Research Article Antibacterial, Antioxidant, and...

Hindawi Publishing CorporationBioMed Research InternationalVolume 2013 Article ID 510736 9 pageshttpdxdoiorg1011552013510736

Research ArticleAntibacterial Antioxidant and Anticholinesterase Activities ofPlant Seed Extracts from Brazilian Semiarid Region

Davi Felipe Farias1 Terezinha Maria Souza2 Martocircnio Ponte Viana1

Bruno Marques Soares3 Arcelina Pacheco Cunha4 Ilka Maria Vasconcelos1

Naacutegila Maria Pontes Silva Ricardo4 Paulo Michel Pinheiro Ferreira56

Vacircnia Maria Maciel Melo7 and Ana Fontenele Urano Carvalho7

1 Departamento de Bioquımica e Biologia Molecular Universidade Federal do Ceara 60440-970 Fortaleza CE Brazil2 Netherlands Toxicogenomics Centre Maastricht University Universiteitssingel 50 6229ER Maastricht The Netherlands3 Departamento de Fisiologia e Farmacologia Faculdade de Medicina Universidade Federal do Ceara60430-270 Fortaleza CE Brazil

4Departamento de Quımica Organica e Inorganica Universidade Federal do Ceara 60440-970 Fortaleza CE Brazil5 Departamento de Ciencias Biologicas Campus Senador Helvıdio Nunes de Barros Universidade Federal do Piauı64600-000 Picos PI Brazil

6 Programa de Pos-Graduacao em Ciencias Farmaceuticas Universidade Federal do Piauı Avenida Universitariasn 64049-550 Teresina PI Brazil

7 Departamento de Biologia Universidade Federal do Ceara Campus do Pici 60451-970 Fortaleza CE Brazil

Correspondence should be addressed to Ana Fontenele Urano Carvalho auranoufcbr

Received 9 July 2013 Accepted 28 October 2013

Academic Editor Kazim Husain

Copyright copy 2013 Davi Felipe Farias 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

The antimicrobial antioxidant and anticholinesterase activities of ethanolic seed extracts of twenty-one plant species fromBraziliansemiarid region were investigated The extracts were tested for antimicrobial activity against six bacteria strains and three yeastsSix extracts presented activity against the Gram (minus) organism Salmonella choleraesuis and the Gram (+) organisms Staphylococcusaureus and Bacillus subtilis The MIC values ranged from 496 to 3732mgmL The Triplaris gardneriana extract presented activityagainst the three species withMIC values 188 1376 and 1115mgmL respectively Five extracts presented antioxidant activity withEC50 values ranging from 6973 120583gmL (T gardneriana) to 48751 120583gmL (Licania rigida) For the anticholinesterase activity elevenextracts were capable of inhibiting the enzyme activity From those T gardneriana Parkia platycephala and Connarus detersuspresented the best activities with inhibition values of 767 715 and 919 respectively The extracts that presented antimicrobialactivity were tested for hemolytic assay against humanA B andOblood types and rabbit blood From those only theMyracrodruonurundeuva extract presented activity (about 20 of hemolysis at the lowest tested concentration 19120583gmL) Infrared spectroscopyof six representative extracts attested the presence of tannins polyphenols and flavonoids which was confirmed by a qualitativephytochemical assay

1 Introduction

Plants are considered natural sources of new compounds ofmedical and biotechnological interest since they synthesizea large variety of bioactive compounds An analysis intothe sources of new drugs from 1981 to 2007 reveals thatalmost half of the drugs approved since 1994 were based on

natural products Among these there are various examplesof development of new drugs from plant sources [1] Manyresearches have dedicated great effort to detect secondarymetabolites with biological activities against cancer microor-ganisms tropical diseases and others [2 3] Well-knownexamples of these compounds include flavonoids phe-nols and phenolic glycosides unsaturated lactones sulphur

2 BioMed Research International

compounds saponins cyanogenic glycosides and glucosino-lates [4ndash6]

In this context the Brazilian semiarid region is a richand underexploited source of bioactive molecules This areais covered mainly by xeric shrublands known as ldquoCaatingardquoThis dry land vegetation grows over an area of 800000 km2in Northeastern Brazil and out of 1000 vascular plant speciesdescribed many are endemic [7 8] Many of these plants areused in folk medicine by local communities to treat severaldiseases pointing out the potential to discover new sourcesof useful compounds for several applications [9 10] Recentlyour team has reported for seed ethanolic extracts of BrazilianNortheastern plants promising results of insecticidal activityagainst Aedes aegypti [11] and antiproliferative effects totumor cell lines [12] In order to extend our knowledgeon the Brazilian semiarid vegetation this work aimed toevaluate the presence of antimicrobial anticholinesteraseand antioxidant activities in ethanolic seed extracts of twenty-one plant species The antimicrobial activity was performedby the agar diffusion assay using pathogenic bacteria andyeast antioxidant activity by the DPPH radical scavengingand anticholinesterase activity were measured In addition ahemolytic activity assay was done for antibacterial samples toassess the effects on nontarget cells

2 Materials and Methods

21 Plant Collection Mature wild seeds of twenty-one plantspecies were harvested (at least 500 g of each species) duringthe dry period (from January 2005 to March 2008) in thesemiarid region of the Ceara State Northeastern BrazilPlants were identified by taxonomist Dr Edson de PaulaNunes and voucher specimens were deposited at HerbariumPrisco Bezerra (EAC) at the Federal University of Ceara(Fortaleza Brazil)The studied species are depicted in Table 1which shows harvest information voucher numbers com-mon names and the traditional medicinal use by localpopulation

22 Preparation of Ethanolic Seed Extracts Seeds of freshlycollected plant material were separated from other plantmaterials air-dried and ground in a laboratory mill to amoderately fine powder (particle size le 05mm) Powderedmaterial (500 g) was submitted to extraction with 99ethanol (15 L) at room temperature (25ndash27∘C) for 3 daysThe supernatant was removed and filtered throughWhatmannumber 1 paper After that the initial powder sample wasstill submitted to two extractions which resulted in a finalvolume of extract near 45 L The ethanolic extracts resultingfrom three consecutive extractions were mixed together thenconcentrated under reduced pressure in rotary evaporatoruntil complete elimination of solvent and stored in a freezerat minus20∘C

A stock solution containing 50000 120583gsdotmLminus1 of eachcrude extract was prepared by suspending 50000mg ofextract in 5mL of pure dimethylsulphoxide (DMSO AldrichMilwaukee WI USA) and mixed by sonication (model RK-100 Bandelin Berlin Germany) for 20min The volume was

adjusted to 1000mL with distilled water to provide assaysolutions as required

23 Microorganisms Six bacterial strains were used Staphy-lococcus aureus (ATCC 25923) Bacillus subtilis (ATCC 6633)Enterobacter aerogenes (ATCC 13048) Salmonella cholerae-suis (ATCC 10708) Klebsiella pneumoniae (ATCC 10031)and Pseudomonas aeruginosa (ATCC 25619)These standardswere obtained from the American Type Culture Collection(ATCC) (Manassas VA USA) The stock culture was main-tained on Mueller Hinton agar (HiMedia Laboratories PvtLtdMumbai India)medium at 4∘CThree pathogenic yeastswere usedCandida albicansC krusei andC tropicalisThesefungal strains were obtained from the Microbial Ecologyand Biotechnology Laboratory Biology Department of theFederal University of Ceara Brazil The stock cultures weremaintained on Sabouraud Dextrose agar (HiMedia Labora-tories Pvt Ltd Mumbai India) medium at 4∘C

24 Antibacterial and Antifungal Activity The disc diffu-sion method was followed for antibacterial and antifungalsusceptibility test as described previously [16] with somemodifications Petri dishes were prepared by pouring 20mLof Mueller-Hinton agar for bacteria and Sabouraud Dextroseagar for fungi and allowed to solidify Dishes were dried and01mL of standardized inoculum suspension (106 CFUmLfor bacteria and absorbance of 0600 at 450 nm for fungi)was poured and uniformly spread The excess inoculum wasdrained and the inoculum was allowed to dry for 5min Thepaper filter discs (7mm of diameter) containing 20120583L ofeach ethanolic extract were then applied and the dishes wereincubated at 37∘C for 18 h for bacterial growth and at thesame temperature for 48 h for fungal growth Tetracycline(30mgdisc) was used as positive control for the antibacterialassay Nystatin solution (100000 IUmL EMS SP Brazil)(20120583Ldisc) was used as positive control for the antifungalassayThe inhibition zone was measured from the edge of thedisc to the inner margin of the surrounding pathogens Alldeterminations were run in triplicate

