anti fungal activity of jatropha curcus

Introduction

CHAPTER – ONE

INTRODUCTION

“Screening of Antifungal Activity of Jatropha curcas oil on Penicillium & Aspergillus” 1

Introduction

Introduction

The development of new and different antimicrobial agents has been a very important

step (Agrafotis, 2002). Much of the research efforts are directed towards the design of

new and available drugs because of the unsatisfactory status of present drugs side

effects and the acquisitum of resistance by the infecting organism to present drugs

(Yalcin et al., 1992; Ertepinar et al., 1995). The resistance of common pathogens to

standard antibiotic therapy is rapidly becoming a major health problem throughout the

world.

A significant portion of the agricultural produce in the country and the world over

become unfit for human consumption due to mycotoxins contamination of grains,

especially those produced by species of Aspergillus (Janardhana et al., 1999; Chandra

and Sarbhoy, 1997; Devi et al., 2001). Eventhough effective and efficient control of

seed borne fungi of seeds can be achieved by the use of synthetic chemical fungicides,

the same cannot be applied to grains for reasons of pesticide toxicity (Ferrer and

Cabral, 1991; Harris et al., 2001; Dukic et al., 2004). Thus, there is a need to search

for alternative approaches to store grains/cereals for human consumption without

toxicity problems that are ecofriendly and not capital intensive. Plant extracts of many

higher plants have been reported to exhibit antibacterial, antifungal and insecticidal

properties under laboratory trails (Satish et al., 1999; Okigbo and Ogbonnaya, 2006;

Shariff et al., 2006; Ergene et al., 2006; Kiran and Raveesha, 2006; Mohana and

Raveesha, 2006). Plant metabolites and plant- based pesticides appear to be one of the

better alternatives as they are known to have minimal environmental impact and

danger to consumers in contrast to the synthetic pesticides (Verma and Dubey, 1999) .

This led the authors to screen in vitro a large number of plants for antifungal activity

against important seed borne Aspergillus species with the ultimate aim of developing

plant based formulations for plant disease management and safe storage of grains.

From time immemorial, use of plants for medicinal purposes has been known through

out the world. Ayurveda, which had developed as a science several centuries, before

the birth of Christ, extensively used raw material of plants in drug formulation. Plants

are vital for the existence of life in the universe they not only synthesize food

necessary for well being of man, but also in manufacturing of different chemicals

necessary for human health. The idea that there are naturally occurring substances


Introduction

useful in the treatment of diseases is very old. And this has been found from the early

second like Egyptian papyri, the ancient Sanskrit writings and many other early

records, including the Bible.

The medicinal plants provide a magnificent variety of natural products responsible for

the medicinal value of plants. The medicinal use of herb represents an important

aspect of the history of medicine as well as contribution to development of modern

pharmatherapeutic. The pharmaceutical preparations derived from these natural

products were in the form of crude drug such as a dried herb.

The essential oils from various plant species are also reported to have multifarious

biological activities like antimicrobial, anti-inflammatory etc. There are reports of

anti-microbial properties of essential oils of various plants against both plant and

human pathogens.

The use of essential oil plants for their pleasant fragrance is an old human civilization.

Incense and myrrh are the oldest known aromatic material. The early use of fragrance

materials is also well documented in Vedic literature. Other classical sources are the

Gilgamesh Epic, the bible and Greek author such as Herodotus and Hippocarates. The

essential oils which were regularly used in ancient Rome, Greece, and Egypt and

throughout the Middle and Far East had, as a common feature, the essence of a plant;

an identifiable aroma, flavour, or other characteristic that was of some practical use.

They were used as perfumes, food flavours, deodorants, pharmaceuticals, and

embalming antiseptics. Usually, plant material was steeped in a fatty oil or wine that

acted as a solvent for the desired flavour or aroma. The extracts (usually impure and

dilute) were used as oils or creams. They were introduced into Europe, without

further development, to become the subject of specialist craftsmen (the English Guild

of Pepperers and the French court perfumers of the 12th Century) and early

publications ("The Book of Nurture", 1430). In Spain and France from the early

1300s, distillation was developed to produce more concentrated essences of rosemary

and sage.


