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Antibacterial Activity of the Crude Extract of a Gastropod Cellana radiata (Born, 1778) from the Visakhapatnam Coast, Andhra Pradesh, India | Duddu | International Journal of Marine Science

Research Report

Antibacterial Activity of the Crude Extract of a Gastropod Cellana radiata (Born, 1778) from the Visakhapatnam Coast, Andhra Pradesh, India  

Sunil Kumar Duddu , Teja Gurugubelli , Krishna Geetha Gandham , Amarnath Dogiparti
Department of Marine Living Resources, Andhra University, Visakhapatnam, India
Author    Correspondence author
International Journal of Marine Science, 2017, Vol. 7, No. 14   doi: 10.5376/ijms.2017.07.0014
Received: 23 Mar., 2017    Accepted: 26 Apr., 2017    Published: 08 May, 2017
© 2017 BioPublisher Publishing Platform
This is an open access article published under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
Preferred citation for this article:

Duddu S.K., Gurugubelli T., Gandham K.G., and Dogiparti A., 2017, Antibacterial activity of the crude extract of a gastropod Cellana radiata (Born, 1778) from the Visakhapatnam coast, Andhra Pradesh, India, International Journal of Marine Science, 7(14): 125-129 (doi: 10.5376/ijms.2017.07.0014)

Abstract

The present study was carried out to evaluate the antibacterial activity of crude extracts of Cellana radiata collected from Visakhapatnam coast, Andhra Pradesh. The whole animal extracts from methanol and ethyl acetate were obtained by both dry and wet methods and tested for their activity against three pathogenic bacteria i.e. Escherichia coli, Staphylococcus aureus and Vibrio harveyi. The extracts obtained from methanol (wet), ethyl acetate (dry) and ethyl acetate (wet) showed inhibition zones of 12 mm, 13 mm and 18 mm respectively against V. harveyi.

Keywords
Cellana radiata; Antibacterial activity; Visakhapatnam coast

Introduction

Molluscs are soft bodied animals and play a major role among marine invertebrate fauna. The marine organisms are exposed to complex ecosystem, tolerating wide range of fluctuations and also defence more than terrestrial animals. So, apart from their use as food, they are also useful as bio indicators and for extracting drugs against several diseases (Whittle, 2009; Tosti and Gallo, 2012). Some marine invertebrates which lack physical defence will be producing certain toxic substances against to the predators for self protection and it contains some compounds will show anti-inflammatory, antimicrobial, cytotoxic and anti-tumour properties (Burkholder and Burkholder, 1958; McCaffrey and Endean, 1985; Koh, 1997; Anand and Edward, 2001; Mayer and Gustafson, 2006). The first attempt was carried out on marine organisms for their antimicrobial activity in 1950’s. Since then large number of marine organisms from a wide range of phyla have been screened for their antimicrobial activity (Shaw et al., 1976). Among the invertebrates, the molluscs are a good source for bio-medically important products (Shenoy, 1998). Many of the molluscs have chemical defence mechanisms and produce secondary metabolites which possess antimicrobial activities (Benkendorff et al., 2001). The bioactive compounds extracted from molluscs were exhibiting antitumor, anti leukemic, antibacterial and antiviral activity (Rajaganapathi et al., 2002). In recent years, the human pathogenic organisms were developing resistance to commercial drugs (Elumalai et al., 2011). The bioactive compounds which are derived naturally have no adverse effect and also low cost than the synthetic drug productions (Grabley and Thiericke, 1999). Some marine gastropods and bivalves have been of great interest to natural products chemists, yielding a diversity of chemical classes and several drug leads currently in clinical trials (Giftson and Patterson, 2014).

 

Cellana radiata is an intertidal rocky shore fouler and showing homing behavior unlike like the other temperate limpets. (Balaparameswara and Ganapati, 1971). It has shown antitumor property in MTT assay (Krishnamoorthi and Yogamoorthi, 2013). So, the present attempt has been made for screening of anti-bacterial activity from natural sources. Because, currently most of the bacteria are rapidly increasing their resistance against the antibiotics (Penesyan et al., 2015) by reason of they are producing the various enzymes which modify or inactivate the antibiotics (Koch, 1981).

 

1 Materials and Methods

Cellana radiate (Figure 1) were collected from inter tidal rocky shore area at VUDA Park (17°43’20.09”N and 83°20’28.23”E) of Visakhapatnam coast, Andhra Pradesh. The samples were collected and brought to the laboratory immediately and cleaned with water to remove sand and other dust particles, if any. The shells were broken with the help of scalps and small hammer and the meat is collected in a clean container. The entire meat collected was divided into two equal parts for extracting by both dry and wet methods.

