International Journal of Medicine and Pharmaceutical Sciences (IJMPS) ISSN 2250-0049 Vol.

3, Issue 2, Jun 2013, 11-18 TJPRC Pvt. Ltd.

SCREENING OF ETHNOMEDICINAL PLANTS FOR ANTIBACTERIAL ACTIVITY

SUBRAMANIYAN VIJAYAKUMAR, SELLAN CHANDRASEKAR & SRINIVASAN PRABHU P.G. and Research Department of Botany and Microbiology, A. V. V. M. Sri Pushpam College (Autonomous), Poondi, Tamil Nadu, India

ABSTRACT
Aim of the Study The present study screening of the ethnobotanical plant is a pre-requisite to evaluate their therapeutic potential and it can lead to the isolation of new bioactive compounds. Methods The crude extracts and fractions of seven medicinal important plants (Abutilon indicum, Cassia tora, Cissampelos pareira, Punica granatum, Terminalia cuneata, Ceropegia tuberosa and Pistacia integerrima) were tested against three gram positive and three gram negative ATCC bacterial species using the agar well diffusion method. Results The crude extracts of Abutilon indicum, Punica granatum and Pistacia integerrima were active against all tested bacterial strains (12-22 mm zone of inhibition). Other four plants crude extract (Cassia tora, Cissampelos pareira, Terminalia cuneata, Ceropegia tuberosa) were active against different bacterial strains. The crude extracts showed varying level of bactericidal activity. The aqueous fractions of Abutilon indicum and Pistacia integerrima crude extract showed maximum activity (18.2 and 16 mm respectively) against B. subtilis, while the chloroform fractions of Cassia tora and Ceropegia tuberosa presented good antibacterial activities (11-16 mm zone of inhibition) against all the bacterial cultures tested. Conclusions The methanol fraction of Abutilon indicum chloroform fractions Ceropegia tuberosa, Punica granatum, Cassia tora and aqueous fraction of Pistacia integerrima are suitable candidates for development of noval antibacterial compounds.

KEYWORDS: Ethnomedicinal Plants, Antibacterial Activity, Western Ghats, Fraction Method INTRODUCTION
Infectious diseases caused by the microbes are health hazards all over the world. Several synthetic antibiotics and drugs are employed in the treatment of microbial infections and communicable disease. But the microbial pathogens develop resistance to the synthetic antibiotics. There is an increasing interest to unlock the secrets of ancient herbal remedies. The synthetic antibiotics have the following limitation; these are costly and are not of range from the patient belonging to developing countries. Secondly with the passage of time, microorganisms develop resistance against antibiotics. Therefore, after some time these antibiotics are not effective against the microbes (Walsh and Amyer, 2004; Alder, 2005). For this purpose, various strategies have been developed eg, biological screening, isolation as well as clinical trials for a variety of plants. Based on the screening methodologies, the therapeutic values of many herbal medicines are

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Subramaniyan Vijayakumar, Sellan Chandrasekar & Srinivasan Prabhu

obtained from natural sources, and considered to be safe for human beings. On the contrary, they would have some adverse affects due to the presence of other active ingredients (Izzo and Ernst, 2009). World wide mortality rate increased due to infectious bacterial diseases (Nathan, 2004). The bacterial organism including gram positive and gram negative like different species of Bacillus, Staphylococcus, Salmonella and Pseudomonas are the main sources which cause severe infections in human beings. Because these organisms have the ability to survive in harsh conditions due to their multiple environmental habitats (Ahameethunisa, 2010). Natural products have got incredible success in serving as a guide post for new antibacterial drug discovery. Moreover, antibiotics obtained in this way have biologically ecofriendly (Walsh, 2003 and Koehn and Carter, 2005). As is well known that the bioactive plant extracts is a promising source of majority of drugs (Bibi et al., 2010). For example, quinine (cinchona) and berberine (Berberis) are the antibiotics obtained from plants which are highly effective against microbes (Staphylococcus aureus and Escherichia coli) (Maridass and Britto, 2008). However, despite enormous amount of the information on the antibacterial properties of plans any yet to be explored. Hence, the present investigation has been taken up with an objective to evaluate the antibacterial potential of seven selected plants used by Tribals in the Western Ghats of Tamil Nadu.

