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Antimicrobial Activity of Extracts of Adenanthera Pavonina and Mussaenda Philippica Against Isolated Bacteria and Fungi

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Antimicrobial Activity of Extracts of Adenanthera Pavonina and Mussaenda Philippica Against Isolated Bacteria and Fungi Research Article ISSN 2250-0480 VOL 5/ ISSUE 4/OCT 2015 ANTIMICROBIAL ACTIVITY OF EXTRACTS OF ADENANTHERA PAVONINA AND MUSSAENDA PHILIPPICA AGAINST ISOLATED BACTERIA AND FUNGI. RENILDA SOPHY A.J 1, ALBIN T. FLEMING *1, B.S.M. RONALD.2, K. GOWRI SHANKAR1, R. VIDHYA1, V. RAJAGOPALAN2, A. SHEEBA2 AND R DURGALAKSHMI2. 1PG & Research Department of Advanced Zoology and Biotechnology, Loyola College, Chennai-600034, India 2Department of Veterinary Microbiology, Veterinary College and Research Institute, Orathanadu – 614625, Thanjavur District, Tamilnadu. ABSTRACT Long before mankind discovered the existence of microbes, the idea of using certain plants for medicinal purpose because of their healing potential due to antimicrobial principles, was well accepted. In the past few decades, the search for new anti-infection agents has occupied many research groups in the field of ethnopharmacology. In the present paper, we analyze two medicinal plants, both as potential antimicrobial crude drugs as well as a source for natural compounds that can act as new anti-infection agents. The antibacterial and antifungal activity of three solvent extracts of Adenanthera pavonina and Mussaenda philippica on microbes isolated from dairy cattle rearing unit was studied. The results revealed that these extracts have antibacterial activity against Salmonella enteritidis, Klebsiella pneumoniae, Bacillus subtilis, Staphylococcus aureus, Escherichia coli and Pseudomonas aeruginosa and hence these crude extracts can be used as anti-infection agents in dairy cattle rearing unit to avoid infections especially to calves which are prone to infections at birth. KEYWORDS: Antibacterial, antifungal, isolated strains, diffusion assay, phytochemicals, disinfectant. INTRODUCTION source of new antibacterial drugs since they are considered as time-tested and comparatively safe 9 Nature has gifted us with a very rich botanical both for human use and for environment . But, wealth and a large number of diverse types of resistance to antimicrobial agents is emerging in a plants grow in different parts of the world. Higher wide variety of pathogens and multiple drug plants produce hundreds to thousands of diverse resistance is becoming common in diverse 10, 11,12 chemical compounds with different biological organisms . Most of the farmers express activities1. There are several reports in the literature desire to learn more about the proper use and regarding the antimicrobial activity of plant crude application of ethnoveterinary practices as these extracts and the bioassay-guided fractionation of were economically, socially and culturally more 13 them to yield active principles2, 3,4,5. These acceptable for marginalized communities . This antimicrobial compounds produced by plants are has necessitated a search for new antimicrobial active against plant and human pathogenic substances from plants. Adenanthera pavonina microorganisms6. From an estimated 250,000 belongs to the family Mimosaceae, commonly higher plants in the world only 5–15% have been known as Anai kundumani or Manjadi in Tamil, is studied for a potential therapeutic value7,8 . Hence an important medicinal plant from the ‘Indian the rest of the plants remain to be investigated. subcontinent’. This species is endemic to Southern Herbal medicines have received much attention as a China and India. Various parts of this plant have L - 21 Life Science Microbiology Research Article ISSN 2250-0480 VOL 5/ ISSUE 4/OCT 2015 also been used in traditional medicine for the from dairy units using standard isolation techniques treatment of asthma, boil, diarrhoea, gout, and identified based on Bergys manual on inflammations, rheumatism, tumour and ulcers, and Determinative Bacteriology25. The bacterial strains as a tonic14,15,16. Traditionally, the ground seed is include Salmonella enteritidis, Staphylococcus widely used for the treatment of various human aureus, Escherichia coli, Bacillus subtilis, ailments such as treatment of boils, inflammation, Klebsiella pneumoniae, Pseudomonas aeruginosa, blood disorders, arthritis, rheumatism, cholera, Enterobacter aerogenes, Proteus vulgaris, and paralysis, epilepsy, convulsion, spasm and Clostridium perfringens. All the strains were indigestion17.Mussaenda philippica of the family confirmed by cultural and biochemical Rubiaceae, commonly known as Flag Bush is found characteristics and maintained in slants for further throughout South East Asia. Due to the colourful use. nature of the plant it is usually used for landscaping. It is used as a medicinal plant in India Fungal Isolates and other South Asian countries. It is