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

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

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

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
*Corresponding author. a transmittance of 25.0 % at 580 nm. That percentage was found spectropho-
Dr. Varaprasad Bobbarala tometrically comparable to 1 McFarland turbidity standard. This level of
Plot No. 92, Phase 1, IDA, Cherlapally turbidity is equivalent to approximately 3.0 × 10 8 CFU/ml. The Shimadzu
Hyderabad, A.P 500051 India
Tel.: + 91-9949129539
E-mail:phytodrugs@gmail.com Journal of Pharmacy Research Vol.3.Issue 10.October 2010 2453-2456
Sunilbabu Koppula et al. / Journal of Pharmacy Research 2010, 3(10),2453-2456
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-
Journal of Pharmacy Research Vol.3.Issue 10.October 2010 2453-2456
Sunilbabu Koppula et al. / Journal of Pharmacy Research 2010, 3(10),2453-2456
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 chebula followed by Morus alba However, minimum activity was shown by
were incubated at 370C for 24 h. Microbial growth was determined by measur- Tinospora cordifolia extract.
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- The antibacterial activity of both aqueous and methanolic extracts of the 15
ment was done in three replicates and the mean values are presented. plants against S. epidermis is shown in (Figure 3). Of the 15 plants, 7 showed
antibacterial activity against this bacterial strain. Maximum activity of
RESULTS AND DISCUSSION methanolic extract was shown by Morus alba followed by Azadirachta
The antibacterial activity of both aqueous and methanolic extracts of the indica, Piper nigrum and Terminalia chebula . Aqueous extracts of only 3 of
different plants against Micrococcus luteus is shown in (Figure 1). Of aqueous the 15 plants showed antibacterial activity against S. epidermis; however,
extracts, only that of Azadirachta indica showed maximum activity against maximum activity was shown by Azadirachta indica, followed by Andrographis
Micrococcus luteus. Methanolic extracts of 6 of the 15 plants, Terminalia paniculata and Terminalia chebula extracts. Antibacterial activity of both
chebula L., Azadirachta indica, Plumbago zeylanica, Piper nigrum , Morus aqueous and methanolic extracts of the 15 plants against K. pneumonia is
alba, and Butea monosperma showed considerable antibacterial activity. Maxi- shown in (Figure 4). Methanolic extracts of all the plants showed considerable
mum activity was hown by Terminalia chebula followed by Piper nigrum and antibacterial activity against this bacterial strain. Maximum activity of
Azadirachta indica. Minimum antibacterial activity was shown by Vernonia methanolic extract was shown by Terminalia chebula and Plumbago zeylanica.
cinerea. However excepting Vernonia cinerea, methanolic extracts of all the remain-
ing plants showed antibacterial activity to a certain extent. Of the 15 plants,
The antibacterial activity of the 15 plants against S. aureus, is shown in aqueous extracts of 3 showed antibacterial activity against K. pneumonia .
(Figure 2). The methanolic extracts of all the plants showed antibacterial Maximum activity of aqueous extract was shown by Terminalia chebula .
activity while the aqueous extracts of only 4 plants showed antibacterial
activity. The antibacterial activity of methanolic extracts was more than that All plants showed greater inhibitory activity in the methanolic when com-
of aqueous extracts. Maximum antibacterial activity was shown by Terminalia pared to aqueous extracts. Aqueous plant extracts of Morus alba and Tinospora
Journal of Pharmacy Research Vol.3.Issue 10.October 2010 2453-2456
Sunilbabu Koppula et al. / Journal of Pharmacy Research 2010, 3(10),2453-2456
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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 2453-2456