Ethnopharmacology vol 107 no 1 pp 99ndash1062006

[35] K M Matthew The Flora of Tamil Nadu Carnatic Part IndashIIIprinted at the Diocesan Press Madras India 1981ndash1983

[36] AW BauerWMKirby J C Sherris andM Turck ldquoAntibioticsusceptibility testing by a standardized single disk methodrdquoAmerican Journal of Clinical Pathology vol 45 no 4 pp 493ndash496 1966

[37] National Committee for Clinical Laboratory Standards Per-formance Standards For Antimicrobial Susceptibility Testing12th Informational Supplement Approved standard M100-S12National Committee for Clinical Laboratory Standards WaynePa USA 2002

[38] E A Elsinghorst ldquoMeasurement of invasion by gentamicinresistancerdquoMethods in Enzymology vol 236 pp 405ndash420 1994

[39] J B Harborne Phytochemical Methods Chapman and HallNew York NY USA 1976

[40] H Wagner S Bladt and E M Zgairski Plant Drug AnalysisSpringer New York NY USA 1984

[41] N Myers R A Mittermeler C G Mittermeler G A B daFonseca and J Kent ldquoBiodiversity hotspots for conservationprioritiesrdquo Nature vol 403 no 6772 pp 853ndash858 2000

[42] R Perumal Samy and S Ignacimuthu ldquoAntibacterial activity ofsome folkloremedicinal plants used by tribals inWestern Ghats

Evidence-Based Complementary and Alternative Medicine 17

of Indiardquo Journal of Ethnopharmacology vol 69 no 1 pp 63ndash712000

[43] K Nampoothiri A Chrispal A Begum S Jasmine K GGopinath and A Zachariah ldquoA clinical study of renal tubulardysfunction inCleistanthus collinus (Oduvanthalai) poisoningrdquoClinical Toxicology vol 48 no 3 pp 193ndash197 2010

[44] M Strathern UsefulKnowledge The Isiah Berlin Lecture Uni-versity of Manchester Brunswick UK

[45] C N Sunil and A K Pradeep ldquo(2001) Another new species ofTripogon (Poaceae) from Indiardquo Sida vol 19 pp 803ndash806 2005

[46] S Biber-Klemm and T Cottier Right To Plant GeneticResources and Traditional Knowledge Basic Issues and Prospec-tive Oxfordshire Nosworthy Way UK 2006

[47] V S Ramachandran S Shijo Joseph and R Aruna ldquoEth-nobotanical studies from amaravathy range of Indira GandhiWildlife SanctuaryWesternGhats CoimbatoreDistrict South-ern Indiardquo Ethnobotanical Leaflets vol 13 pp 1069ndash1087 2009

[48] A H Rajasab and I Mohamad ldquoDocumentation of folkknowledge on edible wild plants of North Karnatakardquo IndianJournal of Traditional Knowledge vol 3 no 4 pp 419ndash4292004

[49] M Rajadurai V G Vidhya M Ramya and A Bhaskar ldquoEthno-medicinal plants used by the traditional healers of pachamalaihills Tamilnadu Indiardquo Studies on Ethno-Medicine vol 3 no 1pp 39ndash41 2009

[50] S K Adesina O Idowu A O Ogundaini et al ldquoAntimicrobialconstituents of the leaves of Acalypha wilkesiana and Acalyphahispidardquo Phytotherapy Research vol 14 no 5 pp 371ndash374 2000

[51] M T Gutierrez-Lugo M P Singh W M Maiese and BN Timmermann ldquoNew antimicrobial cycloartane triterpenesfrom Acalypha communisrdquo Journal of Natural Products vol 65no 6 pp 872ndash875 2002

[52] A K Tyagi andAMalik ldquoAntimicrobial potential and chemicalcomposition of Mentha piperita oil in liquid and vapour phaseagainst food spoiling microorganismsrdquo Food Control vol 22no 11 pp 1707ndash1714 2011

[53] G Devendran and U Balasubramanian ldquoQualitative phyto-chemical screening and GC-MS analysis of Ocimum sanctumL leavesrdquoAsian Journal of Plant Science and Research vol 1 no4 pp 44ndash48 2011

[54] V C Jain N M Patel D P Shah P K Patel and B HJoshi ldquoAntioxidant and antimicrobial activities of Alangiumsalvifolium (Lf) Wang rootrdquo Global Journal of Pharmacologyvol 4 no 1 pp 13ndash18 2010

[55] P Swapna Latha andK Kannabiran ldquoAntimicrobial activity andphytochemicals of Solanum trilobatum LinnrdquoAfrican Journal ofBiotechnology vol 5 no 23 pp 2402ndash2404 2006

[56] M Usha1 S Ragini and S M A Naqvi ldquoAntibacterial activityof acetone and ethanol extracts of Cinnamon (Cinnamomumzeylanicum) and Ajowan (Trachyspermum ammi) on four foodspoilage bacteriardquo International Research Journal of BiologicalSciences vol 1 no 4 pp 7ndash11 2012

[57] S Prabuseenivasan M Jayakumar and S Ignacimuthu ldquoInvitro antibacterial activity of some plant essential oilsrdquo BMCComplementary and Alternative Medicine vol 6 article 39 pp1ndash8 2006

[58] G A Ayoola F M Lawore T Adelowotan et al ldquoChemi-cal analysis and antimicrobial activity of the essential oil ofSyzigium aromaticum (clove)rdquo African Journal of Biotechnologyvol 7 no 13 pp 2227ndash2231 2008

[59] Y J Fu L Chen Y Zu et al ldquoThe antibacterial activity of cloveessential oil against Propionibacterium acnes and its mechanismof actionrdquo Archives of Dermatology vol 145 no 1 pp 86ndash882009

[60] D Venkatesan C M Karrunakarn S Selva Kumar and P TPalani Swamy ldquoIdentification of phytochemical constituents ofAegle marmelos responsible for antimicrobial activity againstselected pathogenic organismsrdquo Ethnobotanical Leaflets vol 13pp 1362ndash1372 2009

[61] P Arun K G Purushotham J Johnsy jayarani and V KumarildquoIn vitro Antibacterial Activity of Oldenlandia umbellata anIndian medicinal Plantrdquo Journal of Pharmaceutical Science andTechnology vol 2 no 4 pp 198ndash201 2010

[62] A Manjamalai R Sardar Sathyajith Singh C Guruvayoorap-pan and V M Berlin Grace ldquoAnalysis of phytochemicalconstituents and antimicrobial activity of some medicinalplants in Taminadu Indiardquo Global Journal of Biotechnology andBiochemistry vol 5 no 2 pp 120ndash128 2010

[63] E Haslam ldquoNatural polyphenols (vegetable tannins) as drugsPossible modes of actionrdquo Journal of Natural Products vol 59no 2 pp 205ndash215 1996

[64] Y Cai Q Luo M Sun and H Corke ldquoAntioxidant activityand phenolic compounds of 112 traditional Chinese medicinalplants associated with anticancerrdquo Life Sciences vol 74 no 17pp 2157ndash2184 2004

[65] B Shan Y Z Cai M Sun and H Corke ldquoAntioxidant capacityof 26 spice extracts and characterization of their phenolicconstituentsrdquo Journal of Agricultural and Food Chemistry vol53 no 20 pp 7749ndash7759 2005

[66] R J W Lambert P N Skandamis P J Coote and G-J ENychas ldquoA study of the minimum inhibitory concentration andmode of action of oregano essential oil thymol and carvacrolrdquoJournal of AppliedMicrobiology vol 91 no 3 pp 453ndash462 2001

[67] M Raccach ldquoThe antimicrobial activity of phenolic antioxi-dants in food a reviewrdquo Journal of Food Safety vol 6 no 3 pp141ndash170 1984

[68] M Taniguchi Y Yano E Tada et al ldquoMode of action of polygo-dial an antifungal sesquiterpene dialdehyderdquo Agricultural andBiological Chemistry vol 52 no 6 pp 1409ndash1414 1988

[69] G J E Nychas ldquoNatural antimicrobials from plantsrdquo in NewMethods of Food Preservation G W Gould Ed pp 58ndash89Blackie Academic London UK 1995

[70] S Burt ldquoEssential oils their antibacterial properties and poten-tial applications in foodsmdasha reviewrdquo International Journal ofFood Microbiology vol 94 no 3 pp 223ndash253 2004

Submit your manuscripts athttpwwwhindawicom

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Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

MEDIATORSINFLAMMATION

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Behavioural Neurology

EndocrinologyInternational Journal of

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Disease Markers

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OncologyJournal of

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Oxidative Medicine and Cellular Longevity

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PPAR Research

The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014

Immunology ResearchHindawi Publishing Corporationhttpwwwhindawicom Volume 2014

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Parkinsonrsquos Disease

Evidence-Based Complementary and Alternative Medicine

Volume 2014Hindawi Publishing Corporationhttpwwwhindawicom

12 Evidence-Based Complementary and Alternative Medicine

(mean plusmn SD n = 3)

0

5

10

15

20

25

30

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn V

d St

Bacteria (+minus)

lowast

lowast

lowast

A vasica

In vitro assay(P

lant

frac

tion)

Inhi

bitio

n zo

nes (

mm

)

(a)

(mean plusmn SD n = 3)

0

5

10

15

20

25

30

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn V

d St

Bacteria (+minus)

lowast

A marmelos

Inhi

bitio

n zo

nes (

mm

)

In vitro assay

(b)

(mean plusmn SD n = 3)

0

5

10

15

20

25

30

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn V

d St

Bacteria (+minus)

lowastlowast

A indica

Inhi

bitio

n zo

nes (

mm

)

Incu

batio

n (37∘C

24 h

)

(c)

(mean plusmn SD n = 3)

0

5

10

15

20

25

30

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn V

d St

Bacteria (+minus)

lowast

lowast lowast

35

C acida

Inhi

bitio

n zo

nes (

mm

)(P

lant

frac

tion)

(d)

(mean plusmn SD n = 3)

0

5

10

15

20

25

30

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn V

d St

Bacteria (+minus)

lowastlowast

lowast

S chamaelea

Inhi

bitio

n zo

nes (

mm

)

(e)

(mean plusmn SD n = 3)

0

5

10

15

20

25

30

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn V

d St

Bacteria (+minus)

lowastlowast

lowastlowast

S indicus

Inhi

bitio

n zo

nes (

mm

)

Incu

batio

n (37∘C

24 h

)

(f)

(mean plusmn SD n = 3)

0

5

10

15

20

25

30

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn V

d St

Bacteria (+minus)

lowast

lowast lowast

E cardamomum

Inhi

bitio

n zo

nes (

mm

)(P

lant

frac

tion)

(g)

(mean plusmn SD n = 3)

0

5

10

15

20

25

30

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn V

d St

Bacteria (+minus)

lowast

lowast

lowast

lowast

Shellsol

35

Inhi

bitio

n zo

nes (

mm

)

Incu

batio

n (37∘C

24 h

)

(h)

Figure 4 In vitro antimicrobial activity of purified fractions from the most active plant extracts tested against bacteria Growth inhibitionzones were measured and analyzed with mean plusmn standard deviation (SD) (119899 = 3) using one way analysis of variance Level of significanceat (lowast119875 gt 001) Most of the fractions exerted a potent inhibitory effect against multidrug resistant Gram-negative bacteria (B pseudomalleistrains KHW and TES) K pneumonia and Gram-positive bacteria S aureus

(T involucrata) andC acida showed an interesting inhibitorypotential against S aureus (MIC of 78 120583gmL) and B pseu-domallei strain of KHW (MIC of 156 120583gmL) A vasicashowed an MIC of 156120583gmL against B pseudomallei(KHW) and an MIC of 3125120583gmL against K pneumo-niae K pneumoniae S pyogenes and V damsela strainsFractions from A marmelos and terpenoid from A indicaexerted bacteriostatic effect with MIC values of 3125120583gmLon some selected bacteria including B pseudomallei ofKHW S aureus and B pseudomallei of TES The MIC of3125 120583gmL was found for E cardamomum against S aureus

K pneumonia and S pyogenes S indicus displayed a verystrong inhibition against MDR K pneumoniae (MIC of156 120583gmL) and against B pseudomallei (KHW and TES)at MIC of 3125 120583gmL When the antimicrobial efficacies ofpurified fractions from aromatic plants were compared theC zeylanicum fraction displayed an important antimicrobialeffect against S aureus (MIC of 78 120583gmL) MDR B pseu-domallei of KHW (MIC of 156 120583gmL) and S pneumoniae(MIC of 3125 120583gmL) The essential oil from M piperateashowed MIC value of 3125120583gmL against K pneumoniae Saureus and B pseudomallei (KHW) respectivelyO sanctum

Evidence-Based Complementary and Alternative Medicine 13

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn

Bacteria (+minus)

0

5

10

15

20

25

30

Inhi

bitio

n zo

nes (

mm

)(P

lant

frac

tion)

lowastlowast

lowast

(mean plusmn SD n = 3)In vitro assay

C citratus

(a)

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn

Bacteria (+minus)

0

5

10

15

20

25

30

Inhi

bitio

n zo

nes (

mm

)

lowast

lowast

lowastlowast

lowast

lowast

(mean plusmn SD n = 3)In vitro assay

35

C zeylanicum

(b)

Bacteria (+minus)

0

5

10

15

20

25

30

Inhi

bitio

n zo

nes (

mm

)

lowast

lowast

lowast

(mean plusmn SD n = 3)

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn V

d St

O sanctum

Incu

batio

n (37∘C

24 h

)

(c)

Bacteria (+minus)

0

5

10

15

20

25

Inhi

bitio

n zo

nes (

mm

)

lowast

lowast

(mean plusmn SD n = 3)

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn V

d St

M piperita

(Pla

nt fr

actio

n)

(d)

Bacteria (+minus)

0

5

10

15

20

25

30

Inhi

bitio

n zo

nes (

mm

)lowast

lowast

lowast

(mean plusmn SD n = 3)

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn V

d St

E caryophyllus

(e)

0

5

10

15

20

25

30

Inhi

bitio

n zo

nes (

mm

) (mean plusmn SD n = 3)

Bacteria (+minus)

KHW

TES

Kp

KprPa Sa

Stp

Spn

E globulus

Incu

batio

n (37∘C

24 h

)

(f)

Bacteria (+minus)

0

5

10

15

20

25

30

Inhi

bitio

n zo

nes (

mm

)(P

lant

frac

tion)

lowast

lowast

lowast

lowast

(mean plusmn SD n = 3)

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn V

d

R officinalis

(g)

0

5

10

15

20

25

30

Inhi

bitio

n zo

nes (

mm

)

lowast

lowast

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn V

d St

Bacteria (+minus)V zizanioides

(mean plusmn SD n = 3)

Incu

batio

n (37∘C

24 h

)

(h)

Figure 5 Comparison of antimicrobial effect of plant compounds obtained from the most popularly used sources of essential oils as assayedby the disc-diffusion method in vitro It displayed a powerful activity against B pseudomallei and S aureus than the other bacteria strainsOther compounds showed only a moderate or weak action against the tested bacteria Values for zone of bacterial growth inhibition werepresented as mean plusmn SD (119899 = 3) with level of significance at (lowast119875 gt 001)

and C citratus fractions also showed antimicrobial activity(MICs of 3125ndash125120583gmL) only at higher concentrationsagainst the tested bacteria In addition to that higher concen-trations (gt250120583gmL) (of Vetiveria fractions) were requiredto inhibit Vibro species and others (including E globulusfractions) failed to show any effect at tested concentrations(78ndash125 120583gmL) However the purified fractions (from mostactive medicinal plants) showed strong bacteriostatic inhibi-tion against the tested organisms (Table 3)

310 Cytotoxic Effects of Plants When the components wereassayed for cytotoxicity against the normal human skinfibroblasts (HEPK) cells the compounds obtained from

E cardamomum T involucrata S indicusC acidaA vasicaA marmelos A indica and A paniculata did not showtoxicity up to 1000120583gmL (see Figures S1 and S2 in Supple-mentaryMaterial available online at httpdxdoiorg1011552013525613) A slight reduction of cell proliferation wasnoted only at higher doses (2000120583gmL) In contrast cellproliferation was markedly reduced after exposure of HEPKcells to O sanctum E globulus V zizanioides C citratusand E globulus compounds There was no gradual reductionin skin cell proliferation seen after exposure to C zeylan-icum R officinalis and M piperita (see Figures S3 and S4)compounds The toxicity was found to be concentration-dependent when the skin fibroblasts (HEPK) cells were

14 Evidence-Based Complementary and Alternative Medicine

Table 3 Minimum inhibitory concentrations (MICs) of purified plant fractions and essential oils against antibiotic resistant bacteria

Botanical name Family Parts used Gram-positive and -negative bacteria (MICs 120583gmL)KHW TES Kp KPr Sa Stp Sp Vd St

