Sunilbabu Koppula et al.

/ Journal of Pharmacy Research 2010, 3(10),2453-2456

Research Article
ISSN: 0974-6943 Available online through
www.jpronline.info
Antibacterial activity screening of few medicinal plants from the Southern Region of India
Sunilbabu Koppula 1, K. Ammani 1, Varaprasad Bobbarala2* and P.V.Bramhachari 3
1
Department of Microbiology, Acharya Nagarjuna University, AP, India
2
Department of Biotechnology, Krishna University, AP, India
3
Vivimed Labs Ltd., Veeranag Towers, Habsiguda, Hyderabad, AP.
Received on: 04-07-2010; Revised on: 09-08-2010; Accepted on:20-09-2010

ABSTRACT
Antibacterial activity of fifteen medicinal plants, Andrographis paniculata Nees, Azadirachta indica A. Juss., Butea monosperma (Lam.) Taub., Calotropis
procera (Ait.) R.Br., Cassia occidentalis Linn., Catharanthus roseus Linn., Hyptis sauveolens (L.) Poit., Morus alba L., Ocimum sanctum L., Physalis minima
Linn., Piper nigrum L., Plumbago zeylanica L., Terminalia chebula L., Tinospora cordifolia (Willd.) and Vernonia cinerea (L.) Less., were screened against
medically important pathogenic bacterial strains, Klebsiella pneumonia MTCC 4032, Staphylococcus aureus MTCC 3160, Staphylococcus epidermis MTCC
3615 and Micrococcus luteus MTCC 7527. The antibacterial activity was determined in aqueous and methanol extracts using both agar disc diffusion and agar well
diffusion methods. The methanol extracts were more potent than aqueous extracts of all the plants studied. Micrococcus luteus was the most resistant strain while
the more susceptible bacterial strain was Staphylococcus epidermis. Terminalia chebula L., Morus alba L. and Azadirachta indica A. Juss., showed strong activity
against all the tested bacterial strains. Hence, these medicinal plants can be used to discern bioactive natural products and new pharmaceutical molecules that serve
in the development of unmet therapeutic needs.

Key words: Antibacterial activity, Methanol extract, Micrococcus luteus,Staphylococcus epidermis, Terminalia chebula L., Morus alba L. and Azadirachta
indica A. Juss.,

INTRODUCTION
Medicinal plants continue to be an important therapeutic aid for alleviating
the ailments of humankind. The search for perpetual health and prolonged
existence for remedies to mitigate pain and discomfort drove early man to
explore his immediate natural surroundings and led to the use of many plants,
animal products, and minerals, etc. and the development of an array of thera-
peutic agents. Today, there is a transformed interest in traditional medicine
and an increasing demand for more drugs from the plant sources. This resur-
gence of interest in plant-derived drugs is mainly due to the current widespread
belief that “green medicine” is safe and more dependable than the costly
synthetic drugs, many of which have adverse side effects. Nature has bestowed
upon us a very rich botanical affluence and a large number of diverse types of
plants grow wild in different parts of our country. In India, the use of different
parts of several medicinal plants to cure specific ailments has been in trend
from ancient times 1. India is rich in medicinal plant diversity. All known types
of agro-climatic, ecologic and edaphic conditions are met within India. India is
rich in all three levels of biodiversity, as species diversity, genetic diversity and
habitat diversity 2.
are well documented15. Large scale evaluation of the local flora exploited in
traditional medicine for various biological activities is a necessary first step in
the isolation and characterization of the active principle and further leading
to drug development. In view of these few medicinal plants were studied ex-
haustively for its potential against important four human pathogenic bacteria.
MATERIALS AND METHODS
Plant material collection:
The plant materials were collected in Jan 2009 from in and around Acharya
Nagarjuna Univesity (ANU), Guntur, A.P, India. The taxonomic identities of
these plants were identified by their vernacular names and later validated by
Prof. T. Pullaiah, Sri Krishna Devaraya University, AP, India. The voucher
specimens were deposited at ANU. The ethno botanical information of the
screened plants is given in (Table 1). A total of fifteen plants were screened
(Table.1) Different parts of the plants were collected, shade dried, and pow-
dered in a homogenizer, and 10g of each plant was used for aqueous and
methanol extraction and finally stored in airtight bottles.