25 Minimum Inhibitory Concentration (MIC) Minimuminhibitory concentrations of the seed ethanolic extracts weretested as described previously by the twofold serial dilutionmethodThe test extract was dissolved in 5DMSO to obtain50mgsdotmLminus1 stock solution Stock solution was diluted withdistilled water to achieve 45 40 35 30 25 20 15 10 and5mgsdotmLminus1 or as required The wet paper discs with dilutionswere placed over the dish and the cultures were incubated inBOD incubators at 37∘C for 18 h (bacteria) and 37∘C for 48 h(yeasts) The lowest concentration which inhibits the visiblegrowth of tested organism after macroscopic evaluation wasdetermined asMICAll determinationswere run in triplicate

26 Antioxidant Activity The capacity to scavenge the 22-diphenyl-1-picrylhydrazyl (DPPH) free radical was moni-tored [17] The diluted crude extracts (01mL) in differ-ent concentrations (5 to 1000 120583gsdotmLminus1) were mixed with39mL of methanolic solution containing DPPH radicals

BioMed Research International 3

Table1Antibacteria

l(MIC)antio

xidantand

hemolyticactiv

ities

ofethano

licseed

extractsof

plantsfro

mBrazilian

semiarid

region

a

Family

Botanicaln

ame

Com

mon

name

(localitydateof

collection)V

oucher

number

Tradition

almedicinaluse

Yield

()b

Antioxidant

activ

ity(EC 5

0120583gmL)

cAnticho

linesterase

activ

ity(

)d

Antibacteria

lactivity

MIC

(mgmL)

e

SCh

oleraesuis

Bsubtilis

Saureus

Anacardiaceae

Myracrodruonurun

deuvaFrA

llAroeira-do-sertao

(Ararip

eNationalForest0107)

EAC34865

Usedto

treatcutaneou

sand

gynecologicalaffections

and

kidn

eyandrespira

tory

prob

lemsantiinfl

ammatory

antiu

lcerandhealing[13]

antih

istam

inicand

antim

icrobial[14

]analgesic

andantid

iarrheal[15]

252

ND

fND

ND

678

ND

Schinopsisbrasiliensis

Engler

Brauna

(Ararip


EAC35643

Usedto

treatnervou

snessa

ndhyste

riaanalgesic[15]

288

ND

ND

3732

ND

ND

Caryocaraceae

Caryocar

coria

ceum

Wittm

Pequ

i(Ararip


EAC35251

Not

describ

ed231

ND

634

ND

ND

ND

Chrysobalana

ceae

Licaniatomentosa

Benth

Oiti(Coastzone010

8)EA

C40215

Not

describ

ed44

21672

139

ND

ND

649

Licaniarig

idaBe

nth

Oiticica

(Coastzone1207)

EAC40216

Usedto

treatdiabetes

anti-inflammatory[15]

281

48751

524

ND

ND

863

Conn

araceae

Conn

arus

detersus

Planch

Cabelode

negro

(Ararip


EAC34733

Not

describ

ed285

ND

919

ND

ND

ND


Table1Con

tinued

Family

Botanicaln

ame

Com

mon

name

(localitydateof

collection)V

oucher

number

Tradition

almedicinaluse

Yield

()b

Antioxidant

activ

ity(EC 5

0120583gmL)

cAnticho

linesterase

activ

ity(

)d

Antibacteria

lactivity

MIC

(mgmL)

e

SCh

oleraesuis

Bsubtilis

Saureus

Fabaceae

Adenantherapavonina

LFalso

-sandalo

(Coastzone0807)

EAC38697

Not

describ

ed612

ND

ND

ND

ND

ND

Amburana

cearensis

(Allemao)A

C

Smith

Cumaruandam

burana

(Ca

atinga

vegetatio

n1107)

EAC39618

Usedto

treatrheumatism

coldand

sinusitis

antispasm

odichealin

gand


analgesic

[13]

643

ND

ND

ND

ND

ND

Anadenantheramacrocarpa

(Benth)Brenan

Ang

icoverm

elho

(Ca

atinga

vegetatio

n0705)

EAC38389

Usedto

treatrespira

tory

prob

lemsgono

rrheaand

diarrheaastrin

gent

antirheum

atic

anti-inflammatorysedativ

eandhealing[15]

935

ND

541

ND

ND

ND

Diocle

amegacarpa

Rolfe

Mucun

aolho

-de-bo

i(Ararip


EAC38110

Not

describ

ed240

ND

ND

ND

ND

ND

Enterolobium

contortisiliquum

(Vell)

Moron

gOrelha-de-negro

(Ararip


EAC38115

Not

describ

ed280

ND

ND

ND

ND

ND

Hym

enaeacourbaril

LJatoba(Ararip


EAC38108

Not

describ

ed95

02479

5ND

ND

ND

ND

Lonchocarpus

seric

eus(Po

iret)Ku

nth

Inga(Coastzone1006)

EAC39615

Not

describ

ed1060

ND

ND

ND

ND

ND

Luetzelburgiaauric

ulata(A

llemao)

Ducke

Pau-moco

(Ca

atinga

vegetatio

n1106)

EAC40369

Not

describ

ed840

ND

ND

ND

ND

ND

BioMed Research International 5Ta

ble1Con

tinued

Family

Botanicaln

ame

Com

mon

name

(localitydateof

collection)V

oucher

number

Tradition

almedicinaluse

Yield

()b

Antioxidant

activ

ity(EC 5

0120583gmL)

cAnticho

linesterase

activ

ity(

)d

Antibacteria

lactivity

MIC

(mgmL)

e

SCh

oleraesuis

Bsubtilis

Saureus

ParkiaplatycephalaBe

nth

Visgueiro

(Ararip


EAC3

8109

Not

describ

ed1750

ND

715

ND

ND

ND

Piptadeniamoniliform

isBe

nth

Catand

uva

(Ca

atinga

vegetatio

n0105)

EAC3

5974

Not

describ

ed618

ND

502

ND

ND

ND

Senn

aobtusifolia(L)HSIrw

inamp

Barneby

Matap

asto

(Ca

atinga

vegetatio

n0705)

EAC3

9320

Not

describ

ed570

ND

ND

ND

496

ND

Polyg

onaceae

Triplaris

gardneria

naWedd

Pajeu

(Ca

atinga

vegetatio

n1106)

EAC3

9600

Not

describ

ed260

6973

798

188

1376

1115

Rham

naceae

Zizip

husjoazeiro

Mart

Juaa

ndjuazeiro

(Coastzone0308)

EAC40366

Expectorantand

antip

yretic

used

totre

atskinblood

sto

machandliver

diseases

ulcerantim

icrobial[15]

722

ND

494

ND

ND

ND

Sapind

aceae

Talisiaesculen

ta(A

St-Hil)

Radlk

Pitomba

(Coastzone0308)

EAC40

685

Not

describ

ed15

0ND

6124

ND

ND

ND

Sapind

ussaponaria

LSabo

nete

(Ca

atinga

vegetatio

n1106)

EAC3

9601

Not

describ

ed107

ND

5860

ND

ND

ND

Controlsg

Quercetin

mdashmdash

5552

mdashmdash

mdashmdash

Vitamin

Cmdash

mdash26027

mdashmdash

mdashmdash

a Thev

aluesa

remeans

ofatrip

licateTh

estand

arddeviationwas

lessthan

5of

them

ean

b (Amou

nt(g)o

fsolid

recoveredfro

mthee

thanolicextracta

mou

nt(g)o

fseedflo

urused)times

100

c Efficientcon

centratio

n(EC)

as120583

gof

sampler

equiredto

decrease

oneg

oftheinitia

l22-diphenyl-1-picrylhydrazyl(DPP

Hsdot)con

centratio

nby

50

d Totalinhibitio

nof

cholinesterase

enzymea

fterc

omparis

onto

theb

lank

Con

centratio

nteste

d500120583

gmL

e Minim

uminhibitory

concentration

thelow

estcon

centratio

nwhich

didno

tsho

wanygrow

thof

teste

dorganism

after

macroscop

icevaluatio

nf N

Dnot

detected

g Sub

stancesthatare

frequ

ently

used

assta

ndardfora

ntioxidant

studies


(65 times 10minus1molsdotLminus1) The reduction of the DPPH radicalwas measured by monitoring continuously the decrease ofabsorption at 515 nm The percentage scavenging of theDPPH radical was calculated according to the followingformula scavenging effect = 100 times [(119860DPPH minus As)119860DPPH]where As is the absorbance of the solution when the samplehas been added at a particular level and 119860DPPH is theabsorbance of the DPPH solution The percentage of theremaining DPPH was plotted against the samplestandardconcentration to obtain the amount of antioxidant necessaryto decrease the initial concentration of DPPH by 50 (EC