Introduction

Essential oils are used in food industry for flavouring, in the cosmetic industry for

fragrance and in pharmaceutical industry for their functional property. The primary

market for essential oils is flavours and fragrance industries which include soft drinks

companies, food companies and perfume companies.

Essential oils produced by plants have been traditionally used for respiratory trait

infections, and are used, now a days, as a medicine for cold as well as urinary trait

infection (Inouye et al., 2001). The most important suggested area of essential oils is

urology, dermatology, sleep and vascular systems, immunomodulating drugs, colds

and coughs.

The inhibitory effect of essential oils obtained by hydrodistillation, from five endemic

Psiadia species of the Asteraceae family on different type of micro organism have

been studied (Goviade et al., 2004). The anti microbial activity of lemon grass was

investigated by Santoyo et al., (2005). After study on anti microbial activity and

minimum inhibitory concentration of the essential oils was done by Effehhar et al.,

(2005) obtained from leaves, root and the seeds of Dioplotaenia damarandica,

Mozaaffarian, an endemic plant of tean. Determination of anti microbial activity of

Chamopodium botrys against selected strains of micro organisms was done by

Maksimoric et al., (2005).

The antimicrobial actions of essential oils are one of the most extensively studied

aspects of botanical medicines. It is well established that some plants contain

compounds that can inhibit the microbial growth (Naqui et al., 1994). These

compounds have different structures and different actions when compared with

antimicrobial conventionally used to control the microbial growth and survival

(Nascimento et al., 2000). The antimicrobial properties of plant depend upon their

ability to synthesize tne various secondary metabolites (Nychas, 1996). These include

alkaloids, flavonoids, isoflavonoids, tannins, cumarins, glycosides, terpenes,

phenlypropanes, and organic acids

CLASSES OF ANTIMICROBIALS

Antibiotics: Antibiotics are generally used to treat bacterial infections. The toxicity

to humans and other animals from antibiotics is generally considered to be low.


http://en.wikipedia.org/wiki/Toxicity

http://en.wikipedia.org/wiki/Infections

http://en.wikipedia.org/wiki/Antibiotics

Introduction

However, prolonged use of certain antibiotics can decrease the number of gut flora,

which can have a negative impact on health.

The term antibiotic originally described only those formulations derived from living

organisms but is now applied also to synthetic antimicrobials, such as the

sulfonamides.

Antiviral: Antiviral drugs are a class of medication used specifically for treating

viral infections. Like antibiotics, specific antivirals are used for specific viruses. They

are relatively harmless to the host, and therefore can be used to treat infections. They

should be distinguished from viricides, which actively deactivate virus particles

outside the body.

Antifungal: An antifungal drug is medication used to treat fungal infections such as

athlete's foot, ringworm, candidiasis (thrush), serious systemic infections such as

cryptococcal meningitis, and others.

Antiparasitic: Antiparasitics are a class of medications which are indicated for the

treatment of infection by parasites such as nematodes, cestodes, trematodes, infectious

protozoa, and amoebas.

Non-pharmaceutical antimicrobials: Traditional healers have long used

plants to prevent or cure infectious disease. Many of these plants have been

investigated scientifically for antimicrobial activity and a large number of plant

products have been shown to inhibit the growth of pathogenic microorganisms. A

number of these agents appear to have structures and modes of action that are distinct

from those of the antibiotics in current use, suggesting that cross-resistance with

agents already in use may be minimal. So, it is worthwhile to study plants and plant

products for activity against resistant bacteria.

The present work includes essential oil from plant Jatropha curcus. Jatropha curcus

oil has remarkable medicinal action. This oil contains some active constituents against

microorganisms.