METHODS
Study Area The plant material was collected from different geographical regions of Tamilnadu, India. Abutilon indicum was collected from Madurai (Western Ghats) Cassia tora was collected from Nilgries, (Western Ghats), Cissampelos pareira was collected from Velliangri (Southern Western Ghats), Punica granatum and Terminalia cuneata were collected from Kancheepuram (Northern Western Ghats), Ceropegia tuberosa was collected from Kolli hills, Namakkal (Eastern Ghats) and Pistacia integerrima was collected from Tirunelveli (Western Ghats). The collected plants were identified by Botanical survey of India (BSI) Southern region, Coimbatore and Voucher no deposited to Dept. of Botany A.V.V.M. Sri Pushpam College Poondi, Thanjavur. Each plant material was thoroughly washed under running tap water and dried under shade. The dried material of Ceropegia tuberosa rhizome (5 kg), Pistacia integerrima stem (2 kg), Terminalia cuneata stem bark (2 kg), Cissampelos Pareira roots (5 kg), Cassia tora leaves (4 kg) and Abutilon indicum leaves (1.5 kg) was ground to powder form for extraction. Soxhelt extraction technique was used for extraction from Ceropegia tuberosa and Cissampelos pareira. Soxhelt apparatus was used to carryout the extraction. For this purpose, plant material was divided into two portions. Each portion was extracted with 200 ml methanol. Quantity of the crude extract obtained was 250 g while, other six plants (Ceropegia tuberosa, Pistacia integerrima, Terminalia cuneata, Cissampelos pareira, Cassia tora and Abutilon indicum were subjected to cold maceration technique for extraction. Powdered material of each plant was soaked in methanol separately at room temperature. After seven days, the extract was filtered under vacuum through whatman filter paper No.1. The residue was again dipped in methanol for additional seven days and filtered thereafter. The filtrates were combined and methanol was evaporated under vacuum, using rotary evaporator (Buchi Rotavapor R-200) at 45C. The quantities of extracts obtained from Ceropegia tuberosa, Pistacia integerrima, Terminalia cuneata, Cissampelos pareira and Cassia tora, Abutilon indicum were 250, 400, 400, 250, 200 and 250 g respectively (Table 1). Fractionation Fractionation was carried out by suspending each extract in 250 mL water separately and partitioning with different organic solvents (hexane, chloroform, ethyl acetate and methanol) in order of increasing polarity by using separating funnel (Figure 1). All the five fractions of each plant extract were dried by evaporating respective solvent using