50)

Based on the parameter EC50 the result was expressed in

terms of 120583g of seed extract per mL of DPPH in the reactionmedium All determinations were run in triplicate

27 Anticholinesterase Activity The anticholinesterase activ-ity of the ethanolic extracts was measured as described by[18] with slight modifications Acetylcholinesterase activitywas measured using a 96-well microplate reader basedon Ellmanrsquos method [19] In this method the enzymehydrolyzes the substrate acetylthiocholine resulting in theproduction of thiocholine which reacts with 551015840-dithiobis(2-nitrobenzoic acid) (DTNB) to produce 2-nitrobenzoate-5-mercaptothiocholine and 5-thio-2-nitrobenzoate which canbe detected at 405 nm In the 96-well plates 25120583L of 15mMacetylthiocholine iodide (ATCI) in water 125 120583L of 3mMDTNB in 50mM Tris-HCl buffer containing 01M NaCland 002M MgCl

2sdot6H2O 50 120583L of 50mM Tris-HCl buffer

containing 01 BSA and 25 120583L of plant extract sample(5mgsdotmLminus1 in MeOH diluted ten times with 50mM Tris-HCl buffer to give a concentration of 05mgsdotmLminus1) wereadded and the absorbance was measured at 405 nm every30 s for three times Then 25120583L of 022UsdotmLminus1 of acetyl-cholinesterase fromelectric eel was added and the absorbancewas again read every 45 s for eight times Any increase inabsorbance due to the spontaneous hydrolysis of the substratewas corrected by subtracting the rate of the reaction beforethe addition of the enzyme from the rate of the enzymereaction The percentage of inhibition was calculated bycomparison of sample rates to a blank

28 Hemolytic Activity Assay The test was performed in15mL microtubes following the method described by [20]with some modifications Firstly a twofold serial dilution ofeach seed ethanolic extract was prepared with 09 NaClranging from 1000 to 19120583gsdotmLminus1 and reservedThen 100120583Lof a 1 red blood cells (A B and O human blood types orrabbit blood) suspension was added to a newmicrotube con-taining 900 120583L of each seed extract dilution which was thentaken to incubator at 37∘C for 1 h After that the tubes werecentrifuged at 3000timesg for 5min The supernatant (200120583L)was placed in a 96-well plate and led to a microplate reader(Epoch BioTek Vermont USA) to measure the absorbanceat 540 nm The cell suspensions of each human blood typeor those of rabbits (100120583L) were mixed with distilled wateror 09 NaCl (900120583L) to obtain the absorbances for 100and 0 of cell lysis respectivelyThe percentage of hemolysiswas calculated as follows hemolysis = AbstestAbs119901119888times 100

where Abstest = Abs540 of the 1 cell suspension treated withsample test and Abs

119901119888= Abs

540of the 1 cell suspension

treated with distilled water To calculate the relation betweenpercentage of hemolysis and seed extract concentration thehemolytic activity was expressed as the lowest seed extractconcentration (120583gsdotmLminus1) capable of causing hemolysis ge20All determinations were run in triplicate

29 Infrared Spectroscopy The infrared spectrum wasrecorded from KBr discs on a FTIR Perkin Elmer model16 PC spectrophotometer (PerkinElmer Waltham USA)Absorptionmaxima (Egraveđmax) were reported in wavenumbers(cmminus1) The spectra were used to determine organic groupsindicative of the main classes of secondary metabolites

210 Phytochemical Study The ethanolic seed extracts werescreened for phytochemical compounds such as phenols andtannins (reaction with ferric chloride) leucoanthocyanidins(heat treatment followed by alkalinization and acidification ofsample) flavonoids and xanthones (reaction of magnesiumgranules with hydrochloric acid) steroids and triterpenes(extraction with chloroform acetic anhydride and sulfuricacid) saponins (foam production after water solubilizationand stirring) and alkaloids (precipitation with Hager Mayerand Dragendorff reagents) [21] These analyses were basedon visual observation of color modification or precipitateformation after addition of specific reagents

3 Results and Discussion

The ethanolic seed extracts did not show activity againstthe Gram (minus) organisms E aerogenes K pneumonia and Paeruginosa nor against the yeastsC albicansC krusei andCtropicalis MIC values obtained against the Gram (+) organ-isms S aureus and B subtilis and the Gram (minus) organism Scholeraesuis are shown in Table 1 Among all samples testedand microorganisms used the seed extract of S obtusifoliapresented the lowest MIC value (496mgsdotmLminus1) against thepathogenic bacteria S aureus However the speciesT gardne-riana showed the broadest spectrum of activity being activeagainst B subtilis S choleraesuis and S aureuswithMIC val-ues of 188 1376 and 1115mgsdotmLminus1 respectively Althoughthere are a great number of studies reporting antimicrobialactivity of plant parts like leaves roots and stems antimi-crobial screenings using specifically seed extracts are veryrare The reason for that is not easy to explain since seedsare admittedly a rich source of compounds involved inplant defense [22] A good example is the seed extract ofBixa orellana that has been described to have antimicrobialactivity against six strains of bacteria among these are Bsubtilis S aureus and P aeruginosa and against the yeastC albicans [23] Likewise guarana (Paullinia cupana) seedextracts presented strong antimicrobial activity against a hugenumber of damaging fungi and bacteria [24] As to thepotency of the antibacterial activities observed these weremuch lower than those described for Syzygium jambolanumseed extracts against the same bacterial strains used in thiswork (MIC values ranging from 006 to 025mgsdotmLminus1) even


when compared to the best result observed as describedabove for S obtusifolia (MIC 496mgsdotmLminus1) In additionthe S jambolanum seed extracts were also effective againstdamaging-fungi strains The low potency of the ethanolicseed extracts could be due to the chemical composition of thesamples with low concentration of antimicrobial substancesIn fact the results of phytochemical study revealed that allseed extracts with activity against bacteria showed in theircomposition classes of secondary metabolites widely knownas efficient antimicrobial compounds (alkaloids flavonesflavonoids phenols saponins and tannins) [25] A fraction-ation of these samples in a polarity gradient could improvetheir activities Therefore the absence or low activity of theseed extracts could be a problem of concentration

The hemolytic activity assay was performed with theseed extracts that were active against the microorganismsused Among six species tested only M urundeuva causedhemolysis in all blood types (1 erythrocytes suspension ofA B and O human blood types and rabbit blood) employedbeing the lowest concentration tested (19120583gsdotmLminus1) capableof causing more than 20 hemolysis These results show thatalthough the antimicrobial extracts are not very potent theyare more specific to bacterial cells except for M urundeuvaseed extract It is well reported that some classes of plantsecondarymetabolites are potent hemolytic compounds [26]such as phenols that were also detected forMurundeuva seedextract in the phytochemical study

The results for antioxidant capacity are also shown inTable 1 Only five seed extracts were able to capture free rad-icals in the concentrations used according to DPPHmethodthese were Hymenaea courbaril (EC

5024795 120583gsdotmLminus1) L

rigida (48751120583gsdotmLminus1) L tomentosa (21672 120583gsdotmLminus1) andT gardneriana (6973120583gsdotmLminus1) The L tomentosa and Tgardneriana seed extracts were more potent than the positivecontrol vitamin C (EC

5026027120583gsdotmLminus1) and the second one

was just a bit less potent than quercetin (EC505552120583gsdotmLminus1)

one of the most potent antioxidant molecules A previousstudy [27] showed that out of 25 plant extracts (hexanedichloromethane and methanol) only 12 presented someradical scavenging activity and the most potent activityshowed an IC

50value of 11199 120583gsdotmLminus1This promising result

for T gardneriana could be attributed to components such astannins flavones flavonoids and especially phenols [28 29]Further steps of fractionation and purification may show themolecule(s) responsible for this excellent activity