OBJECTIVES

In the field of natural medicine, essential oils are important anti infectious and

antimicrobial agents. The importance of essential oils grown as the microbes develop

resistant against presently available antibiotic and antiviral drugs. For their

alternative, essential oils can be used as promising standard antimicrobial drugs. The

present work constitutes an effort in this direction. In this study, the essential oil of


http://en.wikipedia.org/wiki/Cross-resistance

http://en.wikipedia.org/wiki/Amoeba

http://en.wikipedia.org/wiki/Protozoa

http://en.wikipedia.org/wiki/Trematode

http://en.wikipedia.org/wiki/Cestode

http://en.wikipedia.org/wiki/Nematodes

http://en.wikipedia.org/wiki/Parasites

http://en.wikipedia.org/wiki/Meningitis

http://en.wikipedia.org/wiki/Cryptococcus

http://en.wikipedia.org/wiki/Candidiasis

http://en.wikipedia.org/wiki/Ringworm

http://en.wikipedia.org/wiki/Athlete's_foot

http://en.wikipedia.org/wiki/Fungi

http://en.wikipedia.org/wiki/Medication

http://en.wikipedia.org/wiki/Viricide

http://en.wikipedia.org/wiki/Pharmacotherapy

http://en.wikipedia.org/wiki/Virus

http://en.wikipedia.org/wiki/Sulfonamide

http://en.wikipedia.org/wiki/Chemical_synthesis

http://en.wikipedia.org/wiki/Gut_flora

Introduction

Jatropha curcas is employed for its antifungal activity against Penicillium and

Aspergillus Species.


Review of Literature

CHAPTER – TWO

REVIEW OF LITERATURE




Essential oils are chemical products formed odoriferous essences extracted from a

great diversity of plants. They are volatile liquids, insoluble in water highly soluble in

alcohol, ether, vegetable, and minerals oils. Essential oil are complex mixture of

different chemical compounds. However they can be characterized by their main

volatile components (Guenther E, 1948)

Essential oils (also called volatile oils) are aromatic oily liquids obtained from plant

materials (flowers, buds, seeds, leaves, bark, wood, fruits and roots). They can be

obtained by expression, fermentation or extraction but the method of steam distillation

is most commonly used for commercial production. An estimated 3000 essential oils

are known, out of which 300 are commercially important in fragrance market (Vande

Break SAAJ, 1999). Essential oils are complex mixtures comprising many single

compounds. Chemically, they are derived from terpenes and their oxygenated

compounds. Each of these constituents contributes to the beneficial or adverse effects.

Essential oils such as aniseed, calamus, camphor, cedarwood, cinnamon, citronella,

clove, eucalyptus, geranium, lavender, lemon, lemongrass, linmint, mutmeg, orange,

palmarosa, rosemary, basil, vetiver and wintergreen have been traditionally used by

people for various purposes in different parts of the world. Cinnamon, clove and

rosemary oils had shown antimicrobial activity (Quattara et al. 1997). Cinnamon oil

also possesses antidiabetic property. Anti-inflammatory activity has been found in

basil (Singh and Majumdar, 1999).

Chemical Nature of Essential Oil

Essential oil, like all organic compounds, are made up of hydrocarbon molecule and

can further be classified as terpenes, alcohols, ester, aldehydes, ketones and phenols

etc. The essential oil are highly enriched in compounds based on an isoprene structure

called terpenes. Terpenes constitute several thousand compounds with multiple

functionalized molecules. The general chemical structure of terpenes is C10H16 and

they occur as diterpenes and triterpenes and tetraterpens (C20,C30&C40) as well as

hemiterpenes(C5) and sesquiterpenes (C15) (Cowman, 1999).

When the compound contains additional elements, usually oxygen, they are termed as

terpenoids. Terpenoids are synthesized from acetate units, and as such they share

origin with fatty acids. They differ from fatty acid in that they contain extensive



branching and cyclized. Examples of common terpenoids are menthol and camphor

(monoterpenes) and farnesol and artemisin (sesquiterpenoids) (Cowman, 1999)

Anti microbial compounds in essential oils

For safe and effective therapeutic use of essential oils, it is important to have a basic

knowledge of their constituents. Essential oils usually contain several major

compounds in essentials oils fall into two primary groups: Hydrocarbons, which are

mainly terpenes and oxygenated compounds which are phenols, alcohol, esters,

ethers, aldehydes, ketones and oxides.

