Screening of antibacterial drugs from marine gastropod Chicoreus ramosus (Linnaeus, 1758)

Full text

181
Document heading doi:10.12980/JCLM.1.20132013J5 襃 2013 by the Journal of Coastal Life Medicine. All rights reserved.
Screening of antibacterial drugs from marine gastropod Chicoreus ramosus
(Linnaeus, 1758)
Pasiyappazham Ramasamy1, Deepa Padmakumari Krishnan Thampi1, Gurusamy Chelladurai1, Natarajan Gautham1,
Sasirekhamani Mohanraj2, Jeyaraj Mohanraj1*
1Department of Advanced Zoology and Biotechnology, Kamaraj College, Manonmaniam Sundaranar University, Tuticorin-628003, Tamil Nadu,
India
2Centre of Advanced Studies in Botany, University of Madras, Gundy campus, Chennai - 600 025, Tamil Nadu, India
Journal of Coastal Life Medicine 2013; 1(3): 181-185
Journal of Coastal Life Medicine
journal homepage: www.jclmm.com
*Corresponding author: Dr. J. Mohanraj, Principal, Department of Advanced Zoology
and Biotechnology, Kamaraj College, Manonmaniam Sundaranar University, Tuticorin
-628003 Tamil Nadu, India.
Tel: +91461 2376790
E-mail: jmohanrajkathir@gmail.com
Foundation Project: Supported by Centre for Marine Living Resources and Ecology,
Cochin, Ministry of Earth Sciences, New Delhi, India (Grant No. MoES/10-MLR/01/12).
1. Introduction
Marine organisms comprise approximately a half of the total
biodiversity, thus offering infinite source to discover useful
therapeutics. In recent years, a significant number of novel
metabolites with potent pharmacological properties have been
PEE R REVIEW ABSTRACT
KEYWORDS
Antibacterial, Drugs, Marine gastropods, Chicoreus ramosus
Objective: To screen the antibacterial drugs from different solvent extracts of tissue and egg
of marine gastropods Chicoreus ramosus against clinically isolated human pathogenic bacteria.
Methods: Different solvent extracts of Chicoreus ramosus was screened for their activity against
Vibrio parehaemolyticus (J13300), Aeromonus hydrophilla (IDH1585), Salmonella typhi (C6953),
Salmonella paratyphi A (C6915), Vibrio cholerae (IDH5439) and Escherichia coli (H10407) using
standard well diffusion method and its minimum inhibitory concentration.
Results: The study revealed that the acetone and chloroform extract of both the tissues and
egg inhibited the growth of the tested pathogenic bacterial strains. The minimum inhibitory
concentration of both the extract ranged from 4 to 12 mg/mL.
Conclusions: These results suggest that marine gastropods tissue and egg extract contains
comparatively good antibacterial activity.
Peer reviewer
Dr. Jerald Wilson, M.Sc., Ph.D., Assistant
Professor, Department of Marine
Biology, Faculty of Marine Sciences,
King Abdulaziz University, P.O. Box:
80207 Jeddah 21589, Kingdom of Saudi
Arabia
E-mail: jeraldreef@gmail.com
Comments
Numerous pathogenic microorganisms
have developed their resistance against
commonly available antibiotics; hence
the need for developing new virulent
drugs against these harmful pathogens
becomes more important. Chemical
drugs may lead to adverse effects
and researchers now have focused on
pharmacologically active compounds
from natural sources. The present
study revealed that the acetone and
chloroform extract of both the tissues
and egg inhibited the growth of the
tested pathogenic bacterial strains.
The MIC of both extract ranged from
4 to 12 mg/mL. These results suggest
that marine gastropods tissue and egg
extract contains comparatively good
antibacterial activity.
Details on Page 184
Article history:
Received 16 Jul 2013
Received in revised form 22 Jul, 2nd revised form 28 Jul, 3rd revised form 5 Aug 2013
Accepted 13 Sep 2013
Available online 28 Oct 2013

Pasiyappazham Ramasamy et al./Journal of Coastal Life Medicine 2013; 1(3): 181-185
182
discovered from marine organisms. The phylum Mollusca,
which includes soft-bodied invertebrates, the second largest
phylum in the animal kingdom makes up a major part of the
world’s marine invertebrate fauna. Molluscs are the most
successful invertebrate group in occupying different habitats.
