Research Journal of Recent Sciences _________________________________________________ ISSN 2277-2502 Vol. 3(ISC-2013), 275-278 (2014) Res. J. Recent. Sci. International Science Congress Association 275 Antimicrobial Activity of root extract of Phyllanthus fraternus Webster: An Ethnomedicinal plant Kavit Mehta, Patel B.N. and Jain B.K.1,2Mehsana Urban Institute of Science, Ganpat University, Kherwa, Dist. Mehsana, North Gujarat, INDIA M.G. Science Institute, Dadasaheb Mavlankar Campus, Navarangpura, Ahmedabad, Gujarat, INDIAAvailable online at: www.isca.in, www.isca.me Received 28th October 2013, revised 22nd December 2013, accepted 7th February 2014 AbstractPhyllanthus fraternus Webster, a pan tropical weed originated from western India, belongs to family Euphorbiaceae and commonly known as “Bhumyamlaki”. It is a medicinally very useful plant species which is frequently used by tribal of Gujarat to cure certain diseases like asthma, cough, diarrhea, diabetes, skin diseases and scabies. Current study is targeted the on the activity of antimicrobial metabolites from the root. Root were dried and extracted by methanol. This root extract ofPhyllanthus fraternus was screened against eleven clinically important bacterial strains and three antifungal strainby disc diffusion method. Result showed maximum antibacterial activities against P.aeruginosa with zone of inhibition of 25 mm followed by S. typhiB with a zone of inhibition of 11 mm and minimum against S. typhi Awith zone of inhibition of 10 mm. Similarly the maximum antifungal activity was noted against A. niger (10 mm). Absolute methanol was used as control during the experiments. This indicates that antimicrobial activities may be due to presence of secondary metabolites. Hence, the plant can be used to discover bioactive natural products that may serve as leads in the development of new pharmaceuticals research activities. As the antimicrobial work has been not carried out so far on Phyllanthus fraternus, Keyword:Phyllanthys fraternus, root, extract, antimicrobial activities. Introduction The genus Phyllanthus belongs to the family Euphorbiaceae. The family has about 750 -1200 genera. The majority of genera are pan tropical weeds and They are distributed in all tropical and subtropical regions on the earth. Phyllanthus fraturnus Webster is one of the valuable medicinal plants and it has been used for the treatment of various ailments such as flu, dropsy, diabetes and jaundice. India has rich heritage of using medicinal plants as traditional medicines. Although hundreds of plant species have been tested for antimicrobial properties. there is no report on antimicrobial properties of various plant parts like leaves, fruits and roots of Phyllanthus fraternus Webster against the bacterial and fungal microorganisms. The world Health Organization estimates that plant extracts or their active constituents are used as folk medicine in traditional therapies among 80% of the world population. Over 50% of all modern clinical drugs are of natural product origin. The potential of higher plants as a source for new drugs is still largely unexplored. Among the estimated 250,000-500,000 plant species, only a small percentage has been investigated phytochemically and the fraction is submitted to biological or pharmacological screening. The present study is aimed to carry out antimicrobial activity of the root extract against eleven clinically important bacterial strains and three fungal strain by using agar disc diffusion method. Material and methodsPlant material (Figure-1): The roots of Phyllanthus fraternus Webster were collected from Botanical garden of S. K. Pharmaceutical college of education and research; Ganpat University; Ganpat vidyanagar; Kherwa, North Gujarat. The plant was identified by using the Flora of Gujarat by G. L. Shah (1978). The plant is annual having the length of 20-50 cm. It produces pale greenish -yellow colored axillary flowers. The seeds are trigonous. This plant is used by tribals of Gujarat to cure certain diseases like asthama, cough, diarrohea and scabies. Extraction of plant root: The roots of the plants were washed under running tap water and than by distilled water to remove soil particles and dirt. The roots were dried at room temperature in the laboratory. The dried roots were crushed to make powder using grinding machine. Root powder was stored at 4C in tight air container bottle. The extraction of roots was done by methanol using Soxhlet apparatus. The solvent was evaporated by using rotary evaporator at 80C temperature and the extract obtained was cooled and dried under vacuum. Research Journal of Recent Sciences ______________________________________________________________ ISSN 2277-2502Vol. 3(ISC-2013), 275-278 (2014) Res. J. Recent. Sci. International Science Congress Association 276 Figure-1 Root of Phyllanthus