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.
1
Department of Zoology, Acharya Nagarjuna University, Guntur, A.P., India
2
Department of Marine Living Resources, Andhra University, Visakhapatnam, A.P., India
Corresponding Author-E-mail: darwin.chatla@gmail.com

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

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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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International Journal of Current Innovations in Advanced Research ISSN: 2636-6282
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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