Antimicrobial activity of some ethnomedicinal plants used by Paliyar tribe from Tamil Nadu, India
Veeramuthu Duraipandiyan1,
Muniappan Ayyanar1 &
Savarimuthu Ignacimuthu1
BMC Complementary and Alternative Medicine volume 6, Article number: 35 (2006) Cite this article
Antimicrobial activity of 18 ethnomedicinal plant extracts were evaluated against nine bacterial strains (Bacillus subtilis, Staphylococcus aureus, Staphylococcus epidermidis, Enterococcus faecalis, Escherichia coli, Klebsiella pneumonia, Pseudomonas aeruginosa, Ervinia sp, Proteus vulgaris) and one fungal strain (Candida albicans). The collected ethnomedicinal plants were used in folk medicine in the treatment of skin diseases, venereal diseases, respiratory problems and nervous disorders.
Plants were collected from Palni hills of Southern Western Ghats and the ethnobotanical data were gathered from traditional healers who inhabit the study area. The hexane and methanol extracts were obtained by cold percolation method and the antimicrobial activity was found using paper disc diffusion method. All microorganisms were obtained from Christian Medical College, Vellore, Tamil Nadu, India.
The results indicated that out of 18 plants, 10 plants exhibited antimicrobial activity against one or more of the tested microorganisms at three different concentrations of 1.25, 2.5 and 5 mg/disc. Among the plants tested, Acalypha fruticosa, Peltophorum pterocarpum, Toddalia asiatica,Cassia auriculata, Punica granatum and Syzygium lineare were most active. The highest antifungal activity was exhibited by methanol extract of Peltophorum pterocarpum and Punica granatum against Candida albicans.
This study evaluated the antimicrobial activity of the some ethnomedicinal plants used in folkloric medicine. Compared to hexane extract, methanol extract showed significant activity against tested organisms. This study also showed that Toddalia asiatica, Syzygium lineare, Acalypha fruticosa and Peltophorum pterocarpum could be potential sources of new antimicrobial agents.
According to World Health Organization (WHO) more than 80% of the world's population relies on traditional medicine for their primary healthcare needs. Use of herbal medicines in Asia represents a long history of human interactions with the environment. Plants used for traditional medicine contain a wide range of substances that can be used to treat chronic as well as infectious diseases. A vast knowledge of how to use the plants against different illnesses may be expected to have accumulated in areas where the use of plants is still of great importance [1]. The medicinal value of plants lies in some chemical substances that produce a definite physiological action on the human body. The most important of these bioactive compounds of plants are alkaloids, flavanoids, tannins and phenolic compounds [2].
Rural communities, in particular paliyar tribes, depend on plant resources mainly for herbal medicines, food, forage, construction of dwellings, making household implements, sleeping mats, and for fire and shade. The use of medicinal plants as traditional medicines is well known in rural areas of many developing countries [3, 4]. Traditional healers claim that their medicine is cheaper and more effective than modern medicine. In developing countries, low-income people such as farmers, people of small isolate villages and native communities use folk medicine for the treatment of common infections [5].
We chose eighteen plant species used in folk medicine to determine their antimicrobial activity (Table 1). In general, these plants are used in folk medicine in the treatment of skin diseases, venereal diseases, respiratory problems and nervous disorders. Properties of the collected plants are also provided in same table. Evidently, there are not many scientific studies that confirm the antimicrobial properties for most of the plants collected for this study. The phytochemical research based on ethnopharmacological informations is generally considered an effective approach in the discovery of new anti-infective agents from higher plants [6].
Table 1 Uses and properties of ethnomedicinal plants collected for antimicrobial screening
There are a few reports on the use of plants in traditional healing by either tribal people or indigenous communities of Tamil Nadu [7–11]. The development of drug resistance in human pathogens against commonly used antibiotics has necessitated a search for new antimicrobial substances from other sources including plants [12]. Screening of medicinal plants for antimicrobial activities and phytochemicals is important for finding potential new compounds for therapeutic use. This paper reports the results of a survey that was done based on folk uses by traditional practitioners in Palni hills of Tamil Nadu along with bioassay test for antimicrobial activity.