A indica Juss Meliaceae Seed (fraction) 3125 625 125 lt250 3125 mdash mdash mdash mdashA marmelos (L) Rutaceae Root-bark (F) 3125 3125 mdash mdash 3125 625 125 250 mdashA paniculata Nees Acanthaceae Leaf (fraction) 250 mdash 125 mdash 625 mdash mdash 125 mdashA vasica Nees Acanthaceae Fraction (Stem) 156 3125 3125 625 mdash lt250 3125 3125 mdashC acida Roxb Rutaceae Leaf (fraction) 156 625 mdash mdash 78 mdash 3125 mdash mdashE cardamomumWhite et Mason Acanthaceae Fraction (WP) 625 mdash 3125 mdash 3125 250 625 mdash mdashS indicus (L) Euphorbiaceae Whole plant (F) 3125 125 156 mdash 625 mdash mdash lt250 mdashT involucrata (L) Euphorbiaceae Shellosol (leaf) 156 3125 mdash mdash 78 625 mdash mdash mdashCinnamomum zeylanicum (L) Lauraceae Bark (fraction) 156 625 mdash mdash 78 3125 mdash mdash 125Cymbopogon citratus (L) Graminae Leaf (fraction) 250 lt250 625 mdash 625 125 mdash mdash mdashEugenia caryophyllus (L) Myrtaceae Flower buds (F) 625 125 mdash mdash 625 mdash mdash mdash mdashEucalyptus globulus (L) Myrtaceae Fraction (leaf) mdash mdash mdash mdash mdash mdash lt250 mdash lt250Mentha piperita (L) Labiatae Fraction (WP) mdash 625 3125 mdash 3125 mdash lt250 mdash mdashOcimum sanctum (L) Labiatae Leaf (fraction) 625 125 mdash 3125 625 mdash 125 lt250 mdashRosmarinus officinalis (L) Labiatae Rosemary oil 3125 mdash mdash mdash 3125 mdash mdash mdash mdashVetiveria zizanioides (L) Graminae Root (fraction) mdash 125 125 mdash mdash mdash gt250 gt250The bacterial growth inhibitory activity was compared for significant differences within the bacterial strains by broth-dilution method at 250 125 625 3125156 and 78 120583gmL F fractions

exposed to various compounds The cell proliferation wasincreased by the influence of the plant components at thelower concentrations Whereas the oil yielding plant com-pounds showed inhibition of cell proliferation and toxicity athigher doses 250ndash1000120583gmL

4 Discussion

The Western Ghats is considered as one of the richestbiodiversity hotspots in the world [41] In this survey wecollected nearly 78 medicinal plants from Western andEastern Ghats that are edible and popularly used for curingvarious ailments including snakebite Traditional remedieshave a long-standing history in many tribal settlements inTN India and they continue to provide useful and applicabletools for treating ailments [42] The ingredients that makeup the ldquoVishakallurdquo stone which is used as an antidote forsnakebite are different herbs and pebbles available from theriver banks Likewise aqueous paste and decoction obtainedfrom the leaves of A paniculata are widely used for snakebitetreatment by indigenous people [43] Previous studies havereported that ethnomedicine plays major roles in conservingthe disappearing knowledge of tribal communities [44ndash47]The traditional beliefs of reliance on a rich diversity ofethnomedicinal plants located at different settlements havealso been confirmed in another study [48 49] Herein weexplored the various types of traditional practices reported bythe primitive tribal communities with a view to gain furtherknowledge from such studies

In the present investigation potentially rich sources oftribal medicine (71 plants) were scientifically evaluated fortheir antibacterial activity against the MDR bacteria and theaccumulated data was disseminated for the first time to the

scientific community Out of the 71medicinal plants screenedfor the antibacterial activity 10 of them (T involucrataC acida A marmelos A vasica C procera A paniculataand M piperita A indica S indicus and E cardamomum)displayed the highest antibacterial activity against the mul-tidrug resistant B pseudomallei (KHW and TES) and Saureus strains The antibacterial activity of those crude plantextracts was as equally effective as that of the standard drugsOur findings corroborated with the previous reports madeon the antistaphylococcal activity of tribal medicinal plants[34 50 51] On the other hand isolated components from themost active extracts of T involucrata shellsol and C acidaexhibited the most potent action against the antibiotic resis-tant B pseudomallei (KHW) K pneumoniae and S aureusstrainsThese results further confirmed our previous findingson the leaves of T involucrata and its compounds hydrocar-bon ester-like shellsol which displayed a high antibacterialeffect against the different bacterial strains especially thatof S aureus [34] Eugenol and caryophyllene are the activeagents contained in the M piperita [52] and O sanctumplants which are believed to be mainly responsible for theantimicrobial properties of these plants [53] Interestinglythe compounds obtained from the aromatic plants such asC zeylanicum and R officinalis were also found to be veryeffective against the multidrug resistant human pathogen Kpneumoniae S aureus S typhi and B pseudomallei (KHW)that causes melioidosis

The inhibitory potential determined for the shellsol(T involucrata) vaseline (A vasica) C acida and C zey-lanicum indicates that the MIC of 78ndash3125120583gmL foundagainst the B pseudomallei of KHW K pneumoniae Kpneumoniae S pyogenes and V damsel and S pneumoniaewas quite low Similarly lower MIC values were found for

Evidence-Based Complementary and Alternative Medicine 15

M piperita (MIC of 113ndash225 mgmL) against the abovebacterial strains [52] MIC was found for the most activealcohol extracts of A salvifolium (MIC 0034ndash0263mgmL)on S aureus [54] and the MIC for S trilobatum aqueousextracts determined against the tested organisms rangedfrom 006 to 05mgmL [55] C zeylanicum was foundto have an effective antibacterial activity (MIC 64 120583gmL)against P aeruginosa E coli B subtilis and S aureus[56] Previously several investigators have demonstrated thatactive agents exert interesting activity against bacteria evenat lower concentrations tested [57 58] The bacteriostaticmechanism involves damage to the cell walls of bacteriafollowed by inhibition of protein synthesis that ultimatelyleads to bacterial death [59] The most active plants arewidely used by various tribes as traditional treatment (iecut wounds skin infection and scabies) thus indicatingthe potential for further development into promising drugsIn order to ascertain the safety and efficacy of the mostactive compounds their effect on human skin fibroblast cellswas evaluated Chemical constituents of E cardamomumT involucrata S indicus C acida A vasica A marmelosA indica and A paniculata plants failed to produce anynoticeable toxicity up to 1000120583gmL Although some ofthe tested compounds exhibited a slight reduction of cellproliferation and some minor morphological changes suchchanges were insignificant at lower doses and became evidentonly at higher doses However certain aromatic compoundsof O sanctum E globulus V zizanioides C citratus and Eglobulus plants showed reduction of cell proliferation againstHEPK cells

Our phytochemical screening also provides evidenceof the presence of several types of compounds that aremainly responsible for the remarkable antibacterial effectof these plants The differences noted for the bactericidalactivity of various plant extracts in this study appears tobe directly related to the diversity of compounds (shown inparentheses) that are accumulated in the following plants(eg A marmelos and O umbellate) [60ndash62] Compoundslike tannins phenol and polyphenols can bind the Gram-negative bacteria to form a heavy soluble complex on the cellsurface which subsequently disturbs the availability of recep-tor on cells and kills the bacteria [63] Several species havingwide spectra of antimicrobial activity mainly due to theactive constituents such as essential oil phenolic compoundslike thymol carvacrol in oregano and thyme eugenol inclove and cinnamon were also identified previously [64 65]Essential oils degrade the cell wall interact with the cell com-ponents and then disrupt the cytoplasmic membrane [66]The antimicrobial effect of phenolic compounds may involvemultiplemodes of action including damage to themembraneprotein interference with membrane integrated enzymes[67] causing leakage of cellular components coagulation ofcytoplasm depletion of the proton motive force alterationof fatty acid and phospholipid constituents impairmentof enzymatic mechanisms for production and metabolismalteration of nutrient uptake and electron transport [68]influencing the synthesis of DNA and RNA and destroyingprotein translation and the function of the mitochondrionin eukaryotes [69] The mode of action of antimicrobial

agents depends on the type of microorganism and is mainlyrelated to their cell wall structure and the outer membranearrangement Most of the plant spices and herbs containcomplex phenolics (ie phenolic acids flavonoids tanninslignans coumarins quinines) In addition the mechanismsof action of each phenolic compound against various bacteriaare also very complicated [26 70] Further investigation istherefore required to understand the relationship betweenthe antimicrobial action and the chemical structure of everyphenolic compound in the tested extracts The informationavailable from previous pharmacological sources combinedwith the findings herein reported on the medicinal plantextracts may serve as essential data for future drug develop-ment to combat diseases caused by theMDRbacterial strains

Abbreviations

MDR Multidrug resistant bacteriaTES Strain of Burkholderia pseudomalleiMRSA Methicillin-resistant Staphylococcus aureusVRE Vancomycin-resistant enterococciMTB Mycobacterium tuberculosisCH3OH Methanol

CFU Colony forming unitHCl Hydrochloric acidTLC Thin layer chromatographyH2SO4 Sulphuric acids

UV Ultraviolet spectrophotometerHEPK Human skin fibroblasts

Acknowledgments

Theauthors are thankful to the ERI Loyola College ChennaiIndia for his support during the survey The authors alsothank the Department of Microbiology NUHS NUS forthe bacterial cultures used in this investigation The authorsare thankful to the tribal people for sharing their healingknowledge on medicinal aromatic plants

References

[1] M N Alekshun and S B Levy ldquoMolecular mechanisms ofantibacterialmultidrug resistancerdquoCell vol 128 no 6 pp 1037ndash1050 2007

[2] A D Russell ldquoAntibiotic and biocide resistance in bacteriaintroductionrdquo Symposium Series Society for Applied Microbiol-ogy vol 31 no 92 pp 1Sndash3S 2002

[3] D R Guay ldquoTreatment of bacterial skin and skin structureinfectionsrdquo Expert Opinion on Pharmacotherapy vol 4 no 8pp 1259ndash1275 2003

[4] A J Brosnahan and P M Schlievert ldquoGram-positive bac-terial superantigen outside-in signaling causes toxic shocksyndromerdquo FEBS Journal vol 278 no 23 pp 4649ndash4667 2011

[5] K Fellermann J Wehkamp K R Herrlinger and E F StangeldquoCrohnrsquos disease a defensin deficiency syndromerdquo EuropeanJournal of Gastroenterology and Hepatology vol 15 no 6 pp627ndash634 2003

16 Evidence-Based Complementary and Alternative Medicine

[6] L Chen S Lin R Agha-Majzoub L Overbergh C Mathieuand L S Chan ldquoCCL27 is a critical factor for the developmentof atopic dermatitis in the keratin-14 IL-4 transgenic mousemodelrdquo International Immunology vol 18 no 8 pp 1233ndash12422006

[7] J H Song V Thamlikitkul and P R Hsueh ldquoClinical andeconomic burden of community-acquired pneumonia amongstadults in the Asia-Pacific regionrdquo International Journal ofAntimicrobial Agents vol 38 no 2 pp 108ndash117 2011

[8] D A B Dance ldquoMelioidosisrdquo Current Opinion in InfectiousDiseases vol 15 no 2 pp 127ndash132 2002

[9] N J White ldquoMelioidosisrdquo The Lancet vol 361 no 9370 pp1715ndash1722 2003

[10] S Yang ldquoMelioidosis research in Chinardquo Acta Tropica vol 77no 2 pp 157ndash165 2000

[11] P R Hsueh L J Teng L N Lee et al ldquoMelioidosis an emerginginfection in Taiwanrdquo Emerging Infectious Diseases vol 7 no 3pp 428ndash433 2001

[12] C Walsh and S Fanning ldquoAntimicrobial resistance in food-borne pathogensmdasha cause for concernrdquo Current Drug Targetsvol 9 no 9 pp 808ndash815 2008

[13] M Vorachit P Chongtrakool S Arkomsean and S BoonsongldquoAntimicrobial resistance in Burkholderia pseudomalleirdquo ActaTropica vol 74 no 2-3 pp 139ndash144 2000

[14] F M Thibault E Hernandez D R Vidal M Girardet and JD Cavallo ldquoAntibiotic susceptibility of 65 isolates of Burkholde-ria pseudomallei and Burkholderia mallei to 35 antimicrobialagentsrdquo Journal of Antimicrobial Chemotherapy vol 54 no 6pp 1134ndash1138 2004

[15] R L S Sikarwar ldquoEthnogynaecological uses of plants new toIndiardquo Ethnobotany vol 12 no 1-2 pp 112ndash115 2002

[16] R Siva ldquoStatus of natural dyes and dye-yielding plants in IndiardquoCurrent Science vol 92 no 7 pp 916ndash925 2007

[17] P Kumar Singh V Kumar R K Tiwari A Sharma ChV Rao and R H Singh ldquoMedico-ethnobotany of lsquochatararsquoblock of district sonebhadra Uttar Pradesh Indiardquo Advances inBiological Research vol 4 no 1 pp 65ndash80 2010

[18] W H Lewis and M P Elwin-Lewis Medical Botany PlantsAffecting Human Health John Wiley amp Sons New York NYUSA 2003

[19] R Perumal Samy P N Pushparaj and P GopalakrishnakoneldquoA compilation of bioactive compounds fromAyurvedardquo Bioin-formation vol 3 no 3 pp 100ndash110 2008

[20] WorldHealthOrganization (WHO)TraditionalMedicine Strat-egy 2002ndash2005 Genera World Health Organization GenevaSwitzerland 2002

[21] P S Haddad G A Azar S Groom and M Boivin ldquoNaturalhealth products modulation of immune function and preven-tion of chronic diseasesrdquo Evidence Based Complementary andAlternative Medicine vol 2 no 4 pp 513ndash520 2005

[22] K Poonam andG S Singh ldquoEthnobotanical study of medicinalplants used by the Taungya community in Terai Arc LandscapeIndiardquo Journal of Ethnopharmacology vol 123 no 1 pp 167ndash1762009

[23] G B Mahady ldquoMedicinal plants for the prevention and treat-ment of bacterial infectionsrdquo Current Pharmaceutical Designvol 11 no 19 pp 2405ndash2427 2005

[24] V P Kumar N S Chauhan H Padh and M Rajani ldquoSearchfor antibacterial and antifungal agents from selected Indianmedicinal plantsrdquo Journal of Ethnopharmacology vol 107 no2 pp 182ndash188 2006

[25] R Nair T Kalariya and S Chanda ldquoAntibacterial activity ofsome plant extracts used in folk medicinerdquo Journal of HerbalPharmacotherapy vol 7 no 3-4 pp 191ndash201 2007

[26] D Kalemba and A Kunicka ldquoAntibacterial and antifungalproperties of essential oilsrdquo Current Medicinal Chemistry vol10 no 10 pp 813ndash829 2003

[27] A H Ashour ldquoAntibacterial antifungal and anticancer activi-ties of volatile oils and extracts from stems leaves and flowers ofEucalyptus sideroxylon and Eucalyptus torquatardquoCancer BiologyandTherapy vol 7 no 3 pp 399ndash403 2008

[28] M J Abad L M Bedoya L Apaza and P Bermejo ldquoTheArtemisia L genus a review of bioactive essential oilsrdquoMolecules vol 17 no 3 pp 2542ndash2566 2012

[29] H N Marzoug M Romdhane A Lebrihi et al ldquoEucalyptusoleosa essential oils chemical composition and antimicrobialand antioxidant activities of the oils from different plant parts(stems leaves flowers and fruits)rdquoMolecules vol 16 no 2 pp1695ndash1709 2011

[30] C Proestos D Sereli and M Komaitis ldquoDetermination ofphenolic compounds in aromatic plants by RP-HPLC and GC-MSrdquo Food Chemistry vol 95 no 1 pp 44ndash52 2006

[31] S Chao G Young C Oberg and K Nakaoka ldquoInhibition ofmethicillin-resistant Staphylococcus aureus (MRSA) by essentialoilsrdquo Flavour and Fragrance Journal vol 23 no 6 pp 444ndash4492008

[32] M Akin D Oguz and H T Saracoglu ldquoAntibacterial effectsof some plant extracts from Labiatae (Lamiaceae) growingnaturally around Sirnak-Silopi Turkeyrdquo International Journal ofPharmaceutical and Applied Sciences vol 1 no 1 pp 4ndash47 2010

[33] K Bayoub T Baibai D Mountassif A Retmane and ASoukri ldquoAntibacterial activities of the crude ethanol extracts ofmedicinal plants against Listeriamonocytogenes and some otherpathogenic strainsrdquo African Journal of Biotechnology vol 9 no27 pp 4251ndash4258 2010

[34] R P Samy P Gopalakrishnakone M Sarumathi P Houghtonand S Ignacimuthu ldquoPurification of antibacterial agents fromTragia involucratamdasha popular tribal medicine for wound heal-ingrdquo Journal of Ethnopharmacology vol 107 no 1 pp 99ndash1062006

[35] K M Matthew The Flora of Tamil Nadu Carnatic Part IndashIIIprinted at the Diocesan Press Madras India 1981ndash1983

[36] AW BauerWMKirby J C Sherris andM Turck ldquoAntibioticsusceptibility testing by a standardized single disk methodrdquoAmerican Journal of Clinical Pathology vol 45 no 4 pp 493ndash496 1966

[37] National Committee for Clinical Laboratory Standards Per-formance Standards For Antimicrobial Susceptibility Testing12th Informational Supplement Approved standard M100-S12National Committee for Clinical Laboratory Standards WaynePa USA 2002

[38] E A Elsinghorst ldquoMeasurement of invasion by gentamicinresistancerdquoMethods in Enzymology vol 236 pp 405ndash420 1994

[39] J B Harborne Phytochemical Methods Chapman and HallNew York NY USA 1976

[40] H Wagner S Bladt and E M Zgairski Plant Drug AnalysisSpringer New York NY USA 1984

[41] N Myers R A Mittermeler C G Mittermeler G A B daFonseca and J Kent ldquoBiodiversity hotspots for conservationprioritiesrdquo Nature vol 403 no 6772 pp 853ndash858 2000

[42] R Perumal Samy and S Ignacimuthu ldquoAntibacterial activity ofsome folkloremedicinal plants used by tribals inWestern Ghats

Evidence-Based Complementary and Alternative Medicine 17

of Indiardquo Journal of Ethnopharmacology vol 69 no 1 pp 63ndash712000

[43] K Nampoothiri A Chrispal A Begum S Jasmine K GGopinath and A Zachariah ldquoA clinical study of renal tubulardysfunction inCleistanthus collinus (Oduvanthalai) poisoningrdquoClinical Toxicology vol 48 no 3 pp 193ndash197 2010

[44] M Strathern UsefulKnowledge The Isiah Berlin Lecture Uni-versity of Manchester Brunswick UK

