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International Journal of Advanced Research Trends in Engineering and Technology

(IJARTET) Vol. 8, Issue 3, March 2021

Isolation and Characterization of Marine

Bacteria for Potential Antibacterial
activities
Astom Mondal1, Karabi Biswas2 and Sankar Narayan Sinha3
Research Scholar (CSIR-SRF), Department of Botany, University of Kalyani,West Bengal, India 1
Research Scholar, Department of Botany, University of Kalyani,West Bengal, India 2
Professor, Department of Botany, University of Kalyani,West Bengal, India 3

Abstract: Twelve marine bacterial strain were isolated from seawater from coastal areas of Puri, Odisha, India.
The antibacterial activities of these bacteria were investigated. Methanol extracts of marine bacterial
fermentation were screened for antibacterial activities through agar well diffusion method. The results indicated
that out of 12 strains 7 had antibacterial activities. The active marine bacteria were assigned to the genera
Bacillus and Pseudomonas. The TLC autobiographic overlay assay implied that the antibacterial metabolites
produced by two strains with wide antibacterial spectrum were different. These marine bacteria were expected
to be the potential resources of natural antibiotic products. In order to explore more novel structures, new ways
of screening of these compounds should be applied.
Keywords: Marine bacteria, Antibiotic, Seawater,Bacillus, Pseudomonas.

isolation of marine bacterium, investigation of

I. INTRODUCTION antimicrobial properties of these bacterial isolates,
Compared with terrestrial organisms, the characterization and identification of isolated
secondary metabolites produced by marine bacterial strains and investigation of the antibiotic
organisms have more novel and unique structures compounds produced by different active bacteria
owing to the complex living, circumstance and were same or not.
diversity of species and bioactivities are much
stronger. Because of the low content of active II. MATERIALS AND METHODS
compounds in marine animals and plants, as well as Sampling- Sea water samples were
limitation of bioresource supply, more and more collected in the intertidal zone during low tide from
researchers have been focused on marine coastal areas of Puri, Odisha, India. Sample water
microorganisms as sustainable resources. The was collected in clean and sterilized glass water
competition among various microbes for the space bottles and transported to laboratory immediately
and nutrient in marine environment is a powerful for bacteriological study.
selection process which endows marine Isolation of marine bacteria- The sea water
microorganisms to produce many natural products sample were serially diluted with sterile 0.85%
possessing medical as well as industrial values. sodium chloride solution. Aliquots of 0.1ml of
Some antimicrobial, antifouling substances have diluted samples were spreaded onto 1/10 marine
been found among these kinds of bacteria. It is agar plates contain peptone 0.5gm, yeast extract
suggested that the main role of these antibiotic 0.2gm, FeSO4 0.1gm agar 15gm, sea water 1L and
substances might be related to ecological pH was adjusted to 7.0-7.6. All plates were
competition. Secondary metabolites of incubated at 25℃ for two days. Colonies with
microorganisms are used widely in various fields, different morphology was chosen for further
such as medicine, pharmaceuticals, chemical experiment.
industries, agriculture, food processing etc. Antibacterial assay: The antibacterial
However, few industries have been conducted to activity of seven marine bacterial suspension was
study and compare the antibiotic activities of determined using agar well Diffusion method
marine bacteria isolated from different origins. described by Bauer et al (1966) [1] following the
With this prospective in mind, the work has been standard procedure.
under taken with the following objectives are as

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International Journal of Advanced Research Trends in Engineering and Technology

(IJARTET) Vol. 8, Issue 3, March 2021

Identification of the bacterial isolates- and a large number of bacteria could live on it [3].
Bacterial staining, cultural, morphological and These bacterial strains are not actually symbiotic to
biochemical procedures were studied for the the host but can instead be considered as associated
identification of bacterial isolate. Isolated bacteria bacteria [4] with consanguineous relationship with
were identified according to methods described in their hosts. These bacteria, on one hand, could
Bergey’s Manual of Determinative Bacteriology acquire the essential nutrients like sugar, vitamin,
[2]. fatty acids etc. from their plant or. animal hosts
whereas on the other hand they could excrete
III. RESULTS AND DISCUSSION products like antibiotic, amino acid and toxin
Twelve marine bacteria isolated from sea propitious for the metabolism as well as
water samples. The antimicrobial assay showed development of the hosts or. for the improvement
that 7 strains of these isolates that antimicrobial of the chemical defense potential of the hosts [5].
activity against test bacteria (Table 1). After The present results are quite consistent with the
taxonomic study it was found that the bacteria with reported previous investigations. The antimicrobial
antimicrobial activity belonged mainly to the activity of marine bacteria of coastal sea water was
genera Pseudomonas (4 strains) and Bacillus (3 reported previously [,6,7,8,9]. In our work , the
strains). All the seven isolates were then preserved proportion of antimicrobial activity exhibiting
in a slant and broth cultures and were used in bacteria isolated from sea water was near about
studying the colony morphology, microscopical 58% which were tentatively identified as Bacillus
characteristics, gram character, mobility and other and Pseudomonas group though molecular
biochemical assay which shown in the table-2. On identification studyis needed for further
the basis of their morphological, physiological and confirmation, All 7 strains exhibited activity
biochemical anlyses, the twelve strains were against both gram positive and gram-negative
tentatively identified as Pseudomonas putidaand bacteria indicating their broad-spectrum
Bacillus subtilis. Crude extracts of two strains PP2 nature.This result implied that some marine
and BP9 showed greater antimicrobial spectrum (as bacteria could probably release a number of
indicated by larger inhibition zone). In the present antibiotic compounds in provide themselves the
study, we have isolated 12 strains from sea water survival competition superiority.