These are the commonest organisms of Indian sea beaches
and distributed all over the world in almost all types of
habitats. The molluscs have received a considerable amount of
research effort, reflecting both their ecological and economic
importance, and now gaining importance in deriving drugs[1].
The first attempt to the screening of antimicrobial activity
in marine organisms was initiated around 1950’s. Since this
time large numbers of marine organisms from a wide range
of phyla have been screened for antimicrobial activity[2].
From 1960’s to 1990’s approximately 300 bioactive marine
natural products were fields of patent. Approximately
6 500 bioactive compounds were isolated from marine
organisms. Among the invertebrates, the mollusks are highly
delicious seafood because of their nutritive value next to
fin fish and crustaceans. They are also a very good source
for biometrically important products[3]. Many classes of
molluscs with bioactive compounds exhibiting antitumor,
antileukemic, antibacterial and antiviral properties have
been reported worldwide[4]. Many marine molluscs have
evolved chemical defense mechanism for their eggs and
thus producing secondary metabolites which possess
antimicrobial activities[5].
The marine environment is a huge source for discovering
many antibacterial drugs. Apart from the food that is derived
from the marine environment, wide varieties of antibacterial
drugs are being isolated and characterized with great promise
for the treatment of human diseases. Studies on antibacterial
screening provide valuable information for new antibiotic
discoveries and give new insights into the extract of bioactive
compounds from marine molluscs. In the present investigation,
an attempt has been made to screen the antibacterial drugs
from marine molluscs Chicoreus ramosus (C. ramosus) against
clinically isolated human pathogenic bacteria.
2. Materials and methods
2.1. Sampling and preprocessing
The animals were collected from Tuticorin coast located at
southeast coast of Tamil Nadu in the Gulf of Mannar region,
which is situated between India and Sri Lanka (latitude 8°48’N
and longitude 78°09’E). The tissue and egg samples were
washed with tap water until the sand and mud were removed
from the shells. After that, the shells were broken using a
hammer to remove the soft body tissue. The removed tissues
were rinsed with sterile distilled water, cut into small pieces
and kept in Petri dishes and dried at a constant temperature
of 50 °C for 24 h in a hot air oven. The dried material was
powdered thoroughly for solvent extraction.
2.2. Solvent extraction
The powdered tissues and egg mass of C. ramosus were
extracted with eight different solvents like methanol, ethanol,
acetone, acetonitrile, dichloro methane (DCM), choloroform, ethyl
acetate and distilled water with the help of soxlet apparatus, and
the solvents were concentrated by rotary evaporator (VC100A
Lark Rota vapor® at 30 °C) with reduced pressure to give a dark
brown gummy mass. The resultant residues were stored at 4 °C
for further antibacterial screening.
2.3. Bacterial cultures
Six species of bacteria were used for screening the
antibacterial activity, including Vibrio parehaemolyticus
(V. parehaemolyticus) (J13300), Aeromonus hydrophilla (A.
hydrophilla) (IDH1585), Salmonella typhi (S. typhi) (C6953),
Salmonella paratyphi A (S. paratyphi A) (C6915), Vibrio cholerae
(V. cholerae) (IDH5439) and Escherichia coli (E. coli) (H10407). All
the bacterial strains were clinical isolates, obtained from the
Microbial Type Culture Collection & the Gene Bank, Institute of
microbial technology, Chandigarh, India.
2.4. Inoculums preparation for bacteria
Nutrient broth was prepared and sterilized in an autoclave
at 15 pounds pressure for 15 min. All the six bacterial strains
were individually inoculated in the sterilized nutrient broth
and incubated at 37 °C for 24 h. Mueller Hinton agar (Himedia)
was prepared, sterilized in an autoclave at 15 pounds pressure
for 15 min and poured into sterile Petri dishes and incubated
at 37 °C for 24 h. The 24 hours old bacterial broth cultures were
inoculated in the Petri dishes by using a sterile cotton swab.
2.5. Antibacterial screening
The antibacterial screening was investigated against six
human pathogenic bacteria by agar well diffusion method
followed Ramasamy et al[6]. Twenty four hours old nutrient
broth cultures of test bacteria was aseptically swabbed on
sterile Mueller Hinton agar plates. Wells of 5 mm in diameter
were made aseptically using well cutter, and 50 µL of eight
different solvent extracts of tissues and eggs were inoculated.