[45] C N Sunil and A K Pradeep ldquo(2001) Another new species ofTripogon (Poaceae) from Indiardquo Sida vol 19 pp 803ndash806 2005

[46] S Biber-Klemm and T Cottier Right To Plant GeneticResources and Traditional Knowledge Basic Issues and Prospec-tive Oxfordshire Nosworthy Way UK 2006

[47] V S Ramachandran S Shijo Joseph and R Aruna ldquoEth-nobotanical studies from amaravathy range of Indira GandhiWildlife SanctuaryWesternGhats CoimbatoreDistrict South-ern Indiardquo Ethnobotanical Leaflets vol 13 pp 1069ndash1087 2009

[48] A H Rajasab and I Mohamad ldquoDocumentation of folkknowledge on edible wild plants of North Karnatakardquo IndianJournal of Traditional Knowledge vol 3 no 4 pp 419ndash4292004

[49] M Rajadurai V G Vidhya M Ramya and A Bhaskar ldquoEthno-medicinal plants used by the traditional healers of pachamalaihills Tamilnadu Indiardquo Studies on Ethno-Medicine vol 3 no 1pp 39ndash41 2009

[50] S K Adesina O Idowu A O Ogundaini et al ldquoAntimicrobialconstituents of the leaves of Acalypha wilkesiana and Acalyphahispidardquo Phytotherapy Research vol 14 no 5 pp 371ndash374 2000

[51] M T Gutierrez-Lugo M P Singh W M Maiese and BN Timmermann ldquoNew antimicrobial cycloartane triterpenesfrom Acalypha communisrdquo Journal of Natural Products vol 65no 6 pp 872ndash875 2002

[52] A K Tyagi andAMalik ldquoAntimicrobial potential and chemicalcomposition of Mentha piperita oil in liquid and vapour phaseagainst food spoiling microorganismsrdquo Food Control vol 22no 11 pp 1707ndash1714 2011

[53] G Devendran and U Balasubramanian ldquoQualitative phyto-chemical screening and GC-MS analysis of Ocimum sanctumL leavesrdquoAsian Journal of Plant Science and Research vol 1 no4 pp 44ndash48 2011

[54] V C Jain N M Patel D P Shah P K Patel and B HJoshi ldquoAntioxidant and antimicrobial activities of Alangiumsalvifolium (Lf) Wang rootrdquo Global Journal of Pharmacologyvol 4 no 1 pp 13ndash18 2010

[55] P Swapna Latha andK Kannabiran ldquoAntimicrobial activity andphytochemicals of Solanum trilobatum LinnrdquoAfrican Journal ofBiotechnology vol 5 no 23 pp 2402ndash2404 2006

[56] M Usha1 S Ragini and S M A Naqvi ldquoAntibacterial activityof acetone and ethanol extracts of Cinnamon (Cinnamomumzeylanicum) and Ajowan (Trachyspermum ammi) on four foodspoilage bacteriardquo International Research Journal of BiologicalSciences vol 1 no 4 pp 7ndash11 2012

[57] S Prabuseenivasan M Jayakumar and S Ignacimuthu ldquoInvitro antibacterial activity of some plant essential oilsrdquo BMCComplementary and Alternative Medicine vol 6 article 39 pp1ndash8 2006

[58] G A Ayoola F M Lawore T Adelowotan et al ldquoChemi-cal analysis and antimicrobial activity of the essential oil ofSyzigium aromaticum (clove)rdquo African Journal of Biotechnologyvol 7 no 13 pp 2227ndash2231 2008

[59] Y J Fu L Chen Y Zu et al ldquoThe antibacterial activity of cloveessential oil against Propionibacterium acnes and its mechanismof actionrdquo Archives of Dermatology vol 145 no 1 pp 86ndash882009

[60] D Venkatesan C M Karrunakarn S Selva Kumar and P TPalani Swamy ldquoIdentification of phytochemical constituents ofAegle marmelos responsible for antimicrobial activity againstselected pathogenic organismsrdquo Ethnobotanical Leaflets vol 13pp 1362ndash1372 2009

[61] P Arun K G Purushotham J Johnsy jayarani and V KumarildquoIn vitro Antibacterial Activity of Oldenlandia umbellata anIndian medicinal Plantrdquo Journal of Pharmaceutical Science andTechnology vol 2 no 4 pp 198ndash201 2010

[62] A Manjamalai R Sardar Sathyajith Singh C Guruvayoorap-pan and V M Berlin Grace ldquoAnalysis of phytochemicalconstituents and antimicrobial activity of some medicinalplants in Taminadu Indiardquo Global Journal of Biotechnology andBiochemistry vol 5 no 2 pp 120ndash128 2010

[63] E Haslam ldquoNatural polyphenols (vegetable tannins) as drugsPossible modes of actionrdquo Journal of Natural Products vol 59no 2 pp 205ndash215 1996

[64] Y Cai Q Luo M Sun and H Corke ldquoAntioxidant activityand phenolic compounds of 112 traditional Chinese medicinalplants associated with anticancerrdquo Life Sciences vol 74 no 17pp 2157ndash2184 2004

[65] B Shan Y Z Cai M Sun and H Corke ldquoAntioxidant capacityof 26 spice extracts and characterization of their phenolicconstituentsrdquo Journal of Agricultural and Food Chemistry vol53 no 20 pp 7749ndash7759 2005

[66] R J W Lambert P N Skandamis P J Coote and G-J ENychas ldquoA study of the minimum inhibitory concentration andmode of action of oregano essential oil thymol and carvacrolrdquoJournal of AppliedMicrobiology vol 91 no 3 pp 453ndash462 2001

[67] M Raccach ldquoThe antimicrobial activity of phenolic antioxi-dants in food a reviewrdquo Journal of Food Safety vol 6 no 3 pp141ndash170 1984

[68] M Taniguchi Y Yano E Tada et al ldquoMode of action of polygo-dial an antifungal sesquiterpene dialdehyderdquo Agricultural andBiological Chemistry vol 52 no 6 pp 1409ndash1414 1988

[69] G J E Nychas ldquoNatural antimicrobials from plantsrdquo in NewMethods of Food Preservation G W Gould Ed pp 58ndash89Blackie Academic London UK 1995

[70] S Burt ldquoEssential oils their antibacterial properties and poten-tial applications in foodsmdasha reviewrdquo International Journal ofFood Microbiology vol 94 no 3 pp 223ndash253 2004

Submit your manuscripts athttpwwwhindawicom

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Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

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Behavioural Neurology

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Disease Markers

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OncologyJournal of

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Oxidative Medicine and Cellular Longevity

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Immunology ResearchHindawi Publishing Corporationhttpwwwhindawicom Volume 2014

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Parkinsonrsquos Disease

Evidence-Based Complementary and Alternative Medicine

Volume 2014Hindawi Publishing Corporationhttpwwwhindawicom

Evidence-Based Complementary and Alternative Medicine 13

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn

Bacteria (+minus)

0

5

10

15

20

25

30

Inhi

bitio

n zo

nes (

mm

)(P

lant

frac

tion)

lowastlowast

lowast

(mean plusmn SD n = 3)In vitro assay

C citratus

(a)

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn

Bacteria (+minus)

0

5

10

15

20

25

30

Inhi

bitio

n zo

nes (

mm

)

lowast

lowast

lowastlowast

lowast

lowast

(mean plusmn SD n = 3)In vitro assay

35

C zeylanicum

(b)

Bacteria (+minus)

0

5

10

15

20

25

30

Inhi

bitio

n zo

nes (

mm

)

lowast

lowast

lowast

(mean plusmn SD n = 3)

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn V

d St

O sanctum

Incu

batio

n (37∘C

24 h

)

(c)

Bacteria (+minus)

0

5

10

15

20

25

Inhi

bitio

n zo

nes (

mm

)

lowast

lowast

(mean plusmn SD n = 3)

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn V

d St

M piperita

(Pla

nt fr

actio

n)

(d)

Bacteria (+minus)

0

5

10

15

20

25

30

Inhi

bitio

n zo

nes (

mm

)lowast

lowast

lowast

(mean plusmn SD n = 3)

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn V

d St

E caryophyllus

(e)

0

5

10

15

20

25

30

Inhi

bitio

n zo

nes (

mm

) (mean plusmn SD n = 3)

Bacteria (+minus)

KHW

TES

Kp

KprPa Sa

Stp

Spn

E globulus

Incu

batio

n (37∘C

24 h

)

(f)

Bacteria (+minus)

0

5

10

15

20

25

30

Inhi

bitio

n zo

nes (

mm

)(P

lant

frac

tion)

lowast

lowast

lowast

lowast

(mean plusmn SD n = 3)

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn V

d

R officinalis

(g)

0

5

10

15

20

25

30

Inhi

bitio

n zo

nes (

mm

)

lowast

lowast

KHW

TES

Kp

Kpr Pa Sa

Stp

Spn V

d St

Bacteria (+minus)V zizanioides

(mean plusmn SD n = 3)

Incu

batio

n (37∘C

24 h

)

(h)

Figure 5 Comparison of antimicrobial effect of plant compounds obtained from the most popularly used sources of essential oils as assayedby the disc-diffusion method in vitro It displayed a powerful activity against B pseudomallei and S aureus than the other bacteria strainsOther compounds showed only a moderate or weak action against the tested bacteria Values for zone of bacterial growth inhibition werepresented as mean plusmn SD (119899 = 3) with level of significance at (lowast119875 gt 001)

and C citratus fractions also showed antimicrobial activity(MICs of 3125ndash125120583gmL) only at higher concentrationsagainst the tested bacteria In addition to that higher concen-trations (gt250120583gmL) (of Vetiveria fractions) were requiredto inhibit Vibro species and others (including E globulusfractions) failed to show any effect at tested concentrations(78ndash125 120583gmL) However the purified fractions (from mostactive medicinal plants) showed strong bacteriostatic inhibi-tion against the tested organisms (Table 3)

310 Cytotoxic Effects of Plants When the components wereassayed for cytotoxicity against the normal human skinfibroblasts (HEPK) cells the compounds obtained from

E cardamomum T involucrata S indicusC acidaA vasicaA marmelos A indica and A paniculata did not showtoxicity up to 1000120583gmL (see Figures S1 and S2 in Supple-mentaryMaterial available online at httpdxdoiorg1011552013525613) A slight reduction of cell proliferation wasnoted only at higher doses (2000120583gmL) In contrast cellproliferation was markedly reduced after exposure of HEPKcells to O sanctum E globulus V zizanioides C citratusand E globulus compounds There was no gradual reductionin skin cell proliferation seen after exposure to C zeylan-icum R officinalis and M piperita (see Figures S3 and S4)compounds The toxicity was found to be concentration-dependent when the skin fibroblasts (HEPK) cells were

14 Evidence-Based Complementary and Alternative Medicine

Table 3 Minimum inhibitory concentrations (MICs) of purified plant fractions and essential oils against antibiotic resistant bacteria

Botanical name Family Parts used Gram-positive and -negative bacteria (MICs 120583gmL)KHW TES Kp KPr Sa Stp Sp Vd St

A indica Juss Meliaceae Seed (fraction) 3125 625 125 lt250 3125 mdash mdash mdash mdashA marmelos (L) Rutaceae Root-bark (F) 3125 3125 mdash mdash 3125 625 125 250 mdashA paniculata Nees Acanthaceae Leaf (fraction) 250 mdash 125 mdash 625 mdash mdash 125 mdashA vasica Nees Acanthaceae Fraction (Stem) 156 3125 3125 625 mdash lt250 3125 3125 mdashC acida Roxb Rutaceae Leaf (fraction) 156 625 mdash mdash 78 mdash 3125 mdash mdashE cardamomumWhite et Mason Acanthaceae Fraction (WP) 625 mdash 3125 mdash 3125 250 625 mdash mdashS indicus (L) Euphorbiaceae Whole plant (F) 3125 125 156 mdash 625 mdash mdash lt250 mdashT involucrata (L) Euphorbiaceae Shellosol (leaf) 156 3125 mdash mdash 78 625 mdash mdash mdashCinnamomum zeylanicum (L) Lauraceae Bark (fraction) 156 625 mdash mdash 78 3125 mdash mdash 125Cymbopogon citratus (L) Graminae Leaf (fraction) 250 lt250 625 mdash 625 125 mdash mdash mdashEugenia caryophyllus (L) Myrtaceae Flower buds (F) 625 125 mdash mdash 625 mdash mdash mdash mdashEucalyptus globulus (L) Myrtaceae Fraction (leaf) mdash mdash mdash mdash mdash mdash lt250 mdash lt250Mentha piperita (L) Labiatae Fraction (WP) mdash 625 3125 mdash 3125 mdash lt250 mdash mdashOcimum sanctum (L) Labiatae Leaf (fraction) 625 125 mdash 3125 625 mdash 125 lt250 mdashRosmarinus officinalis (L) Labiatae Rosemary oil 3125 mdash mdash mdash 3125 mdash mdash mdash mdashVetiveria zizanioides (L) Graminae Root (fraction) mdash 125 125 mdash mdash mdash gt250 gt250The bacterial growth inhibitory activity was compared for significant differences within the bacterial strains by broth-dilution method at 250 125 625 3125156 and 78 120583gmL F fractions

exposed to various compounds The cell proliferation wasincreased by the influence of the plant components at thelower concentrations Whereas the oil yielding plant com-pounds showed inhibition of cell proliferation and toxicity athigher doses 250ndash1000120583gmL

4 Discussion

The Western Ghats is considered as one of the richestbiodiversity hotspots in the world [41] In this survey wecollected nearly 78 medicinal plants from Western andEastern Ghats that are edible and popularly used for curingvarious ailments including snakebite Traditional remedieshave a long-standing history in many tribal settlements inTN India and they continue to provide useful and applicabletools for treating ailments [42] The ingredients that makeup the ldquoVishakallurdquo stone which is used as an antidote forsnakebite are different herbs and pebbles available from theriver banks Likewise aqueous paste and decoction obtainedfrom the leaves of A paniculata are widely used for snakebitetreatment by indigenous people [43] Previous studies havereported that ethnomedicine plays major roles in conservingthe disappearing knowledge of tribal communities [44ndash47]The traditional beliefs of reliance on a rich diversity ofethnomedicinal plants located at different settlements havealso been confirmed in another study [48 49] Herein weexplored the various types of traditional practices reported bythe primitive tribal communities with a view to gain furtherknowledge from such studies

In the present investigation potentially rich sources oftribal medicine (71 plants) were scientifically evaluated fortheir antibacterial activity against the MDR bacteria and theaccumulated data was disseminated for the first time to the

scientific community Out of the 71medicinal plants screenedfor the antibacterial activity 10 of them (T involucrataC acida A marmelos A vasica C procera A paniculataand M piperita A indica S indicus and E cardamomum)displayed the highest antibacterial activity against the mul-tidrug resistant B pseudomallei (KHW and TES) and Saureus strains The antibacterial activity of those crude plantextracts was as equally effective as that of the standard drugsOur findings corroborated with the previous reports madeon the antistaphylococcal activity of tribal medicinal plants[34 50 51] On the other hand isolated components from themost active extracts of T involucrata shellsol and C acidaexhibited the most potent action against the antibiotic resis-tant B pseudomallei (KHW) K pneumoniae and S aureusstrainsThese results further confirmed our previous findingson the leaves of T involucrata and its compounds hydrocar-bon ester-like shellsol which displayed a high antibacterialeffect against the different bacterial strains especially thatof S aureus [34] Eugenol and caryophyllene are the activeagents contained in the M piperita [52] and O sanctumplants which are believed to be mainly responsible for theantimicrobial properties of these plants [53] Interestinglythe compounds obtained from the aromatic plants such asC zeylanicum and R officinalis were also found to be veryeffective against the multidrug resistant human pathogen Kpneumoniae S aureus S typhi and B pseudomallei (KHW)that causes melioidosis

The inhibitory potential determined for the shellsol(T involucrata) vaseline (A vasica) C acida and C zey-lanicum indicates that the MIC of 78ndash3125120583gmL foundagainst the B pseudomallei of KHW K pneumoniae Kpneumoniae S pyogenes and V damsel and S pneumoniaewas quite low Similarly lower MIC values were found for

Evidence-Based Complementary and Alternative Medicine 15

M piperita (MIC of 113ndash225 mgmL) against the abovebacterial strains [52] MIC was found for the most activealcohol extracts of A salvifolium (MIC 0034ndash0263mgmL)on S aureus [54] and the MIC for S trilobatum aqueousextracts determined against the tested organisms rangedfrom 006 to 05mgmL [55] C zeylanicum was foundto have an effective antibacterial activity (MIC 64 120583gmL)against P aeruginosa E coli B subtilis and S aureus[56] Previously several investigators have demonstrated thatactive agents exert interesting activity against bacteria evenat lower concentrations tested [57 58] The bacteriostaticmechanism involves damage to the cell walls of bacteriafollowed by inhibition of protein synthesis that ultimatelyleads to bacterial death [59] The most active plants arewidely used by various tribes as traditional treatment (iecut wounds skin infection and scabies) thus indicatingthe potential for further development into promising drugsIn order to ascertain the safety and efficacy of the mostactive compounds their effect on human skin fibroblast cellswas evaluated Chemical constituents of E cardamomumT involucrata S indicus C acida A vasica A marmelosA indica and A paniculata plants failed to produce anynoticeable toxicity up to 1000120583gmL Although some ofthe tested compounds exhibited a slight reduction of cellproliferation and some minor morphological changes suchchanges were insignificant at lower doses and became evidentonly at higher doses However certain aromatic compoundsof O sanctum E globulus V zizanioides C citratus and Eglobulus plants showed reduction of cell proliferation againstHEPK cells

