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Aeruginosa, Bacillus Subtilis, E.Coli, Klebsiella Pneumoniae, and Fungi Like Aspergillus Copyright ISSN: 2636-6282 Volume 1, Issue 1, May-2018 : Page No: 13-18 International Journal of Current Innovations in Advanced Research ANTIMICROBIAL ACTIVITY OF MARINE GASTRIPOD PURFURA BUFO FROM VISAKHAPATNAM, EAST COAST OF INDIA Darwin, Ch., Kavitha, K. and Suneetha, K. Department of Zoology, Acharya Nagarjuna University, Guntur Andhra Pradesh, India For Correspondence-E-mail: [email protected] Abstract: The present study was aimed to provide basic information regarding antimicrobial activity of marine gastropod i.e. Purfura bufo. The antimicrobial activity was estimated in extracts of body tissues of Purfura bufo against Pseudomonas aeruginosa, Bacillus subtilis, E.coli, Klebsiella pneumoniae, and fungi like Aspergillus flavus and Penicillium notatum. The maximum inhibition zone was observed in Klebsiella pneumonia (26 mm) and minimum in Penicillium notatum (11 mm). It is evident from the current study that significant reduction in growth of bacteria and fungi was observed with methanolic extracts only. Keywords: Purfura bufo, Antimicrobial activity, Inhibition zone, Methanol extract. Cite this article as: Darwin, Ch., Kavitha, K. and Suneetha, K. 2018. Antimicrobial activity of marine gastripod Purfura bufo from Visakhapatnam, East Coast of India. Int. J. Curr. Innov. Adv. Res., 1(1):13-18 Copyright: All rights reserved for the International Journal of Current Innovations in Advanced Research (IJCIAR). Copyright © 2018; Darwin, Ch., Kavitha, K. and Suneetha, K. (2018). Introduction Antimicrobial activity The first attempt to locate the antimicrobial activity in marine organisms was initiated around 1950’s (Berkholder and Burkholder, 1958) since this time many marine organisms from a wide range of phyla have been screened for antimicrobial activity. The marine compounds possess many biological activities like anti-helminthic, antibacterial, anticoagulant, antifungal, antimalarial, antiplatelet, antiprotozoal, anti-tuberculosis and antiviral properties (Mayer et al., 2007). A few drugs have already found a place in therapy like the antibiotic cephalotin from the marine fungus Cephalosporium acremonium and the anticancer agent arobinoside from the gorgonian Eunicella covaloni (Narahashi, 1988). Mollusks are a highly diverse group, in size, in anatomical structure, in behavior and in habitat (Haszprunar, 2001). The marine environment comprises of complex ecosystem with a plethora of organisms and many of these organisms are known to possess compounds as a common means of defense (Indap and Pathare, 1998). In the most of the publications concerning antimicrobial activity in mollusk, either single body compartment alone, like haemolymph and egg masses or extracts of whole bodies have been tested for activity (Haug et al ., 2003). The marine gastropods are expected to serve as source of novel chemicals for pharmacological and other applications. The bioactive compounds isolated from the gastropods are considered to have a role in the chemical defense of the animals against their predators (Avila et al., 2000). Rajaganapathi (2001) reported the antimicrobial activities of marine mollusks. Mollusks and soft corals are the prolific sources of structurally unique and diverse natural products and, they www.ijciar.com 13 ISSN: 2636-6282 Volume 1, Issue 1, May-2018 : Page No: 13-18 provide a large proportion of bioactive compounds with different biological activities (Ocky Karna and Agus, 2009). Acetone extract of the egg mass of Rapana rapiformis showed broad spectrum of antibacterial activity (Santhana Ramasamy and Murugan, 2005). The overall objective of the current study was capability of antibacterial and antifungal activity of methanol extracts of the body tissue of a gastropod snail Purpura bufo. Material and Methods Antimicrobial activity Solvent extraction The extraction method was followed by Chellaram et al., (2004). The powdered tissue of Purpura bufo was soaked in methanol for 3 days. Methanol was evaporated by using rota evaporator and the resultant residue was weighed and dissolved in Dimethyl sulfoxide (DMSO) at concentrations 0.5mg/ml, 1mg/ml DMSO. 1mg/ml Erythromycin was used as positive control. DMSO (1 ml) used as negative control. The antibacterial activity was carried out by using standard disc diffusion method (Sri Kumaran et al., (2011). The test bacterial strains collected for the present study are Pseudomonas aeruginosa, Bacillus subtilis, E.coli, Klebsiella pneumoniae, and fungi like Aspergillus flavus and Penicillium notatum respectively, sub cultured before use. The antibacterial activity was performed on nutrient agar plates (Figure 1). In vitro antifungal activity was determined against potato dextrose agar. 0.5 mm wells were prepared in the petriplates