Terpenes: Essentials oils are highly enriched in compounds based on isoprene’s

structures called as terpenes. Terpenes constituents several thousands compounds with

multiple functionalized molecules. The general chemical structure of terpenes is C10

H16 and they occur as di-terpenes, tri-terpenes and tetra-terpenes (C20 , C30 and C40) as

well as hemi terpenes (C5) and sesqui terpenes (C15) (Cowan,1999).

When the compounds contains additional elements usually oxygen, they are termed as

terpenoid. Example of common terpenoid is menthol and camphor (monoterpenes)

and farnesol and artemisin (sesqui terpenes) (Cowan, 1999). Different terpenes have

anti-inflammatory, anti-septic, anti-viral and anti-bacterial properties.

Terpenes alcohols: Terpenes alcohols as are a group among the most beneficial and

versatile of the essential oils compounds they have broad spectrum anti infectious,

anti bacterial, anti fungal and anti viral properties. In general they are non irritating

and relatively safe. Two of the more common terpenic alcohols found in anti

microbial oils are terpinen-4-ol in Melaleuca alternifolia (tea tree) and Origanum

majorana (sweet majoram).

Phenols: Phenols are among the most potent of the anit microbial compounds.

Phenols have powerful broad spectrum anti infectious, anti bacterial functions, anit

septic and disinfectant and have strong anit parasitic properties. They have moderately

strong tonic, stimulant, anti-viral, anti –fungal and immune enhancing properties.

Three of the most important from essential oils are thymol, carvacrol and eugenol.

Aldehydes: Monoterpenes aldehydes are found primarily in the lemon scented oils.

This group contains oils that have anti inflammatory, anti infectious, anti fungal, anti

bacterial and disinfectant powers. These must be used with cautions as they can cause

skin irritation. Monoterpenes aldehydes include citrol (bergamot, lemon, lime and

lemon grass), geranial (orange, mellisa) and citronellal (citronella, lemon grass).



Ketones: Ketones are some of the most toxic compounds found in essential oils.

However some ketones-containing oils have excellent therapeutic value, although

they are not generally considered strong anti microbial. Some oils containing ketones

aid in wound healing and dissolving mucous, some are immune system stimulants and

some are anti fungal. They can be used effectively in conjunction with stronger anti

microbial oils. Oils such as hyssop, eucalyptus, and rosemary have moderate amounts

of ketones.

Esters: Esters are not major anti microbial, but they can be used in stronger anti

microbial oils. Esters found in essential oils are normally very fragrant with a fruity

aroma. Their therapeutic effects are balancing to the nervous system, calming, anti-

inflammatory and anti –spasmodic. An example of a well known ester is lineally

acetate which is found in lavender, clary, sage and petit grain.

Antimicrobial Literature

The application of plant extract for the control of plant disease was first attempted by

Democritus as early as 470 BC. With the passage of time, various active fraction or

constituents isolated from higher plants in the form of essential oils have been

extensively investigated for their activity against the number of stored fungi, plant

pathogens, bacterial and other harmful organisms. The potential antimicrobial

properties of plant have been related to their ability to synthesise by the secondary

metabolites, several chemical compounds of relatively complex structured with

antimicrobial activity, including alkanoids, flavonoids, isoflavonoids, tannins,

cumarins, glycosides, terprens, phenylpropanes, organic acid (Nychas, 1996).

Hammer et al., 1999, investigated the 52 plant oils and extracts for activity against

Acinetobacter baumanii, Aeromonas veronii, Candida albicans, Enterococcus

faecalis, Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa,

Salmonella enterica subsp. enterica, Serratia marcescens and Staphylococcus aureus,

using an agar dilution method and twenty of the plant oils and extracts were

investigated, using a broth microdilution method, for activity against C. albicans, S.

aureus and E. coli. From this study ,they reported plant essential oils and extracts may

have a role as pharmaceuticals and preservatives.