Our phytochemical screening also provides evidenceof the presence of several types of compounds that aremainly responsible for the remarkable antibacterial effectof these plants The differences noted for the bactericidalactivity of various plant extracts in this study appears tobe directly related to the diversity of compounds (shown inparentheses) that are accumulated in the following plants(eg A marmelos and O umbellate) [60ndash62] Compoundslike tannins phenol and polyphenols can bind the Gram-negative bacteria to form a heavy soluble complex on the cellsurface which subsequently disturbs the availability of recep-tor on cells and kills the bacteria [63] Several species havingwide spectra of antimicrobial activity mainly due to theactive constituents such as essential oil phenolic compoundslike thymol carvacrol in oregano and thyme eugenol inclove and cinnamon were also identified previously [64 65]Essential oils degrade the cell wall interact with the cell com-ponents and then disrupt the cytoplasmic membrane [66]The antimicrobial effect of phenolic compounds may involvemultiplemodes of action including damage to themembraneprotein interference with membrane integrated enzymes[67] causing leakage of cellular components coagulation ofcytoplasm depletion of the proton motive force alterationof fatty acid and phospholipid constituents impairmentof enzymatic mechanisms for production and metabolismalteration of nutrient uptake and electron transport [68]influencing the synthesis of DNA and RNA and destroyingprotein translation and the function of the mitochondrionin eukaryotes [69] The mode of action of antimicrobial

agents depends on the type of microorganism and is mainlyrelated to their cell wall structure and the outer membranearrangement Most of the plant spices and herbs containcomplex phenolics (ie phenolic acids flavonoids tanninslignans coumarins quinines) In addition the mechanismsof action of each phenolic compound against various bacteriaare also very complicated [26 70] Further investigation istherefore required to understand the relationship betweenthe antimicrobial action and the chemical structure of everyphenolic compound in the tested extracts The informationavailable from previous pharmacological sources combinedwith the findings herein reported on the medicinal plantextracts may serve as essential data for future drug develop-ment to combat diseases caused by theMDRbacterial strains

Abbreviations

MDR Multidrug resistant bacteriaTES Strain of Burkholderia pseudomalleiMRSA Methicillin-resistant Staphylococcus aureusVRE Vancomycin-resistant enterococciMTB Mycobacterium tuberculosisCH3OH Methanol

CFU Colony forming unitHCl Hydrochloric acidTLC Thin layer chromatographyH2SO4 Sulphuric acids

UV Ultraviolet spectrophotometerHEPK Human skin fibroblasts

Acknowledgments

Theauthors are thankful to the ERI Loyola College ChennaiIndia for his support during the survey The authors alsothank the Department of Microbiology NUHS NUS forthe bacterial cultures used in this investigation The authorsare thankful to the tribal people for sharing their healingknowledge on medicinal aromatic plants

References

[1] M N Alekshun and S B Levy ldquoMolecular mechanisms ofantibacterialmultidrug resistancerdquoCell vol 128 no 6 pp 1037ndash1050 2007

[2] A D Russell ldquoAntibiotic and biocide resistance in bacteriaintroductionrdquo Symposium Series Society for Applied Microbiol-ogy vol 31 no 92 pp 1Sndash3S 2002

[3] D R Guay ldquoTreatment of bacterial skin and skin structureinfectionsrdquo Expert Opinion on Pharmacotherapy vol 4 no 8pp 1259ndash1275 2003

[4] A J Brosnahan and P M Schlievert ldquoGram-positive bac-terial superantigen outside-in signaling causes toxic shocksyndromerdquo FEBS Journal vol 278 no 23 pp 4649ndash4667 2011

[5] K Fellermann J Wehkamp K R Herrlinger and E F StangeldquoCrohnrsquos disease a defensin deficiency syndromerdquo EuropeanJournal of Gastroenterology and Hepatology vol 15 no 6 pp627ndash634 2003

16 Evidence-Based Complementary and Alternative Medicine

[6] L Chen S Lin R Agha-Majzoub L Overbergh C Mathieuand L S Chan ldquoCCL27 is a critical factor for the developmentof atopic dermatitis in the keratin-14 IL-4 transgenic mousemodelrdquo International Immunology vol 18 no 8 pp 1233ndash12422006

[7] J H Song V Thamlikitkul and P R Hsueh ldquoClinical andeconomic burden of community-acquired pneumonia amongstadults in the Asia-Pacific regionrdquo International Journal ofAntimicrobial Agents vol 38 no 2 pp 108ndash117 2011

[8] D A B Dance ldquoMelioidosisrdquo Current Opinion in InfectiousDiseases vol 15 no 2 pp 127ndash132 2002

[9] N J White ldquoMelioidosisrdquo The Lancet vol 361 no 9370 pp1715ndash1722 2003

[10] S Yang ldquoMelioidosis research in Chinardquo Acta Tropica vol 77no 2 pp 157ndash165 2000

[11] P R Hsueh L J Teng L N Lee et al ldquoMelioidosis an emerginginfection in Taiwanrdquo Emerging Infectious Diseases vol 7 no 3pp 428ndash433 2001

[12] C Walsh and S Fanning ldquoAntimicrobial resistance in food-borne pathogensmdasha cause for concernrdquo Current Drug Targetsvol 9 no 9 pp 808ndash815 2008

[13] M Vorachit P Chongtrakool S Arkomsean and S BoonsongldquoAntimicrobial resistance in Burkholderia pseudomalleirdquo ActaTropica vol 74 no 2-3 pp 139ndash144 2000

[14] F M Thibault E Hernandez D R Vidal M Girardet and JD Cavallo ldquoAntibiotic susceptibility of 65 isolates of Burkholde-ria pseudomallei and Burkholderia mallei to 35 antimicrobialagentsrdquo Journal of Antimicrobial Chemotherapy vol 54 no 6pp 1134ndash1138 2004

[15] R L S Sikarwar ldquoEthnogynaecological uses of plants new toIndiardquo Ethnobotany vol 12 no 1-2 pp 112ndash115 2002

[16] R Siva ldquoStatus of natural dyes and dye-yielding plants in IndiardquoCurrent Science vol 92 no 7 pp 916ndash925 2007

[17] P Kumar Singh V Kumar R K Tiwari A Sharma ChV Rao and R H Singh ldquoMedico-ethnobotany of lsquochatararsquoblock of district sonebhadra Uttar Pradesh Indiardquo Advances inBiological Research vol 4 no 1 pp 65ndash80 2010

[18] W H Lewis and M P Elwin-Lewis Medical Botany PlantsAffecting Human Health John Wiley amp Sons New York NYUSA 2003

[19] R Perumal Samy P N Pushparaj and P GopalakrishnakoneldquoA compilation of bioactive compounds fromAyurvedardquo Bioin-formation vol 3 no 3 pp 100ndash110 2008

[20] WorldHealthOrganization (WHO)TraditionalMedicine Strat-egy 2002ndash2005 Genera World Health Organization GenevaSwitzerland 2002

[21] P S Haddad G A Azar S Groom and M Boivin ldquoNaturalhealth products modulation of immune function and preven-tion of chronic diseasesrdquo Evidence Based Complementary andAlternative Medicine vol 2 no 4 pp 513ndash520 2005

[22] K Poonam andG S Singh ldquoEthnobotanical study of medicinalplants used by the Taungya community in Terai Arc LandscapeIndiardquo Journal of Ethnopharmacology vol 123 no 1 pp 167ndash1762009

[23] G B Mahady ldquoMedicinal plants for the prevention and treat-ment of bacterial infectionsrdquo Current Pharmaceutical Designvol 11 no 19 pp 2405ndash2427 2005

[24] V P Kumar N S Chauhan H Padh and M Rajani ldquoSearchfor antibacterial and antifungal agents from selected Indianmedicinal plantsrdquo Journal of Ethnopharmacology vol 107 no2 pp 182ndash188 2006

[25] R Nair T Kalariya and S Chanda ldquoAntibacterial activity ofsome plant extracts used in folk medicinerdquo Journal of HerbalPharmacotherapy vol 7 no 3-4 pp 191ndash201 2007

[26] D Kalemba and A Kunicka ldquoAntibacterial and antifungalproperties of essential oilsrdquo Current Medicinal Chemistry vol10 no 10 pp 813ndash829 2003

[27] A H Ashour ldquoAntibacterial antifungal and anticancer activi-ties of volatile oils and extracts from stems leaves and flowers ofEucalyptus sideroxylon and Eucalyptus torquatardquoCancer BiologyandTherapy vol 7 no 3 pp 399ndash403 2008

[28] M J Abad L M Bedoya L Apaza and P Bermejo ldquoTheArtemisia L genus a review of bioactive essential oilsrdquoMolecules vol 17 no 3 pp 2542ndash2566 2012

[29] H N Marzoug M Romdhane A Lebrihi et al ldquoEucalyptusoleosa essential oils chemical composition and antimicrobialand antioxidant activities of the oils from different plant parts(stems leaves flowers and fruits)rdquoMolecules vol 16 no 2 pp1695ndash1709 2011

[30] C Proestos D Sereli and M Komaitis ldquoDetermination ofphenolic compounds in aromatic plants by RP-HPLC and GC-MSrdquo Food Chemistry vol 95 no 1 pp 44ndash52 2006

[31] S Chao G Young C Oberg and K Nakaoka ldquoInhibition ofmethicillin-resistant Staphylococcus aureus (MRSA) by essentialoilsrdquo Flavour and Fragrance Journal vol 23 no 6 pp 444ndash4492008

[32] M Akin D Oguz and H T Saracoglu ldquoAntibacterial effectsof some plant extracts from Labiatae (Lamiaceae) growingnaturally around Sirnak-Silopi Turkeyrdquo International Journal ofPharmaceutical and Applied Sciences vol 1 no 1 pp 4ndash47 2010

[33] K Bayoub T Baibai D Mountassif A Retmane and ASoukri ldquoAntibacterial activities of the crude ethanol extracts ofmedicinal plants against Listeriamonocytogenes and some otherpathogenic strainsrdquo African Journal of Biotechnology vol 9 no27 pp 4251ndash4258 2010

[34] R P Samy P Gopalakrishnakone M Sarumathi P Houghtonand S Ignacimuthu ldquoPurification of antibacterial agents fromTragia involucratamdasha popular tribal medicine for wound heal-ingrdquo Journal of Ethnopharmacology vol 107 no 1 pp 99ndash1062006

[35] K M Matthew The Flora of Tamil Nadu Carnatic Part IndashIIIprinted at the Diocesan Press Madras India 1981ndash1983

[36] AW BauerWMKirby J C Sherris andM Turck ldquoAntibioticsusceptibility testing by a standardized single disk methodrdquoAmerican Journal of Clinical Pathology vol 45 no 4 pp 493ndash496 1966

[37] National Committee for Clinical Laboratory Standards Per-formance Standards For Antimicrobial Susceptibility Testing12th Informational Supplement Approved standard M100-S12National Committee for Clinical Laboratory Standards WaynePa USA 2002

[38] E A Elsinghorst ldquoMeasurement of invasion by gentamicinresistancerdquoMethods in Enzymology vol 236 pp 405ndash420 1994

[39] J B Harborne Phytochemical Methods Chapman and HallNew York NY USA 1976

[40] H Wagner S Bladt and E M Zgairski Plant Drug AnalysisSpringer New York NY USA 1984

[41] N Myers R A Mittermeler C G Mittermeler G A B daFonseca and J Kent ldquoBiodiversity hotspots for conservationprioritiesrdquo Nature vol 403 no 6772 pp 853ndash858 2000

[42] R Perumal Samy and S Ignacimuthu ldquoAntibacterial activity ofsome folkloremedicinal plants used by tribals inWestern Ghats

Evidence-Based Complementary and Alternative Medicine 17

of Indiardquo Journal of Ethnopharmacology vol 69 no 1 pp 63ndash712000

[43] K Nampoothiri A Chrispal A Begum S Jasmine K GGopinath and A Zachariah ldquoA clinical study of renal tubulardysfunction inCleistanthus collinus (Oduvanthalai) poisoningrdquoClinical Toxicology vol 48 no 3 pp 193ndash197 2010

[44] M Strathern UsefulKnowledge The Isiah Berlin Lecture Uni-versity of Manchester Brunswick UK

[45] C N Sunil and A K Pradeep ldquo(2001) Another new species ofTripogon (Poaceae) from Indiardquo Sida vol 19 pp 803ndash806 2005

[46] S Biber-Klemm and T Cottier Right To Plant GeneticResources and Traditional Knowledge Basic Issues and Prospec-tive Oxfordshire Nosworthy Way UK 2006

[47] V S Ramachandran S Shijo Joseph and R Aruna ldquoEth-nobotanical studies from amaravathy range of Indira GandhiWildlife SanctuaryWesternGhats CoimbatoreDistrict South-ern Indiardquo Ethnobotanical Leaflets vol 13 pp 1069ndash1087 2009

[48] A H Rajasab and I Mohamad ldquoDocumentation of folkknowledge on edible wild plants of North Karnatakardquo IndianJournal of Traditional Knowledge vol 3 no 4 pp 419ndash4292004

[49] M Rajadurai V G Vidhya M Ramya and A Bhaskar ldquoEthno-medicinal plants used by the traditional healers of pachamalaihills Tamilnadu Indiardquo Studies on Ethno-Medicine vol 3 no 1pp 39ndash41 2009

[50] S K Adesina O Idowu A O Ogundaini et al ldquoAntimicrobialconstituents of the leaves of Acalypha wilkesiana and Acalyphahispidardquo Phytotherapy Research vol 14 no 5 pp 371ndash374 2000

[51] M T Gutierrez-Lugo M P Singh W M Maiese and BN Timmermann ldquoNew antimicrobial cycloartane triterpenesfrom Acalypha communisrdquo Journal of Natural Products vol 65no 6 pp 872ndash875 2002

[52] A K Tyagi andAMalik ldquoAntimicrobial potential and chemicalcomposition of Mentha piperita oil in liquid and vapour phaseagainst food spoiling microorganismsrdquo Food Control vol 22no 11 pp 1707ndash1714 2011

[53] G Devendran and U Balasubramanian ldquoQualitative phyto-chemical screening and GC-MS analysis of Ocimum sanctumL leavesrdquoAsian Journal of Plant Science and Research vol 1 no4 pp 44ndash48 2011

[54] V C Jain N M Patel D P Shah P K Patel and B HJoshi ldquoAntioxidant and antimicrobial activities of Alangiumsalvifolium (Lf) Wang rootrdquo Global Journal of Pharmacologyvol 4 no 1 pp 13ndash18 2010

[55] P Swapna Latha andK Kannabiran ldquoAntimicrobial activity andphytochemicals of Solanum trilobatum LinnrdquoAfrican Journal ofBiotechnology vol 5 no 23 pp 2402ndash2404 2006

[56] M Usha1 S Ragini and S M A Naqvi ldquoAntibacterial activityof acetone and ethanol extracts of Cinnamon (Cinnamomumzeylanicum) and Ajowan (Trachyspermum ammi) on four foodspoilage bacteriardquo International Research Journal of BiologicalSciences vol 1 no 4 pp 7ndash11 2012

[57] S Prabuseenivasan M Jayakumar and S Ignacimuthu ldquoInvitro antibacterial activity of some plant essential oilsrdquo BMCComplementary and Alternative Medicine vol 6 article 39 pp1ndash8 2006

[58] G A Ayoola F M Lawore T Adelowotan et al ldquoChemi-cal analysis and antimicrobial activity of the essential oil ofSyzigium aromaticum (clove)rdquo African Journal of Biotechnologyvol 7 no 13 pp 2227ndash2231 2008

[59] Y J Fu L Chen Y Zu et al ldquoThe antibacterial activity of cloveessential oil against Propionibacterium acnes and its mechanismof actionrdquo Archives of Dermatology vol 145 no 1 pp 86ndash882009

[60] D Venkatesan C M Karrunakarn S Selva Kumar and P TPalani Swamy ldquoIdentification of phytochemical constituents ofAegle marmelos responsible for antimicrobial activity againstselected pathogenic organismsrdquo Ethnobotanical Leaflets vol 13pp 1362ndash1372 2009

[61] P Arun K G Purushotham J Johnsy jayarani and V KumarildquoIn vitro Antibacterial Activity of Oldenlandia umbellata anIndian medicinal Plantrdquo Journal of Pharmaceutical Science andTechnology vol 2 no 4 pp 198ndash201 2010

[62] A Manjamalai R Sardar Sathyajith Singh C Guruvayoorap-pan and V M Berlin Grace ldquoAnalysis of phytochemicalconstituents and antimicrobial activity of some medicinalplants in Taminadu Indiardquo Global Journal of Biotechnology andBiochemistry vol 5 no 2 pp 120ndash128 2010

[63] E Haslam ldquoNatural polyphenols (vegetable tannins) as drugsPossible modes of actionrdquo Journal of Natural Products vol 59no 2 pp 205ndash215 1996

[64] Y Cai Q Luo M Sun and H Corke ldquoAntioxidant activityand phenolic compounds of 112 traditional Chinese medicinalplants associated with anticancerrdquo Life Sciences vol 74 no 17pp 2157ndash2184 2004

[65] B Shan Y Z Cai M Sun and H Corke ldquoAntioxidant capacityof 26 spice extracts and characterization of their phenolicconstituentsrdquo Journal of Agricultural and Food Chemistry vol53 no 20 pp 7749ndash7759 2005

[66] R J W Lambert P N Skandamis P J Coote and G-J ENychas ldquoA study of the minimum inhibitory concentration andmode of action of oregano essential oil thymol and carvacrolrdquoJournal of AppliedMicrobiology vol 91 no 3 pp 453ndash462 2001

[67] M Raccach ldquoThe antimicrobial activity of phenolic antioxi-dants in food a reviewrdquo Journal of Food Safety vol 6 no 3 pp141ndash170 1984

[68] M Taniguchi Y Yano E Tada et al ldquoMode of action of polygo-dial an antifungal sesquiterpene dialdehyderdquo Agricultural andBiological Chemistry vol 52 no 6 pp 1409ndash1414 1988