Ethnobotanical survey
Plants were selected for this study based on their medicinal use. Fresh plant parts were collected from the tribal villages (Paliyar tribe) in Palni hills of Tamil Nadu, India in Jan – April 2005. The tribal villages were approximately lie between 10° 12' – 10° 15' N longitudes and 77° 26' – 77° 33' latitude. The ethnobotanical data (local name, mode of preparation, medicinal uses) were collected through questionnaire, interviews and discussions among the tribal practitioners in their local language (Tamil). The voucher specimens in duplicate were deposited in the herbarium of Entomology Research Institute, Loyola College, Chennai (India).
Preparation of hexane and methanol extract
Plant extracts were prepared by cold percolation method. The plant materials were dried under shade and ground into fine powder using electric blender. 50 g of dried powder was soaked in 300 ml hexane for 48 hours with intermittent shaking. The plant extracts were filtered through Whatman No. 1 filter paper into pill vials. The filtrates were dried until a constant dry weight of each extract was obtained. The residues were stored at 4°C for further use. The remaining plant residue was dried and soaked in 300 ml of methanol as above and the extract was collected as described earlier.
Antimicrobial screening
The hexane and methanol extracts of 18 plants were screened against a total of 9 bacterial strains and one fungal strain. The test organisms were Bacillus subtilis (ATCC 441), Staphylococcus aureus (ATCC 25923), Staphylococcus epidermidis (MTCC 3615), Enterococcus faecalis (ATCC 29212), Escherichia coli (ATCC 25922), Klebsiella pneumonia (ATCC 15380), Pseudomonas aeruginosa (ATCC 27853), Ervinia sp (MTCC 2760), Proteus vulgaris (MTCC 1771) and one fungal strain Candida albicans (MTCC 227) obtained from the Christian Medical College, Vellore, Tamil Nadu.
Preparation of inoculum
Stock cultures were maintained at 4°C on slopes of nutrient agar. Active cultures for experiments were prepared by transferring a loopful of cells from the stock cultures to test tubes of Mueller-Hinton broth (MHB) for bacteria and Sabouraud dextrose broth (SDB) for fungi that were incubated without agitation for 24 hrs at 37°C and 25°C respectively. The cultures were diluted with fresh Mueller-Hinton and Sabouraud dextrose broth to achieve optical densities corresponding to 2.0·106 colony forming units (CFU/ml) for bacteria and 2.0·105 spore/ml for fungal strains.
Antimicrobial susceptibility test
The disc diffusion method [13] was used to screen the antimicrobial activity. In vitro antimicrobial activity was screened by using Mueller Hinton Agar (MHA) obtained from Himedia (Mumbai). The MHA plates were prepared by pouring 15 ml of molten media into sterile petriplates. The plates were allowed to solidify for 5 minutes and 0.1 % inoculum suspension was swabbed uniformly and the inoculum was allowed to dry for 5 minutes. The different concentrations of extracts (1.25, 2.5 and 5 mg/disc) were loaded on 6 mm sterile disc. The loaded disc was placed on the surface of medium and the compound was allowed to diffuse for 5 minutes and the plates were kept for incubation at 37°C for 24 hrs. At the end of incubation, inhibition zones formed around the disc were measured with transparent ruler in millimeter. The same procedure was followed for the fungus also. These studies were performed in triplicate.
Results and discussions
Table 1 provides the botanical name, family, local name, plant parts used together with their traditional therapeutic uses and properties for the 18 ethnomedicinal plants collected from Palni hills of Tamil Nadu. Out of the 18 plants tested for antimicrobial activity, 10 plant species showed antimicrobial activity by inhibiting one or more microorganisms. The results of the antimicrobial screening of the crude extracts of all species of plants are shown in Table 2. Among the plants screened, Acalypha fruticosa, Peltophorum pterocarpum, Toddalia asiatica,Cassia auriculata, Syzygium cumini and Syzygium lineare showed promising activity against tested microorganisms. The tested plant extracts were most active against gram-positive microorganisms than gram-negative microorganisms. This is in agreement with previous reports by the several workers [14–19].