TABLE I
Results of antibacterial activity of marine bacteria using agar well diffusion assay (‘-’ no inhibition zone, ‘+’
inhibition zone, ‘+++’ large inhibition zone, PP- Pseudomonas sp. and BP- Bacillus sp.).
Isolated Antibacterial Activity
Strain Escherichi Vibrio Shigella Shigella Staphylococcus Micrococus
a coli cholerae flexneri dysenteriae aureus luteus
PP1 + + + + + +
PP2 + + + + + +
PP3 - - - - - -
PP4 + + + + + +
PP5 - - - - - -
PP6 + + + + + +
BP7 + + + + + +
BP8 - - - - - -
BP9 +++ +++ +++ +++ +++ +++
BP10 + + + + + +
BP11 - - - - - -
BP12 - - - - - -

TABLE II
Table-2: Characterization of bacterial isolates (+ presence or positive reaction, - absent or negative reaction, O
Oxidation, F Fermentation)
Test Bacterial isolates

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PP1 PP2 PP4 PP6 BP7 BP9 BP10

Cell shape Rod Rod Rod Rod Rod Rod Rod
Gram reaction - - - - + + +
Motility + + + + + + +
Growth at 5% NaCl + + + + + + +
Catalase + + + + + + +
Oxidase + + + + + + +
IMVIC test
Indole - - - - - - -
Methyl red - - - - - - -
VogesProskauer - - - - + + +
Citrate utilization + + + + + + +
Urease - - - - - - -
H2S production - - - - - - -
NO3 production - - - - - - -
Gelatine liquefaction + + + + + + +
Starch hydrolysis - - - - + + +
Huge leifson (O/F) O/F O/F O/F O/F F F F
reaction
Utilization of carbon
source + + + + + + +
Glucose + + + + + + +
Fructose + + + + - - -
Xylose + + + + + + +
Sucrose + + + + + + +
Lactose - - - - + + +
Cellobiose - - - - + + +
Raffinose - - - - + + +
Mannitol - - - - + + +

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International Journal of Advanced Research Trends in Engineering and Technology (IJARTET)

Vol. 8, Issue 3, March 2021

[7]. S.Harounabadi, P.E. Shamsabad and S.M. Meybodi,

Isolation and characterization of bacteria with
IV. CONCLUSION potential antimicrobial activity from the Caspian Sea,
Marine microorganisms as model systems offer Russian J. of Marine Biol. (2016), 42,36-41
the potential to understand and develop treatments for [8]. E. Rodrigues, S. Tilvi and C.G. Naik, Antimicrobial
disease on the basis of normal physiological role of their activity of marine bacteria organisms collected of the
secondary metabolites and are currently being applied to coast of South East India, Journal of Experimental
the development of new drugs. It can be concluded that marine Biology and Ecology (2004), 309 (1) 121-127
marine water is a potential source of large number of
bacterial strains for sources of new biomolecules that can [9]. K. Kumari, R.J. Madhuri and C.D. Reddy,
be exploited to produce antibiotics. In order to find more Antimicrobial potential of marine bacterial isolates
novel structures new ways of screening of these from different Coastal regions of Andhra Pradesh and
compounds should be applied. The separation and Tamil Nadu, India International Journal of Current
identification of bioactive compounds with wide Microbiology and Applied Science Int. J. Curr.
antibacterial spectrum from these marine bacteria is in Microbiol. App. Sci (2013), 2(10) 230-237
progress
BIOGRAPHY
ACKNOWLEDGMENT Astom Mondal is currently working
Our sincere thanks to CSIR and DST PURSE-II for as Senior Research Fellow (CSIR-
research funding and also University of Kalyani for SRF) in the Department of Botany,
providing research laboratory. University of Kalyani, Kalyani,
Nadia, West Bengal, India. He has
REFERENCES authored over5 research papers in
[1]. Bauer AW, Kirby WM, Sherris JC, TurckM international/national journals. His
Antibiotic susceptibility testing by a standardized research interests include
single diskmethod. Am J Clin Pathol(1966) 45(4): environmental research microbiology.
493-496.
Karabi Biswas is currently
[2]. Bergey D.H., Holt J.G., Bergey’s manual of working as Senior Research
determinative bacteriology, Baltimore:Williams and Scholar in the Department of
Wilkins (1994) Botany, University of Kalyani,
[3]. J.M. Shewan, G. Hobbs and W. Hodgkiss, A Kalyani, Nadia, West Bengal,
determinative scheme for the identification of certain India. She has authored over 10
genera of gram-negative bacteria with special research papers in
reference to the Pseudomonadaceae, J. Appl. Bact. international/national journals.
(1960), 23(3), 379-390 His research interests include
environmental research microbiology.
[4]. Bultel-Ponce V, Berge JP, Debitus C. Metabolites
from the sponge associated bacterium Pseudomonas Dr. Sankar Narayan Sinha is
species. Mar. Biotechnol. (1999) 1(4), 384-390. currently working as senior most
[5]. G. Bowler, B.I. Duerden and D.G. Armstrong, Wound Professor in the Department of
microbiology and associated approaches to wound Botany, University of Kalyani,
management, ClinMicrobiol Rev. (2001) 14(2), 244- Kalyani, Nadia, West Bengal, India.
269 He has authored over 100 research
papers in international/national
[6]. B. Flemer, J. Kennedy, L. M. Margassery, J. P. journals. His research interests
Morrissey, F. O.Gara and A. D. W. Dobson, Diversity include environmental research microbiology.
and antimicrobial activities of microbes from two
Irish marine sponges, Suberitescarnosus and
Leucosoleniasp Affiliations Expand J ApplMicrobiol .
(2012),112 (2):289-301.

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