[69] G J E Nychas ldquoNatural antimicrobials from plantsrdquo in NewMethods of Food Preservation G W Gould Ed pp 58ndash89Blackie Academic London UK 1995

[70] S Burt ldquoEssential oils their antibacterial properties and poten-tial applications in foodsmdasha reviewrdquo International Journal ofFood Microbiology vol 94 no 3 pp 223ndash253 2004

Submit your manuscripts athttpwwwhindawicom

Stem CellsInternational

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

MEDIATORSINFLAMMATION

of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Behavioural Neurology

EndocrinologyInternational Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Disease Markers

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

BioMed Research International

OncologyJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Oxidative Medicine and Cellular Longevity

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

PPAR Research

The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014

Immunology ResearchHindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Journal of

ObesityJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Computational and Mathematical Methods in Medicine

OphthalmologyJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Diabetes ResearchJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Research and TreatmentAIDS

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Gastroenterology Research and Practice

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Parkinsonrsquos Disease

Evidence-Based Complementary and Alternative Medicine

Volume 2014Hindawi Publishing Corporationhttpwwwhindawicom

14 Evidence-Based Complementary and Alternative Medicine

Table 3 Minimum inhibitory concentrations (MICs) of purified plant fractions and essential oils against antibiotic resistant bacteria

Botanical name Family Parts used Gram-positive and -negative bacteria (MICs 120583gmL)KHW TES Kp KPr Sa Stp Sp Vd St

A indica Juss Meliaceae Seed (fraction) 3125 625 125 lt250 3125 mdash mdash mdash mdashA marmelos (L) Rutaceae Root-bark (F) 3125 3125 mdash mdash 3125 625 125 250 mdashA paniculata Nees Acanthaceae Leaf (fraction) 250 mdash 125 mdash 625 mdash mdash 125 mdashA vasica Nees Acanthaceae Fraction (Stem) 156 3125 3125 625 mdash lt250 3125 3125 mdashC acida Roxb Rutaceae Leaf (fraction) 156 625 mdash mdash 78 mdash 3125 mdash mdashE cardamomumWhite et Mason Acanthaceae Fraction (WP) 625 mdash 3125 mdash 3125 250 625 mdash mdashS indicus (L) Euphorbiaceae Whole plant (F) 3125 125 156 mdash 625 mdash mdash lt250 mdashT involucrata (L) Euphorbiaceae Shellosol (leaf) 156 3125 mdash mdash 78 625 mdash mdash mdashCinnamomum zeylanicum (L) Lauraceae Bark (fraction) 156 625 mdash mdash 78 3125 mdash mdash 125Cymbopogon citratus (L) Graminae Leaf (fraction) 250 lt250 625 mdash 625 125 mdash mdash mdashEugenia caryophyllus (L) Myrtaceae Flower buds (F) 625 125 mdash mdash 625 mdash mdash mdash mdashEucalyptus globulus (L) Myrtaceae Fraction (leaf) mdash mdash mdash mdash mdash mdash lt250 mdash lt250Mentha piperita (L) Labiatae Fraction (WP) mdash 625 3125 mdash 3125 mdash lt250 mdash mdashOcimum sanctum (L) Labiatae Leaf (fraction) 625 125 mdash 3125 625 mdash 125 lt250 mdashRosmarinus officinalis (L) Labiatae Rosemary oil 3125 mdash mdash mdash 3125 mdash mdash mdash mdashVetiveria zizanioides (L) Graminae Root (fraction) mdash 125 125 mdash mdash mdash gt250 gt250The bacterial growth inhibitory activity was compared for significant differences within the bacterial strains by broth-dilution method at 250 125 625 3125156 and 78 120583gmL F fractions

exposed to various compounds The cell proliferation wasincreased by the influence of the plant components at thelower concentrations Whereas the oil yielding plant com-pounds showed inhibition of cell proliferation and toxicity athigher doses 250ndash1000120583gmL

4 Discussion

The Western Ghats is considered as one of the richestbiodiversity hotspots in the world [41] In this survey wecollected nearly 78 medicinal plants from Western andEastern Ghats that are edible and popularly used for curingvarious ailments including snakebite Traditional remedieshave a long-standing history in many tribal settlements inTN India and they continue to provide useful and applicabletools for treating ailments [42] The ingredients that makeup the ldquoVishakallurdquo stone which is used as an antidote forsnakebite are different herbs and pebbles available from theriver banks Likewise aqueous paste and decoction obtainedfrom the leaves of A paniculata are widely used for snakebitetreatment by indigenous people [43] Previous studies havereported that ethnomedicine plays major roles in conservingthe disappearing knowledge of tribal communities [44ndash47]The traditional beliefs of reliance on a rich diversity ofethnomedicinal plants located at different settlements havealso been confirmed in another study [48 49] Herein weexplored the various types of traditional practices reported bythe primitive tribal communities with a view to gain furtherknowledge from such studies

In the present investigation potentially rich sources oftribal medicine (71 plants) were scientifically evaluated fortheir antibacterial activity against the MDR bacteria and theaccumulated data was disseminated for the first time to the

scientific community Out of the 71medicinal plants screenedfor the antibacterial activity 10 of them (T involucrataC acida A marmelos A vasica C procera A paniculataand M piperita A indica S indicus and E cardamomum)displayed the highest antibacterial activity against the mul-tidrug resistant B pseudomallei (KHW and TES) and Saureus strains The antibacterial activity of those crude plantextracts was as equally effective as that of the standard drugsOur findings corroborated with the previous reports madeon the antistaphylococcal activity of tribal medicinal plants[34 50 51] On the other hand isolated components from themost active extracts of T involucrata shellsol and C acidaexhibited the most potent action against the antibiotic resis-tant B pseudomallei (KHW) K pneumoniae and S aureusstrainsThese results further confirmed our previous findingson the leaves of T involucrata and its compounds hydrocar-bon ester-like shellsol which displayed a high antibacterialeffect against the different bacterial strains especially thatof S aureus [34] Eugenol and caryophyllene are the activeagents contained in the M piperita [52] and O sanctumplants which are believed to be mainly responsible for theantimicrobial properties of these plants [53] Interestinglythe compounds obtained from the aromatic plants such asC zeylanicum and R officinalis were also found to be veryeffective against the multidrug resistant human pathogen Kpneumoniae S aureus S typhi and B pseudomallei (KHW)that causes melioidosis

The inhibitory potential determined for the shellsol(T involucrata) vaseline (A vasica) C acida and C zey-lanicum indicates that the MIC of 78ndash3125120583gmL foundagainst the B pseudomallei of KHW K pneumoniae Kpneumoniae S pyogenes and V damsel and S pneumoniaewas quite low Similarly lower MIC values were found for

Evidence-Based Complementary and Alternative Medicine 15

M piperita (MIC of 113ndash225 mgmL) against the abovebacterial strains [52] MIC was found for the most activealcohol extracts of A salvifolium (MIC 0034ndash0263mgmL)on S aureus [54] and the MIC for S trilobatum aqueousextracts determined against the tested organisms rangedfrom 006 to 05mgmL [55] C zeylanicum was foundto have an effective antibacterial activity (MIC 64 120583gmL)against P aeruginosa E coli B subtilis and S aureus[56] Previously several investigators have demonstrated thatactive agents exert interesting activity against bacteria evenat lower concentrations tested [57 58] The bacteriostaticmechanism involves damage to the cell walls of bacteriafollowed by inhibition of protein synthesis that ultimatelyleads to bacterial death [59] The most active plants arewidely used by various tribes as traditional treatment (iecut wounds skin infection and scabies) thus indicatingthe potential for further development into promising drugsIn order to ascertain the safety and efficacy of the mostactive compounds their effect on human skin fibroblast cellswas evaluated Chemical constituents of E cardamomumT involucrata S indicus C acida A vasica A marmelosA indica and A paniculata plants failed to produce anynoticeable toxicity up to 1000120583gmL Although some ofthe tested compounds exhibited a slight reduction of cellproliferation and some minor morphological changes suchchanges were insignificant at lower doses and became evidentonly at higher doses However certain aromatic compoundsof O sanctum E globulus V zizanioides C citratus and Eglobulus plants showed reduction of cell proliferation againstHEPK cells

Our phytochemical screening also provides evidenceof the presence of several types of compounds that aremainly responsible for the remarkable antibacterial effectof these plants The differences noted for the bactericidalactivity of various plant extracts in this study appears tobe directly related to the diversity of compounds (shown inparentheses) that are accumulated in the following plants(eg A marmelos and O umbellate) [60ndash62] Compoundslike tannins phenol and polyphenols can bind the Gram-negative bacteria to form a heavy soluble complex on the cellsurface which subsequently disturbs the availability of recep-tor on cells and kills the bacteria [63] Several species havingwide spectra of antimicrobial activity mainly due to theactive constituents such as essential oil phenolic compoundslike thymol carvacrol in oregano and thyme eugenol inclove and cinnamon were also identified previously [64 65]Essential oils degrade the cell wall interact with the cell com-ponents and then disrupt the cytoplasmic membrane [66]The antimicrobial effect of phenolic compounds may involvemultiplemodes of action including damage to themembraneprotein interference with membrane integrated enzymes[67] causing leakage of cellular components coagulation ofcytoplasm depletion of the proton motive force alterationof fatty acid and phospholipid constituents impairmentof enzymatic mechanisms for production and metabolismalteration of nutrient uptake and electron transport [68]influencing the synthesis of DNA and RNA and destroyingprotein translation and the function of the mitochondrionin eukaryotes [69] The mode of action of antimicrobial

agents depends on the type of microorganism and is mainlyrelated to their cell wall structure and the outer membranearrangement Most of the plant spices and herbs containcomplex phenolics (ie phenolic acids flavonoids tanninslignans coumarins quinines) In addition the mechanismsof action of each phenolic compound against various bacteriaare also very complicated [26 70] Further investigation istherefore required to understand the relationship betweenthe antimicrobial action and the chemical structure of everyphenolic compound in the tested extracts The informationavailable from previous pharmacological sources combinedwith the findings herein reported on the medicinal plantextracts may serve as essential data for future drug develop-ment to combat diseases caused by theMDRbacterial strains

Abbreviations

MDR Multidrug resistant bacteriaTES Strain of Burkholderia pseudomalleiMRSA Methicillin-resistant Staphylococcus aureusVRE Vancomycin-resistant enterococciMTB Mycobacterium tuberculosisCH3OH Methanol

CFU Colony forming unitHCl Hydrochloric acidTLC Thin layer chromatographyH2SO4 Sulphuric acids

UV Ultraviolet spectrophotometerHEPK Human skin fibroblasts

Acknowledgments

Theauthors are thankful to the ERI Loyola College ChennaiIndia for his support during the survey The authors alsothank the Department of Microbiology NUHS NUS forthe bacterial cultures used in this investigation The authorsare thankful to the tribal people for sharing their healingknowledge on medicinal aromatic plants

References

[1] M N Alekshun and S B Levy ldquoMolecular mechanisms ofantibacterialmultidrug resistancerdquoCell vol 128 no 6 pp 1037ndash1050 2007

[2] A D Russell ldquoAntibiotic and biocide resistance in bacteriaintroductionrdquo Symposium Series Society for Applied Microbiol-ogy vol 31 no 92 pp 1Sndash3S 2002

[3] D R Guay ldquoTreatment of bacterial skin and skin structureinfectionsrdquo Expert Opinion on Pharmacotherapy vol 4 no 8pp 1259ndash1275 2003

[4] A J Brosnahan and P M Schlievert ldquoGram-positive bac-terial superantigen outside-in signaling causes toxic shocksyndromerdquo FEBS Journal vol 278 no 23 pp 4649ndash4667 2011

[5] K Fellermann J Wehkamp K R Herrlinger and E F StangeldquoCrohnrsquos disease a defensin deficiency syndromerdquo EuropeanJournal of Gastroenterology and Hepatology vol 15 no 6 pp627ndash634 2003

16 Evidence-Based Complementary and Alternative Medicine

[6] L Chen S Lin R Agha-Majzoub L Overbergh C Mathieuand L S Chan ldquoCCL27 is a critical factor for the developmentof atopic dermatitis in the keratin-14 IL-4 transgenic mousemodelrdquo International Immunology vol 18 no 8 pp 1233ndash12422006

[7] J H Song V Thamlikitkul and P R Hsueh ldquoClinical andeconomic burden of community-acquired pneumonia amongstadults in the Asia-Pacific regionrdquo International Journal ofAntimicrobial Agents vol 38 no 2 pp 108ndash117 2011

[8] D A B Dance ldquoMelioidosisrdquo Current Opinion in InfectiousDiseases vol 15 no 2 pp 127ndash132 2002

[9] N J White ldquoMelioidosisrdquo The Lancet vol 361 no 9370 pp1715ndash1722 2003

[10] S Yang ldquoMelioidosis research in Chinardquo Acta Tropica vol 77no 2 pp 157ndash165 2000

[11] P R Hsueh L J Teng L N Lee et al ldquoMelioidosis an emerginginfection in Taiwanrdquo Emerging Infectious Diseases vol 7 no 3pp 428ndash433 2001

[12] C Walsh and S Fanning ldquoAntimicrobial resistance in food-borne pathogensmdasha cause for concernrdquo Current Drug Targetsvol 9 no 9 pp 808ndash815 2008

[13] M Vorachit P Chongtrakool S Arkomsean and S BoonsongldquoAntimicrobial resistance in Burkholderia pseudomalleirdquo ActaTropica vol 74 no 2-3 pp 139ndash144 2000

[14] F M Thibault E Hernandez D R Vidal M Girardet and JD Cavallo ldquoAntibiotic susceptibility of 65 isolates of Burkholde-ria pseudomallei and Burkholderia mallei to 35 antimicrobialagentsrdquo Journal of Antimicrobial Chemotherapy vol 54 no 6pp 1134ndash1138 2004

[15] R L S Sikarwar ldquoEthnogynaecological uses of plants new toIndiardquo Ethnobotany vol 12 no 1-2 pp 112ndash115 2002

[16] R Siva ldquoStatus of natural dyes and dye-yielding plants in IndiardquoCurrent Science vol 92 no 7 pp 916ndash925 2007

[17] P Kumar Singh V Kumar R K Tiwari A Sharma ChV Rao and R H Singh ldquoMedico-ethnobotany of lsquochatararsquoblock of district sonebhadra Uttar Pradesh Indiardquo Advances inBiological Research vol 4 no 1 pp 65ndash80 2010

[18] W H Lewis and M P Elwin-Lewis Medical Botany PlantsAffecting Human Health John Wiley amp Sons New York NYUSA 2003

[19] R Perumal Samy P N Pushparaj and P GopalakrishnakoneldquoA compilation of bioactive compounds fromAyurvedardquo Bioin-formation vol 3 no 3 pp 100ndash110 2008

[20] WorldHealthOrganization (WHO)TraditionalMedicine Strat-egy 2002ndash2005 Genera World Health Organization GenevaSwitzerland 2002

[21] P S Haddad G A Azar S Groom and M Boivin ldquoNaturalhealth products modulation of immune function and preven-tion of chronic diseasesrdquo Evidence Based Complementary andAlternative Medicine vol 2 no 4 pp 513ndash520 2005

[22] K Poonam andG S Singh ldquoEthnobotanical study of medicinalplants used by the Taungya community in Terai Arc LandscapeIndiardquo Journal of Ethnopharmacology vol 123 no 1 pp 167ndash1762009

[23] G B Mahady ldquoMedicinal plants for the prevention and treat-ment of bacterial infectionsrdquo Current Pharmaceutical Designvol 11 no 19 pp 2405ndash2427 2005

[24] V P Kumar N S Chauhan H Padh and M Rajani ldquoSearchfor antibacterial and antifungal agents from selected Indianmedicinal plantsrdquo Journal of Ethnopharmacology vol 107 no2 pp 182ndash188 2006

[25] R Nair T Kalariya and S Chanda ldquoAntibacterial activity ofsome plant extracts used in folk medicinerdquo Journal of HerbalPharmacotherapy vol 7 no 3-4 pp 191ndash201 2007

[26] D Kalemba and A Kunicka ldquoAntibacterial and antifungalproperties of essential oilsrdquo Current Medicinal Chemistry vol10 no 10 pp 813ndash829 2003

[27] A H Ashour ldquoAntibacterial antifungal and anticancer activi-ties of volatile oils and extracts from stems leaves and flowers ofEucalyptus sideroxylon and Eucalyptus torquatardquoCancer BiologyandTherapy vol 7 no 3 pp 399ndash403 2008

[28] M J Abad L M Bedoya L Apaza and P Bermejo ldquoTheArtemisia L genus a review of bioactive essential oilsrdquoMolecules vol 17 no 3 pp 2542ndash2566 2012

[29] H N Marzoug M Romdhane A Lebrihi et al ldquoEucalyptusoleosa essential oils chemical composition and antimicrobialand antioxidant activities of the oils from different plant parts(stems leaves flowers and fruits)rdquoMolecules vol 16 no 2 pp1695ndash1709 2011

[30] C Proestos D Sereli and M Komaitis ldquoDetermination ofphenolic compounds in aromatic plants by RP-HPLC and GC-MSrdquo Food Chemistry vol 95 no 1 pp 44ndash52 2006

[31] S Chao G Young C Oberg and K Nakaoka ldquoInhibition ofmethicillin-resistant Staphylococcus aureus (MRSA) by essentialoilsrdquo Flavour and Fragrance Journal vol 23 no 6 pp 444ndash4492008

[32] M Akin D Oguz and H T Saracoglu ldquoAntibacterial effectsof some plant extracts from Labiatae (Lamiaceae) growingnaturally around Sirnak-Silopi Turkeyrdquo International Journal ofPharmaceutical and Applied Sciences vol 1 no 1 pp 4ndash47 2010