Table 2 Antimicrobial activity of the hexane and methanol extracts of collected ethnomedicinal plants
Methanol extracts exhibited a higher degree of antimicrobial activity as compared with hexane extracts. Methanol extracts of Albizia procera,Cassia auriculata (leaves and flowers), Peltophorum pterocarpum, Punica granatum and Syzygium cumini showed activity. Punica granatum possesed 25% of tannin [20] and the antibacterial activity may be indicative of the presence of some metabolic toxins or broad-spectrum antibiotic compounds. Also ethanolic extract of Punica granatum was most active against E. coli. Prasanth et al [21] reported that, different extracts of Punica granatum fruit showed some antibacterial activity against P.vulgaris and B.subtilis. Rajakaruna et al [22] reported that Syzygium cumini showed good activity against Staphylococcus aureus and Bacillus subtilis.
Both hexane and methanol extracts of Syzygium lineare and Toddalia asiatica showed antimicrobial activity. The essential oils from the leaves of Toddalia asiatica were most active against E. coli, K. pneumoniae, P. aeruginosa and S. aureus [23]. Peltophorum pterocarpum and Syzigium lineare had the highest inhibitory activity against both gram positive and gram negative bacteria. On the other hand, Cassia alata showed only slight activity against bacteria such as S. aureus and B. subtilis. In the previous findings leaves, flowers, root and stem barks of Cassia alata showed a range of activity against several bacteria and protozoa [24]. In this study methanol extract of leaves of Cassia alata showed antibacterial activity against Bacillus subtilis and Staphylococcus aureus. Somchit et al [25] also tested the whole plant parts of Cassia alata and showed activity in the leaves against Staphylococcus aureus.
Syzygium lineare, Punica granatum, Syzygium cumini and Toddalia asiatica produced the largest zones of inhibition against Bacillus subtilis, Staphylococcus aureus and Staphylococcus epidermidis. Voravuthikunchai et al [20] reported good antibacterial activity in P. pterocarpum and P. granatum against Escherichia coli using aqueous and methanol extracts. Methanol extracts of Peltophorum pterocarpum and Punica granatum showed activity against Candida albicans. In general, among the tested microbial strains, bacteria were found to be more sensitive to many of the test agents than fungi.
The most sensitive bacterium was Bacillus subtilis, which was inhibited by methanol or hexane of 10 plants. On the other hand, no inhibition was observed in the Eruvinia sps. Some organisms exhibited only slight susceptibility. E. coli was inhibited by methanol extract of flowers of Cassia auriculata and hexane extract of Punica granatum. Proteus vulgaris was inhibited by methanol extract of Peltophorum pterocarpum and Syzigium lineare. Klebsiella pneumonia was inhibited by hexane extract of Olax scandens, methanol extracts of Peltophorum pterocarpum and Syzigium cumini.
The processing of the plants performed in this study was not comparable to the traditional approach when the Paliyar tribe used water for extracts whereas we have used hexane and methanol for extraction. In this sense it is not an exact replication of the traditional knowledge. All the same, given that methanol extracts were more effective then hexane extract, it is likely that water extracts were will be effective as well and possibly mort so.
The antibacterial activity of Syzigium lineare, Olax scandens, Albizia procera and Acalypha fruticosa are reported for the first time. No previous report on the antibacterial activity of these species could be found in the literature. Among the medicinal plants tested in this work, Peltophorum pterocarpum and Punica granatum showed the most promising antimicrobial properties indicating the potential for discovery of antibacterial principles. Further phytochemical studies are required to determine the types of compounds responsible for the antibacterial effects of these species. The results also indicate that scientific studies carried out on medicinal plants having traditional claims of effectiveness might warrant fruitful results. Several plants used by Paliyar tribe exhibit some degree of antibacterial activity towards gram-positive bacteria such as, Bacillus subtilis, Staphylococcus aureus and Staphylococcus epidermidis. These plants could serve as useful sources for new antimicrobial agents.