Screening of Ethnomedicinal Plants for Antibacterial Activity

13

rotary evaporator. All extracts were stored at 4C till further analysis. (Location for Figure 1) Antibacterial Activity The agar well diffusion method was performed to exploit antibacterial potential of used extracts (Bibi et al., 2010). Each extract (200 mg) was dissolved in 10 mL of 99.9 % dimethyl sulfoxide (DMSO) (Sigma-Aldrich USA) to get 20 mg/mL concentration. Cefotaxime (2 mg/mL) in DMSO was prepared as positive control. Pure DMSO (99.9%) was used as negative control. Test Microorganism In the present study, total six bacterial strains were obtained from Microbial Type Culture Collection (MTCC), Institute of Microbial Technology, Chandigarh, India. Three of them were gram positive, i.e., Bacillus subtilis (MTCC 6633), Stephylococcus aureus (MTCC 6538) and Micrococcus luteus (MTCC 10240). Out of six bacteria strains, three were gram negative namely Klebsiella pneumoniae (MTCC 15380), Pseudomonas aeruginosa (MTCC 25668), Salmonella typhi (MTCC 19430). The nutrient broth (8 g/L) was prepared by dissolving nutrient broth (Merck) in distilled water. At pH = 7.0, 50 ml of media was dispensed in a 100 ml flask and then autoclaved. The bacterial cultures were inoculated individually and kept at 37C overnight in a shaker incubator at 150 rpm. Turbidity Standard and Preparation of Inocula The turbidity standard was prepared by mixing 0.5 ml of BaCl2 (0.048 M) in 99.5 ml 0.3 6N H2SO4BaSO4. The standard was taken in screw cap test tube to compare the turbidity. The bacterial culture of selected strains were grown overnight, and subsequently mixed with physiological saline. Turbidity was corrected by adding sterile saline until McFarland 0.5 BaSO4 turbidity standard 108 Colony Forming Unit (CFU) per ml was achieved. These inocula were used for seeding of the nutrient agar. Procedure Nutrient agar medium as prepared by suspending nutrient agar (Merck) 20 g/L in distilled water. The pH value of the media was adjusted to 7.0, autoclaved, and allowed to cool up to 45C. The media was seeded with 10 ml prepared inocula. Subsequently, the seeded medium (75-80 ml) and allowed to solidify. Required numbers of wells per plate (six wells for extracts, one for positive and negative control each) were made with 8 mm sterile cork borer. These wells were sealed by pouring 20 m of liquid nutrient agar medium in each well. With the help of micropipette, 100 l of test solution was poured into respective well. Sample of extracts, one positive control (Cefotaxime), and the negative control (DMSO) were applied to each Petri plate. Then the plates were incubated at 37C. After 24 hr of incubation period, diameter of clear zones around each well was measured, showing no bacterial growth. In this study triplicate plates were prepared for each extract and bacterial strain. The mean zone of inhibition was calculated with standard deviation procedure. The percentage growth inhibition was calculated by Percentage inhibition = (TS-SC)/PC 100 Where TS/SC and PC represents test sample, solvent control and positive control, respectively. Statistical Analysis Analysis of variance (ANOVA) and Least Significant Difference (LSD) test at P < 0.05 was carried out using MSTATC to determine the significance of percentage inhibition values between the extracts against bacterial strains.

RESULTS AND DISCUSSIONS

Totally seven medicinal plants used for different remedies by local communities were tested against MTCC bacterial cultures to determine and investigate their antibacterial potential. We observed that the crude methanol extract of Abutilon indicum showed significant antibacterial activities against all the tested bacterial strains. Maximum activity was

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Subramaniyan Vijayakumar, Sellan Chandrasekar & Srinivasan Prabhu