[33] K Bayoub T Baibai D Mountassif A Retmane and ASoukri ldquoAntibacterial activities of the crude ethanol extracts ofmedicinal plants against Listeriamonocytogenes and some otherpathogenic strainsrdquo African Journal of Biotechnology vol 9 no27 pp 4251ndash4258 2010

[34] R P Samy P Gopalakrishnakone M Sarumathi P Houghtonand S Ignacimuthu ldquoPurification of antibacterial agents fromTragia involucratamdasha popular tribal medicine for wound heal-ingrdquo Journal of Ethnopharmacology vol 107 no 1 pp 99ndash1062006

[35] K M Matthew The Flora of Tamil Nadu Carnatic Part IndashIIIprinted at the Diocesan Press Madras India 1981ndash1983

[36] AW BauerWMKirby J C Sherris andM Turck ldquoAntibioticsusceptibility testing by a standardized single disk methodrdquoAmerican Journal of Clinical Pathology vol 45 no 4 pp 493ndash496 1966

[37] National Committee for Clinical Laboratory Standards Per-formance Standards For Antimicrobial Susceptibility Testing12th Informational Supplement Approved standard M100-S12National Committee for Clinical Laboratory Standards WaynePa USA 2002

[38] E A Elsinghorst ldquoMeasurement of invasion by gentamicinresistancerdquoMethods in Enzymology vol 236 pp 405ndash420 1994

[39] J B Harborne Phytochemical Methods Chapman and HallNew York NY USA 1976

[40] H Wagner S Bladt and E M Zgairski Plant Drug AnalysisSpringer New York NY USA 1984

[41] N Myers R A Mittermeler C G Mittermeler G A B daFonseca and J Kent ldquoBiodiversity hotspots for conservationprioritiesrdquo Nature vol 403 no 6772 pp 853ndash858 2000

[42] R Perumal Samy and S Ignacimuthu ldquoAntibacterial activity ofsome folkloremedicinal plants used by tribals inWestern Ghats

Evidence-Based Complementary and Alternative Medicine 17

of Indiardquo Journal of Ethnopharmacology vol 69 no 1 pp 63ndash712000

[43] K Nampoothiri A Chrispal A Begum S Jasmine K GGopinath and A Zachariah ldquoA clinical study of renal tubulardysfunction inCleistanthus collinus (Oduvanthalai) poisoningrdquoClinical Toxicology vol 48 no 3 pp 193ndash197 2010

[44] M Strathern UsefulKnowledge The Isiah Berlin Lecture Uni-versity of Manchester Brunswick UK

[45] C N Sunil and A K Pradeep ldquo(2001) Another new species ofTripogon (Poaceae) from Indiardquo Sida vol 19 pp 803ndash806 2005

[46] S Biber-Klemm and T Cottier Right To Plant GeneticResources and Traditional Knowledge Basic Issues and Prospec-tive Oxfordshire Nosworthy Way UK 2006

[47] V S Ramachandran S Shijo Joseph and R Aruna ldquoEth-nobotanical studies from amaravathy range of Indira GandhiWildlife SanctuaryWesternGhats CoimbatoreDistrict South-ern Indiardquo Ethnobotanical Leaflets vol 13 pp 1069ndash1087 2009

[48] A H Rajasab and I Mohamad ldquoDocumentation of folkknowledge on edible wild plants of North Karnatakardquo IndianJournal of Traditional Knowledge vol 3 no 4 pp 419ndash4292004

[49] M Rajadurai V G Vidhya M Ramya and A Bhaskar ldquoEthno-medicinal plants used by the traditional healers of pachamalaihills Tamilnadu Indiardquo Studies on Ethno-Medicine vol 3 no 1pp 39ndash41 2009

[50] S K Adesina O Idowu A O Ogundaini et al ldquoAntimicrobialconstituents of the leaves of Acalypha wilkesiana and Acalyphahispidardquo Phytotherapy Research vol 14 no 5 pp 371ndash374 2000

[51] M T Gutierrez-Lugo M P Singh W M Maiese and BN Timmermann ldquoNew antimicrobial cycloartane triterpenesfrom Acalypha communisrdquo Journal of Natural Products vol 65no 6 pp 872ndash875 2002

[52] A K Tyagi andAMalik ldquoAntimicrobial potential and chemicalcomposition of Mentha piperita oil in liquid and vapour phaseagainst food spoiling microorganismsrdquo Food Control vol 22no 11 pp 1707ndash1714 2011

[53] G Devendran and U Balasubramanian ldquoQualitative phyto-chemical screening and GC-MS analysis of Ocimum sanctumL leavesrdquoAsian Journal of Plant Science and Research vol 1 no4 pp 44ndash48 2011

[54] V C Jain N M Patel D P Shah P K Patel and B HJoshi ldquoAntioxidant and antimicrobial activities of Alangiumsalvifolium (Lf) Wang rootrdquo Global Journal of Pharmacologyvol 4 no 1 pp 13ndash18 2010

[55] P Swapna Latha andK Kannabiran ldquoAntimicrobial activity andphytochemicals of Solanum trilobatum LinnrdquoAfrican Journal ofBiotechnology vol 5 no 23 pp 2402ndash2404 2006

[56] M Usha1 S Ragini and S M A Naqvi ldquoAntibacterial activityof acetone and ethanol extracts of Cinnamon (Cinnamomumzeylanicum) and Ajowan (Trachyspermum ammi) on four foodspoilage bacteriardquo International Research Journal of BiologicalSciences vol 1 no 4 pp 7ndash11 2012

[57] S Prabuseenivasan M Jayakumar and S Ignacimuthu ldquoInvitro antibacterial activity of some plant essential oilsrdquo BMCComplementary and Alternative Medicine vol 6 article 39 pp1ndash8 2006

[58] G A Ayoola F M Lawore T Adelowotan et al ldquoChemi-cal analysis and antimicrobial activity of the essential oil ofSyzigium aromaticum (clove)rdquo African Journal of Biotechnologyvol 7 no 13 pp 2227ndash2231 2008

[59] Y J Fu L Chen Y Zu et al ldquoThe antibacterial activity of cloveessential oil against Propionibacterium acnes and its mechanismof actionrdquo Archives of Dermatology vol 145 no 1 pp 86ndash882009

[60] D Venkatesan C M Karrunakarn S Selva Kumar and P TPalani Swamy ldquoIdentification of phytochemical constituents ofAegle marmelos responsible for antimicrobial activity againstselected pathogenic organismsrdquo Ethnobotanical Leaflets vol 13pp 1362ndash1372 2009

[61] P Arun K G Purushotham J Johnsy jayarani and V KumarildquoIn vitro Antibacterial Activity of Oldenlandia umbellata anIndian medicinal Plantrdquo Journal of Pharmaceutical Science andTechnology vol 2 no 4 pp 198ndash201 2010

[62] A Manjamalai R Sardar Sathyajith Singh C Guruvayoorap-pan and V M Berlin Grace ldquoAnalysis of phytochemicalconstituents and antimicrobial activity of some medicinalplants in Taminadu Indiardquo Global Journal of Biotechnology andBiochemistry vol 5 no 2 pp 120ndash128 2010

[63] E Haslam ldquoNatural polyphenols (vegetable tannins) as drugsPossible modes of actionrdquo Journal of Natural Products vol 59no 2 pp 205ndash215 1996

[64] Y Cai Q Luo M Sun and H Corke ldquoAntioxidant activityand phenolic compounds of 112 traditional Chinese medicinalplants associated with anticancerrdquo Life Sciences vol 74 no 17pp 2157ndash2184 2004

[65] B Shan Y Z Cai M Sun and H Corke ldquoAntioxidant capacityof 26 spice extracts and characterization of their phenolicconstituentsrdquo Journal of Agricultural and Food Chemistry vol53 no 20 pp 7749ndash7759 2005

[66] R J W Lambert P N Skandamis P J Coote and G-J ENychas ldquoA study of the minimum inhibitory concentration andmode of action of oregano essential oil thymol and carvacrolrdquoJournal of AppliedMicrobiology vol 91 no 3 pp 453ndash462 2001

[67] M Raccach ldquoThe antimicrobial activity of phenolic antioxi-dants in food a reviewrdquo Journal of Food Safety vol 6 no 3 pp141ndash170 1984

[68] M Taniguchi Y Yano E Tada et al ldquoMode of action of polygo-dial an antifungal sesquiterpene dialdehyderdquo Agricultural andBiological Chemistry vol 52 no 6 pp 1409ndash1414 1988

[69] G J E Nychas ldquoNatural antimicrobials from plantsrdquo in NewMethods of Food Preservation G W Gould Ed pp 58ndash89Blackie Academic London UK 1995

[70] S Burt ldquoEssential oils their antibacterial properties and poten-tial applications in foodsmdasha reviewrdquo International Journal ofFood Microbiology vol 94 no 3 pp 223ndash253 2004

Submit your manuscripts athttpwwwhindawicom

Stem CellsInternational

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

MEDIATORSINFLAMMATION

of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Behavioural Neurology

EndocrinologyInternational Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Disease Markers

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

BioMed Research International

OncologyJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Oxidative Medicine and Cellular Longevity

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

PPAR Research

The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014

Immunology ResearchHindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Journal of

ObesityJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Computational and Mathematical Methods in Medicine

OphthalmologyJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Diabetes ResearchJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Research and TreatmentAIDS

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Gastroenterology Research and Practice

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Parkinsonrsquos Disease

Evidence-Based Complementary and Alternative Medicine

Volume 2014Hindawi Publishing Corporationhttpwwwhindawicom

Evidence-Based Complementary and Alternative Medicine 15

M piperita (MIC of 113ndash225 mgmL) against the abovebacterial strains [52] MIC was found for the most activealcohol extracts of A salvifolium (MIC 0034ndash0263mgmL)on S aureus [54] and the MIC for S trilobatum aqueousextracts determined against the tested organisms rangedfrom 006 to 05mgmL [55] C zeylanicum was foundto have an effective antibacterial activity (MIC 64 120583gmL)against P aeruginosa E coli B subtilis and S aureus[56] Previously several investigators have demonstrated thatactive agents exert interesting activity against bacteria evenat lower concentrations tested [57 58] The bacteriostaticmechanism involves damage to the cell walls of bacteriafollowed by inhibition of protein synthesis that ultimatelyleads to bacterial death [59] The most active plants arewidely used by various tribes as traditional treatment (iecut wounds skin infection and scabies) thus indicatingthe potential for further development into promising drugsIn order to ascertain the safety and efficacy of the mostactive compounds their effect on human skin fibroblast cellswas evaluated Chemical constituents of E cardamomumT involucrata S indicus C acida A vasica A marmelosA indica and A paniculata plants failed to produce anynoticeable toxicity up to 1000120583gmL Although some ofthe tested compounds exhibited a slight reduction of cellproliferation and some minor morphological changes suchchanges were insignificant at lower doses and became evidentonly at higher doses However certain aromatic compoundsof O sanctum E globulus V zizanioides C citratus and Eglobulus plants showed reduction of cell proliferation againstHEPK cells

Our phytochemical screening also provides evidenceof the presence of several types of compounds that aremainly responsible for the remarkable antibacterial effectof these plants The differences noted for the bactericidalactivity of various plant extracts in this study appears tobe directly related to the diversity of compounds (shown inparentheses) that are accumulated in the following plants(eg A marmelos and O umbellate) [60ndash62] Compoundslike tannins phenol and polyphenols can bind the Gram-negative bacteria to form a heavy soluble complex on the cellsurface which subsequently disturbs the availability of recep-tor on cells and kills the bacteria [63] Several species havingwide spectra of antimicrobial activity mainly due to theactive constituents such as essential oil phenolic compoundslike thymol carvacrol in oregano and thyme eugenol inclove and cinnamon were also identified previously [64 65]Essential oils degrade the cell wall interact with the cell com-ponents and then disrupt the cytoplasmic membrane [66]The antimicrobial effect of phenolic compounds may involvemultiplemodes of action including damage to themembraneprotein interference with membrane integrated enzymes[67] causing leakage of cellular components coagulation ofcytoplasm depletion of the proton motive force alterationof fatty acid and phospholipid constituents impairmentof enzymatic mechanisms for production and metabolismalteration of nutrient uptake and electron transport [68]influencing the synthesis of DNA and RNA and destroyingprotein translation and the function of the mitochondrionin eukaryotes [69] The mode of action of antimicrobial

agents depends on the type of microorganism and is mainlyrelated to their cell wall structure and the outer membranearrangement Most of the plant spices and herbs containcomplex phenolics (ie phenolic acids flavonoids tanninslignans coumarins quinines) In addition the mechanismsof action of each phenolic compound against various bacteriaare also very complicated [26 70] Further investigation istherefore required to understand the relationship betweenthe antimicrobial action and the chemical structure of everyphenolic compound in the tested extracts The informationavailable from previous pharmacological sources combinedwith the findings herein reported on the medicinal plantextracts may serve as essential data for future drug develop-ment to combat diseases caused by theMDRbacterial strains

Abbreviations

MDR Multidrug resistant bacteriaTES Strain of Burkholderia pseudomalleiMRSA Methicillin-resistant Staphylococcus aureusVRE Vancomycin-resistant enterococciMTB Mycobacterium tuberculosisCH3OH Methanol

CFU Colony forming unitHCl Hydrochloric acidTLC Thin layer chromatographyH2SO4 Sulphuric acids

UV Ultraviolet spectrophotometerHEPK Human skin fibroblasts

Acknowledgments

Theauthors are thankful to the ERI Loyola College ChennaiIndia for his support during the survey The authors alsothank the Department of Microbiology NUHS NUS forthe bacterial cultures used in this investigation The authorsare thankful to the tribal people for sharing their healingknowledge on medicinal aromatic plants

References

[1] M N Alekshun and S B Levy ldquoMolecular mechanisms ofantibacterialmultidrug resistancerdquoCell vol 128 no 6 pp 1037ndash1050 2007

[2] A D Russell ldquoAntibiotic and biocide resistance in bacteriaintroductionrdquo Symposium Series Society for Applied Microbiol-ogy vol 31 no 92 pp 1Sndash3S 2002

[3] D R Guay ldquoTreatment of bacterial skin and skin structureinfectionsrdquo Expert Opinion on Pharmacotherapy vol 4 no 8pp 1259ndash1275 2003

[4] A J Brosnahan and P M Schlievert ldquoGram-positive bac-terial superantigen outside-in signaling causes toxic shocksyndromerdquo FEBS Journal vol 278 no 23 pp 4649ndash4667 2011

[5] K Fellermann J Wehkamp K R Herrlinger and E F StangeldquoCrohnrsquos disease a defensin deficiency syndromerdquo EuropeanJournal of Gastroenterology and Hepatology vol 15 no 6 pp627ndash634 2003

16 Evidence-Based Complementary and Alternative Medicine

[6] L Chen S Lin R Agha-Majzoub L Overbergh C Mathieuand L S Chan ldquoCCL27 is a critical factor for the developmentof atopic dermatitis in the keratin-14 IL-4 transgenic mousemodelrdquo International Immunology vol 18 no 8 pp 1233ndash12422006

[7] J H Song V Thamlikitkul and P R Hsueh ldquoClinical andeconomic burden of community-acquired pneumonia amongstadults in the Asia-Pacific regionrdquo International Journal ofAntimicrobial Agents vol 38 no 2 pp 108ndash117 2011

[8] D A B Dance ldquoMelioidosisrdquo Current Opinion in InfectiousDiseases vol 15 no 2 pp 127ndash132 2002

[9] N J White ldquoMelioidosisrdquo The Lancet vol 361 no 9370 pp1715ndash1722 2003

[10] S Yang ldquoMelioidosis research in Chinardquo Acta Tropica vol 77no 2 pp 157ndash165 2000

[11] P R Hsueh L J Teng L N Lee et al ldquoMelioidosis an emerginginfection in Taiwanrdquo Emerging Infectious Diseases vol 7 no 3pp 428ndash433 2001

[12] C Walsh and S Fanning ldquoAntimicrobial resistance in food-borne pathogensmdasha cause for concernrdquo Current Drug Targetsvol 9 no 9 pp 808ndash815 2008

[13] M Vorachit P Chongtrakool S Arkomsean and S BoonsongldquoAntimicrobial resistance in Burkholderia pseudomalleirdquo ActaTropica vol 74 no 2-3 pp 139ndash144 2000

[14] F M Thibault E Hernandez D R Vidal M Girardet and JD Cavallo ldquoAntibiotic susceptibility of 65 isolates of Burkholde-ria pseudomallei and Burkholderia mallei to 35 antimicrobialagentsrdquo Journal of Antimicrobial Chemotherapy vol 54 no 6pp 1134ndash1138 2004

[15] R L S Sikarwar ldquoEthnogynaecological uses of plants new toIndiardquo Ethnobotany vol 12 no 1-2 pp 112ndash115 2002

[16] R Siva ldquoStatus of natural dyes and dye-yielding plants in IndiardquoCurrent Science vol 92 no 7 pp 916ndash925 2007

[17] P Kumar Singh V Kumar R K Tiwari A Sharma ChV Rao and R H Singh ldquoMedico-ethnobotany of lsquochatararsquoblock of district sonebhadra Uttar Pradesh Indiardquo Advances inBiological Research vol 4 no 1 pp 65ndash80 2010

[18] W H Lewis and M P Elwin-Lewis Medical Botany PlantsAffecting Human Health John Wiley amp Sons New York NYUSA 2003

[19] R Perumal Samy P N Pushparaj and P GopalakrishnakoneldquoA compilation of bioactive compounds fromAyurvedardquo Bioin-formation vol 3 no 3 pp 100ndash110 2008

[20] WorldHealthOrganization (WHO)TraditionalMedicine Strat-egy 2002ndash2005 Genera World Health Organization GenevaSwitzerland 2002

[21] P S Haddad G A Azar S Groom and M Boivin ldquoNaturalhealth products modulation of immune function and preven-tion of chronic diseasesrdquo Evidence Based Complementary andAlternative Medicine vol 2 no 4 pp 513ndash520 2005

