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ISSN 0973 - 8207

Research Article

EV
ALUA
TION OF WITHANIA OBTUSIFOLIA T
ACKH. (SOLANACEAE) LEAF AND
EVALUA
ALUATION
TACKH.
ROOT EXTRACTS AS AN ANTIMICROBIAL AGENT
M. Senthil Kumar, A. Saravana Kumar, A. Ramachandran* and A. Shajahan**
Plant Molecular Biology Lab, PG and Research Dept of Botany Jamal Mohamed College (Autonomous)
Tiruchirappalli- 620 020
*Director in Centre for Climate Change and Adaptation, Anna University, Chennai
Abstract: Leaf and root samples of Withania obtusifolia were used to examine their antimicrobial
potential against some human pathogenic bacteria (E. coli, Bacillus and Shigella) fungi (A. niger and
Trichophyton rubrum) growth inhibition was observed in different volumetric concentrations of this
extract. Phytochemical analysis showed the presence of nitrogenous compound (8-Azabicyclo [3.2.1]
octan-3-ol, 8-methyl-, endo-) as a major constituent of the plant leaf and roots which are commonly
known to be used in antimicrobial treatments. Root sample showed higher antimicrobial activity
than the leaf sample. These results confirm the antimicrobial property of W. obtusifolia leaf and
root support the traditional use of the plant in therapeutic use against microbial infections.
Key words: Withania obtusifolia, medicinal plants, antimicrobial activity, Solanaceae

Introduction
Some bacteria and fungi are extremely
pathogenic causing serious human infections.
The discovery of antibiotics to combat these
pathogens marked a revolution in the twentieth
century (Evan, 1992). Unfortunately, because
of the inappropriate usage of antibiotics in
human and veterinary medicine, certain strains
of bacteria and fungi have developed the ability
to resist their antibiotics or change their
infection process to penetrate the host cells (Ali
et al., 1995). Therefore, disease causing microbes
that have become resistant to antibiotic therapy
are becoming a public health problem. The large
part of the problem is the increased use and
abuse of existing antibiotics in human and
veterinary medicine (Muhammad and
Muhammad, 2005). To substitute synthetic
antibiotics, many of the modern and effective
drugs have their origin in traditional folk
medicine (Natarajan et al., 2003). Plants have
been used to treat human, animals and plant

disease from time immemorial, also herbal


medicines have been known to man for centuries
(Goun et al., 2003; Misra and Sahu, 1977).
Therapeutic efficacy of many indigenous plants
for many disorders have been described by
practitioners of traditional medicine (Al
Maqbool et al., 1985, 1988; Iqbal et al., 2002;
Khattak et al., 1985). Withania somnifera belongs
to the family Solanceae grows widely in the
Eastern cape and used for the treatment of
arthritis, tuberculosis, cancer, and STIs (Sexual
Transmitted Infections) (Devi, 1996; Van Wyk
et al., 1997; Singh and Kumar, 1998). Extensive
field survey and critical perusal of literature
revealed the presence of additional diploid
species W. obtusifolia in the natural population
(Tirupura Sundari et al., 1999). The
ethanobotanical data of W. obtusifolia plant
prompted an investigation into its antimicrobial
activity. Therefore, the present study was
planned to find out the antimicrobial potential
of W. obtusifolia leaves and roots for their
efficacy against various human pathogenic
fungal and bacterial strains.
1

Journal of Basic and Applied Biology, 4, 2010, pp. 1-4


2009, by the Centre for Biological Research, Puthalam, 629 602, TN, India

Senthil Kumar et al., 2010

Phytochemical analysis: Phytochemical


analysis was carried out for W. obtusifolia root
methanolic extract using the GCMS method
to analysis the phytochemical constituents.