conferred against Bacillus subtilis (22 mm) while minimum was observed against Staphylococcus aureus and Klebsiella pneumoniae with mean inhibition zone diameter 14 and 14.3 mm respectively (Table 2). The fractions of Abutilon indicum extract showed activity to different extent against different bacterial strains. However, Bacillus subtilis was found more susceptible among other tested strains as all fractions except methanol fraction exhibited activity against it. This observation contradicts previous findings that Bacillus subtilis was found least sensitive among other strains against different plant extracts (Papp, 2004). Bacillus subtilis has also been reported most sensitive among different stains (Duraipandiyan et al., 2006). The aqueous fraction showed maximum inhibition value (18.2 mm) among other fractions against Bacillus subtilis. The crude extract and all fractions except aqueous fraction were active against Micrococcus luteus. Crude extract also exhibit remarkable activity against Salmonella typhi with mean zone of inhibition (19.66 mm). Abutilon indicum contains -pinene and Geraniol has been reported for antimicrobial properties (Milind and Pates, 2012). The crude extract of Cassia tora leaves was significantly active against all bacterial strains except Pseudomonas aeruginosa. Maximum zone of inhibition (19 mm) was observed against Staphylococcus aureus. All five fractions were active, but the chloroform fraction was observed as most active fraction. It showed considerable active against all bacterial strains. Maximum inhibition zone (16.8 mm) was shown against Klebsiella pneumoniae. Previous studies indicate the presence of chrysophanol, rhein and d-mannitol in the Cassia tora plant extract that might be responsible for anticancer and purgative activity (Seth and Sharma, 2004). The crude extract of Cissampelos pareira showed least activity against Bacillus subtilis (11.2 mm). The extract was inactive against other tested bacterial strains. The chloroform and ethyl acetate fractions also active against Bacillus subtilis slightly more than the crude extract, representing zones of inhibition 13 and 15 mm, respectively (Table 2). Amritpal et al. (2010) reported that Cissampelos pareira extract contain a large portion of Cissampareine and Hayline which might be responsible for antileukamic and antifertility activity. No other fraction was active against any bacterial strain used in the present study. Punica granatum crude extract showed moderate activity with rang of inhibition zone 13-16.5 mm. Maximum inhibition was observed against Klebsiella pneumoniae and minimum inhibition against Bacillus subtilis. Extract also showed good inhibition zone (14 mm) against Pseudomonas aeruginosa, Staphylococcus aureus and Salmonella typhi were moderately inhibited showing zone of inhibition 15 mm and 14 mm respectively (Table 2). Hexane fraction of Punica granatum did not show any activity against any tested bacterial strains. The chloroform fraction showed almost same level of activity (11-15 mm) similar to crude extract against all bacterial strains. Ethyl acetate and methanol fraction showed moderate inhibition, however, aqueous fraction proved most active among all fractions except Pseudomonas aeruginosa and Micrococcus luteus showing maximum inhibition zone 17 mm against Bacillus subtilis. Maximum activity by aqueous fraction might be due to presence of compounds which are polar in nature. It is evident form the traditional use of water decoctions to treat ailments (Kamuhabwa et al., 2000). 2-phenoxy, 2-methoxy, 1, 3, 4-trimethyl, and 9, 2 octade cadinic acid were isolated from Punica granatum have shown antivirus and immune stimulatory activities (Lali et al., 2012). Terminalia cuneata crude extract was active against all bacterial strains except Pseudomonas aeruginosa, Salmonella typhi. Maximum zone of inhibition (13.7 mm) was observed against Micrococcus luteus. An average zone of 10-11 mm was observed against Bacillus subtilis, Staphylococcus aureus and Klebsiella pneumoniae. Chloroform and methanol fractions of Punica granatum showed moderate activity against three strains while ethyl acetate fraction showed mild inhibition (9.6 mm) of Klebsiella pneumoniae. Rochfort et al. (2008) reported that Terminalia cuneata contain saponin and tannin activity against rheumatism and seminal defects.