and the wells were loaded with erythromycin, DMSO and 0.5 mg, 1 mg concentrations of methanolic extracts. Plates were sealed with parafilm and incubated at 370C for 24 hrs. After incubation, zone of inhibition for extracts were measured in millimeters using Vernier calipers Figure 1. Preparing Plates for microbial analysis www.ijciar.com 14 ISSN: 2636-6282 Volume 1, Issue 1, May-2018 : Page No: 13-18 Results and Discussion Antimicrobial activity The antimicrobial activity of Purpura bufo extracts was assayed. The data revealed that significant reduction in growth of bacteria and fungi was observed with methanolic extracts. Methanol extracts showed broad spectrum inhibition zone against the bacteria Pseudomonas aeruginosa, Bacillus subtilis, E.coli, Klebsiella pneumoniae, in fungi Aspergillus flavus and Penicillium notatum were the species respectively. The inhibition zone was observed only with 0.5 mg where as 1 mg concentration it was absent. Overall the inhibition zone ranged from 11 mm to 15 mm with 0.5 mg concentrations of methanol extract. The lowest (11 mm) inhibition zone was recorded in Penicillium notatum whereas the highest was observed in Bacillus subtilis (15 mm). Erythromycin as a positive control showed 16 mm inhibition zone and DMSO did not show any inhibition zone. The maximum inhibition zone was observed in Klebsiella pneumonia (26 mm) and minimum in Penicillium notatum (11 mm) (Plate 1; Figures A-F) Plate 1. Antimicrobial activity-Inhibition zones Figure A. Bacillus subtilis Figure B. Escherichia coli Figure C. Pseudomonas aeruginosa Figure D. Klebsiella pneumoniae www.ijciar.com 15 ISSN: 2636-6282 Volume 1, Issue 1, May-2018 : Page No: 13-18 Figure E. Aspergillus flavus Figure F. Pencillium notatum In recent years, great attention has been paid towards the bioactivity of natural products because of their potential pharmacological utilization. Most of homeopathic medicines are either of plant or animal origin. Thousands of bioactive compounds identified in marine organisms reveal that sea creatures constitute a large reservoir for pharmacologically active drug recently reviewed (Mayer et al., 2011). Some of the molecules responsible for antimicrobial activities have been identified and characterized. Molluscs are widely used in world research institution for various studies, but recently they have been recognized as potential sources of antibacterial and antifungal substances. Antibacterial and antiviral activities have been previously described in the haemolymph of several molluscan species such as, sea hares, sea slug, oysters, and mussels (Maktoob and Ronald, 1997; Mitta et al., 1999; Nakamura et al., 1998; Zasloff, 2002; Olicard et al., 2005; Roch et al., 2008). In the present study the inhibition zone ranged from 11 mm to 15 mm with 0.5 mg concentrations of methanol extract. The microbial strains were more resistance at 1 mg concentration than 0.5 mg. Hence the results were absent at 1 mg concentration. The lowest (11 mm) inhibition zone was recorded in Penicillium notatum whereas the highest was observed in Bacillus subtilis (15 mm). Erythromycin as a positive control showed the maximum inhibition zone in Klebsiella pneumonia (26 mm) and minimum in Penicillium notatum (16 mm) and DMSO did not show any inhibition zone. The antibacterial activity of ethanol extracts of gastropod B. spirata and Turbo brunneus was observed maximum activities against E. coli, K. pneumoniae, P. vulgaris and S. typhi respectively (Anand et al., 1997). The crude methanol extracts of Cypraea errones exhibited higher antibacterial and antifungal activities (Anand and Patterson Edward, 2002). These results lend support to the present findings of the antimicrobial activity of Purpura bufo. The knowledge of the self-defense mechanism of molluscs is extremely limited compared to that of vertebrates and arthropods. Marine molluscs have been found to produce a great diversity of novel bioactive secondary metabolites and to be a potential source for new drug discovery. There has been a remarkable progress in the prevention; control and even eradication of infectious diseases with improved hygiene and development of antimicrobial compounds and vaccines. www.ijciar.com 16 ISSN: 2636-6282 Volume 1, Issue 1, May-2018 : Page No: 13-18 Conclusions The present study revealed that the species of Purpura bufo showed potential antimicrobial activity against pathogenic microorganisms. The vast variety of marine compounds and micro-organisms feature a wide array of new properties that could open avenues for innovative products from the seas, especially pharmaceuticals. However, efforts now need to focus on closing the gap between frontier research and clinical applications. Two promising areas in this sector are new antibiotics to tackle the antibiotic resistance crisis worldwide
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