Concerning the anticholinesterase activity (Table 1)among the 21 ethanolic extracts tested 11 presented inhibitionof the cholinesterase enzyme T gardneriana P platycephalaand C detersus extracts presented the strongest activitywith inhibition values of respectively 767 715 and 919at the final concentration of 500120583gmL These values aresignificant when compared to those obtained in previousstudies [18] which have shown inhibition values of 17 plantextracts ranging from 9 to 81 using a single concentrationof 1mgmL Several plant-derived drugs such as galantamineand rivastigmine are used to inhibit AChE and thus increaseendogenous levels of acetylcholine to improve cholinergictransmission [30] Many studies described alkaloids as the

So

Pp

Lr

Tg

Hc

Lt

Tran

smitt

ance

()

4000 3500 3000 2500 2000 1500 1000 500

Wavenumber (cmminus1)

Pp

Lr

Tg

Hc

Lt

Figure 1 Infrared absorption spectra of some ethanolic extractstested in this study So Senna obtusifolia Pp Parkia platycephala LrLicania rigida Tg Triplaris gardneriana Hc Hymenaea courbarilLt Licania tomentosa

main compounds capable of inhibiting AChE enzyme [31]but recent studies have pointed out several new classes ofsecondarymetabolites as potent inhibitors such as flavonoids[32] flavones [33] and xanthones [34] as well as steroids ter-penoids oils and other phenolic compounds [35] Thus thephytochemical analysis data in the present study corroboratepreviousmolecular findings since polyphenols were detectedin all extracts Additionally the extract of T gardnerianashowed the presence of chalcones and aurones Thereforefurther studies should be conducted to assess the potential ofthese extracts and their respective molecules to inhibit AChEenzyme and thus offer a potential to fight neurodegenerativediseases such as Alzheimerrsquos

After running all the assays described above infraredspectroscopywith themost promising ethanolic seed extractswas also performed (S obtusifolia P platycephala L rigida Ltomentosa and T gardneriana) to confirm the results of phy-tochemical qualitative studyThe phytochemical studies withthese ethanolic extracts showed the presence of tannins andflavonoids which were confirmed by infrared spectroscopy(Figure 1)The spectra obtained between 1800 and 800 cmminus1are present at the fingerprint zone of aromatic compoundswhich can be found in tannins polyphenols and flavonoids[36] Further steps of fractionation andpurificationmay showthe molecule(s) responsible for this excellent activity

4 Conclusions

This study revealed antibacterial antioxidant and anti-cholinesterase activities in some seed ethanolic extracts ofBrazilian semiarid region plants emphasizing the antioxidantactivity presented by T gardneriana seeds This showeda potent capacity to catch free radicals by DPPH assayThis promising result could be attributed to its composition


featuring phenols tannins flavones and flavonoidsNewphy-tochemical and molecular studies are underway to identifythe bioactive compound(s) responsible for this activity

Conflict of Interests

The authors declare that there is no conflict of interestsregarding the publication of this paper

Acknowledgment

The authors thank theMinisterio da Saude Departamento deCiencias e Tecnologia (DECIT) Secretaria de Ciencias Tec-nologia e Insumos estrategicos (SCTIE) Conselho Nacionalde Desenvolvimento Cientıfico e Tecnologico (CNPq) andFundacao Cearense deApoioCientıfico e Tecnologico (FUN-CAP) for financial support of this research

References

[1] C Katiyar A Gupta S Kanjilal and S Katiyar ldquoDrug discoveryfromplant sources an integrated approachrdquoAyu vol 33 pp 10ndash19 2012

[2] P S Luize T S Tiuman L G Morello et al ldquoEffects of medic-inal plant extracts on growth of Leishmania (L) amazonensisand Trypanosoma cruzirdquo Brazilian Journal of PharmaceuticalSciences vol 41 no 1 pp 85ndash94 2005

[3] A E Edris ldquoAnti-cancer properties of Nigella spp essential oilsand their major constituents thymoquinone and 120573-ElemenerdquoCurrent Clinical Pharmacology vol 4 no 1 pp 43ndash46 2009

[4] R N Bennett and R M Wallsgrove ldquoSecondary metabolites inplant defence mechanismsrdquo New Phytologist vol 127 no 4 pp617ndash633 1994

[5] R J Grayer and J B Harborne ldquoA survey of antifungal com-pounds from higher plants 1982-1993rdquo Phytochemistry vol 37no 1 pp 19ndash42 1994

[6] A E Osbourn ldquoPreformed antimicrobial compounds and plantdefense against fungal attackrdquo Plant Cell vol 8 no 10 pp 1821ndash1831 1996

[7] E V S B Sampaio ldquoVegetacao e flora da Caatingardquo APNECNIP Recife 2000

[8] C F C B R De Almeida T C De Lima E Silva E L CDe Amorim M B D S Maia and U P De AlbuquerqueldquoLife strategy and chemical composition as predictors of theselection of medicinal plants from the caatinga (NortheastBrazil)rdquo Journal of Arid Environments vol 62 no 1 pp 127ndash1422005

[9] U P de Albuquerque P M de Medeiros A L S de Almeida etal ldquoMedicinal plants of the caatinga (semi-arid) vegetation ofNE Brazil a quantitative approachrdquo Journal of Ethnopharmacol-ogy vol 114 no 3 pp 325ndash354 2007

[10] U P de Albuquerque V A da Silva M D C Cabral NLeal Alencar and L D H C Andrade ldquoComparisons betweenthe use of medicinal plants in indigenous and rural caatinga(dryland) communities in NE Brazilrdquo Boletin Latinoamericanoy del Caribe de Plantas Medicinales y Aromaticas vol 7 no 3pp 156ndash170 2008

[11] T M Souza D F Farias B M Soares et al ldquoToxicity ofBrazilian plant seed extracts to two strains of Aedes aegypti(Diptera Culicidae) andNontargetAnimalsrdquo Journal ofMedicalEntomology vol 48 no 4 pp 846ndash851 2011

[12] P M P Ferreira D F Farias M P Viana et al ldquoStudy of theantiproliferative potential of seed extracts from NortheasternBrazilian plantsrdquo Anais da Academia Brasileira de Ciencias vol83 no 3 pp 1045ndash1058 2011

[13] F J AMatosM EOMatosM P SousaM I LMachado andA A Craveiro Constituintes Quımicos Ativos e PropriedadesBiologicas de Plantas Medicinais Brasileiras UFC FortalezaBrazil 1st edition 2004

[14] F J A Matos Plantas Medicinais Guia de Selecao e Empregode Plantas Usadas em Fitoterapia no Nordeste do Brasil IUFortaleza Brazil 2nd edition 2000

[15] G N Maia Caatinga Arvores e Arbustos e Suas Utilidades D ampZ Computacao Grafica e Editora Sao Paulo Brazil 1st edition2004

[16] M Chandrasekaran and V Venkatesalu ldquoAntibacterial andantifungal activity of Syzygium jambolanum seedsrdquo Journal ofEthnopharmacology vol 91 no 1 pp 105ndash108 2004

[17] B Yepez M Espinosa S Lopez and G Bolaos ldquoProducingantioxidant fractions from herbaceousmatrices by supercriticalfluid extractionrdquo Fluid Phase Equilibria vol 194ndash197 pp 879ndash884 2002

[18] E M Cardoso-Lopes J A Maier M R Da Silva et alldquoAlkaloids from stems of esenbeckia leiocarpa Engl (Rutaceae)as potential treatment for alzheimer diseaserdquoMolecules vol 15no 12 pp 9205ndash9213 2010

[19] G L Ellman K D Courtney V Andres Jr and R MFeatherstone ldquoA new and rapid colorimetric determination ofacetylcholinesterase activityrdquo Biochemical Pharmacology vol 7no 2 pp 88ndash95 1961

[20] A W Bernheimer ldquoAssay of hemolytic toxinsrdquo Methods inEnzymology vol 165 pp 213ndash217 1988

[21] F J A Matos Introducao A Fitoquımica Experimental EditoraUFC Ceara Brazil 1st edition 2009

[22] J Xavier-Filho ldquoSementes e suas defesas contra insetos ProjetoMultinacional de Biotecnologia e Alimentosrdquo Organizacao dosEstados Americanos (OEA) pp1ndash31 1993

[23] T C Fleischer E P K Ameade M L K Mensah and I KSawer ldquoAntimicrobial activity of the leaves and seeds of Bixaorellanardquo Fitoterapia vol 74 no 1-2 pp 136ndash138 2003

[24] L Majhenic M Skerget and Z Knez ldquoAntioxidant and antimi-crobial activity of guarana seed extractsrdquo Food Chemistry vol104 no 3 pp 1258ndash1268 2007