Aureli et al (1992) studied the antimicrobial activity of 32 plants essential oils against

Listeria innocua and L.monocytogenes. The cinnamon, clove oraginum, pimeto and

thyme oil showed antilisterial activity even at lower concentration (1:50 v/v).


http://www.ncbi.nlm.nih.gov/sites/entrez?Db=pubmed&Cmd=Search&Term=%22Hammer%20KA%22%5BAuthor%5D&itool=EntrezSystem2.PEntrez.Pubmed.Pubmed_ResultsPanel.Pubmed_DiscoveryPanel.Pubmed_RVAbstractPlus


Scientific studies have also shown that essential oils contain chemical components

that can exact specific effects on the mind and body. Their chemistry is complex but

generally includes alcohols, esters, ketones, aldehydes and terpenes. Each essential oil

contains as much as 100 chemical components which together exert a strong effect on

the body. Depending on which component is predominating in an oil, the oils act

differently. For example, some oils are relaxing, some soothes down, some relieves

pain etc. there are also oils such as lemon and lavender, which adapt to the needs of

body, and adapt to particular situation, these are called “adaptogenic”. The

mechanism in which these essential oils acts on body is not very well understood.

They affect mind and emotion, leave no harmful residues, enter into the body either

by absorption or inhalation.



Jatropha curcus L.

Source: Indian Council of Forestry Research and Education, Dehradun.

Kingdom: Plantae

Division: Magnoliophyta

Class: Magnoliopsida

Order: Malpighiales

Family: Euphorbiaceae

Genus: Jatropha

Species: curcus


http://en.wikipedia.org/wiki/Euphorbiaceae

http://en.wikipedia.org/wiki/Malpighiales

http://en.wikipedia.org/wiki/Magnoliopsida

http://en.wikipedia.org/wiki/Flowering_plant

http://en.wikipedia.org/wiki/Plant


Jatropha curcus, Linn of family Euphorbiaceae is commonly called Physic nut.

It is a monoceiuous shrub or small tree up to 6-8m high; bark pale brown, papery,

peeling slash exudes a copious watery latex, soapy to touch but soon becoming brittle

and brownish when dry ; branches glabrous, ascending, stout (FAO,1986).

Leaves alternate, palmate, petiolate, stipulate; stipules minute; petiole 2-20cm long,

blade 3-7 lobed, 12.5-18cm long, 11-16cm wide, lobes acute or shortly acuminate at

the apex, margins entire or undulating, leaf base deeply cordate, glaborous or only

pubescent on the nervous below, basal nerves 7-9, prominent, venation reticulate

(FAO,1986).

Chemical composition of Jatropha curcus

Curcin - a phytotoxin (toxalbumin), found mainly in the seeds and also in the fruit

and sap.

Purgative oil - the seed yields 40% oil, known as hell oil, pinheon oil, oleum

infernale or oleum ricini majoris, which contains small amounts of an irritant

curcanoleic acid, which is related to ricinoleic acid and crotonoleic acid, the principle

active ingredients of castor oil and croton oil respectively (Joubert et al., 1984). This

genus also may contain hydrocyanic acid (CRC Critical Reviews in Toxicology

1977). There may be a dermatitis producing resin (Lampe & Fagerstrom, 1968).

There may be an alkaloid, and a glycoside which produce cardiovascular and

respiratory depression.

The fixed oil (41.5%) from Jatropha curcus seeds was consisted of palmitic acid

(14.16%), stearic acid (7.68%), oleic acid (46.72%) and linoleic acid (30.31%). In

India, oil contents were found to be rich in oleic and linoleic acids.

Uses

Jatropha is an ornamental plant naturalized in many tropical areas. The roots, stems,

leaves seeds and fruits of the plant have been widely used in traditional folk medicine

in many parts of West Africa. The seeds of J. curcas have been used as a purgative,

antihelminthic and abortifacient as well as for treating ascites, gout, paralysis and

skin diseases. The seed oil of the plant has been used as an ingredient in the treatment

of rheumatic conditions, itch and parasitic skin diseases, and in the treatment of

fever, jaundice and gonorrhoea, as a diuretic agent, and a mouth-wash. The leaf has

been used as a haemostatic agent and the bark as a fish poison. In certain African

countries people are accustomed to chewing these seeds when in need of a laxative.