In order to promote the use of medicinal plants as potential sources of anti-
bacterial compounds, it is pertinent to thoroughly investigate their bioactiv-
ity and thus validate their potential use 3-5. Natural products are known to play
an important role in both drug discovery and chemical biology. In fact, many
of the current drugs either mimic naturally occurring molecules or have struc-
tures that are fully or in part derived from natural motifs6. Natural Antibacte-
rials can be derived from barks, stems, leaves, flowers and fruits of plants,
various animal tissues or from microorganisms7-9.
Preparation of plant extracts:
Plant parts were air dried at room temperature at constant weights. The dried
plant material was separately ground to powder. The powdered was then weighed
and extracted with aquaous and methanol with reflux on water bath at 40° C
for three consecutive days. The extract was filtered and evaporated under
vacuum to dryness with rotary evaporator and then placed in and over at 60°
C until constant weight was obtained and the solutions were prepared with
concentrations 1mg/ml using DMSO.

Some phytochemicals produced by plants have antibacterial activity allowing
these plants to be studied and used for the development of new Antibacterial
drugs10. The effectiveness of phytochemicals in the treatment of various
dreadful diseases may lie in their antioxidant effects 11. In the ancient times,
plant metabolites were generally considered as sources of antinutritional fac-
tors. According to World Health Organization, medicinal plants are the best
sources to obtain a variety of newer herbal drugs. In many developing coun-
tries, traditional medicine is one of the primary health care systems12-14. Herbs
are widely exploited in the traditional medicine and their curative potentials
Determination of Antibacterial activity:
The test organisms used were obtained from Microbial Type Culture Collec-
tion & Gene Bank (MTCC), (Institute of Microbial Technology (IMTECH),
Chandigarh, India. The tested organisms used in this study were as follows:
Micrococcus luteus (MTCC 5198), Staphylococcus aureus (MTCC B96), Kleb-
siella pneumonia (MTCC 109) and Staphylococcus epidermis (MTCC 435).
The medium used for the activation of the microorganisms was soybean casein
broth (SBCB). All the culture media were prepared and treated according to the
manufacturer guidelines (Himedia laboratories, Mumbai, India).

*Corresponding author.
Dr. Varaprasad Bobbarala
Plot No. 92, Phase 1, IDA, Cherlapally
Hyderabad, A.P 500051 India
Tel.: + 91-9949129539
E-mail:phytodrugs@gmail.com
The microorganisms were inoculated into SBCB and incubated at 35 ± 2°C for
4 h. The turbidity of the resulting suspensions was diluted with SBCB to obtain
a transmittance of 25.0 % at 580 nm. That percentage was found spectropho-
tometrically comparable to 1 McFarland turbidity standard. This level of
turbidity is equivalent to approximately 3.0 × 10 8 CFU/ml. The Shimadzu
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Table 1. Ethnobotanical information of some medicinal plant species of Southern India selected for antibacterial activity.