 

 
Figure 1 Cellana radiate dorsal and ventral views

 

1.1 Wet method

The whole animal is removed and cut into small pieces and ground in a pestle-mortar until the mixer becomes as homogenous mass. The whole content was soaked in 1:5 ratio (w/v) of methanol: chloroform (9:1) and ethyl acetate: chloroform (9:1) for 72 hours (Sunil et al., 2016). After soaking the samples were concentrated to 100 mg/ml in methanol and ethyl acetate by rota-evaporator.

 

1.2 Dry method

The known amount of meat was kept in hot air oven at 55°C for 72 hours. The dried samples were powdered and dissolved in 1:5 ratio (w/v) of methanol: chloroform (9:1) and ethyl acetate: chloroform (9:1) for 72 hours (Sunil et al., 2016). After soaking the samples they were concentrated to 100 mg/ml in methanol and ethyl acetate.

 

1.3 Preparation of microbial cultures

Three pathogens namely, Escherichia coli, Staphylococcus aureus, Vibrio harveyi were selected to know the antagonistic activity. 2.5 gms of nutrient broth (HIMEDIA) was dissolved in 100 ml of distilled water and autoclaved at 121°C, 15 lbs. for 15 min. The broth was cooled down to room temperature and bacterial cultures were inoculated in sterilized nutrient broth and were incubated at 37°C for 24 hrs.

 

1.4 Antagonistic activity

The extracts were tested for their anti-bacterial activity by agar well diffusion method (Perez et al., 1990). The 18 hrs. young culture broths were taken and the pathogenic strains were spread evenly over sterile Mueller Hinton Agar plates. The 6 mm diameter wells were prepared on agar plates by using sterile cork borer. The wells were filled with 100 µL extract and controls were arranged. The plates were incubated at 37°C for 24 hours. Antibacterial activities were evaluated by measuring the zone of inhibition showed in millimeters (mm).

 

2 Results

The two different extracts of Cellana radiata were screened for their antibacterial activity against three different pathogenic bacteria. The methanolic dry extracts did not show any inhibitory zones but the wet extracts have shown antibacterial activity against Vibrio harveyi (Figure 2; Figure 3; Table 1). The ethyl acetate extracts (from both wet and dry methods) have shown inhibitory zones against Vibrio harveyi (Figure 2; Figure 3; Table 1).

 

 
Figure 2 Agar well diffusion method against V. harveyi, S. aureus and E.coli

 

 

Figure 3 Antibacterial activity of crude extracts of Cellana radiata against three different pathogens

 

 

Table 1 Antibacterial activity against three different Pathogenic Bacteria

 

3 Discussion

From ancient times the molluscs are useful organisms which support the coastal communities as food and also economically. These are used in medical applications and as bio-indicators in recent trends (Asta, 2011). The molluscs, particularly gastropods were showing antibacterial properties. In the present study three different pathogenic bacteria viz. Escherichia coli (human), Staphylococcus aureus (Fish) and Vibrio harveyi (Shrimp) were tested to know the antibacterial activity against them to methanolic and ethyl acetate crude extracts. The wet extract of methanol has shown antibacterial activity (12 mm) against V. harveyi. The ethyl acetate extracts by both dry and wet methods have shown antibacterial activity against V. harveyi, the dry extract 13 mm and wet extract 18 mm.

 

V. harveyi showing severe effect on commercially important resources like fish, shrimp and also pearl oysters and causes mass mortality. The pearl oyster Pinctada maxima died and damaged heavily by V. harveyi infection (Lester, 1989). Some gastropods have shown antibacterial activity against V. harveyi and in near future these metabolites may be helpful to aquaculture industries. The Donax faba shell extract has shown inhibitory zone of 3 mm to V. harveyi from ethanol extract (Giftson and Patterson, 2014). The extracts of crude peptides shown zone of inhibition to V. harveyi from pelecypods, Crassostrea madrasensis, Saccostrea cucullata, Meretrix meretrix, Perna viridis, Polymesodaerosa and from gastropods, Ficus gracilis, Hemifusus tuba, Bursa rana, Xenophora mekranensis and from cephalopods, Sepeilla inermis, Octopus dolfusii, Vampyroteuthis infernalis, Loligosp, Ancirtroclieinix leriieii (Naveen et al., 2014). The ethanolic extract of Chicerous ramous shown inhibitory zone of 6 mm to V. harveyi (Giftson et al., 2015).

 

4 Conclusion

The present study has shown that Cellana radiata is having antimicrobial properties against shrimp pathogen i.e., Vibrio harveyi.

 

Authors contributions

This work was carried out in all collaboration between authors. All authors read and approved the final manuscript.

 

Acknowledgments

Authors are grateful to the Department of Science and technology (DST), New Delhi, Govt. of India for the financial support in the form of INSPIRE Fellowship to carry out this work.

 

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