The stock solutions were prepared at a concentration of 20 mg/
mL. Positive control well containing 50 µL of tetracycline (1
mg/mL) and negative control containing 50 µL of appropriate
solvents were used. The result was calculated by measuring the
zone of inhibition in millimeters. For each concentration tested,
triplicates were maintained for the confirmation of activity.
2.6. Determination of the minimum inhibitory concentration
(MIC)
The solvent extracts of marine gastropods C. ramosus which
showed significant antibacterial activity were selected for the

Pasiyappazham Ramasamy et al./Journal of Coastal Life Medicine 2013; 1(3): 181-185 183
determination of MIC followed by the method of Ramasamy et
al[7]. A stock solution of 20 mg/mL was prepared and serially
diluted to obtain various ranges of concentrations between 4
mg/mL and 20 mg/mL. About 0.5 mL of each dilution of different
concentrations was transferred into a sterile test tube containing
2 mL of nutrient broth. To the test tubes, 0.5 mL of test organism
previously adjusted to a concentration of 105 cells/mL was
then introduced. A set of test tubes containing broth alone
was used as a control. All the test tubes and control were then
incubated at 37 °C for 24 h. After the period of incubation, the
tube containing the least concentration of extract showing no
visible sign of growth was taken as the minimum inhibitory
concentration.
2.7. Statistical analysis
Data on the inhibitory effects of solvent extracts of C. ramosus
was analyzed by One-way analysis of variance (ANOVA) using
SPSS-16 version software followed by Duncun’s multiple range
test and standard errors依SEM. P<0.05 were considered for
describing the significant levels.
3. Results
3.1. Antibacterial screening
The inhibition zone in different solvent tissue extracts of C.
ramosus against clinical isolate human pathogenic bacteria was
shown in Table 1. Among the various strains, the maximum zone
of inhibition [(26.00依1.53) mm] was recorded in acetone extracts
against S. paratyphi A, followed by (25.00依1.53) mm in E. coli
strain and the minimum zone of inhibition (7 mm) was noticed
in ethanol, petroleum ether, chloroform and water extracts. In
that similar way the inhibition zone in different solvent egg
extracts of C. ramosus against clinical isolate human pathogenic
bacteria was described in Table 1. Among the various strains,
the maximum zone of inhibition [(26.00依1.53) mm] was recorded
in acetone extracts against V. cholerae, followed by (25.00依
1.53) mm in E. coli strain and the minimum zone of inhibition
(7mm) was noticed in DCM and acetonitrile. The positive control
(tetracycline) was active against all the bacterial strains tested.
3.2. MIC of the active extract against the test organisms
MIC values of acetone tissue and egg extracts against bacterial
strains such as A. hydrophilla, S. typhi, S. paratyphi A, V.
cholerae and E. coli were reported as 12, 12, 8, 8, 4 mg/mL and
8, 8, 12, 4, 4 mg/mL respectively. Likewise in chloroform tissue
extracts against bacterial strains such as A. hydrophilla, S.
typhi, S. paratyphi A and V. cholerae was reported as 20, 20, 20
and 20 mg/mL respectively. The chloroform egg extracts against
bacterial strains such as A. hydrophilla, S. typhi, S. paratyphi A,
V. cholerae and E. coli were reported as 16, 12, 16, 16 and 20 mg/
mL respectively.
4. Discussion
Numerous pathogenic microorganisms have developed
their resistance to commonly available antibiotics; hence the
need for developing new virulent drugs against these harmful
pathogens becomes more important. Chemical drugs may
lead to adverse effects and recent researchers have focused
on pharmacologically active compounds from natural sources.
Marine organisms contain many undiscovered bioactive
Table 2
MIC of tissue and egg extracts of C. ramosus against clinically isolated human pathogens.
Bacterial strains/solvents
Acetone extract Chloroform extract
Tissue (mg/mL)Egg (mg/mL)Tissue (mg/mL)Egg (mg/mL)
20 16 12 8 4 20 16 12 8 4 20 16 12 8 4 20 16 12 8 4
V. parahaemolyticus +++ +++ +++ +++ +++ +++ +++ +++ +++ +++ +++ +++ +++ +++ +++ +++ +++ +++ +++ +++
A. hydrophilla - - * + ++ - - - * +*+ ++ +++ +++ - * + ++ +++
S. typhi - - * + ++ - - - * +*+ ++ +++ +++ - - * + ++
S. paratyphi A- - - * +- - * + ++ *+ ++ +++ +++ - * + ++ +++
V. cholerae - - - * +- - - - * * + ++ +++ +++ - * + ++ +++
E. coli - - - - * - - - - * +++ +++ +++ +++ +++ *+ ++ +++ +++
* MIC concentration; - No growth; + Cloudy solution (slight growth); ++ Turbid solution (strong growth); +++ Highly turbid solution (dense growth).