[25] MM Cowan ldquoPlant products as antimicrobial agentsrdquoClinicalMicrobiology Reviews vol 12 no 4 pp 564ndash582 1999

[26] A Rafat K Philip and SMuniandy ldquoAntioxidant potential andcontent of phenolic compounds in ethanolic extracts of selectedparts of Andrographis paniculatardquo Journal of Medicinal PlantResearch vol 4 no 3 pp 197ndash202 2010

[27] O M Mosquera Y M Correa D C Buitrago and J NinoldquoAntioxidant activity of twenty five plants from ColombianbiodiversityrdquoMemorias do Instituto Oswaldo Cruz vol 102 no5 pp 631ndash634 2007

[28] N Cotelle J-L Bernier J-P Catteau J Pommery J-C Wal-let and E M Gaydou ldquoAntioxidant properties of hydroxy-flavonesrdquo Free Radical Biology and Medicine vol 20 no 1 pp35ndash43 1996

[29] H Oh D-H Kim J-H Cho and Y-C Kim ldquoHepatoprotectiveand free radical scavenging activities of phenolic petrosinsand flavonoids isolated from Equisetum arvenserdquo Journal ofEthnopharmacology vol 95 no 2-3 pp 421ndash424 2004


[30] S Lopez J Bastida F Viladomat and C Codina ldquoAcetyl-cholinesterase inhibitory activity of some Amaryllidaceae alka-loids and Narcissus extractsrdquo Life Sciences vol 71 no 21 pp2521ndash2529 2002

[31] M G Ortega A M Agnese and J L Cabrera ldquoAnti-cholinesterase activity in an alkaloid extract ofHuperzia sauru-rusrdquo Phytomedicine vol 11 no 6 pp 539ndash543 2004

[32] B J Hillhouse D S Ming C J French and G H NTowers ldquoAcetylcholine esterase inhibitors in Rhodiola roseardquoPharmaceutical Biology vol 42 no 1 pp 68ndash72 2004

[33] P Sawasdee C Sabphon D Sitthiwongwanit and U KokpolldquoAnticholinesterase activity of 7-methoxyflavones isolated fromKaempferia parviflorardquo Phytotherapy Research vol 23 no 12pp 1792ndash1794 2009

[34] M-J R Howes N S L Perry and P J Houghton ldquoPlants withtraditional uses and activities relevant to the management ofAlzheimerrsquos disease and other cognitive disordersrdquo Phytother-apy Research vol 17 no 1 pp 1ndash18 2003

[35] H-F Ji andH-Y Zhang ldquoMultipotent natural agents to combatAlzheimerrsquos disease Functional spectrum and structural fea-turesrdquo Acta Pharmacologica Sinica vol 29 no 2 pp 143ndash1512008

[36] K Fernandez and E Agosin ldquoQuantitative analysis of red winetannins using Fourier-transform mid-infrared spectrometryrdquoJournal of Agricultural and Food Chemistry vol 55 no 18 pp7294ndash7300 2007

Submit your manuscripts athttpwwwhindawicom

PainResearch and TreatmentHindawi Publishing Corporationhttpwwwhindawicom Volume 2014

The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom

Volume 2014

ToxinsJournal of

VaccinesJournal of

Hindawi Publishing Corporation httpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

AntibioticsInternational Journal of

ToxicologyJournal of


StrokeResearch and TreatmentHindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Drug DeliveryJournal of



Advances in Pharmacological Sciences

Tropical MedicineJournal of


Medicinal ChemistryInternational Journal of


AddictionJournal of



BioMed Research International

Emergency Medicine InternationalHindawi Publishing Corporationhttpwwwhindawicom Volume 2014


Autoimmune Diseases


Anesthesiology Research and Practice

ScientificaHindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Journal of


Pharmaceutics


MEDIATORSINFLAMMATION

of


compounds saponins cyanogenic glycosides and glucosino-lates [4ndash6]

In this context the Brazilian semiarid region is a richand underexploited source of bioactive molecules This areais covered mainly by xeric shrublands known as ldquoCaatingardquoThis dry land vegetation grows over an area of 800000 km2in Northeastern Brazil and out of 1000 vascular plant speciesdescribed many are endemic [7 8] Many of these plants areused in folk medicine by local communities to treat severaldiseases pointing out the potential to discover new sourcesof useful compounds for several applications [9 10] Recentlyour team has reported for seed ethanolic extracts of BrazilianNortheastern plants promising results of insecticidal activityagainst Aedes aegypti [11] and antiproliferative effects totumor cell lines [12] In order to extend our knowledgeon the Brazilian semiarid vegetation this work aimed toevaluate the presence of antimicrobial anticholinesteraseand antioxidant activities in ethanolic seed extracts of twenty-one plant species The antimicrobial activity was performedby the agar diffusion assay using pathogenic bacteria andyeast antioxidant activity by the DPPH radical scavengingand anticholinesterase activity were measured In addition ahemolytic activity assay was done for antibacterial samples toassess the effects on nontarget cells

2 Materials and Methods

21 Plant Collection Mature wild seeds of twenty-one plantspecies were harvested (at least 500 g of each species) duringthe dry period (from January 2005 to March 2008) in thesemiarid region of the Ceara State Northeastern BrazilPlants were identified by taxonomist Dr Edson de PaulaNunes and voucher specimens were deposited at HerbariumPrisco Bezerra (EAC) at the Federal University of Ceara(Fortaleza Brazil)The studied species are depicted in Table 1which shows harvest information voucher numbers com-mon names and the traditional medicinal use by localpopulation

22 Preparation of Ethanolic Seed Extracts Seeds of freshlycollected plant material were separated from other plantmaterials air-dried and ground in a laboratory mill to amoderately fine powder (particle size le 05mm) Powderedmaterial (500 g) was submitted to extraction with 99ethanol (15 L) at room temperature (25ndash27∘C) for 3 daysThe supernatant was removed and filtered throughWhatmannumber 1 paper After that the initial powder sample wasstill submitted to two extractions which resulted in a finalvolume of extract near 45 L The ethanolic extracts resultingfrom three consecutive extractions were mixed together thenconcentrated under reduced pressure in rotary evaporatoruntil complete elimination of solvent and stored in a freezerat minus20∘C

A stock solution containing 50000 120583gsdotmLminus1 of eachcrude extract was prepared by suspending 50000mg ofextract in 5mL of pure dimethylsulphoxide (DMSO AldrichMilwaukee WI USA) and mixed by sonication (model RK-100 Bandelin Berlin Germany) for 20min The volume was

adjusted to 1000mL with distilled water to provide assaysolutions as required

23 Microorganisms Six bacterial strains were used Staphy-lococcus aureus (ATCC 25923) Bacillus subtilis (ATCC 6633)Enterobacter aerogenes (ATCC 13048) Salmonella cholerae-suis (ATCC 10708) Klebsiella pneumoniae (ATCC 10031)and Pseudomonas aeruginosa (ATCC 25619)These standardswere obtained from the American Type Culture Collection(ATCC) (Manassas VA USA) The stock culture was main-tained on Mueller Hinton agar (HiMedia Laboratories PvtLtdMumbai India)medium at 4∘CThree pathogenic yeastswere usedCandida albicansC krusei andC tropicalisThesefungal strains were obtained from the Microbial Ecologyand Biotechnology Laboratory Biology Department of theFederal University of Ceara Brazil The stock cultures weremaintained on Sabouraud Dextrose agar (HiMedia Labora-tories Pvt Ltd Mumbai India) medium at 4∘C

24 Antibacterial and Antifungal Activity The disc diffu-sion method was followed for antibacterial and antifungalsusceptibility test as described previously [16] with somemodifications Petri dishes were prepared by pouring 20mLof Mueller-Hinton agar for bacteria and Sabouraud Dextroseagar for fungi and allowed to solidify Dishes were dried and01mL of standardized inoculum suspension (106 CFUmLfor bacteria and absorbance of 0600 at 450 nm for fungi)was poured and uniformly spread The excess inoculum wasdrained and the inoculum was allowed to dry for 5min Thepaper filter discs (7mm of diameter) containing 20120583L ofeach ethanolic extract were then applied and the dishes wereincubated at 37∘C for 18 h for bacterial growth and at thesame temperature for 48 h for fungal growth Tetracycline(30mgdisc) was used as positive control for the antibacterialassay Nystatin solution (100000 IUmL EMS SP Brazil)(20120583Ldisc) was used as positive control for the antifungalassayThe inhibition zone was measured from the edge of thedisc to the inner margin of the surrounding pathogens Alldeterminations were run in triplicate