S.No. Botanical Name Family Parts used Uses / Ailments treated

1. Andrographis paniculata Nees Acanthaceae Whole plant, leaves Anti-biotic, anti-viral, anti-parasitic and immune System stimulant
2. Azadirachta indica A. Juss. Meliaceae Fruits and leaves Effective against fungi, bacteria, viral, antimalarial and antihelmintic.
3. Butea monosperma (Lam.) Taub. Fabaceae Bark, flowers &leaves Leprosy, antihelmintic and tumerous
4. Calotropis procera (Ait.) R.Br. Asclepiadaceae Root bark Leprosy, diarrhea, dysentery, cough and asthma.
5. Cassia occidentalis Linn. Caesalpinaceae Whole plant Boils, Spasm. Hysteria, Whooping cough
6. Catharanthus roseus Linn. Apocynaceae Leaves and roots Anti-mitotic and Anti-microtubule agents
7. Hyptis sauveolens (L.) Poit. Lamiaceae Leaves Stimulant, carminative and used for healing wounds.
8. Morus alba L Moraceae Bark and fruit Asthma, bronchitis, cold and constipation
9. Ocimum sanctum L. Lamiaceae Leaves Anthelmintic, bronchitis and genitor-urinary diseases
10. Physalis minima Linn. Solanaceae Leaves, fruit & flower Laxative, abdominal troubles and spleen enlargement.
11. Piper nigrum L. Piperaceae Fruits Asthma, fever, arthritis and hemorrhoids.
12. Plumbago zeylanica L. Plumbaginaceae Roots Ulcers, piles, skin diseases and influenza.
13. Terminalia chebula Retz. Combritaceae Fruit Antibacterial, digestive problems, mouthwash/gargle, astringent and douche for vaginitis.
14. Tinospora cordifolia (Willd.) Asclepiadaceae Stem Analgesic and anti-inflammatory.
15. Vernonia cinerea (L.) Less Asteraceae Leaves Elephantiasis, cough, asthma, bronchitis and gastrointestinal disorders.

Figure.1. Antibacterial activity of selected medicinal plant extracts (aqueous and methanol) against Micrococcus luteus

Figure.2. Antibacterial activity of selected medicinal plant extracts (aqueous and methanol) against Staphylococcus aureus

spectrophotometer was used to adjust the transmittance of the working sus- the strain in the SBC broth (30 ml) and incubated on a rotary shaker. Then
pensions. bacterial cultures was inoculated into the molten Muller Hinton agar media and
Antibacterial Activity: after proper homogenization it was poured into the Petri plate (Hi-media)
The antibacterial activity of different plant species was evaluated by agar disc using pour plate technique. For agar disc diffusion method, the test compound
diffusion method 16-17 for aqueous extracts and agar well diffusion method18-19 (0.1 ml) was introduced on the disc (0.7 cm) (Hi-Media) and then allowed to
for solvent extract using Mueller Hinton agar No. 2 medium for the antibac- dry. Then the disc was impregnated on the seeded agar plate. For agar well
terial assay. The test microorganism was activated by inoculating a loop full of diffusion method, a well was made in the seeded plates with the help of a cup-
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Figure.3. Antibacterial activity of selected medicinal plant extracts (aqueous and methanol) against Staphylococcus epidermis

Figure.4. Antibacterial activity of selected medicinal plant extracts (aqueous and methanol) against Klebsiella pneumonia.