Table 1
Antibacterial activity of tissue and egg extracts of C. ramosus against clinically isolated human pathogens.
Bacterial strains/solvents
Methanol
(mm)
Ethanol
(mm)
Acetone
(mm)
DCM
(mm)
Petroleum
ether (mm)
Acetonitrile
(mm)
Chloroform
(mm)
Water
(mm)
Tissue Egg Tissue Egg Tissue Egg Tissue Egg Tissue Egg Tissue Egg Tissue Egg Tissue Egg
V. parahaemolyticus - - - - - - - 7.00依0.58 - - - - - - - -
A. hydrophilla - - 8.00依0.52 -16.00依0.82 21.00依1.25 - - 7.00依0.58 - - - 8.00依0.58 14.00依0.82 - -
S. typhi - - - 10.00依0.58 18.00依0.82 20.00依1.25 - - 7.00依0.52 - - 7.00依0.58 8.00依0.58 15.00依1.25 - -
S. paratyphi A- - 8.00依0.58 11.00依0.82 26.00依1.53 22.00依1.25 9.00依0.58 7.00依0.58 9.00依0.58 - - 7.00依0.52 7.00依0.58 14.00依0.82 7.00依0.58 -
V. cholerae - - 7.00依0.52 9.00依0.58 20.00依1.25 26.00依1.53 - - - - - - 7.00依0.58 11.00依0.82 - -
E. coli - - 9.00依0.58 12.00依0.82 25.00依1.53 25.00依1.53 - - 11.00依0.58 - - 7.00依0.58 -9.00依0.58 - -

Pasiyappazham Ramasamy et al./Journal of Coastal Life Medicine 2013; 1(3): 181-185
184
compounds; the number of new active compounds isolated
from marine organisms are estimated at 10 000[8]. Molluscs
are considered as one of the important natural sources to
derive many bioactive compounds that exhibit antitumor,
antimicrobial, anti-inflammatory and antioxidant activities[9].
Molluscs also contain highly rich nutrients, which are beneficial
to humans of all ages[10]. Compounds isolated from marine
molluscs were also used in the treatment of rheumatoid arthritis
and osteoarthritis[11]. Marine mollusc extracts also exhibited
antibacterial and antiviral activity against fish pathogenic
bacteria and the extract also may be applied in aquaculture[12].
In the present investigation among the various strains tested
maximum zone of inhibition [(26.00依1.53) mm] was recorded
in acetone extracts against S. paratyphi A and minimum zone
of inhibition (7 mm) was noticed in ethanol, petroleum ether,
chloroform and water extracts. In egg the maximum zone of
inhibition [(26依1.53) mm] was recorded in acetone extracts
against V. cholera, and minimum zone of inhibition (7mm)
was noticed in DCM and acetonitrile. Similar observation was
made by Suresh et al[13]. Lactobacillus vulagaris, Pseudomonas
aeruginosa and S. typhi were resistant to a crude methanolic
extract of gastropod Hemifusus pugilinus with highest activity
against E. coli (6 mm), followed by Klebsiella pneumoniae (4
mm), and the lowest activity against S. paratyphi (1 mm), V.
paraheamolyticus (1 mm).
Suresh et al. also reported that the maximum inhibition zone
[(10.13依0.13) mm] was observed against Klebsiella pneumonia
in the ethanol extract of Babylonia zeylanica and the minimum
inhibition zone (1 mm) was observed against V. cholerae[13]. In
the case of Harpa conoidalis, the maximum inhibition zone
[(9.16依0.13) mm] was observed against S. paratyphi in ethanol
extract and the minimum inhibition zone [(1.03依0.05) mm] was
noticed against Proteus mirabilis. In the similar way Babylonia
spirata exhibited the antibacterial activity of ethanol, acetone,
methanol, chloroform and water extracts; the maximum
inhibition zone (12 mm) was observed against Pseudomonas
aeruginosa in the crude ethanol extract of Babylonia spirata
and the minimum inhibition zone (2 mm) was noticed against
Staphylococcus aureus bacterial strains[14].