[22] K Poonam andG S Singh ldquoEthnobotanical study of medicinalplants used by the Taungya community in Terai Arc LandscapeIndiardquo Journal of Ethnopharmacology vol 123 no 1 pp 167ndash1762009

[23] G B Mahady ldquoMedicinal plants for the prevention and treat-ment of bacterial infectionsrdquo Current Pharmaceutical Designvol 11 no 19 pp 2405ndash2427 2005

[24] V P Kumar N S Chauhan H Padh and M Rajani ldquoSearchfor antibacterial and antifungal agents from selected Indianmedicinal plantsrdquo Journal of Ethnopharmacology vol 107 no2 pp 182ndash188 2006

[25] R Nair T Kalariya and S Chanda ldquoAntibacterial activity ofsome plant extracts used in folk medicinerdquo Journal of HerbalPharmacotherapy vol 7 no 3-4 pp 191ndash201 2007

[26] D Kalemba and A Kunicka ldquoAntibacterial and antifungalproperties of essential oilsrdquo Current Medicinal Chemistry vol10 no 10 pp 813ndash829 2003

[27] A H Ashour ldquoAntibacterial antifungal and anticancer activi-ties of volatile oils and extracts from stems leaves and flowers ofEucalyptus sideroxylon and Eucalyptus torquatardquoCancer BiologyandTherapy vol 7 no 3 pp 399ndash403 2008

[28] M J Abad L M Bedoya L Apaza and P Bermejo ldquoTheArtemisia L genus a review of bioactive essential oilsrdquoMolecules vol 17 no 3 pp 2542ndash2566 2012

[29] H N Marzoug M Romdhane A Lebrihi et al ldquoEucalyptusoleosa essential oils chemical composition and antimicrobialand antioxidant activities of the oils from different plant parts(stems leaves flowers and fruits)rdquoMolecules vol 16 no 2 pp1695ndash1709 2011

[30] C Proestos D Sereli and M Komaitis ldquoDetermination ofphenolic compounds in aromatic plants by RP-HPLC and GC-MSrdquo Food Chemistry vol 95 no 1 pp 44ndash52 2006

[31] S Chao G Young C Oberg and K Nakaoka ldquoInhibition ofmethicillin-resistant Staphylococcus aureus (MRSA) by essentialoilsrdquo Flavour and Fragrance Journal vol 23 no 6 pp 444ndash4492008

[32] M Akin D Oguz and H T Saracoglu ldquoAntibacterial effectsof some plant extracts from Labiatae (Lamiaceae) growingnaturally around Sirnak-Silopi Turkeyrdquo International Journal ofPharmaceutical and Applied Sciences vol 1 no 1 pp 4ndash47 2010

[33] K Bayoub T Baibai D Mountassif A Retmane and ASoukri ldquoAntibacterial activities of the crude ethanol extracts ofmedicinal plants against Listeriamonocytogenes and some otherpathogenic strainsrdquo African Journal of Biotechnology vol 9 no27 pp 4251ndash4258 2010

[34] R P Samy P Gopalakrishnakone M Sarumathi P Houghtonand S Ignacimuthu ldquoPurification of antibacterial agents fromTragia involucratamdasha popular tribal medicine for wound heal-ingrdquo Journal of Ethnopharmacology vol 107 no 1 pp 99ndash1062006

[35] K M Matthew The Flora of Tamil Nadu Carnatic Part IndashIIIprinted at the Diocesan Press Madras India 1981ndash1983

[36] AW BauerWMKirby J C Sherris andM Turck ldquoAntibioticsusceptibility testing by a standardized single disk methodrdquoAmerican Journal of Clinical Pathology vol 45 no 4 pp 493ndash496 1966

[37] National Committee for Clinical Laboratory Standards Per-formance Standards For Antimicrobial Susceptibility Testing12th Informational Supplement Approved standard M100-S12National Committee for Clinical Laboratory Standards WaynePa USA 2002

[38] E A Elsinghorst ldquoMeasurement of invasion by gentamicinresistancerdquoMethods in Enzymology vol 236 pp 405ndash420 1994

[39] J B Harborne Phytochemical Methods Chapman and HallNew York NY USA 1976

[40] H Wagner S Bladt and E M Zgairski Plant Drug AnalysisSpringer New York NY USA 1984

[41] N Myers R A Mittermeler C G Mittermeler G A B daFonseca and J Kent ldquoBiodiversity hotspots for conservationprioritiesrdquo Nature vol 403 no 6772 pp 853ndash858 2000

[42] R Perumal Samy and S Ignacimuthu ldquoAntibacterial activity ofsome folkloremedicinal plants used by tribals inWestern Ghats

Evidence-Based Complementary and Alternative Medicine 17

of Indiardquo Journal of Ethnopharmacology vol 69 no 1 pp 63ndash712000

[43] K Nampoothiri A Chrispal A Begum S Jasmine K GGopinath and A Zachariah ldquoA clinical study of renal tubulardysfunction inCleistanthus collinus (Oduvanthalai) poisoningrdquoClinical Toxicology vol 48 no 3 pp 193ndash197 2010

[44] M Strathern UsefulKnowledge The Isiah Berlin Lecture Uni-versity of Manchester Brunswick UK

[45] C N Sunil and A K Pradeep ldquo(2001) Another new species ofTripogon (Poaceae) from Indiardquo Sida vol 19 pp 803ndash806 2005

[46] S Biber-Klemm and T Cottier Right To Plant GeneticResources and Traditional Knowledge Basic Issues and Prospec-tive Oxfordshire Nosworthy Way UK 2006

[47] V S Ramachandran S Shijo Joseph and R Aruna ldquoEth-nobotanical studies from amaravathy range of Indira GandhiWildlife SanctuaryWesternGhats CoimbatoreDistrict South-ern Indiardquo Ethnobotanical Leaflets vol 13 pp 1069ndash1087 2009

[48] A H Rajasab and I Mohamad ldquoDocumentation of folkknowledge on edible wild plants of North Karnatakardquo IndianJournal of Traditional Knowledge vol 3 no 4 pp 419ndash4292004

[49] M Rajadurai V G Vidhya M Ramya and A Bhaskar ldquoEthno-medicinal plants used by the traditional healers of pachamalaihills Tamilnadu Indiardquo Studies on Ethno-Medicine vol 3 no 1pp 39ndash41 2009

[50] S K Adesina O Idowu A O Ogundaini et al ldquoAntimicrobialconstituents of the leaves of Acalypha wilkesiana and Acalyphahispidardquo Phytotherapy Research vol 14 no 5 pp 371ndash374 2000

[51] M T Gutierrez-Lugo M P Singh W M Maiese and BN Timmermann ldquoNew antimicrobial cycloartane triterpenesfrom Acalypha communisrdquo Journal of Natural Products vol 65no 6 pp 872ndash875 2002

[52] A K Tyagi andAMalik ldquoAntimicrobial potential and chemicalcomposition of Mentha piperita oil in liquid and vapour phaseagainst food spoiling microorganismsrdquo Food Control vol 22no 11 pp 1707ndash1714 2011

[53] G Devendran and U Balasubramanian ldquoQualitative phyto-chemical screening and GC-MS analysis of Ocimum sanctumL leavesrdquoAsian Journal of Plant Science and Research vol 1 no4 pp 44ndash48 2011

[54] V C Jain N M Patel D P Shah P K Patel and B HJoshi ldquoAntioxidant and antimicrobial activities of Alangiumsalvifolium (Lf) Wang rootrdquo Global Journal of Pharmacologyvol 4 no 1 pp 13ndash18 2010

[55] P Swapna Latha andK Kannabiran ldquoAntimicrobial activity andphytochemicals of Solanum trilobatum LinnrdquoAfrican Journal ofBiotechnology vol 5 no 23 pp 2402ndash2404 2006

[56] M Usha1 S Ragini and S M A Naqvi ldquoAntibacterial activityof acetone and ethanol extracts of Cinnamon (Cinnamomumzeylanicum) and Ajowan (Trachyspermum ammi) on four foodspoilage bacteriardquo International Research Journal of BiologicalSciences vol 1 no 4 pp 7ndash11 2012

[57] S Prabuseenivasan M Jayakumar and S Ignacimuthu ldquoInvitro antibacterial activity of some plant essential oilsrdquo BMCComplementary and Alternative Medicine vol 6 article 39 pp1ndash8 2006

[58] G A Ayoola F M Lawore T Adelowotan et al ldquoChemi-cal analysis and antimicrobial activity of the essential oil ofSyzigium aromaticum (clove)rdquo African Journal of Biotechnologyvol 7 no 13 pp 2227ndash2231 2008

[59] Y J Fu L Chen Y Zu et al ldquoThe antibacterial activity of cloveessential oil against Propionibacterium acnes and its mechanismof actionrdquo Archives of Dermatology vol 145 no 1 pp 86ndash882009

[60] D Venkatesan C M Karrunakarn S Selva Kumar and P TPalani Swamy ldquoIdentification of phytochemical constituents ofAegle marmelos responsible for antimicrobial activity againstselected pathogenic organismsrdquo Ethnobotanical Leaflets vol 13pp 1362ndash1372 2009

[61] P Arun K G Purushotham J Johnsy jayarani and V KumarildquoIn vitro Antibacterial Activity of Oldenlandia umbellata anIndian medicinal Plantrdquo Journal of Pharmaceutical Science andTechnology vol 2 no 4 pp 198ndash201 2010

[62] A Manjamalai R Sardar Sathyajith Singh C Guruvayoorap-pan and V M Berlin Grace ldquoAnalysis of phytochemicalconstituents and antimicrobial activity of some medicinalplants in Taminadu Indiardquo Global Journal of Biotechnology andBiochemistry vol 5 no 2 pp 120ndash128 2010

[63] E Haslam ldquoNatural polyphenols (vegetable tannins) as drugsPossible modes of actionrdquo Journal of Natural Products vol 59no 2 pp 205ndash215 1996

[64] Y Cai Q Luo M Sun and H Corke ldquoAntioxidant activityand phenolic compounds of 112 traditional Chinese medicinalplants associated with anticancerrdquo Life Sciences vol 74 no 17pp 2157ndash2184 2004

[65] B Shan Y Z Cai M Sun and H Corke ldquoAntioxidant capacityof 26 spice extracts and characterization of their phenolicconstituentsrdquo Journal of Agricultural and Food Chemistry vol53 no 20 pp 7749ndash7759 2005

[66] R J W Lambert P N Skandamis P J Coote and G-J ENychas ldquoA study of the minimum inhibitory concentration andmode of action of oregano essential oil thymol and carvacrolrdquoJournal of AppliedMicrobiology vol 91 no 3 pp 453ndash462 2001

[67] M Raccach ldquoThe antimicrobial activity of phenolic antioxi-dants in food a reviewrdquo Journal of Food Safety vol 6 no 3 pp141ndash170 1984

[68] M Taniguchi Y Yano E Tada et al ldquoMode of action of polygo-dial an antifungal sesquiterpene dialdehyderdquo Agricultural andBiological Chemistry vol 52 no 6 pp 1409ndash1414 1988

[69] G J E Nychas ldquoNatural antimicrobials from plantsrdquo in NewMethods of Food Preservation G W Gould Ed pp 58ndash89Blackie Academic London UK 1995

[70] S Burt ldquoEssential oils their antibacterial properties and poten-tial applications in foodsmdasha reviewrdquo International Journal ofFood Microbiology vol 94 no 3 pp 223ndash253 2004

Submit your manuscripts athttpwwwhindawicom

Stem CellsInternational

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

MEDIATORSINFLAMMATION

of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Behavioural Neurology

EndocrinologyInternational Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Disease Markers

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

BioMed Research International

OncologyJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Oxidative Medicine and Cellular Longevity

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

PPAR Research

The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014

Immunology ResearchHindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Journal of

ObesityJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Computational and Mathematical Methods in Medicine

OphthalmologyJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Diabetes ResearchJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Research and TreatmentAIDS

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Gastroenterology Research and Practice

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Parkinsonrsquos Disease

Evidence-Based Complementary and Alternative Medicine

Volume 2014Hindawi Publishing Corporationhttpwwwhindawicom

16 Evidence-Based Complementary and Alternative Medicine

[6] L Chen S Lin R Agha-Majzoub L Overbergh C Mathieuand L S Chan ldquoCCL27 is a critical factor for the developmentof atopic dermatitis in the keratin-14 IL-4 transgenic mousemodelrdquo International Immunology vol 18 no 8 pp 1233ndash12422006

[7] J H Song V Thamlikitkul and P R Hsueh ldquoClinical andeconomic burden of community-acquired pneumonia amongstadults in the Asia-Pacific regionrdquo International Journal ofAntimicrobial Agents vol 38 no 2 pp 108ndash117 2011

[8] D A B Dance ldquoMelioidosisrdquo Current Opinion in InfectiousDiseases vol 15 no 2 pp 127ndash132 2002

[9] N J White ldquoMelioidosisrdquo The Lancet vol 361 no 9370 pp1715ndash1722 2003

[10] S Yang ldquoMelioidosis research in Chinardquo Acta Tropica vol 77no 2 pp 157ndash165 2000

[11] P R Hsueh L J Teng L N Lee et al ldquoMelioidosis an emerginginfection in Taiwanrdquo Emerging Infectious Diseases vol 7 no 3pp 428ndash433 2001

[12] C Walsh and S Fanning ldquoAntimicrobial resistance in food-borne pathogensmdasha cause for concernrdquo Current Drug Targetsvol 9 no 9 pp 808ndash815 2008

[13] M Vorachit P Chongtrakool S Arkomsean and S BoonsongldquoAntimicrobial resistance in Burkholderia pseudomalleirdquo ActaTropica vol 74 no 2-3 pp 139ndash144 2000

[14] F M Thibault E Hernandez D R Vidal M Girardet and JD Cavallo ldquoAntibiotic susceptibility of 65 isolates of Burkholde-ria pseudomallei and Burkholderia mallei to 35 antimicrobialagentsrdquo Journal of Antimicrobial Chemotherapy vol 54 no 6pp 1134ndash1138 2004

[15] R L S Sikarwar ldquoEthnogynaecological uses of plants new toIndiardquo Ethnobotany vol 12 no 1-2 pp 112ndash115 2002

[16] R Siva ldquoStatus of natural dyes and dye-yielding plants in IndiardquoCurrent Science vol 92 no 7 pp 916ndash925 2007

[17] P Kumar Singh V Kumar R K Tiwari A Sharma ChV Rao and R H Singh ldquoMedico-ethnobotany of lsquochatararsquoblock of district sonebhadra Uttar Pradesh Indiardquo Advances inBiological Research vol 4 no 1 pp 65ndash80 2010

[18] W H Lewis and M P Elwin-Lewis Medical Botany PlantsAffecting Human Health John Wiley amp Sons New York NYUSA 2003

[19] R Perumal Samy P N Pushparaj and P GopalakrishnakoneldquoA compilation of bioactive compounds fromAyurvedardquo Bioin-formation vol 3 no 3 pp 100ndash110 2008

[20] WorldHealthOrganization (WHO)TraditionalMedicine Strat-egy 2002ndash2005 Genera World Health Organization GenevaSwitzerland 2002

[21] P S Haddad G A Azar S Groom and M Boivin ldquoNaturalhealth products modulation of immune function and preven-tion of chronic diseasesrdquo Evidence Based Complementary andAlternative Medicine vol 2 no 4 pp 513ndash520 2005

[22] K Poonam andG S Singh ldquoEthnobotanical study of medicinalplants used by the Taungya community in Terai Arc LandscapeIndiardquo Journal of Ethnopharmacology vol 123 no 1 pp 167ndash1762009

[23] G B Mahady ldquoMedicinal plants for the prevention and treat-ment of bacterial infectionsrdquo Current Pharmaceutical Designvol 11 no 19 pp 2405ndash2427 2005

[24] V P Kumar N S Chauhan H Padh and M Rajani ldquoSearchfor antibacterial and antifungal agents from selected Indianmedicinal plantsrdquo Journal of Ethnopharmacology vol 107 no2 pp 182ndash188 2006

[25] R Nair T Kalariya and S Chanda ldquoAntibacterial activity ofsome plant extracts used in folk medicinerdquo Journal of HerbalPharmacotherapy vol 7 no 3-4 pp 191ndash201 2007

[26] D Kalemba and A Kunicka ldquoAntibacterial and antifungalproperties of essential oilsrdquo Current Medicinal Chemistry vol10 no 10 pp 813ndash829 2003

[27] A H Ashour ldquoAntibacterial antifungal and anticancer activi-ties of volatile oils and extracts from stems leaves and flowers ofEucalyptus sideroxylon and Eucalyptus torquatardquoCancer BiologyandTherapy vol 7 no 3 pp 399ndash403 2008

[28] M J Abad L M Bedoya L Apaza and P Bermejo ldquoTheArtemisia L genus a review of bioactive essential oilsrdquoMolecules vol 17 no 3 pp 2542ndash2566 2012

[29] H N Marzoug M Romdhane A Lebrihi et al ldquoEucalyptusoleosa essential oils chemical composition and antimicrobialand antioxidant activities of the oils from different plant parts(stems leaves flowers and fruits)rdquoMolecules vol 16 no 2 pp1695ndash1709 2011

[30] C Proestos D Sereli and M Komaitis ldquoDetermination ofphenolic compounds in aromatic plants by RP-HPLC and GC-MSrdquo Food Chemistry vol 95 no 1 pp 44ndash52 2006

[31] S Chao G Young C Oberg and K Nakaoka ldquoInhibition ofmethicillin-resistant Staphylococcus aureus (MRSA) by essentialoilsrdquo Flavour and Fragrance Journal vol 23 no 6 pp 444ndash4492008

