Antimicrobial Activity of Marine Gastropod Purfura Bufo from Visakhapatnam, East Coast of India

Volume 1, Issue 1, May-2018: 12-17 International Journal of Current Innovations in Advanced Research ISSN: 2636-6282

Antimicrobial Activity of Marine Gastropod Purfura bufo from Visakhapatnam, East Coast of India

*Darwin Chatla1, Joseph Uday Ranjan, T2. and Ramesh Babu, K2.

1Department of Zoology, Acharya Nagarjuna University, Guntur, A.P., India 2Department of Marine Living Resources, Andhra University, Visakhapatnam, A.P., India Corresponding Author-E-mail: [email protected]

Abstract: The present study was aimed to investigate the 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, Escherichia 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. Keywords: Purfura bufo, Antimicrobial activity, Inhibition Zone, Methanol extract.

Citation: Darwin Chatla, Joseph Uday Ranjan, T. and Ramesh Babu, K. 2018. Antimicrobial activity of marine gastropod Purfura bufo from Visakhapatnam, East Coast of India. International Journal of Current Innovations in Advanced Research, 1(1): 12-17. Copyright: This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Copyright©2018; Darwin Chatla, Joseph Uday Ranjan, T. and Ramesh Babu, K.

Introduction The marine environment comprises of complex ecosystem with a plethora of organisms (Indap and Pathare, 1998). These organisms have been found possessing a vast array of new pharmaceutical compound with novel activities that will provide new drug leads to compact microbial pathogens currently developing resistance to conventional antibiotic therapies. The recent research on multi-drug-resistant bacteria proposes that animals living in unsanitary conditions have developed ways of protecting themselves against pathogenic microorganisms (Yoneyama and Katsumata, 2006). Among the marine phylum, the invertebrates especially sponges, ascidians, bryozoans and molluscans (Proksch et al., 2002) are the potential source for antimicrobial drugs (Bazes et al., 2009).

Marine molluscs are of one of the largest and enigmatic groups of marine organisms. These molluscans 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. With these positive progressions the marine molluscans have attracted the attention of biologists and chemists worldwide (Laith et al., 2017).

The majority of research on natural products from the phylum mollusca has been focused on primarily soft-bodies or shell-less mollusks, particularly nudibranches and opisthobranches (Karuso, 1987; Faulkner, 1992). However, some studies have also been reported biological www.ijciaropenaccess.com 12 Volume 1, Issue 1, May-2018: 12-17 International Journal of Current Innovations in Advanced Research ISSN: 2636-6282 activity from shelled molluscs (Kumar, 2011; Kumaran et al., 2011). 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). Keeping in view of the significance of marine mollusks, the present study was aimed to investigate the capability of antibacterial and antifungal activity of a marine gastropod Purpura bufo.

Material and Methods Live specimens of Purpura bufo were collected during low tide from Visakhapatnam, East coast of India (17º.41’18” N 83º13’07” E). The specimens were brought to the laboratory in ziploc bags and identified up to species level by standard methods (Fernando and Fernando, 2000). The extraction method was followed by Chellaram et al., (2004). In brief, 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 Kumaran et al., (2011). The test bacterial strains collected for the present study are Pseudomonas aeruginosa, Bacillus subtilis, Escherichia 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.

Results and Discussion 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, Escherichia coli, Klebsiella pneumoniae, in fungi Aspergillus flavus and Penicillium notatum were the species respectively.

The inhibition zone was observed only with 0.5 mg whereas 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).

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Plate 1. Antimicrobial activity-Inhibition zones

Figure A. Bacillus subtilis Figure B. Escherichia coli

Figure C. Pseudomonas aeruginosa Figure D. Klebsiella pneumoniae

Figure E. Aspergillus flavus Figure F. Penicillium notatum

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Mollusks are widely used in world research institution for various studies, but recently they have been recognized as potential sources of antibacterial and antifungal substances. The marine gastropods are expected to serve as source of novel chemicals for pharmacological and other applications (Darwin and Padmavathi, 2018). 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). 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; Rajaganapathi, 2001; Zasloff, 2002; Olicard et al., 2005; Roch et al., 2008). 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). Acetone extract of the egg mass of Rapana rapiformis showed broad spectrum of antibacterial activity (Santhana Ramasamy and Murugan, 2005).

Conclusions These results lend support to the present findings of the antimicrobial activity of Purpura bufo. 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. Studies of antimicrobial from natural resources would be the alternative to overcome the resistance problems. It is promising that the tested gastropods species synthesis novel antibiotics for bacterial infections and fungal infections. Mechanisms and compounds in mollusks may provide valuable information for new antibiotic discoveries and give new insights into bioactive compounds. Further investigations intending to purify these active compounds should be considered to clarify their chemical nature.

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