Diallo D, Hveem B, Mahmoud MA, Betge G, Paulsen BS, Maiga A: An ethnobotanical survey of herbal drugs of Gourma district, Mali. Pharmaceutical Biology. 1999, 37: 80-91.
Edeoga HO, Okwu DE, Mbaebie BO: Phytochemical constituents of some Nigerian medicinal plants. African Journal of Biotechnology. 2005, 4: 685-688.
Sandhu DS, Heinrich M: The use of health foods, spices and other botanicals in the Sikh community in London. Phytotherapy Research. 2005, 19: 633-42. 10.1002/ptr.1714.
Gupta MP, Solis PN, Calderon AI, Guionneau-Sinclair F, Correa M, Galdames C, Guerra C, Espinosa A, Alvenda GI, Robles G, Ocampo R: Medical ethnobotany of the Teribes of Bocas del Toro, Panama. Journal of Ethnopharmacology. 2005, 96: 389-401. 10.1016/j.jep.2004.08.032.
Rojas JJ, Ochoa VJ, Ocampo SA, Muñoz JF: Screening for antimicrobial activity of ten medicinal plants used in Colombian folkloric medicine: A possible alternative in the treatment of non-nosocomial infections. BMC Compelmentary and Alternative Medicine. 2006, 6: 2-10.1186/1472-6882-6-2. [http://www.biomedcentral.com/1472-6882/6/2]
Kloucek P, Polesny Z, Svobodova B, Vlkova E, Kokoska L: Antibacterial screening of some Peruvian medicinal plants used in Callería District. Journal of Ethnopharmacology. 2005, 99: 309-312. 10.1016/j.jep.2005.01.062.
Sandhya B, Thomas S, Isabel W, Shenbagarathai R: Ethnomedicinal plants used by the valaiyan community of Piranmalai hills (Reserved forest), Tamil Nadu, India – A pilot study. African journal of Traditional Complementary and Alternative Medicines. 2006, 3: 101-114.
Ayyanar M, Ignacimuthu S: Traditional Knowledge of Kani tribals in Kouthalai of Tirunelveli hills, Tamil Nadu, India. Journal of Ethnopharmacology. 2005, 102: 246-255. 10.1016/j.jep.2005.06.020.
Rajan S, Sethuraman M, Mukherjee PK: Ethnobiology of the Nilgiri Hills, India. Phytotherapy – Research. 2002, 16: 98-116. 10.1002/ptr.1098.
Natarajan B, Paulsen BS, Pushpangadan P: An Ethnopharmacological Study from the Coimbatore District, Tamil Nadu, India: Traditional Knowledge Compared With Modern Biological Science. Pharmaceutical Biology. 1999, 37: 378-390.
Ignacimuthu S, Sankarasivaraman K, Kesavan L: Medico-ethnobotanical survey among Kanikar tribals of Mundanthurai Sanctuary. Fitoterapia. 1998, 69: 409-414.
Erdogrul OT: Antibacterial activities of some plant extracts used in folk medicine. Pharmaceutical Biology. 2002, 40: 269-273.
Bauer RW, Kirby MDK, Sherris JC, Turck M: Antibiotic susceptibility testing by standard single disc diffusion method. American Journal of Clinical Pathology. 1966, 45: 493-496.
Buwa LV, Staden JV: Antibacterial and antifungal activity of traditional medicinal plants used against venereal diseases in South Africa. Journal of Ethnopharmacology. 2006, 103: 139-142. 10.1016/j.jep.2005.09.020.
Valsaraj R, Pushpangadan P, Smitt UW, Adsersen A, Nyman U: Antimicrobial screening of selected medicinal plants from India. Journal of Ethnopharmacology. 1997, 58: 75-83. 10.1016/S0378-8741(97)00085-8.