J. curcas seeds have been found to be highly effective against Strongyloides apillosus



infection in goats (Adam & Magzoub, in press). It has also been suggested that J.

curcas seeds could be a useful chemotherapeutic agent provided that it is active at a

non-lethal dose (Adam, 1974). This may be because of it's reported antihelminthic

activity. Jatropha oil is an environmentally safe, cost-effective renewable source of

non-conventional energy and a promising substitute for diesel, kerosene and other

fuels. Physic nut oil was used in engines in Segou, Mali, during World War II. The oil

burns without smoke and has been employed for street lighting near Rio de Janeiro.

Medicinal Use

The plant is used as a medicine in treatment of more than 32 cases in different parts of

the world. The different parts of the plant is used to treat many diseases like anasarca,

ascites, burns, carbuncles, caries, cataplasm, circumcision, collapse, convulsions,

cough, dentition, depurative, diarrhoea, dropsy, dysentery, eczema, erysipelas, fever,

gonorrhoea, guinea worms, hernia, incontinence, itch, jaundice, neuralgia, parturition,

piles, pleurisy, pneumonia, rash, rheumatism, roundworms, sore, sprains, stomach

ache, syphilis, tetanus, thrush, toothache, tumour, wounds, yaws, yellow fever ,

enema, homicide, pesticide, raticide, medicine, mouthwash, poison. In Southern

Sudan, the seed as well as the fruits is used as a contraceptive.

Emmanuel and Gabriel (1996) reported that it is used to cure jaundice. The leaves are

ground to a thin material; shea butter is added to ground leaves. The mixture is used

to rub anybody suffering from measles (Thomas, 1981).

The fresh latex of the plant is applied to ringworm, bleeding wounds, eczema, itch,

scabies and decayed teeth. Adecoction of the leaves is used as a remedy for fevers and

a mouthwash for strengthing the gums. The poultice of the leaves is applied over

ulcers, boils and abscesses to promote suppuration, as galactagogue, it is applied to

the breasts of nursing mothers. The ash of the leaves or their hot infusion is used for

extracting guinea-worms (Gill, 1992).


Materials & Methods

CHAPTER – 3

MATERIALS AND METHODS


Materials & Methods

Materials & Methods

Anti microbial activity of oil

The anti microbial activity of oil of Jatropha curcus was carried out. The essential oil

was screened for anti fungal activities.

Anti fungal activity of oil

In this study, the anti fungal activity was studied against the micro organism viz.

Aspergillus niger and Penicillium chrysogenum. These cultures are obtained from the

standard culture maintained in the department of biotechnology, D.I.B.N.S, Dehradun.

Fresh culture were obtained by transferring a loopful of culture into sabourand

dextrose broth and then incubated at 270C for 72 hrs. To test antifungal activity, the

plate diffusion method was used.

Seed oil

Jatropha curcus seed oil which was used in experiments was obtained from Forest

Research Institute, Dehradun.

Culture media preparation

The microbiological media were prepared as standard instructions provided by the HI-

MEDIA laboratories Pvt Ltd, Mumbai. The medium used for antifungal activity was

S.D.S which was prepared and sterilized at 121oC at 15 psi for 15-30 minutes in

autoclave.

Plate preparation

25 ml of pre autoclaved S.D.A was poured into 90 mm diameter pre sterilized

petriplates and was allowed to solidify at room temperature.

Well diffusion method

After the plate solidify, the freshly prepared 72 hrs fungal broth culture suspensions

(about 0.1 ml) was spreaded over the S.D.A media using L shaped sterilized glass

spreader separately under aseptic condition using laminar air flow. A well of 5 mm


Materials & Methods

diameter was made in the centre of petriplates with the help of a puncher. 100 µl of

neat oil as well as oil of different dilutions was poured in the well in different

petriplates. This method depends upon the diffusion of oil through well in the

solidified agar layer of petriplate to such an extent that the growth of added micro

organism is prevented entirely in the circular area or zone around the well containing

oil.

Incubation

Petri plates were incubated for 72 hrs at 28oc in the incubator.