extract was active against both bacteria (E. coli,


Bacillus and Shigella) and fungi (A. niger and
Trichophyton rubrum) strains. As a general rule,
plant is considered highly active against both
fungi and bacteria when the zone of inhibition
is greater than 6 mm (Muhammad and
Muhammad, 2005). But our samples the
maximum inhibition zone was found to be 3.1
mm in 100 l, for the root sample against an
isolated human pathogen E. coli and the
moderate inhibition zone 3.0 mm was found in
100 l, against an isolated pathogen Shigella
species. The minimum inhibition zone 2.8 mm
was found in 100 l, against B. subtilis. The
fungal isolate of the pathogen Trichophyton
rubrum, has the least inhibition zone found in
root when compared to leaf for the same
concentration. The minimum inhibition zone in
25 l concentrations was observed in T. rubrum
with 1.2 mm in the root and 0.2 mm in leaf
extracts. But the maximum inhibition zone was
3.0 mm in 100 l for the root when compared
to leaf for the same concentration. There was
an enhanced of maximum microbial activity
found in the root when compared to leaf of W.
obtusifolia. The phytochemical analyses showed
the presence of nitrogen compound (8-Azabicyclo
[3.2.1] octan-3-ol, 8-methyl-, endo-) as major
constituents of the plant leaves and root are
commonly known to posses antimicrobial
activity. The present study identifies that W.
obtusifolia acts as a potential source for
biological antimicrobial activity against
selective fungi and bacteria, which enable only
human pathogenic fungi and bacteria to be
controlled without any side effects. The tribal
people are using Withania species since time
immemorial for treating many ailments, back
pain, nerve disorder, diabetes, cancer and female
disorders properties and/or bacterial resistance
as current synthetic antibiotics are doing and
this specifically appears as additional point in
the natural antibiotics research.

Results and Discussion

References

Materials and Methods


Plant material: This is an erect,
herbaceous, evergreen, tomentose shrub. The
leaf is obtuse, its base is cuneate and is densely
hairy beneath. The flowers are yellow and the
berry fruit which is orange red in colour. The
roots have a long and even structure, a strong
odour, bitter and acrid taste. W. obtusifolia leaves
and roots samples were collected from
Karudamangalam, Thiruchirappalli District in
Tamilnadu. The fresh leaves and roots were
dried in shade condition and powdered.
Preparation of extracts: Ten grams of
the coarsely powdered plant material were
successively Soxhlet extracted with methanol
for 24 hrs. The extracts were evaporated under
vacuum and the residues were separately
dissolved in the same solvent.
Medium for microbial growth: The
nutrient agar and Rose Bengal agar medium
were prepared and sterilized for bacteria and
fungi respectively. Two fungal species namely
Aspergillus niger and Trichophyton rubrum and
three bacterial species namely Escherichia coli,
Shigella and Bacillus subtilis were obtained from
Eumic Analytic Lab Thiruchirappalli, Tamil
Nadu. Both bacterial and fungal strains were
streaked on the respective temperature at 370C
for 48 to 72 hours. Methonalic extracts of both
leaves and roots were taken separately at
various concentrations of 25, 50, 75 and 100
l. They were kept under incubation. After
incubation the plates were observed for the zone
formation and the length (mm) of the zone was
measured and tabulated.

The results of antimicrobial activity of


methanolic extracts of W. obtusifolia on tested
against microorganisms using disc diffusion
method are presented in table 1. Methanolic

Ali, B. H., Bashir, A.K. and Tanira, M.O.M.


1995. Anti inflammatory, antipyretic and
analgesic effect of Lawsonia inermis L.
(henna) in rats. Pharmacology, 51: 356 363.

Senthil Kumar et al., 2010

Table 1. Bioactivity of W. obtusifolia leaf and root methanolic extracts against fungal and
bacterial isolates

Organism
A. niger
T. rubrum
E. coli
Bacillus subtilis
Shigella

Zone formation (mm) at volume of


leaf sample loaded (l)
25l
1.4
0.2
1.1

50 l
1.6
0.4
1.1
1.1
1.2

75 l
1.8
0.7
1.2
1.3
1.3

100 l
2.4
1.8
1.4
1.5
1.5

Zone formation (mm) at volume of


root sample loaded (l)
25 l
2.1
1.2
2.2
2.2
1.9

50 l
2.3
1.4
2.4
2.4
2.2

75 l
2.8
1.6
3
2.6
2.8

100 l
3
1.8
3.1
2.8
3

Graph 1. Bioactivity of W. obtusifolia leaf methanolic extracts against fungal isolates

Graph 2. Bioactivity of W. obtusifolia root methanolic extracts against bacterial isolates

Senthil Kumar et al., 2010

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* Author for correspondence

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