[32] M Akin D Oguz and H T Saracoglu ldquoAntibacterial effectsof some plant extracts from Labiatae (Lamiaceae) growingnaturally around Sirnak-Silopi Turkeyrdquo International Journal ofPharmaceutical and Applied Sciences vol 1 no 1 pp 4ndash47 2010

[33] K Bayoub T Baibai D Mountassif A Retmane and ASoukri ldquoAntibacterial activities of the crude ethanol extracts ofmedicinal plants against Listeriamonocytogenes and some otherpathogenic strainsrdquo African Journal of Biotechnology vol 9 no27 pp 4251ndash4258 2010

[34] R P Samy P Gopalakrishnakone M Sarumathi P Houghtonand S Ignacimuthu ldquoPurification of antibacterial agents fromTragia involucratamdasha popular tribal medicine for wound heal-ingrdquo Journal of Ethnopharmacology vol 107 no 1 pp 99ndash1062006

[35] K M Matthew The Flora of Tamil Nadu Carnatic Part IndashIIIprinted at the Diocesan Press Madras India 1981ndash1983

[36] AW BauerWMKirby J C Sherris andM Turck ldquoAntibioticsusceptibility testing by a standardized single disk methodrdquoAmerican Journal of Clinical Pathology vol 45 no 4 pp 493ndash496 1966

[37] National Committee for Clinical Laboratory Standards Per-formance Standards For Antimicrobial Susceptibility Testing12th Informational Supplement Approved standard M100-S12National Committee for Clinical Laboratory Standards WaynePa USA 2002

[38] E A Elsinghorst ldquoMeasurement of invasion by gentamicinresistancerdquoMethods in Enzymology vol 236 pp 405ndash420 1994

[39] J B Harborne Phytochemical Methods Chapman and HallNew York NY USA 1976

[40] H Wagner S Bladt and E M Zgairski Plant Drug AnalysisSpringer New York NY USA 1984

[41] N Myers R A Mittermeler C G Mittermeler G A B daFonseca and J Kent ldquoBiodiversity hotspots for conservationprioritiesrdquo Nature vol 403 no 6772 pp 853ndash858 2000

[42] R Perumal Samy and S Ignacimuthu ldquoAntibacterial activity ofsome folkloremedicinal plants used by tribals inWestern Ghats

Evidence-Based Complementary and Alternative Medicine 17

of Indiardquo Journal of Ethnopharmacology vol 69 no 1 pp 63ndash712000

[43] K Nampoothiri A Chrispal A Begum S Jasmine K GGopinath and A Zachariah ldquoA clinical study of renal tubulardysfunction inCleistanthus collinus (Oduvanthalai) poisoningrdquoClinical Toxicology vol 48 no 3 pp 193ndash197 2010

[44] M Strathern UsefulKnowledge The Isiah Berlin Lecture Uni-versity of Manchester Brunswick UK

[45] C N Sunil and A K Pradeep ldquo(2001) Another new species ofTripogon (Poaceae) from Indiardquo Sida vol 19 pp 803ndash806 2005

[46] S Biber-Klemm and T Cottier Right To Plant GeneticResources and Traditional Knowledge Basic Issues and Prospec-tive Oxfordshire Nosworthy Way UK 2006

[47] V S Ramachandran S Shijo Joseph and R Aruna ldquoEth-nobotanical studies from amaravathy range of Indira GandhiWildlife SanctuaryWesternGhats CoimbatoreDistrict South-ern Indiardquo Ethnobotanical Leaflets vol 13 pp 1069ndash1087 2009

[48] A H Rajasab and I Mohamad ldquoDocumentation of folkknowledge on edible wild plants of North Karnatakardquo IndianJournal of Traditional Knowledge vol 3 no 4 pp 419ndash4292004

[49] M Rajadurai V G Vidhya M Ramya and A Bhaskar ldquoEthno-medicinal plants used by the traditional healers of pachamalaihills Tamilnadu Indiardquo Studies on Ethno-Medicine vol 3 no 1pp 39ndash41 2009

[50] S K Adesina O Idowu A O Ogundaini et al ldquoAntimicrobialconstituents of the leaves of Acalypha wilkesiana and Acalyphahispidardquo Phytotherapy Research vol 14 no 5 pp 371ndash374 2000

[51] M T Gutierrez-Lugo M P Singh W M Maiese and BN Timmermann ldquoNew antimicrobial cycloartane triterpenesfrom Acalypha communisrdquo Journal of Natural Products vol 65no 6 pp 872ndash875 2002

[52] A K Tyagi andAMalik ldquoAntimicrobial potential and chemicalcomposition of Mentha piperita oil in liquid and vapour phaseagainst food spoiling microorganismsrdquo Food Control vol 22no 11 pp 1707ndash1714 2011

[53] G Devendran and U Balasubramanian ldquoQualitative phyto-chemical screening and GC-MS analysis of Ocimum sanctumL leavesrdquoAsian Journal of Plant Science and Research vol 1 no4 pp 44ndash48 2011

[54] V C Jain N M Patel D P Shah P K Patel and B HJoshi ldquoAntioxidant and antimicrobial activities of Alangiumsalvifolium (Lf) Wang rootrdquo Global Journal of Pharmacologyvol 4 no 1 pp 13ndash18 2010

[55] P Swapna Latha andK Kannabiran ldquoAntimicrobial activity andphytochemicals of Solanum trilobatum LinnrdquoAfrican Journal ofBiotechnology vol 5 no 23 pp 2402ndash2404 2006

[56] M Usha1 S Ragini and S M A Naqvi ldquoAntibacterial activityof acetone and ethanol extracts of Cinnamon (Cinnamomumzeylanicum) and Ajowan (Trachyspermum ammi) on four foodspoilage bacteriardquo International Research Journal of BiologicalSciences vol 1 no 4 pp 7ndash11 2012

[57] S Prabuseenivasan M Jayakumar and S Ignacimuthu ldquoInvitro antibacterial activity of some plant essential oilsrdquo BMCComplementary and Alternative Medicine vol 6 article 39 pp1ndash8 2006

[58] G A Ayoola F M Lawore T Adelowotan et al ldquoChemi-cal analysis and antimicrobial activity of the essential oil ofSyzigium aromaticum (clove)rdquo African Journal of Biotechnologyvol 7 no 13 pp 2227ndash2231 2008

[59] Y J Fu L Chen Y Zu et al ldquoThe antibacterial activity of cloveessential oil against Propionibacterium acnes and its mechanismof actionrdquo Archives of Dermatology vol 145 no 1 pp 86ndash882009

[60] D Venkatesan C M Karrunakarn S Selva Kumar and P TPalani Swamy ldquoIdentification of phytochemical constituents ofAegle marmelos responsible for antimicrobial activity againstselected pathogenic organismsrdquo Ethnobotanical Leaflets vol 13pp 1362ndash1372 2009

[61] P Arun K G Purushotham J Johnsy jayarani and V KumarildquoIn vitro Antibacterial Activity of Oldenlandia umbellata anIndian medicinal Plantrdquo Journal of Pharmaceutical Science andTechnology vol 2 no 4 pp 198ndash201 2010

[62] A Manjamalai R Sardar Sathyajith Singh C Guruvayoorap-pan and V M Berlin Grace ldquoAnalysis of phytochemicalconstituents and antimicrobial activity of some medicinalplants in Taminadu Indiardquo Global Journal of Biotechnology andBiochemistry vol 5 no 2 pp 120ndash128 2010

[63] E Haslam ldquoNatural polyphenols (vegetable tannins) as drugsPossible modes of actionrdquo Journal of Natural Products vol 59no 2 pp 205ndash215 1996

[64] Y Cai Q Luo M Sun and H Corke ldquoAntioxidant activityand phenolic compounds of 112 traditional Chinese medicinalplants associated with anticancerrdquo Life Sciences vol 74 no 17pp 2157ndash2184 2004

[65] B Shan Y Z Cai M Sun and H Corke ldquoAntioxidant capacityof 26 spice extracts and characterization of their phenolicconstituentsrdquo Journal of Agricultural and Food Chemistry vol53 no 20 pp 7749ndash7759 2005

[66] R J W Lambert P N Skandamis P J Coote and G-J ENychas ldquoA study of the minimum inhibitory concentration andmode of action of oregano essential oil thymol and carvacrolrdquoJournal of AppliedMicrobiology vol 91 no 3 pp 453ndash462 2001

[67] M Raccach ldquoThe antimicrobial activity of phenolic antioxi-dants in food a reviewrdquo Journal of Food Safety vol 6 no 3 pp141ndash170 1984

[68] M Taniguchi Y Yano E Tada et al ldquoMode of action of polygo-dial an antifungal sesquiterpene dialdehyderdquo Agricultural andBiological Chemistry vol 52 no 6 pp 1409ndash1414 1988

[69] G J E Nychas ldquoNatural antimicrobials from plantsrdquo in NewMethods of Food Preservation G W Gould Ed pp 58ndash89Blackie Academic London UK 1995

[70] S Burt ldquoEssential oils their antibacterial properties and poten-tial applications in foodsmdasha reviewrdquo International Journal ofFood Microbiology vol 94 no 3 pp 223ndash253 2004

Submit your manuscripts athttpwwwhindawicom

Stem CellsInternational

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

MEDIATORSINFLAMMATION

of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Behavioural Neurology

EndocrinologyInternational Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Disease Markers

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

BioMed Research International

OncologyJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Oxidative Medicine and Cellular Longevity

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

PPAR Research

The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014

Immunology ResearchHindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Journal of

ObesityJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Computational and Mathematical Methods in Medicine

OphthalmologyJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Diabetes ResearchJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Research and TreatmentAIDS

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Gastroenterology Research and Practice

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Parkinsonrsquos Disease

Evidence-Based Complementary and Alternative Medicine

Volume 2014Hindawi Publishing Corporationhttpwwwhindawicom

Evidence-Based Complementary and Alternative Medicine 17

of Indiardquo Journal of Ethnopharmacology vol 69 no 1 pp 63ndash712000

[43] K Nampoothiri A Chrispal A Begum S Jasmine K GGopinath and A Zachariah ldquoA clinical study of renal tubulardysfunction inCleistanthus collinus (Oduvanthalai) poisoningrdquoClinical Toxicology vol 48 no 3 pp 193ndash197 2010

[44] M Strathern UsefulKnowledge The Isiah Berlin Lecture Uni-versity of Manchester Brunswick UK

[45] C N Sunil and A K Pradeep ldquo(2001) Another new species ofTripogon (Poaceae) from Indiardquo Sida vol 19 pp 803ndash806 2005

[46] S Biber-Klemm and T Cottier Right To Plant GeneticResources and Traditional Knowledge Basic Issues and Prospec-tive Oxfordshire Nosworthy Way UK 2006

[47] V S Ramachandran S Shijo Joseph and R Aruna ldquoEth-nobotanical studies from amaravathy range of Indira GandhiWildlife SanctuaryWesternGhats CoimbatoreDistrict South-ern Indiardquo Ethnobotanical Leaflets vol 13 pp 1069ndash1087 2009

[48] A H Rajasab and I Mohamad ldquoDocumentation of folkknowledge on edible wild plants of North Karnatakardquo IndianJournal of Traditional Knowledge vol 3 no 4 pp 419ndash4292004

[49] M Rajadurai V G Vidhya M Ramya and A Bhaskar ldquoEthno-medicinal plants used by the traditional healers of pachamalaihills Tamilnadu Indiardquo Studies on Ethno-Medicine vol 3 no 1pp 39ndash41 2009

[50] S K Adesina O Idowu A O Ogundaini et al ldquoAntimicrobialconstituents of the leaves of Acalypha wilkesiana and Acalyphahispidardquo Phytotherapy Research vol 14 no 5 pp 371ndash374 2000

[51] M T Gutierrez-Lugo M P Singh W M Maiese and BN Timmermann ldquoNew antimicrobial cycloartane triterpenesfrom Acalypha communisrdquo Journal of Natural Products vol 65no 6 pp 872ndash875 2002

[52] A K Tyagi andAMalik ldquoAntimicrobial potential and chemicalcomposition of Mentha piperita oil in liquid and vapour phaseagainst food spoiling microorganismsrdquo Food Control vol 22no 11 pp 1707ndash1714 2011

[53] G Devendran and U Balasubramanian ldquoQualitative phyto-chemical screening and GC-MS analysis of Ocimum sanctumL leavesrdquoAsian Journal of Plant Science and Research vol 1 no4 pp 44ndash48 2011

[54] V C Jain N M Patel D P Shah P K Patel and B HJoshi ldquoAntioxidant and antimicrobial activities of Alangiumsalvifolium (Lf) Wang rootrdquo Global Journal of Pharmacologyvol 4 no 1 pp 13ndash18 2010

[55] P Swapna Latha andK Kannabiran ldquoAntimicrobial activity andphytochemicals of Solanum trilobatum LinnrdquoAfrican Journal ofBiotechnology vol 5 no 23 pp 2402ndash2404 2006

[56] M Usha1 S Ragini and S M A Naqvi ldquoAntibacterial activityof acetone and ethanol extracts of Cinnamon (Cinnamomumzeylanicum) and Ajowan (Trachyspermum ammi) on four foodspoilage bacteriardquo International Research Journal of BiologicalSciences vol 1 no 4 pp 7ndash11 2012

[57] S Prabuseenivasan M Jayakumar and S Ignacimuthu ldquoInvitro antibacterial activity of some plant essential oilsrdquo BMCComplementary and Alternative Medicine vol 6 article 39 pp1ndash8 2006

[58] G A Ayoola F M Lawore T Adelowotan et al ldquoChemi-cal analysis and antimicrobial activity of the essential oil ofSyzigium aromaticum (clove)rdquo African Journal of Biotechnologyvol 7 no 13 pp 2227ndash2231 2008

[59] Y J Fu L Chen Y Zu et al ldquoThe antibacterial activity of cloveessential oil against Propionibacterium acnes and its mechanismof actionrdquo Archives of Dermatology vol 145 no 1 pp 86ndash882009

[60] D Venkatesan C M Karrunakarn S Selva Kumar and P TPalani Swamy ldquoIdentification of phytochemical constituents ofAegle marmelos responsible for antimicrobial activity againstselected pathogenic organismsrdquo Ethnobotanical Leaflets vol 13pp 1362ndash1372 2009

[61] P Arun K G Purushotham J Johnsy jayarani and V KumarildquoIn vitro Antibacterial Activity of Oldenlandia umbellata anIndian medicinal Plantrdquo Journal of Pharmaceutical Science andTechnology vol 2 no 4 pp 198ndash201 2010

[62] A Manjamalai R Sardar Sathyajith Singh C Guruvayoorap-pan and V M Berlin Grace ldquoAnalysis of phytochemicalconstituents and antimicrobial activity of some medicinalplants in Taminadu Indiardquo Global Journal of Biotechnology andBiochemistry vol 5 no 2 pp 120ndash128 2010

[63] E Haslam ldquoNatural polyphenols (vegetable tannins) as drugsPossible modes of actionrdquo Journal of Natural Products vol 59no 2 pp 205ndash215 1996

[64] Y Cai Q Luo M Sun and H Corke ldquoAntioxidant activityand phenolic compounds of 112 traditional Chinese medicinalplants associated with anticancerrdquo Life Sciences vol 74 no 17pp 2157ndash2184 2004

[65] B Shan Y Z Cai M Sun and H Corke ldquoAntioxidant capacityof 26 spice extracts and characterization of their phenolicconstituentsrdquo Journal of Agricultural and Food Chemistry vol53 no 20 pp 7749ndash7759 2005

[66] R J W Lambert P N Skandamis P J Coote and G-J ENychas ldquoA study of the minimum inhibitory concentration andmode of action of oregano essential oil thymol and carvacrolrdquoJournal of AppliedMicrobiology vol 91 no 3 pp 453ndash462 2001

[67] M Raccach ldquoThe antimicrobial activity of phenolic antioxi-dants in food a reviewrdquo Journal of Food Safety vol 6 no 3 pp141ndash170 1984

[68] M Taniguchi Y Yano E Tada et al ldquoMode of action of polygo-dial an antifungal sesquiterpene dialdehyderdquo Agricultural andBiological Chemistry vol 52 no 6 pp 1409ndash1414 1988

[69] G J E Nychas ldquoNatural antimicrobials from plantsrdquo in NewMethods of Food Preservation G W Gould Ed pp 58ndash89Blackie Academic London UK 1995

[70] S Burt ldquoEssential oils their antibacterial properties and poten-tial applications in foodsmdasha reviewrdquo International Journal ofFood Microbiology vol 94 no 3 pp 223ndash253 2004

Submit your manuscripts athttpwwwhindawicom

Stem CellsInternational

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

MEDIATORSINFLAMMATION

of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Behavioural Neurology

EndocrinologyInternational Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Disease Markers

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

BioMed Research International

OncologyJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Oxidative Medicine and Cellular Longevity

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

PPAR Research

The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014

Immunology ResearchHindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Journal of

ObesityJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Computational and Mathematical Methods in Medicine

OphthalmologyJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Diabetes ResearchJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Research and TreatmentAIDS

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Gastroenterology Research and Practice

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Parkinsonrsquos Disease

Evidence-Based Complementary and Alternative Medicine

Volume 2014Hindawi Publishing Corporationhttpwwwhindawicom

Submit your manuscripts athttpwwwhindawicom

Stem CellsInternational

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

MEDIATORSINFLAMMATION

of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Behavioural Neurology

EndocrinologyInternational Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Disease Markers

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

BioMed Research International

OncologyJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Oxidative Medicine and Cellular Longevity

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

PPAR Research

The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014

Immunology ResearchHindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Journal of

ObesityJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Computational and Mathematical Methods in Medicine

OphthalmologyJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Diabetes ResearchJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Research and TreatmentAIDS

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Gastroenterology Research and Practice

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Parkinsonrsquos Disease

Evidence-Based Complementary and Alternative Medicine

Volume 2014Hindawi Publishing Corporationhttpwwwhindawicom