borer (0.6 cm). The test compound was introduced into the well and the plates
were incubated at 370C for 24 h. Microbial growth was determined by measur-
ing the diameter of zone of inhibition. For each bacterial strain, controls were
maintained in which pure solvents were used instead of extract. The experi-
ment was done in three replicates and the mean values are presented.
RESULTS AND DISCUSSION
The antibacterial activity of both aqueous and methanolic extracts of the
different plants against Micrococcus luteus is shown in (Figure 1). Of aqueous
extracts, only that of Azadirachta indica showed maximum activity against
Micrococcus luteus. Methanolic extracts of 6 of the 15 plants, Terminalia
chebula L., Azadirachta indica, Plumbago zeylanica, Piper nigrum , Morus
alba, and Butea monosperma showed considerable antibacterial activity. Maxi-
mum activity was hown by Terminalia chebula followed by Piper nigrum and
Azadirachta indica. Minimum antibacterial activity was shown by Vernonia
cinerea.
The antibacterial activity of the 15 plants against S. aureus, is shown in
(Figure 2). The methanolic extracts of all the plants showed antibacterial
activity while the aqueous extracts of only 4 plants showed antibacterial
activity. The antibacterial activity of methanolic extracts was more than that
of aqueous extracts. Maximum antibacterial activity was shown by Terminalia
Journal of Pharmacy Research Vol.3.Issue 10.October 2010
chebula followed by Morus alba However, minimum activity was shown by
Tinospora cordifolia extract.
The antibacterial activity of both aqueous and methanolic extracts of the 15
plants against S. epidermis is shown in (Figure 3). Of the 15 plants, 7 showed
antibacterial activity against this bacterial strain. Maximum activity of
methanolic extract was shown by Morus alba followed by Azadirachta
indica, Piper nigrum and Terminalia chebula . Aqueous extracts of only 3 of
the 15 plants showed antibacterial activity against S. epidermis; however,
maximum activity was shown by Azadirachta indica, followed by Andrographis
paniculata and Terminalia chebula extracts. Antibacterial activity of both
aqueous and methanolic extracts of the 15 plants against K. pneumonia is
shown in (Figure 4). Methanolic extracts of all the plants showed considerable
antibacterial activity against this bacterial strain. Maximum activity of
methanolic extract was shown by Terminalia chebula and Plumbago zeylanica.
However excepting Vernonia cinerea, methanolic extracts of all the remain-
ing plants showed antibacterial activity to a certain extent. Of the 15 plants,
aqueous extracts of 3 showed antibacterial activity against K. pneumonia .
Maximum activity of aqueous extract was shown by Terminalia chebula .
All plants showed greater inhibitory activity in the methanolic when com-
pared to aqueous extracts. Aqueous plant extracts of Morus alba and Tinospora
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cordifolia had no antibacterial activity against any of the four bacterial strains
investigated. Of the investigated 4 bacterial strains, Micrococcus luteus was
the most resistant strain, whereas the more susceptible bacterial strain was
Staphylococcus epidermis. Terminalia chebula , Morus alba and Azadirachta
indica showed strong activity against all the tested bacterial strains. Based on
our results, it is concluded that plant extracts have great potential as Antibac-
terial compounds against microorganisms and they can be used in the treat-
ment of infectious diseases caused by resistant microorganisms. Several pub-
lished reports show the effectiveness of traditional herbs against microorgan-
isms, as a result, plants are one of the bedrocks for modern medicine to attain
new principles20. Acquired resistance to various antibiotics in bacteria over 25
years now constitutes a serious threat to public health21-22. Nearly all of the
identified components from plants active against microorganisms are aro-
matic or saturated organic compounds, they are often obtained through initial
ethanol or methanol extraction 23. Terminalia chebula showed stronger activ-
ity than the other plants against all the tested bacterial strains. Therefore,
Terminalia chebula could be selected for further analysis. Similar results were
reported in the ethanolic extracts of T. chebula fruit, which showed highest
activity against various human pathogens such as Salmonella typhi SSFP 4S,
Staphylococcus epidermidis MTCC 3615, Staphylococcus aureus ATCC 25923,
Bacillus subtilis MTCC 441 and Pseudomonas aeruginosa ATCC 27853.
Conversely, the alcoholic extracts of black myrobalan (Teminalia chebula
Retz) on Helicobactor pylori also showed significant antibacterial activity24.
Based on these results, it can be concluded that plant extracts have a great
potential Antibacterial compounds against microorganisms that can be used in
treatment of infectious diseases caused by a range of resistant microorgan-
isms. Therefore screening of various natural organic compounds and identifi-
cation of active agents is the need of the hour, because successful prediction of
lead molecules and drug-like properties at the inception of drug discovery will
pay off later in drug development. It is vital that research strategies be ori-
ented towards discovery and development of Antibacterial agents urgently
required in the future.
ACKNOWLEDGEMENTS:
The authors acknowledge the assistance of the technicians Prasad and Nagraju.
We also acknowledge Professor T. Pullaiah for assisting us with the identifica-
tion of plant materials. This work was supported by grants from the Rajiv
Gandhi National fellowship as Senior Research Fellowship, UGC Grants, India.
Source of support: Nil, Conflict of interest: None Declared
Journal of Pharmacy Research Vol.3.Issue 10.October 2010
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