Screening of Ethnomedicinal Plants for Antibacterial Activity

15

The crude methanol extract of Ceropegia tuberosa stem was active against two bacterial strains. Zones of inhibition; 14 mm and 13.3 mm; were observed against Bacillus subtilis and Micrococcus luteus, respectively. The fractions of Ceropegia tuberosa indicated the distribution of active constituents in chloroform and ethyl acetate fraction. Chloroform fraction was most active fraction as it showed considerable activity against all bacterial strains. Maximum zone of inhibition was observed against Micrococcus luteus that is 17 mm (Table 2). Distribution of activity in less polar fractions indicates that active compounds of the plant are non polar to slightly polar in nature. Aqueous fraction was mildly active only against Pseudomonas aeruginosa with mean zone of inhibition 12.6 mm. Flavonoids and phenols isolated from Ceropegia tuberosa have shown leuchorroeal properties (Kessler et al., 2003). The fractions of P. integerima extract showed activity of different extent against different bacterial strains. However, Bacillus subtilis was found more susceptible among other tested strains as all fractions except methanol fraction exhibited activity against it. This observation contradicts previous findings that Bacillus subtilis was found least sensitive among other strains against different plant extracts (Papp, 2004). Bacillus subtilis has also been reported most sensitive among different strains (Duraipandiyan et al., 2006). The aqueous fraction showed maximum inhibition value (16 mm) among other fractions against B. subtilis. The crude extract also exhibit remarkable activity against P. aeruginosa with mean zone of inhibition 22 mm. Pistacia integerrima contains beta sitosterol has been reported for antimicrobial properties (Ahmed et al., 2010). The comparative percent inhibition by all plant extracts were represented in Figure 2. The crude extract of Abutilon indicum showed maximum inhibition of Bacillus subtilis (23.2%). Same extract also showed 19.5% inhibition of Salmonella typhi while, chloroform fraction of Cassia tora showed good inhibition of Staphylococcus aureus (18.4%). A number of extract showed good activities against many strains in a range 18.24% inhibition. Other scientist working in the same field has also been reported on the variable antibacterial activity by plants extracts and their fractions. Such studies inspired the scientist to identify other bioactive compound through isolation (Janicsak et al., 2011; Yin et al., 2010 and Nenaah, 2010). Figure 2 also describes that in few cases crude extract exhibits moderate activity while their respective fractions showed pronounced activity. This might be due to distribution of active component in specific fraction depending upon its solubility nature. While, in few cases the crude extract is more active then fractions. The crude extract of Abutilon indicum exhibited more activity against all tested bacterial strains, as compared to its fractions. In such cases more than one active component might be present in crude extract which distributed into different fractions depending on their solubility. The clinical isolates were inhibited by plant extracts at varying degree. The extract presenting > 18 inhibition provides a good reason to use these plants for isolation of new antibacterial compound(s). (Location for Figure 2)

CONCLUSIONS
In summary, we have described antibacterial properties of aqueous fraction of Abutilon indicum and Pistacia integerrima, Chloroform fraction of Cassia tora and Ceropegia tuberosa against the tested microbes. Therefore, the extracts of these plants were considered as suitable candidates for antibacterial drug discovery. The other extract showed lower activity which might suggest lack of bio-active components and/or insufficient quantities in the extract. Based on our findings, we envision that the discovery of novel antibacterial agent from natural sources (plants) will help to minimize the adverse effects of synthetic drugs.

ACKNOWLEDGEMENTS
The authors are thankful to the management of A.V.V.M. Sri Pushpam College (Autonomous), Poondi, for providing them necessary facilities and support to carry out this work.

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Subramaniyan Vijayakumar, Sellan Chandrasekar & Srinivasan Prabhu

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21. Pant, S., and Samant, S.S., (2010). Ethnobotanical observations in the Moranula reserve forest of Kumoun West Himalaya. Ethnobotanical Leaflets, 14: 193-217. 22. Papp, N., (2004). Antimicrobial activity of extracts of five Hungarian Euphorbia species and some plant metabolites. Acta Botanica. Hungarica, 463: 363-371. 23. Rao, V.H., Gohil, T.G., and Thakor, A.B., (2012). Study of ethnobotanical plants of Kaparada Taluka and their significance to the tribes. Life Science. Leaflets, 11: 73-85. 24. Rouchfort, S., Anthony, J., and Frank, R., (2008). Plant bioactives for ruminant health and productivity. J. Phytochemistry, 69: 299-322. 25. Rudrapal, M., Sridhar, N., and Raghavendra, M., (2011). Ethnomedicinal plants used by traditional healers in East Godavari district of Andhra Pradesh, India. 3(3): 426-431. 26. Rufeng, W.Y.D., Ruining, L., Lan, X., and Lijun, D.,(2010). Pomegranate: constituents. Bioactivities and pharmacokinetics. J. Fruit Vegetables and Cereal Science and Biotechnology, 4(2): 77-87. 27. Samanta, J., and Bhattacharya, S., (2011). Cissampelos pareira: A promising antifertlity agent. Inter. J. Research Ayurveda and Pharmacy, 2(2): 439-442. 28. Samydurai, P., Thangapandiyan, V., and Aravinthan, V., (2012). Wild habits of Kolli hills being stable food of inhabitant of eastern ghats, Tamil Nadu, India. Indian J. Natural Prodoucts Resesrch., 3(3): 432-437. 29. Seth, S.D., and Sharma, B., (2004). Medicinal plants of India. Indian J. Medical Research., 120: 9-11. 30. Singh, A., Singh, G.S., and Singh, P.K., (2012). Medico ethnobotanical inventory of Renukoot forest division of district Sonbhadra, Uttar Pradesh, India. Indian J. Natural Prodoucts Research., 3(3): 448-457. 31. Walsh, C., (2003). Where will new antibiotics come from? Nature Reviews Microbiology, 1: 65-70. 32. Walsh, F.M., and Amyes, S.G.B., (2004). Microbiology and drug resistance mechanisms of fully resistant pathogens. Current. Opinion Microbiology., 7: 439-444. 33. Yin, S., Sykes, M.L., Davis, R.A., Shelper, T., Avery, V.M., Camp, D., and Quinn, R.J., (2010). New Galloylated Flavonoids from the Australian Plant Glochidion sumatranum. Panta Medica, 16: 1877-1881.