Measurement of zone of inhibition

After incubation the diameter of clear zone of inhibition around the disc were

measured in mm.


Results

CHAPTER – 4

RESULTS

Results


Results

Table 4.1: Physico-chemical characteristics of Jatropha Oil

Property Jatropha curcas Oil

Viscosity (cp) (30°C) 5.51

Speciflc gravity (15°C/4°C) 0.917/ 0.923(0.881)

Solidfying Point (°C) 2.0

Cetane Value 51.0

Flash Point (°C) 110 / 340

Carbon Residue (%) 0.64

Distillation (°C) 284 to 295

Sulfur (%) 0.13 to 0.16

Acid Value 1.0 to 38.2

Saponification Value 188 to 198

Iodine Value 90.8 to 112.5

Refractive Index (30°C) 1.47

Source: Environmental Intermediaries & Trading Group Limited

Colony Morphology of Fungal Cultures

Penicillium Sp.

Colour – Yellow

Mycelium: - It is well developed and copiously branched. The hyphae constituting

the mycelium are septate and the cells are short. The septa between the cells have

each a central pore. Through the pores, the protoplasm flows from cell to cell.

Conidiophores: - A conidiophores arise as an erect, tubuler hyphal outgrowth from

any cell of the mycelium and not from, specialized cell (foot cell) as in Aspergillus.

Conidiophores are septate. Reaching a certain height the septate conidiophores

branches once or twice or even more times. Asexual spores ‘The conidia’ are

produced exogenously at the tips of the conidiophores.

Aspergillus Sp.

Colour – Yellow


Results

Mycelium: - It is well developed and made up of a loosely interwoven mass of

hyaline, bright or pale coloured, extensively branched, septate hyphae . The hyphae

are freely branched and form dense mats on the substratum.

Conidiophores: - Conidiophores are developed from the chrematistic T- Shaped cell

called foot cells. Conidiophores grow up to a length of about 2.5 m and swells at its

tip to form vesicle. From the surface of vesicle sterigmata are formed.

Table 4.2: Antifungal activity of Jatropha curcus oil

(Diameter of Inhibition zone)

Name of bacteria Neat 1:2 1:4

Penicillium sp. 1.8mm 1.4mm 0.9mm

Aspergillus sp. 2.25mm 1.3mm 1.1mm

Fig. 4.1: Diameter of Zone of Inhibition at different oil concentrations.

0

0.5

1

1.5

2

2.5

Neat 1:20 1:40

Concentration of oil

Zo

ne

of

Inh

ibit

ion

(m

m)

Penicillium sp.

Aspergillus sp.

Plate 4.1. Inhibitory effect of essential oil on Penicillium sp. at different oil concentrations.


Results

A. Neat

B. 1:2

C. 1:4

Plate 4. 2. Inhibitory effect of essential oil on Aspergillus sp. at different oil concentrations.


Results

A. Neat

B. 1:2

C. 1:4


Discussion

CHAPTER – 5

DISCUSSION


Discussion

Discussion

Essential oils are volatile substances derived from plants (Cowan, 1999) and contain

compounds that can inhibit the microbial growth (Naqui et al., 1994). The most

important compounds are linalool, methyl chevicol, α – pinene, β-pinene, geraniol,

citrol, citronellal, menthol etc. (Guenther, 1949; Simon et al., 1984). The present

work involves the antibacterial activity of Jatropha curcus L. seed oil.

In the field of natural medicine, essential oils are important anti-infectious and

antimicrobial agents, whose importance grows as microbial resistant to antibiotics and

antiviral drugs increases.

Two strains of fungus namely Penicillium sp. and Aspergillus sp. were taken for the

study of antifungal activity. The purpose of this study was to examine the antifungal

effects of essential oil on Penicillium sp. and Aspergillus sp.

In the present study the antifungal effect of essential oil was screened against the

isolates of fungus using well-diffusion agar method. The diameters of zone of

inhibition were measured at two dilutions 1:2 and 1:4. The neat (no solvent, only oil)

had maximum effect against fungal strains. The maximum diameter of zone of

inhibition was measured at dilution 1:2 (Fig. 4.1). The essential oil showed more

inhibition against Aspergillus sp. in comparison to that of Penicillium sp.

