Introduction

CHAPTER – ONE

INTRODUCTION

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 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

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 Introduction

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
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.

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 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.

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 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

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 Introduction

Jatropha curcas is employed for its antifungal activity against Penicillium and
Aspergillus Species.

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 Review of Literature

CHAPTER – TWO

REVIEW OF LITERATURE

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 Review of Literature

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

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 Review of Literature

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).

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 Review of Literature

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).

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 Review of Literature

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.

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 Review of Literature

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

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 Review of Literature

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

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 Review of Literature

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).

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 Materials & Methods

CHAPTER – 3
MATERIALS AND METHODS

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 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

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 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.

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 Results

CHAPTER – 4

RESULTS

Results

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 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
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.

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 Results

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.

2.5
Zone of Inhibition (mm)

1.5
Penicillium sp.
Aspergillus sp.
1

0.5

0
Neat 1:20 1:40
Concentration of oil

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

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 Results

A. Neat

B. 1:2

C. 1:4

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

concentrations.

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 Results

A. Neat

B. 1:2

C. 1:4

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 Discussion

CHAPTER – 5

DISCUSSION

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 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

“Screening of Antifungal Activity of Jatropha curcas oil on Penicillium & Aspergillus” 24

 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).

“Screening of Antifungal Activity of Jatropha curcas oil on Penicillium & Aspergillus” 25

 References

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“Screening of Antifungal Activity of Jatropha curcas oil on Penicillium & Aspergillus” 30

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

Supervisor
Dr. Vidit Tyagi
Lecturer
Dept of Biosciences
D.I.B.N.S. Dehradun

Submitted by
Varun Jaglan

Department Of Biotechnology
Dolphin (PG) Institute of Biomedical&
Natural Sciences, Dehradun
Acknowledgement
 CONTENTS

Page no.

Acknowledgement i
Abstract
List of Tables

Introduction
Review of Literature
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 concentrations
4.2 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.