APPENDICES
Table 1: Ethnomedicinal Plants Used by Traditional Healers of Different Regions in Tamil Nadu
S. N o. 1. Plant Family Local Name Duvven aku Occurrence in Tamilnadu Collection Site Western Ghats Madurai Part Used (Kg) Leaves (1.5) Traditional Use Bleeding piles (Rudrapal et al., 2012) Edema, Leprosy skin disease (Singh et al., 2012) heart disease urinary problem (Rudrapal et al., 2012) intestinal worms (Rudrapal et al., 2012) asthma, cold, fever (Rao et al., 2012) Diabetic, health tonic (Samydurai et al., 2012) Properties Anti-inflammation (Khabdai et al., 2009) Anti-Malarial (Abdul et al., 2008) Anti-cancer ophthalmic and purgative (Choudhary et al., 2011) Antileukameic, antifertility (Samanta and Bhattachara, 2011) Immuno-stimulatory, Antivirus (Rufeng et al., 2010) Rheumatism, seminal defects (Rockfort et al., 2008) Leuchorroea (Kessler et al., 2003) Method of Extraction Cold maceration Extract Obtaine d (g) 250

Abutilon indicum L.

Malvace ae

2.

Cassia tora L.

Caeasal piniacea e

Thagara sa

Wester Ghats Nilgris, Ooty

Leaves (4)

Cold maceration

200

3.

Cissampel os pareira L. Punica granatum L. Terminali a cuneata Roth Ceropegia tuberosa Dalz

Menispe rmaceae

Chirubo ddi Dhanem ma chettu Tellama ddi

Southern western Ghats, Velliangiri, hills Coimbatore Northern Western Ghats, Kancheepuram Coimbatore Northern Western Ghats, Kancheepuram Eastern Ghats, Kolli hills, Namakkal

Roots (5)

Soxheit extraction

250

4.

Punicac eae

Roots (4) Stem bark (2) Rhizo me (5)

Cold maceration

200

5.

Combret aceae

Cold maceration

400

6.

Asclepia daceae

Manchi mandai

Soxhelt extraction

250

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Subramaniyan Vijayakumar, Sellan Chandrasekar & Srinivasan Prabhu

7.