reported to Fungal isolates used in this study were Candida have analgesic18, anticonvulsant19 and antitumor albicans and Aspergillus niger. They were isolated activity20. The antibacterial activity of the leaves of from dairy cattle rearing unit and both were M. roxburghii has been studied21. Antimicrobial identified by staining and cultural characteristics in activity of the extracts of these two plants against sabouraud dextrose agar. very few microbial species have been studied22,23,24. In this study, the antimicrobial and antifungal Agar disc diffusion method activity of chloroform, ethyl acetate and ethanol Agar disc-diffusion method was carried to extracts of the leaves of M. philippica and A. determine the antimicrobial and antifungal activity. pavonina were assessed against bacteria and fungi Brain Heart Infusion agar (BHI) plates for bacteria isolated from dairy units. and Potato Dextrose agar plates for fungi were swabbed with 8 hour old - broth culture of the MATERIALS AND METHODS isolates. Stock solution of chloroform, ethyl acetate, and ethanol extracts of A.pavonina and Collection M.philippica were prepared at a concentration of Fresh leaves of Adenanthera pavonina and 100mg/ml in DMSO. About 20 µl of different Mussaenda philippica were collected from the concentrations (2 mg, 1mg and 0.5 mg ) of suburbs of Chennai, Tamil Nadu, India. The chloroform, ethyl acetate, and ethanol extracts of taxonomic authentication was done by Dr. P. A.pavonina and M.philippica and DMSO (20ul) Jayaraman, Plant Anatomy Research Centre, were added to the disc and allowed to dry at room Tambaram, Chennai. The voucher specimen temperature. Then the discs were placed over the number for A. pavonina is PARC/2014/2022 and swabbed plates. DMSO was used as negative M. philippica is PARC/2015/3131. The collected control and Enrofloxacin (30mcg/disc) for bacterial plants were washed with running tap water, again strains and Amphotericin B (20mcg/disc) for fungal washed with distilled water, air dried, homogenized strains were used as positive control. The plates 0 to a fine powder and stored in air-tight bottles . were incubated overnight at 37°C/28 C for bacterial / fungal isolates. The diameter of the inhibition Preparation of crude extracts. zone (mm) was measured. Dried plant material of both A. pavonina and M. Philippica were extracted with chloroform, ethyl RESULTS acetate and ethanol separately. They were kept on a rotary shaker for 9 days, changing the solvents once Chloroform, ethyl acetate, and ethanol extracts of in three days to elute maximum amount of extracts both M.philippica and A.pavonina showed from the plants. The supernatant was collected by antibacterial activity against few isolates but did not filtration using Whatman no.1 filter paper and the show any activity against fungal isolates. The filtrate was evaporated at room temperature to details of the microbial activity of A.pavonina are retain thermo labile compounds in the extract. The shown in Table1 and M.philippica is shown in o extract was stored at 4 C in airtight sterile vials for Table 2. In the present study, the extracts of both further studies. the plants exhibited promising anti-bacterial activity against Staphylococcus aureus and Bacterial Isolates Klebsialla pneumoniae. Pseudomonas aeruginosa Bacterial isolates used in this study were isolated was resistant to all the three extracts of A.pavonina L - 22 Life Science Microbiology Research Article ISSN 2250-0480 VOL 5/ ISSUE 4/OCT 2015 and ethyl acetate extract of M.philippica. Both the and Enterobacter aerogenes. The zones of plant extracts showed no activity against Proteus inhibition of all the extracts were comparatively vulgaris, Enterobacter aerogenes, and Clostridium lesser than the positive control Enrofloxacin (Table perfringens. Ethanol extract of M.philippica 1). DMSO which was used as a carrier (negative showed antibacterial activity against all the control) did not show any growth inhibition. bacterial species studied except Proteus vulgaris Table 1 Antimicrobial activity of A.pavonina. Test organism Diameter of inhibition zone(mm) Bacteria CL EA ET EX Concentration per disc 2mg 1mg 0.5mg 2mg 1mg 0.5mg 2mg 1mg 0.5mg 120mcg Staphylococcus aureus 12 12 12 18 14 10 19 18 12 40 Escherichia coli 14 13 11 10 10 10 8 - - 24 Klebsiella pneumoniae 8 8 8 10 8 8 12 10 8 13 Salmonella enteritidis - - - 10 8 8 8 8 8 35 Pseudomonas aeruginosa - - - - - - - - - 24 Bacillus subtilis 12 12 8 8 8 8 8 8 - 23 Proteus vulgaris - - - - - - - - - 26 Enterobacter aerogenes - - - - - - - - - 23 Clostridium perfringens - - - - - - - - - 40 Fungi AB Candida albicans - - - - - - - - - 10 Aspergillus niger - - - - - - - - - 10 CL-Chloroform, EA-Ethyl Acetate, ET-Ethanol, EX-Enrofloxacin, AB-Amphotericin B. Both the fungal strains Candida albicans and mm diameter. DMSO showed no activity against Asperagillus niger were resistant against extracts of the two
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