fraternus plant Bacterial and fungal strains used: Microbial strainsEscherichia coli, Bacillus megaterium, Bacillus cereus, Bacillus subtilis, Cornynebacterium glutamicum, Staphylococcus aureus, Salmonella typhi, Salmonella typhi A, Salmonella typhi B, Pseudomonas aeruginosa, Proteus vulgaris, and fungal strain Aspergillus niger, penicillium chryosogenus and candida were obtained from MUIS ,Ganpat University, Mehsana and M. G. Science Institute , Ahmedabad (Gujarat). Antimicrobial assay for disc diffusion method: Antimicrobial assay of solvent extracts were performed by Disc diffusion method. For bacteria Nutrient broth and Fungal Potato dextrose broth, pH was adjusted to 7.2 and 7.0. respectively. Suspensions of the different strains of the bacteria were added to sterile Nutrient broth and that of fungal strains were added to sterile Potato dextrose broth in separate petridishes . Bacterial and fungal strains were swabbed separately on the Nutrient agar plate and Potato dextrose agar plate respectively aseptically. The sterile disc, 5mm in diameter, is saturated with 10µL to 50 µL /10ml test culture of methanolic extracts separately. The discs with absolute methanol is used as control. The bacterial plates were incubated at 37C for 24 hr while fungal plates were incubated at 28C for 24-48 hr . The sterile impregnated discs with root extract were placed on the agar surface with flamed forceps and gently pressed down to ensure complete contact of the disc with the agar surface. After the incubation, the size of the inhibition zone was measured. Antimicrobial activities were determined by measuring the diameter of the zone of inhibition surrounding microbial growth. For each strain, control performed that comprised pure solvent instead of the root extract. Results and Discussion In the present investigation antimicrobial activities of methanol extracts of root of Phyllanthus fraternus, in terms of zone of inhibition was presented in table-1. Table-1 Zone of inhibitory activity (in millimeter) of methanol root extracts against microorganisms Sl. no. OrganismZone of inhibition (mm) 10 20 30 40 50 1. Escherichia coli10 No No No No 2. Bacillus megaterium 16 No 10 11 No 3. Bacillus cereus 11 10 14 11 13 4. Bacillus subtilis 19 12 10 10 12 5. Cornynebacterium glutamicum 11 10 12 10 10 6. Stephylococcus aureus 17 10 12 13 10 7. Salmonella typhi 21 12 11 10 13 8. Salmonella typhi A 11 12 10 13 11 9. Salmonella typhi B 11 11 11 11 10 10. Pseudomonas aeruginosa 25 20 23 22 20 11. Proteus vulgaris 19 10 11 10 11 12. Aspergillus niger No No No 10 No 13. P.chrysogenus 11 12 13 11 11 14. Candida 10 11 11 11 11 The results obtained in the present study revealed that tested root extracts of Phyllanthus fraternus possess potential antibacterial activity against all 11 bacterial strains and 3 fungal strain. When tested by disc diffusion method , the methanol root extract showed significant activity where inhibition zone ranged between 10 mm and 25 mm against all the microorganisms. The highest antibacterial activity of 25 mm in Pseudomonas aeroginosa and least activity of 11 mm in Salmonella typhi A were recorded in methanol extract Similarly, Bacillus megaterium and Salmonella typhi revealed maximum activity of 16 mm and 21 mm respectively in methanol extract. It is also examined from the present data that methanolic root extract shows significant higher rate of sensitivity against fungal strain Aspergillus niger and Penicillium chryosogenus. Discussion: The medicinal properties of plants are due to the presence of different complex chemical substances which are exclusively accumulated in different parts of the plants and produce marked healing action on human body6,7. The most important of these agents are alkaloids, flavanoids and tannins.8,9. These compounds have been associated with antimicrobial effects in various studies using plant extracts10Antibacterial compounds pf plant origin have enormous therapeutical potential as they can serve the purpose without side effects that are often associated with synthetic antimicrobials11,12. In the present work the methanol root extracts of Phyllanthus fraturnus were used for antimicrobial activity against 11 bacterial and 3 fungal strains.The test organisms used in the study are associated with various forms of human infections. Apart from Salmonella typhi Research Journal of Recent Sciences ______________________________________________________________ ISSN 2277-2502Vol. 3(ISC-2013), 275-278 (2014) Res. J. Recent. Sci. International Science Congress Association 277 infection, Salmonella paratyphi A and B also widely persist in Indian population13. In the present study methanolic root extracts of Phyllanthus fraternus showed considerable inhibitory activity against both enteric isolates of Salmonella typhi and Salmonella paratyphi12 have also shown that