Pistacia integerrim a galls

Anacard iaeae

Villanay / Vellatch y

Western Ghats, Thirunelveli

Stem (2)

Cough, phthisis, asthma, dysentery (Pant and Samant, 2010)

Antioxidant, radical scarvenging, xanthine oxidase inhibitory activities (Ahmad et al., 2008)

Cold maceration

400

Table 2: Zone of Inhibition (mm: In Diameter) Against Different Bacterial Strains by Plants Extracts
Plant Extract Crude Hexane Chloroform acetate Ethyl acetate Methanol Aqueous Cefotaxime DMSO Crude Hexane Chloroform Ethyl acetate Methanol Aqueous Cefotaxime DMSO Crude Hexane Chloroform Ethyl acetate Methanol Aqueous Cefotaxime DMSO Crude Hexane Chloroform Ethyl acetate Methanol Aqueous Cefotaxime DMSO Crude Hexane Chloroform Ethyl acetate Methanol Aqueous Cefotaxime DMSO Crude Hexane Chloroform Ethyl acetate Methanol Aqueous Cefotaxime DMSO Crude Hexane Chloroform Ethyl acetate Methanol Aqueous Cefotaxime DMSO Bacillus Subtilis 22.42 0.41 15.2 0.12 12 0 14 0 18.2 0.6 33.44 0 13.2 0.06 10.8 0.21 16.3 0.21 18 0.06 38 0.6 11.2 0.07 13 0.03 15 0.04 32.7 0.9 13 0 11.5 0.8 14 0.05 12 0.5 17 0.1 30 0.5 10.3 0.61 13.2 0.8 13 0.05 38 0.05 14 0.02 16 0.2 14.7 0.26 37 0.5 20 0.41 14 0.01 12 0 15 0 16 0.61 40 0.21 Staphylococcus Aureus 14 0 12.3 0.5 11 0 12.55 0.5 10 0.01 31 0.56 18 0.41 17 0 19 0.6 14.7 0.1 39 0.21 38 0.7 15 0.5 13 0 11 0.1 15 0.5 30 0.2 10.8 0.07 14 0 40 0.21 15 0.2 39 0.02 18 0 13.6 0.05 11 0 13.44 0.5 14 0.20 36 0.05 Microorganisms Used Klebsiella P. Pneumoniae Aeruginosa 14.23 0.36 19 0.31 12.3 0.2 10 0.2 11 0 10.26 0 13.3 0.7 12.2 0.2 10 0.3 11.6 0 30 0.23 8 0.26 18 0.2 16.8 0.4 15 0.02 34 0.04 34 0.2 13 0.64 36 0.08 39 0.62 16.5 0.01 16 0 12.6 0.5 15 0.8 13 0.5 10 0.1 11 0.06 16 0.5 30 0 32 0.1 10.6 0.21 9.6 0.06 13 0.02 32.3 0.61 39 0.2 13 0.29 12 0.21 12.6 0.8 32 0.05 38 0.06 18.36 0.4 22 0.4 13 0 12 0.2 11 0 10.66 0 14.42 0.37 13.3 0.2 10 0.01 10 0.23 15 0.11 14 0.21 33 0.5 33 0.2 Salmonella Typhi 19.66 0.04 9 0.6 10 0.23 8 0.23 11.3 0.2 28 0 15 0.11 13 0.21 30 0.12 103 0.5 35 0.6 14 0.08 14 0.1 12 0.2 10 0.26 13 0.06 32 0 13.6 0.21 27 0.21 15 0.21 11.3 0.1 29 0.26 19 0.05 11 0.05 13 0 13 0.11 30 0 M. Luteus 18 0.24 8.6 0.2 9.6 0.3 27 0.2 14.2 0.1 14 0.02 13 0.11 30 0 9 0.7 12 0.61 31 0.6 14 0 13 0.21 12 0.05 10 0 31 0.61 13.7 0.61 11 0.61 29 0.61 13.3 0.6 17 0.21 13 0.7 30 0.26 20 0 10 0.05 12 0.01 12 0.51 32 0.61 -

Abutilon indicum

Cassia tora

Cissampelos pareira

Punica granatum

Terminalia cuneata

Ceropegia tuberosa

Pistacia integerrima

Mean Standard Deviation of Triplicate, (-) No Zone of Inhibition Observed, Cefotaxime as Positive Control, DMSO as Negative Control