The hypobranchial gland extracts of C. ramosus was found to
inhibit the growth of bacterial strains; among these the broad
inhibition zone was formed against Streptococcus faecalis and
Staphylococcus aureus[15]. The ethanol extracts of hypobranchial
gland of Chicoreus virgineus showed 10 mm of inhibition zone
against S. typhi, 7 mm against Shigella flexineri, 6 mm against V.
cholerae, 5 mm against Klebsiella pnemoniae and 4 mm against
Bacillus subtilis and E. coli, but methanolic extracts exhibited
inhibition against Streptococcus pyogenes[16]. Although different
species and experimental procedure were followed in different
studies, they indicated the high degree of antimicrobial activity
in marine molluscs. These results encourage the idea that
marine molluscs are potent sources for antibacterial drug
development.
MIC methods are widely used in the comparative testing of
new drugs. In clinical laboratories they are used to establish
the susceptibility of organisms that give equivocal results in
disk or well tests, for tests on organisms where disk or well
tests may be unreliable and when a more accurate result is
required for clinical management. The present study MIC values
of acetone tissue and egg extracts against bacterial strains
such as A. hydrophilla, S. typhi, S. paratyphi A, V. cholerae
and E.coli were reported as 12, 12, 8, 8, 4 mg/mL and 8, 8, 12, 4,
4 mg/mL respectively. Likewise in chloroform tissue extracts
against bacterial strains such as A. hydrophilla, S. typhi, S.
paratyphi A and V. cholerae was reported as 20, 20, 20 and
20 mg/mL respectively. The chloroform egg extracts against
bacterial strains such as A. hydrophilla, S. typhi, S. paratyphi
A, V. cholerae and E. coli were reported as 16, 12, 16, 16 and 20
mg/mL respectively. The study revealed that the acetone and
chloroform extract of both the tissues and egg inhibited the
growth of the tested pathogenic bacterial strains. Hence the
present study indicated that the C. ramosus extracts contain
compounds with the broad antibacterial activity. However,
further investigations involving purification of the active
extracts as drugs for humans are needed.
Conflict of interest statement
We declare that we have no conflict of interest.
Acknowledgements
Authors are thankful to Dr. T. Ramamurthy, Scientist F,
National Institute of Cholera & Enteric Diseases, Kolkata
with necessary facilities. The authors are also thankful to
the Centre for Marine Living Resources and Ecology, Cochin,
Ministry of Earth Sciences, New Delhi, India for providing
the financial support (Grant No. MoES/10-MLR/01/12).
Comments
Background
The marine environment is a huge source for discovering
many antimicrobial drugs. Apart from the food that is
derived from the marine environment, wide varieties of
antimicrobial drugs are being isolated and characterized
with great promise for the treatment of human diseases.
Studies on antimicrobial screening provide valuable
information for new antibiotic discoveries and give new
insights into the extraction of bioactive compounds from
marine molluscs.
Research frontiers
To screen the antibacterial drugs from different solvent
extraction of tissue and egg of marine gastropods C. ramosus

Pasiyappazham Ramasamy et al./Journal of Coastal Life Medicine 2013; 1(3): 181-185 185
against clinically isolated human pathogenic bacteria like V.
parehaemolyticus (J13300), A. hydrophilla (IDH1585), S. typhi
(C6953), S. paratyphi A (C6915), V. cholerae (IDH5439) and E.
coli (H10407).
Related reports
Marine molluscs contain many undiscovered bioactive
compounds; the number of new active compounds isolated
from marine organisms is estimated at 10 000. Molluscs
are considered as one of the important natural sources to
derive many bioactive compounds that exhibit antitumor,
antimicrobial, anti-inflammatory and antioxidant activities.
Compounds isolated from marine molluscs were also used
in the treatment of rheumatoid arthritis and osteoarthritis.
Marine mollusc extracts also exhibited antibacterial and
antiviral activity against fish pathogenic bacteria and the
extract also may be applied in aquaculture.
Innovations and breakthroughs
The innovative outcome of this research paper is to
screen the antibacterial drugs in eight different solvent
extractions of tissue and egg of marine molluscs.
Applications
The maximum zone of inhibition [(26±1.53) mm] was
recorded in acetone extracts against S. paratyphi A and the
minimum zone of inhibition (7mm) was noticed in ethanol,
petroleum ether, chloroform and water extracts. In egg the
maximum zone of inhibition [(26±1.53) mm] was recorded in
acetone extracts against V. cholera and minimum zone of
inhibition (7 mm) was noticed in DCM and acetonitrile.