In the present study, anti bacterial activity of Jatropha oil and sensitivity of micro

organism can be determined. Research in present study can not be directly co related

to other studies. The results of effect of dilution on anti microbial activity can be

compared with the study of Yonzon et al., (2005), in which the inhibitory effect of

some Nepalese oil on five microbes using paper dics method was determined. These

oils were Anthopagon oil, Chamonile oil, Curcuma zedoaria oil, French basil oil,

Cornmint oil and Palmorosa oil. The result indicated that the activity of essential oil is

decrease with dilution.

The anti fungal activity of oil decreased with dilution. Diameter of zone of inhibition

was more in case of neat than that of zone at 1:2 and 1:4 dilutions. The anti fungal

activity of Jatropha oil was found to be more against Aspergillus sp, the diameter of

zone inhibition at neat is 2.25 mm and is decreased with dilution at 1:2 and 1:4

(1.3mm and 1.1mm respectively). The anti fungal activity of Jatropha oil against


Discussion

Penicllium sp is also more at neat as compared to zones at dilutions 1:2 and 1:4. The

diameter of zone of inhibition at neat is 1.8 mm. The diameter zone of inhibition is

decreases with dilutions at 1:2 and 1:4 (1.4mm and 0.9mm respectively).


References

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Media composition

All compositions are in gram/ litre.

Muller Hinton Agar

Casein acid hydrolysate 17.5 gm

Beef hurt infusion 2.00 gm

Starch soluble 1.50 gm

Distilled water 1000ml

pH 7.5

Potato Dextrose Agar

Potato infusion 200 gm

Dextrose 10 gm

Agar 15 gm

pH 7

D.W 1000ml

Sabouraud Dextrose Broth

Special peptone 10.00 gm

Dextrose 20.00 gm

D.W 1000ml

pH 5.6

Screening of Antifungal activity of Jatropha curcus L. oil against Penicillium and Aspergillus Species

Dissertation submitted to the

H.N.B. Garhwal University,Srinagar, (Uttrakhand)

In partial fulfillment for the award of the degree of

BACHELOR OF SCIENCE

IN

BIOTECHNOLOGY

2008

SupervisorDr. Vidit Tyagi

LecturerDept of BiosciencesD.I.B.N.S. Dehradun

Submitted by Varun Jaglan

Department Of BiotechnologyDolphin (PG) Institute of Biomedical&

Natural Sciences, Dehradun

Acknowledgement

CONTENTS

Page no.

Acknowledgement i

AbstractList of Tables

Introduction


Materials and Methods

Result and Discussion

References

Appendix ii

List of Tables

Table No. Subject 4.1 Physico-chemical characteristics of Jatropha Oil 4.2 Antifungal activity of Jatropha curcus oil

List of Figures

Figure No. Subject 4.1 Diameter of Zone of Inhibition at different oil

Concentrations.

List of Plates

Plate No. Subject 4.1 Inhibitory effect of essential oil on Penicillium sp. at different oil concentrations4.2 Inhibitory effect of essential oil on Aspergillus sp. at different oil concentrations

ABSTRACT

The development of new and different antimicrobial agents has been a very important

step. From time immemorial, use of plants for medicinal purposes has been known

through out the world. The essential oils from various plant species are also reported

to have multifarious biological activities like antimicrobial, anti-inflammatory etc.

The antimicrobial action of essential oils is one of the most extensively studied

aspects of botanical medicine. The objective of the present study was to screen the

antibacterial activity of essential oil against bacteria. The antifungal activity of the

seed oil of Jatropha curcus L. was carried out. The seed oil was screened for

antifungal activity by well diffusion technique. The diameters of zone of inhibition

were measured at two dilutions 1:2 and 1:4. The neat (no solvent, only oil) had

maximum effect against fungal strains. The essential oil showed more inhibition

against Aspergillus sp. in comparison to that of Penicillium sp.

anti fungal activity of jatropha curcus

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