25 Minimum Inhibitory Concentration (MIC) Minimuminhibitory concentrations of the seed ethanolic extracts weretested as described previously by the twofold serial dilutionmethodThe test extract was dissolved in 5DMSO to obtain50mgsdotmLminus1 stock solution Stock solution was diluted withdistilled water to achieve 45 40 35 30 25 20 15 10 and5mgsdotmLminus1 or as required The wet paper discs with dilutionswere placed over the dish and the cultures were incubated inBOD incubators at 37∘C for 18 h (bacteria) and 37∘C for 48 h(yeasts) The lowest concentration which inhibits the visiblegrowth of tested organism after macroscopic evaluation wasdetermined asMICAll determinationswere run in triplicate

26 Antioxidant Activity The capacity to scavenge the 22-diphenyl-1-picrylhydrazyl (DPPH) free radical was moni-tored [17] The diluted crude extracts (01mL) in differ-ent concentrations (5 to 1000 120583gsdotmLminus1) were mixed with39mL of methanolic solution containing DPPH radicals


Table1Antibacteria

l(MIC)antio

xidantand

hemolyticactiv

ities

ofethano

licseed

extractsof

plantsfro

mBrazilian

semiarid

region

a

Family

Botanicaln

ame

Com

mon

name

(localitydateof

collection)V

oucher

number

Tradition

almedicinaluse

Yield

()b

Antioxidant

activ

ity(EC 5

0120583gmL)

cAnticho

linesterase

activ

ity(

)d

Antibacteria

lactivity

MIC

(mgmL)

e

SCh

oleraesuis

Bsubtilis

Saureus

Anacardiaceae

Myracrodruonurun

deuvaFrA

llAroeira-do-sertao

(Ararip


EAC34865

Usedto

treatcutaneou

sand


and

kidn

eyandrespira

tory

prob

lemsantiinfl

ammatory

antiu

lcerandhealing[13]

antih

istam

inicand

antim

icrobial[14

]analgesic

andantid

iarrheal[15]

252

ND

fND

ND

678

ND


Engler

Brauna

(Ararip


EAC35643

Usedto

treatnervou

snessa

ndhyste

riaanalgesic[15]

288

ND

ND

3732

ND

ND

Caryocaraceae

Caryocar

coria

ceum

Wittm

Pequ

i(Ararip


EAC35251

Not

describ

ed231

ND

634

ND

ND

ND

Chrysobalana

ceae

Licaniatomentosa

Benth

Oiti(Coastzone010

8)EA

C40215

Not

describ

ed44

21672

139

ND

ND

649

Licaniarig

idaBe

nth

Oiticica

(Coastzone1207)

EAC40216

Usedto

treatdiabetes


281

48751

524

ND

ND

863

Conn

araceae

Conn

arus

detersus

Planch

Cabelode

negro

(Ararip


EAC34733

Not

describ

ed285

ND

919

ND

ND

ND


Table1Con

tinued

Family

Botanicaln

ame

Com

mon

name

(localitydateof

collection)V

oucher

number

Tradition

almedicinaluse

Yield

()b

Antioxidant

activ

ity(EC 5

0120583gmL)

cAnticho

linesterase

activ

ity(

)d

Antibacteria

lactivity

MIC

(mgmL)

e

SCh

oleraesuis

Bsubtilis

Saureus

Fabaceae

Adenantherapavonina

LFalso

-sandalo

(Coastzone0807)

EAC38697

Not

describ

ed612

ND

ND

ND

ND

ND

Amburana

cearensis

(Allemao)A

C

Smith

Cumaruandam

burana

(Ca

atinga

vegetatio

n1107)

EAC39618

Usedto

treatrheumatism

coldand

sinusitis

antispasm

odichealin

gand


analgesic

[13]

643

ND

ND

ND

ND

ND


(Benth)Brenan

Ang

icoverm

elho

(Ca

atinga

vegetatio

n0705)

EAC38389

Usedto

treatrespira

tory

prob

lemsgono

rrheaand

diarrheaastrin

gent

antirheum

atic


eandhealing[15]

935

ND

541

ND

ND

ND

Diocle

amegacarpa

Rolfe

Mucun

aolho

-de-bo

i(Ararip


EAC38110

Not

describ

ed240

ND

ND

ND

ND

ND

Enterolobium

contortisiliquum

(Vell)

Moron

gOrelha-de-negro

(Ararip


EAC38115

Not

describ

ed280

ND

ND

ND

ND

ND

Hym

enaeacourbaril

LJatoba(Ararip


EAC38108

Not

describ

ed95

02479

5ND

ND

ND

ND

Lonchocarpus

seric

eus(Po

iret)Ku

nth

Inga(Coastzone1006)

EAC39615

Not

describ

ed1060

ND

ND

ND

ND

ND

Luetzelburgiaauric

ulata(A

llemao)

Ducke

Pau-moco

(Ca

atinga

vegetatio

n1106)

EAC40369

Not

describ

ed840

ND

ND

ND

ND

ND


ble1Con

tinued

Family

Botanicaln

ame

Com

mon

name

(localitydateof

collection)V

oucher

number

Tradition

almedicinaluse

Yield

()b

Antioxidant

activ

ity(EC 5

0120583gmL)

cAnticho

linesterase

activ

ity(

)d

Antibacteria

lactivity

MIC

(mgmL)

e

SCh

oleraesuis

Bsubtilis

Saureus


nth

Visgueiro

(Ararip


EAC3

8109

Not

describ

ed1750

ND

715

ND

ND

ND


isBe

nth

Catand

uva

(Ca

atinga

vegetatio

n0105)

EAC3

5974

Not

describ

ed618

ND

502

ND

ND

ND

Senn


inamp

Barneby

Matap

asto

(Ca

atinga

vegetatio

n0705)

EAC3

9320

Not

describ

ed570

ND

ND

ND

496

ND

Polyg

onaceae

Triplaris

gardneria

naWedd

Pajeu

(Ca

atinga

vegetatio

n1106)

EAC3

9600

Not

describ

ed260

6973

798

188

1376

1115

Rham

naceae

Zizip

husjoazeiro

Mart

Juaa

ndjuazeiro

(Coastzone0308)

EAC40366

Expectorantand

antip

yretic

used

totre

atskinblood

sto

machandliver

diseases

ulcerantim

icrobial[15]

722

ND

494

ND

ND

ND

Sapind

aceae

Talisiaesculen

ta(A

St-Hil)

Radlk

Pitomba

(Coastzone0308)

EAC40

685

Not

describ

ed15

0ND

6124

ND

ND

ND

Sapind

ussaponaria

LSabo

nete

(Ca

atinga

vegetatio

n1106)

EAC3

9601

Not

describ

ed107

ND

5860

ND

ND

ND

Controlsg

Quercetin

mdashmdash

5552

mdashmdash

mdashmdash

Vitamin

Cmdash

mdash26027

mdashmdash

mdashmdash

a Thev

aluesa

remeans

ofatrip

licateTh

estand

arddeviationwas

lessthan

5of

them

ean

b (Amou

nt(g)o

fsolid

recoveredfro

mthee

thanolicextracta

mou

nt(g)o

fseedflo

urused)times

100

c Efficientcon

centratio

n(EC)

as120583

gof

sampler

equiredto

decrease

oneg

oftheinitia


Hsdot)con

centratio

nby

50

d Totalinhibitio

nof

cholinesterase

enzymea

fterc

omparis

onto

theb

lank

Con

centratio

nteste

d500120583

gmL

e Minim

uminhibitory

concentration

thelow

estcon

centratio

nwhich

didno

tsho

wanygrow

thof

teste

dorganism

after

macroscop

icevaluatio

nf N

Dnot

detected

g Sub

stancesthatare

frequ

ently

used

assta

ndardfora

ntioxidant

studies



50)








119901119888= Abs



















So

Pp

Lr

Tg

Hc

Lt

Tran

smitt

ance

()