Srinivasan D, Nathan S, Suresh T, Lakshmanaperumalsamy P: Antimicrobial activity of certain Indian medicinal plants used in folkloric medicine. Journal of Ethnopharmacology. 2001, 74: 217-20. 10.1016/S0378-8741(00)00345-7.
Samy RP, Ignacimuthu S: Antibacterial activity of some folklore medicinal plants used by tribals in Western Ghats of India. Journal of Ethnopharmacology. 2000, 69: 63-71. 10.1016/S0378-8741(98)00156-1.
Perumalsamy R, Ignacimuthu S, Raja DP: Preliminary screening of ethnomedicinal plants from India. Journal of Ethnopharmacology. 1999, 66: 235-40. 10.1016/S0378-8741(99)00038-0.
Parekh J, Jadeja S, Chanda S: Efficacy of Aqueous and Methanol Extracts of Some Medicinal Plants for Potential Antibacterial Activity. Turkish Journal of Biology. 2005, 29: 203-210.
Voravuthikunchai S, Lortheeranuwat A, Jeeju W, Sririrak T, Phongpaichit S, Supawita T: Effective medicinal plants against Enterohaemorrhagic Escherichia coli O157: H7. Journal of Ethnopharmacology. 2004, 94: 49-54. 10.1016/j.jep.2004.03.036.
Prashanth D, Asha MK, Amit A: Antibacterial activity of Punica granatum. Fitoterapia. 2001, 72: 171-173. 10.1016/S0367-326X(00)00270-7.
Rajakaruna N, Harris CS, Towers GHN: Antimicrobial activity of plants from Serpentine outcrops in Sri Lanka. Pharmaceutical Biology. 2002, 40: 235-244.
Saxena VK, Sharma RN: Antimicrobial activity of the essential oil of Toddalia asiatica. Fitoterapia. 1999, 70: 64-66. 10.1016/S0367-326X(98)00002-1.
Khan MR, Kihara M, Omoloso AD: Antimicrobial activity of Cassia alata. Fitoterapia. 2001, 72: 561-564. 10.1016/S0367-326X(00)00335-X.
Somchit MN, Reezal I, Nur IE, Mutalib AR: In vitro antimicrobial activity of ethanol and water extracts of Cassia alata. Journal of Ethnopharmacology. 2003, 84: 1-4. 10.1016/S0378-8741(02)00146-0.
Yoha Narasimhan SN: Medicinal plants of India. Tamil Nadu Cyber Media Bangalore India. 2003, II:
Nadkarni KM: Indian Materia Medica. Popular Prakashan Bombay India. 1976
Rastogi RP, Mehrotra BN: Compendium of Indian Medicinal Plants. Central Drug Research Institute Lucknow and National Institute of Science Communication New Delhi India. 1990, I-V: –1994
The pre-publication history for this paper can be accessed here:http://www.biomedcentral.com/1472-6882/6/35/prepub
The authors are thankful to the tribal people for sharing their knowledge on plants.
Entomology Research Institute, Loyola College, Chennai, 600 034, India
Veeramuthu Duraipandiyan, Muniappan Ayyanar & Savarimuthu Ignacimuthu
Veeramuthu Duraipandiyan
Muniappan Ayyanar
Savarimuthu Ignacimuthu
Correspondence to Savarimuthu Ignacimuthu.
VD has carried out the experimental part such as extraction, inoculum preparation and antimicrobial evaluation. MA collected the ethnomedicinal plants from the Palni hills of Tamil Nadu with the help of tribal practitioners. SI supervised the work, evaluated the results and corrected the manuscript for publication. All authors read and approved the final manuscript.
Duraipandiyan, V., Ayyanar, M. & Ignacimuthu, S. Antimicrobial activity of some ethnomedicinal plants used by Paliyar tribe from Tamil Nadu, India. BMC Complement Altern Med 6, 35 (2006). https://doi.org/10.1186/1472-6882-6-35
Antimicrobial Activity
Methanol Extract
Hexane Extract
Mueller Hinton Agar