Peer review
Numerous pathogenic microorganisms have developed
their resistance against commonly available antibiotics;
hence the need for developing new virulent drugs against
these harmful pathogens becomes more important.
Chemical drugs may lead to adverse effects and researchers
now have focused on pharmacologically active compounds
from natural sources. The present study revealed that the
acetone and chloroform extract of both the tissues and egg
inhibited the growth of the tested pathogenic bacterial
strains. The MIC of both extracts ranged from 4 to 12 mg/
mL. These results suggest that marine gastropods tissue
and egg extract contains comparatively good antibacterial
activity.
References
[1] Huges RN. A functional biology of marine gastropods. London:
Croom Helm Ltd; 1986, p. 245.
[2] Shaw PD, McLure WO, Van Blaricom G, Sims J, Fenical W, Rude
J. Antimicrobial activities from marine organisms. In: Webber
HH, Ruggieri GD, editors. Food-drug from sea proceedings.
1974; Washington DC: Marine Technology Society; 1976, p. 55-
60.
[3] Kamboj VP. Bioactive agent from the Ocean Biota. In: Somayajulu
BL, editor. Ocean science: trends and future directions. New Delhi,
India: Indian National Science Academy; 1999.
[4] Rajaganapathi J, Kathiresan K, Singh TP. Purification of anti-
HIV protein from purple fluid of the sea hare Bursatella leachii
de Blainville. Mar Biotechnol (NY) 2002; 4(5): 447-453.
[5] Benkendorff K, Davis AR , Bremner JB. Chemical defense
in the egg masses of benthic invertebrates: an assessment
of antibacterial activity in 39 mollusks and 4 polychaetes. J
Invertebr Pathol 2001; 78(2): 109-118.
[6] Ramasamy P, Subhapradha N, Srinivasan A, Shanmugam
V, Krishnamoorthy J, Shanmugam A. In vitro evaluation of
antimicrobial activity of methanolic extract from selected
species of cephalopods on clinical isolates. Afri J Microbiol Res
2011; 5(23): 3884-3889.
[7] Ramasamy P, Barwin Vino A, Saravanan R, Subhapradha
N, Vairamani S. Shanmugam A. Screening of antimicrobial
potential of polysaccharide from cuttlebone and methanolic
extract from body tissue of Sepia prashadi Winkworth, 1936.
Asian Paci J Trop Biomed 2011; 1(2): S244-248.
[8] Kelecom A. Secondary metabolites from marine microorganisms.
An Acad Bras Cienc 2002; 74(1): 151-170.
[9] Benkendorff K, McIver CM, Abott CA. Bioactivity of the murex
homeopathic remedy and ofextracts from an Australian murcid
mollusc against human cancer cells. Evid-Based Compl Altern
Med 2011; 1: 12-16.
[10] Anand PT, Chellaram C, Kumaran S, Shanthini CF. Biochemical
composition and antioxidant activity of Pleuroploca trapezium. J
Chem Pharma Res 2010; 2: 526-535.
[11] Chellaram C, Edward JK. In vivo anti-inflammatory bustle of
reef associated mollusc, Trochus tentorium. Adv Biotech 2009;
8(12): 32 -34.
[12] Defer D, Bourgnon N, Fleury Y. Screening for antibacterial and
antiviral activities in three bivalve and two gastropod marine
molluscs. Aquaculture 2009; 293(1-2): 1-7.
[13] Suresh M, Arularasan S, Srikumaran N. Screening on
antimicrobial activity of marine gastropods Babylonia zeylanica
(Bruguière, 1789) and Harpa conoidalis (Lamarck, 1822) from
Mudasalodai, southeast coast of India. Int J Pharm Pharm Sci
2012; 4(4): 552-556.
[14] Periyasamy N, Srinivasan M, Balakrishnan S. Antimicrobial
activities of the tissue extracts of Babylonia spirata Linnaeus,
1758 (Mollusca: Gastropoda) from Thazhanguda, southeast coast
of India. Asian Pac J Trop Biomed 2012; 2(1): 36-40.
[15] Kagoo IK, Ayyakkannu K. Bioactive components from Chicoreus
ramosus antibacterial activity in vivo. Phuket Mar Biol Cent
Publ 1992; 11: 147-150.
[16] Rajaganapathi J. Studies on antibacterial activity on five marine
molluscs[D]. India: Annamalai University; 1996, p. 43.