4000 3500 3000 2500 2000 1500 1000 500


Pp

Lr

Tg

Hc

Lt




4 Conclusions






Acknowledgment


References










































Volume 2014

ToxinsJournal of

VaccinesJournal of















AddictionJournal of






Autoimmune Diseases




Journal of


Pharmaceutics



of


Table1Antibacteria

l(MIC)antio

xidantand

hemolyticactiv

ities

ofethano

licseed

extractsof

plantsfro

mBrazilian

semiarid

region

a

Family

Botanicaln

ame

Com

mon

name

(localitydateof

collection)V

oucher

number

Tradition

almedicinaluse

Yield

()b

Antioxidant

activ

ity(EC 5

0120583gmL)

cAnticho

linesterase

activ

ity(

)d

Antibacteria

lactivity

MIC

(mgmL)

e

SCh

oleraesuis

Bsubtilis

Saureus

Anacardiaceae

Myracrodruonurun

deuvaFrA

llAroeira-do-sertao

(Ararip


EAC34865

Usedto

treatcutaneou

sand


and

kidn

eyandrespira

tory

prob

lemsantiinfl

ammatory

antiu

lcerandhealing[13]

antih

istam

inicand

antim

icrobial[14

]analgesic

andantid

iarrheal[15]

252

ND

fND

ND

678

ND


Engler

Brauna

(Ararip


EAC35643

Usedto

treatnervou

snessa

ndhyste

riaanalgesic[15]

288

ND

ND

3732

ND

ND

Caryocaraceae

Caryocar

coria

ceum

Wittm

Pequ

i(Ararip


EAC35251

Not

describ

ed231

ND

634

ND

ND

ND

Chrysobalana

ceae

Licaniatomentosa

Benth

Oiti(Coastzone010

8)EA

C40215

Not

describ

ed44

21672

139

ND

ND

649

Licaniarig

idaBe

nth

Oiticica

(Coastzone1207)

EAC40216

Usedto

treatdiabetes


281

48751

524

ND

ND

863

Conn

araceae

Conn

arus

detersus

Planch

Cabelode

negro

(Ararip


EAC34733

Not

describ

ed285

ND

919

ND

ND

ND


Table1Con

tinued

Family

Botanicaln

ame

Com

mon

name

(localitydateof

collection)V

oucher

number

Tradition

almedicinaluse

Yield

()b

Antioxidant

activ

ity(EC 5

0120583gmL)

cAnticho

linesterase

activ

ity(

)d

Antibacteria

lactivity

MIC

(mgmL)

e

SCh

oleraesuis

Bsubtilis

Saureus

Fabaceae

Adenantherapavonina

LFalso

-sandalo

(Coastzone0807)

EAC38697

Not

describ

ed612

ND

ND

ND

ND

ND

Amburana

cearensis

(Allemao)A

C

Smith

Cumaruandam

burana

(Ca

atinga

vegetatio

n1107)

EAC39618

Usedto

treatrheumatism

coldand

sinusitis

antispasm

odichealin

gand


analgesic

[13]

643

ND

ND

ND

ND

ND


(Benth)Brenan

Ang

icoverm

elho

(Ca

atinga

vegetatio

n0705)

EAC38389

Usedto

treatrespira

tory

prob

lemsgono

rrheaand

diarrheaastrin

gent

antirheum

atic


eandhealing[15]

935

ND

541

ND

ND

ND

Diocle

amegacarpa

Rolfe

Mucun

aolho

-de-bo

i(Ararip


EAC38110

Not

describ

ed240

ND

ND

ND

ND

ND

Enterolobium

contortisiliquum

(Vell)

Moron

gOrelha-de-negro

(Ararip


EAC38115

Not

describ

ed280

ND

ND

ND

ND

ND

Hym

enaeacourbaril

LJatoba(Ararip


EAC38108

Not

describ

ed95

02479

5ND

ND

ND

ND

Lonchocarpus

seric

eus(Po

iret)Ku

nth

Inga(Coastzone1006)

EAC39615

Not

describ

ed1060

ND

ND

ND

ND

ND

Luetzelburgiaauric

ulata(A

llemao)

Ducke

Pau-moco

(Ca

atinga

vegetatio

n1106)

EAC40369

Not

describ

ed840

ND

ND

ND

ND

ND


ble1Con

tinued

Family

Botanicaln

ame

Com

mon

name

(localitydateof

collection)V

oucher

number

Tradition

almedicinaluse

Yield

()b

Antioxidant

activ

ity(EC 5

0120583gmL)

cAnticho

linesterase

activ

ity(

)d

Antibacteria

lactivity

MIC

(mgmL)

e

SCh

oleraesuis

Bsubtilis

Saureus


nth

Visgueiro

(Ararip


EAC3

8109

Not

describ

ed1750

ND

715

ND

ND

ND


isBe

nth

Catand

uva

(Ca

atinga

vegetatio

n0105)

EAC3

5974

Not

describ

ed618

ND

502

ND

ND

ND

Senn


inamp

Barneby

Matap

asto

(Ca

atinga

vegetatio

n0705)

EAC3

9320

Not

describ

ed570

ND

ND

ND

496

ND

Polyg

onaceae

Triplaris

gardneria

naWedd

Pajeu

(Ca

atinga

vegetatio

n1106)

EAC3

9600

Not

describ

ed260

6973

798

188

1376

1115

Rham

naceae

Zizip

husjoazeiro

Mart

Juaa

ndjuazeiro

(Coastzone0308)

EAC40366

Expectorantand

antip

yretic

used

totre

atskinblood

sto

machandliver

diseases

ulcerantim

icrobial[15]

722

ND

494

ND

ND

ND

Sapind

aceae

Talisiaesculen

ta(A

St-Hil)

Radlk

Pitomba

(Coastzone0308)

EAC40

685

Not

describ

ed15

0ND

6124

ND

ND

ND

Sapind

ussaponaria

LSabo

nete

(Ca

atinga

vegetatio

n1106)

EAC3

9601

Not

describ

ed107

ND

5860

ND

ND

ND

Controlsg

Quercetin

mdashmdash

5552

mdashmdash

mdashmdash

Vitamin

Cmdash

mdash26027

mdashmdash

mdashmdash

a Thev

aluesa

remeans

ofatrip

licateTh

estand

arddeviationwas

lessthan

5of

them

ean

b (Amou

nt(g)o

fsolid

recoveredfro

mthee

thanolicextracta

mou

nt(g)o

fseedflo

urused)times

100

c Efficientcon

centratio

n(EC)

as120583

gof

sampler

equiredto

decrease

oneg

oftheinitia


Hsdot)con

centratio

nby

50

d Totalinhibitio

nof

cholinesterase

enzymea

fterc

omparis

onto

theb

lank

Con

centratio

nteste

d500120583

gmL

e Minim

uminhibitory

concentration

thelow

estcon

centratio

nwhich

didno

tsho

wanygrow

thof

teste

dorganism

after

macroscop

icevaluatio

nf N

Dnot

detected

g Sub

stancesthatare

frequ

ently

used

assta

ndardfora

ntioxidant

studies



50)








119901119888= Abs



















So

Pp

Lr

Tg

Hc

Lt

Tran

smitt

ance

()

4000 3500 3000 2500 2000 1500 1000 500


Pp

Lr

Tg

Hc

Lt




4 Conclusions






Acknowledgment


References










































Volume 2014

ToxinsJournal of

VaccinesJournal of















AddictionJournal of






Autoimmune Diseases




Journal of


Pharmaceutics



of


Table1Con

tinued

Family

Botanicaln

ame

Com

mon

name

(localitydateof

collection)V

oucher

number

Tradition

almedicinaluse

Yield

()b

Antioxidant

activ

ity(EC 5

0120583gmL)

cAnticho

linesterase

activ

ity(

)d

Antibacteria

lactivity

MIC

(mgmL)

e

SCh

oleraesuis

Bsubtilis

Saureus

Fabaceae

Adenantherapavonina

LFalso

-sandalo

(Coastzone0807)

EAC38697

Not

describ

ed612

ND

ND

ND

ND

ND

Amburana

cearensis

(Allemao)A

C

Smith

Cumaruandam

burana

(Ca

atinga

vegetatio

n1107)

EAC39618

Usedto

treatrheumatism

coldand

sinusitis

antispasm

odichealin

gand


analgesic

[13]

643

ND

ND

ND

ND

ND


(Benth)Brenan

Ang

icoverm

elho

(Ca

atinga

vegetatio

n0705)

EAC38389

Usedto

treatrespira

tory

prob

lemsgono

rrheaand

diarrheaastrin

gent

antirheum

atic


eandhealing[15]

935

ND

541

ND

ND

ND

Diocle

amegacarpa

Rolfe

Mucun

aolho

-de-bo

i(Ararip


EAC38110

Not

describ

ed240

ND

ND

ND

ND

ND

Enterolobium

contortisiliquum

(Vell)