reports of such similar work on enteric Salmonella paratyphi (A and B). Conclusion From the results of present investigation it is reported that root extract prepared in methanol shows significant higher antimicrobial activity against all test microorganisms revealing inhibition zones between 10 mm and 25 mm The present study ascertains the value of solvents used in the drug preparation, which could be of considerable interest to the development of new drugs. The fact that the root extract of Phyllanthus fraternus was active against all the tested microorganisms is also an indication that it can be a source of very potent antibiotic substances that can be used against drug resistant microorganisms. As there is no report on antimicrobial activity of root extract of Phyllanthus fraternus, further studies are needed to isolate and characterize the bioactive principles to develop new antimicrobial drugs. Figure-2 Zone of inhibition A: Methanol root extract and B: Control against P.aeroginosa Figure-3 Zone of inhibition A & B: Methanol root extract against c.glutamicum and B.subtillusReferences 1.Prasannabalaji N., Muralitharan G., Sivanandan R.N., Kumaran S. and Pugazhvendan S.R., Antibacterial activities of some Indian traditional plant extracts, Asian Pacific J. tropical Disease, S291-S295, (2012)2.Anonynous, World Health Organisation summary of WHO guidelines for the assessment of herbal mediocines,Gram., 28, 13-14 (1993) 3.Baker J.T., Borris R.P. and Carte B., Natural product drug discovery and development. New perspective on international collaboration, J. Nat. Prod., 58, 1325-1327 (1995) 4.Pavithra P.S., Janani V.S., Charumathi K.H., Indumathy R., Sirisha Potala and Rama S. Verma; Antibacterial activity of plants used in herbal medicine, International Journal of pharmacy, 24-27 (2010) 5.Parekh J. and Chanda S., In vitro screening of antibacterial activity of aqueous and alcoholic extracts of various Indian plant species against selected pathogens from Enteriobacteriaceac, Afr. J. Microbiol. Res.,1(6), 92-99, (2007) Research Journal of Recent Sciences ______________________________________________________________ ISSN 2277-2502Vol. 3(ISC-2013), 275-278 (2014) Res. J. Recent. Sci. International Science Congress Association 278 6.Bashir Sajid, Erum Alia, Rizwana Kausar, Saleem Uzma, Umme Ruqia-Tulain and Alamgeer, Antimicrobacterial activity of some ethno-medicinal plants used in Pakistan, Res in Pharm, 2(1), 42-45 (2012)7.Manal Ismail A., Musa H.A., Yousif K.H. and Sabahelkhier M.K., Investigation Antibacterial Activity Extraction from two Medicinal Plants Available in Sudan, Res. J. Recent Sci.,2(10), 6-9 (2013)8.Edeoga H.O., Okwu D.E. and Mbaebie B.O., Phytochemical constituents of some Nigerian medicinal plants. Afr. J. Biotech, 4, 685-688 (2005)9.Mehta Kavit, Patel B.N. and Jain B.K., Phytochemical analysis of leaf extract of Phyllanthus fraternus, Res. J. Recent Sci.,2(1), 12-15 (2013) 10.Abo K.A., Ogunleye V.O. and Ashidi J.S., Antimicrobial potential of Spondias mombin, Croton zambesicus and Zygotritonia crocea, Phytotherapy Research, 13, 494-497 (1999) 11.Jain Monica, Rajput Ritika and Mishra Anamika, Enhancement of secondary Metabolite Biosynthesis in Bacopa monnieri: An in vitro Study, Res. J. Recent Sci., 2(1), 13-16 (2013)12.Minuta L., Journal of public health and biological sciences,1(3), 74-78 (2012) 13.Javed Ahmad, Iffat Khan, Ashfaq Ahmad and kaushar Imam, In Vitro Antioxidant and Antimicrobial Activity of Methanolic root Extracts of Hyptis suaveolens, Res. J. Recent Sci.,2(1), 41-46 (2013)14.Lubna Tahir and Naeem Khan, Antibacterial potential of crude leaf ,fruit and flower extracts of Tagetes, Afr.J.Biotech,1(2),(2009) 15.More S.M., Shinde V.A., Khan Saiqua, Girde A.V. and Pawar V.N., Antimicrobial Activity of Phospholipid Compound Produced by Acidophilic Bacillus subtilis Isolated from Lonar Lake, Buldhana, India Res. J. Recent Sci.,1(11), 22-26 (2012)16.Sujatha S. Complementary and alternative therapies in palliative care:A transtition from modern medicine to traditional medicine in india, J Cancer Pain Symptom Palliation,1, 25-9 (2005)17.Sukanya S.L., Sudisha J., Hariprasad P., Niranjana S.R., Prakash H.S. and Fathima S.K., Antimicrobial activity of leaf extracts of Indian medicinal plants against clinical and phytopathogenic bacteria, Afr. J. Biotech.,8(23), 6677-6682 (2009) 18.Uzun Y., Keles A., Imali A., Ogun E and Kaya A., Antimicrobial activity of Urtica dioica L. and Rheum ribesL., Bioscience Res. Bulle, 18, 43-50 (2002)19.Vashist H. and Jindal A., Antimicrobial activities of medicinal plants- review. Inter, J. Res. Pharm. Biomed. Sci., 3(1), 222-224 (2012)20.Veeramuthu D., Muniappan A. and Savarimuthu I., Antimicrobial activity of some ethno medicinal plants used by Paliyar tribe from Tamil nadu, India, BMC Complementary and Alternative Medicine, 6, 35, (2006)