Moron

gOrelha-de-negro

(Ararip


EAC38115

Not

describ

ed280

ND

ND

ND

ND

ND

Hym

enaeacourbaril

LJatoba(Ararip


EAC38108

Not

describ

ed95

02479

5ND

ND

ND

ND

Lonchocarpus

seric

eus(Po

iret)Ku

nth

Inga(Coastzone1006)

EAC39615

Not

describ

ed1060

ND

ND

ND

ND

ND

Luetzelburgiaauric

ulata(A

llemao)

Ducke

Pau-moco

(Ca

atinga

vegetatio

n1106)

EAC40369

Not

describ

ed840

ND

ND

ND

ND

ND


ble1Con

tinued

Family

Botanicaln

ame

Com

mon

name

(localitydateof

collection)V

oucher

number

Tradition

almedicinaluse

Yield

()b

Antioxidant

activ

ity(EC 5

0120583gmL)

cAnticho

linesterase

activ

ity(

)d

Antibacteria

lactivity

MIC

(mgmL)

e

SCh

oleraesuis

Bsubtilis

Saureus


nth

Visgueiro

(Ararip


EAC3

8109

Not

describ

ed1750

ND

715

ND

ND

ND


isBe

nth

Catand

uva

(Ca

atinga

vegetatio

n0105)

EAC3

5974

Not

describ

ed618

ND

502

ND

ND

ND

Senn


inamp

Barneby

Matap

asto

(Ca

atinga

vegetatio

n0705)

EAC3

9320

Not

describ

ed570

ND

ND

ND

496

ND

Polyg

onaceae

Triplaris

gardneria

naWedd

Pajeu

(Ca

atinga

vegetatio

n1106)

EAC3

9600

Not

describ

ed260

6973

798

188

1376

1115

Rham

naceae

Zizip

husjoazeiro

Mart

Juaa

ndjuazeiro

(Coastzone0308)

EAC40366

Expectorantand

antip

yretic

used

totre

atskinblood

sto

machandliver

diseases

ulcerantim

icrobial[15]

722

ND

494

ND

ND

ND

Sapind

aceae

Talisiaesculen

ta(A

St-Hil)

Radlk

Pitomba

(Coastzone0308)

EAC40

685

Not

describ

ed15

0ND

6124

ND

ND

ND

Sapind

ussaponaria

LSabo

nete

(Ca

atinga

vegetatio

n1106)

EAC3

9601

Not

describ

ed107

ND

5860

ND

ND

ND

Controlsg

Quercetin

mdashmdash

5552

mdashmdash

mdashmdash

Vitamin

Cmdash

mdash26027

mdashmdash

mdashmdash

a Thev

aluesa

remeans

ofatrip

licateTh

estand

arddeviationwas

lessthan

5of

them

ean

b (Amou

nt(g)o

fsolid

recoveredfro

mthee

thanolicextracta

mou

nt(g)o

fseedflo

urused)times

100

c Efficientcon

centratio

n(EC)

as120583

gof

sampler

equiredto

decrease

oneg

oftheinitia


Hsdot)con

centratio

nby

50

d Totalinhibitio

nof

cholinesterase

enzymea

fterc

omparis

onto

theb

lank

Con

centratio

nteste

d500120583

gmL

e Minim

uminhibitory

concentration

thelow

estcon

centratio

nwhich

didno

tsho

wanygrow

thof

teste

dorganism

after

macroscop

icevaluatio

nf N

Dnot

detected

g Sub

stancesthatare

frequ

ently

used

assta

ndardfora

ntioxidant

studies



50)








119901119888= Abs



















So

Pp

Lr

Tg

Hc

Lt

Tran

smitt

ance

()

4000 3500 3000 2500 2000 1500 1000 500


Pp

Lr

Tg

Hc

Lt




4 Conclusions






Acknowledgment


References










































Volume 2014

ToxinsJournal of

VaccinesJournal of















AddictionJournal of






Autoimmune Diseases




Journal of


Pharmaceutics



of


ble1Con

tinued

Family

Botanicaln

ame

Com

mon

name

(localitydateof

collection)V

oucher

number

Tradition

almedicinaluse

Yield

()b

Antioxidant

activ

ity(EC 5

0120583gmL)

cAnticho

linesterase

activ

ity(

)d

Antibacteria

lactivity

MIC

(mgmL)

e

SCh

oleraesuis

Bsubtilis

Saureus


nth

Visgueiro

(Ararip


EAC3

8109

Not

describ

ed1750

ND

715

ND

ND

ND


isBe

nth

Catand

uva

(Ca

atinga

vegetatio

n0105)

EAC3

5974

Not

describ

ed618

ND

502

ND

ND

ND

Senn


inamp

Barneby

Matap

asto

(Ca

atinga

vegetatio

n0705)

EAC3

9320

Not

describ

ed570

ND

ND

ND

496

ND

Polyg

onaceae

Triplaris

gardneria

naWedd

Pajeu

(Ca

atinga

vegetatio

n1106)

EAC3

9600

Not

describ

ed260

6973

798

188

1376

1115

Rham

naceae

Zizip

husjoazeiro

Mart

Juaa

ndjuazeiro

(Coastzone0308)

EAC40366

Expectorantand

antip

yretic

used

totre

atskinblood

sto

machandliver

diseases

ulcerantim

icrobial[15]

722

ND

494

ND

ND

ND

Sapind

aceae

Talisiaesculen

ta(A

St-Hil)

Radlk

Pitomba

(Coastzone0308)

EAC40

685

Not

describ

ed15

0ND

6124

ND

ND

ND

Sapind

ussaponaria

LSabo

nete

(Ca

atinga

vegetatio

n1106)

EAC3

9601

Not

describ

ed107

ND

5860

ND

ND

ND

Controlsg

Quercetin

mdashmdash

5552

mdashmdash

mdashmdash

Vitamin

Cmdash

mdash26027

mdashmdash

mdashmdash

a Thev

aluesa

remeans

ofatrip

licateTh

estand

arddeviationwas

lessthan

5of

them

ean

b (Amou

nt(g)o

fsolid

recoveredfro

mthee

thanolicextracta

mou

nt(g)o

fseedflo

urused)times

100

c Efficientcon

centratio

n(EC)

as120583

gof

sampler

equiredto

decrease

oneg

oftheinitia


Hsdot)con

centratio

nby

50

d Totalinhibitio

nof

cholinesterase

enzymea

fterc

omparis

onto

theb

lank

Con

centratio

nteste

d500120583

gmL

e Minim

uminhibitory

concentration

thelow

estcon

centratio

nwhich

didno

tsho

wanygrow

thof

teste

dorganism

after

macroscop

icevaluatio

nf N

Dnot

detected

g Sub

stancesthatare

frequ

ently

used

assta

ndardfora

ntioxidant

studies



50)








119901119888= Abs



















So

Pp

Lr

Tg

Hc

Lt

Tran

smitt

ance

()

4000 3500 3000 2500 2000 1500 1000 500


Pp

Lr

Tg

Hc

Lt




4 Conclusions






Acknowledgment


References










































Volume 2014

ToxinsJournal of

VaccinesJournal of















AddictionJournal of






Autoimmune Diseases




Journal of


Pharmaceutics



of



50)








119901119888= Abs



















So

Pp

Lr

Tg

Hc

Lt

Tran

smitt

ance

()

4000 3500 3000 2500 2000 1500 1000 500


Pp

Lr

Tg

Hc

Lt




4 Conclusions






Acknowledgment


References










































Volume 2014

ToxinsJournal of

VaccinesJournal of















AddictionJournal of






Autoimmune Diseases




Journal of


Pharmaceutics



of













So

Pp

Lr

Tg

Hc

Lt

Tran

smitt

ance

()

4000 3500 3000 2500 2000 1500 1000 500


Pp

Lr

Tg

Hc

Lt




4 Conclusions






Acknowledgment


References










































Volume 2014

ToxinsJournal of

VaccinesJournal of















AddictionJournal of






Autoimmune Diseases




Journal of


Pharmaceutics



of





Acknowledgment


References










































Volume 2014

ToxinsJournal of

VaccinesJournal of















AddictionJournal of






Autoimmune Diseases




Journal of


Pharmaceutics



of













Volume 2014

ToxinsJournal of

VaccinesJournal of















AddictionJournal of






Autoimmune Diseases




Journal of


Pharmaceutics



of

Research Article Antibacterial, Antioxidant, and...

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Transcript of Research Article Antibacterial, Antioxidant, and...