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16S Rdna Sequence Analysis of Culturable Marine Biofilm Forming Bacteria from a Ship's Hull D See discussions, stats, and author profiles for this publication at: http://www.researchgate.net/publication/280736395 paper inbakandan 08927014%2E2010%2E530347 DATASET · AUGUST 2015 DOWNLOADS VIEWS 11 14 7 AUTHORS, INCLUDING: Ramasamy Venkatesan G. Latha National Institute of Ocean Technology Natioanl institute of ocean technology 103 PUBLICATIONS 654 CITATIONS 49 PUBLICATIONS 88 CITATIONS SEE PROFILE SEE PROFILE Simi Mathew Rokkam Rao National Institute of Ocean Technology Indian Institute of Tropical Meteorology 8 PUBLICATIONS 104 CITATIONS 40 PUBLICATIONS 821 CITATIONS SEE PROFILE SEE PROFILE Available from: Ramasamy Venkatesan Retrieved on: 15 September 2015 This article was downloaded by: [Nat Institute of Ocean Technology] On: 20 July 2015, At: 03:15 Publisher: Taylor & Francis Informa Ltd Registered in England and Wales Registered Number: 1072954 Registered office: 5 Howick Place, London, SW1P 1WG Biofouling: The Journal of Bioadhesion and Biofilm Research Publication details, including instructions for authors and subscription information: http://www.tandfonline.com/loi/gbif20 16S rDNA sequence analysis of culturable marine biofilm forming bacteria from a ship's hull D. Inbakandan a b , P. Sriyutha Murthy c , R. Venkatesan d & S. Ajmal Khan b a Centre for Ocean Research , Sathyabama University , Chennai , 600 119 , India b Centre of Advanced Study in Marine Biology , Annamalai University , Port Nova , 608502 , India c Biofouling and Biofilm Processes Section , WSCL, BARC Facilities, IGCAR , Kalpakkam , 603 102 , India d Ocean Science and Technology for Islands , National Institute of Ocean Technology , Chennai , 600 100 , India Published online: 27 Oct 2010. To cite this article: D. Inbakandan , P. Sriyutha Murthy , R. Venkatesan & S. Ajmal Khan (2010) 16S rDNA sequence analysis of culturable marine biofilm forming bacteria from a ship's hull, Biofouling: The Journal of Bioadhesion and Biofilm Research, 26:8, 893-899, DOI: 10.1080/08927014.2010.530347 To link to this article: http://dx.doi.org/10.1080/08927014.2010.530347 PLEASE SCROLL DOWN FOR ARTICLE Taylor & Francis makes every effort to ensure the accuracy of all the information (the “Content”) contained in the publications on our platform. However, Taylor & Francis, our agents, and our licensors make no representations or warranties whatsoever as to the accuracy, completeness, or suitability for any purpose of the Content. Any opinions and views expressed in this publication are the opinions and views of the authors, and are not the views of or endorsed by Taylor & Francis. The accuracy of the Content should not be relied upon and should be independently verified with primary sources of information. Taylor and Francis shall not be liable for any losses, actions, claims, proceedings, demands, costs, expenses, damages, and other liabilities whatsoever or howsoever caused arising directly or indirectly in connection with, in relation to or arising out of the use of the Content. This article may be used for research, teaching, and private study purposes. Any substantial or systematic reproduction, redistribution, reselling, loan, sub-licensing, systematic supply, or distribution in any form to anyone is expressly forbidden. Terms & Conditions of access and use can be found at http:// www.tandfonline.com/page/terms-and-conditions Biofouling Vol. 26, No. 8, November 2010, 893–899 16S rDNA sequence analysis of culturable marine biofilm forming bacteria from a ship’s hull D. Inbakandana,b*, P. Sriyutha Murthyc, R. Venkatesand and S. Ajmal Khanb aCentre for Ocean Research, Sathyabama University, Chennai 600 119, India; bCentre of Advanced Study in Marine Biology, Annamalai University, Port Nova 608502, India; cBiofouling and Biofilm Processes Section, WSCL, BARC Facilities, IGCAR, Kalpakkam 603 102, India; dOcean Science and Technology for Islands, National Institute of Ocean Technology, Chennai 600 100, India (Received 27 April 2010; final version received 29 September 2010) Marine bacteria from the hull of a ship in the form of biofilms or microfouling were isolated, cultured, and identified by phylogenetic analysis using 16S rDNA sequences. With an average length of 946 bp, all the 16 sequences were classified using the Ribosomal database project (RDP) and were submitted to the National Center for Biotechnology Information. Phylogenetic analysis using 16S rDNA sequences indicated that the 16 strains belonged to the Firmicutes (IK-MB6 Exiguobacterium aurantiacum, IK-MB7 Exiguobacterium arabatum, IK-MB8 Exiguobacterium arabatum, IK-MB9 Jeotgalibacillus alimentarius, IK-MB10 Bacillus megaterium, IK-MB11 Bacillus pumilus, IK-MB12 Bacillus pumilus, IK-MB13 Bacillus pumilus, IK-MB14 Bacillus megaterium), High GC, Gram- positive bacteria (IK-MB2 Micrococcus luteus, IK-MB5 Micrococcus luteus, IK-MB16 Arthrobacter mysorens), G-Proteobacteria (IK-MB3 Halomonas aquamarina, IK-MB15 Halotalea alkalilenta), CFB group bacteria (IK-MB1 Myroides odoratimimus), and Enterobacteria (IK-MB4 Proteus mirabilis). Among the 16 strains, representatives of the Firmicutes were dominant (56.25%) compared to the high GC, Gram-positive bacteria (18.75%), G-Proteobacteria (12.5%), CFB group bacteria (6.25%), and Enterobacteria (6.25%). Analysis revealed that majority of marine species found in marine biofilm are of anthropogenic origin. Keywords: marine bacteria; biofilm; 16S rDNA sequence; phylogenetic analysis; anthropogenic origin Introduction metamorphosis of fouling organisms (Callow and Marine biofouling is the biotic progression by which Callow 2002). Bacteria and other colonizing micro- prokaryotic and eukaryotic organisms adhere to solid organisms secrete extracellular polysaccharides which surfaces immersed in seawater. Biofilm formation anchor them to the substratum thereby altering the occurs through a sequence of processes, beginning local surface chemistry which can stimulate further with the adsorption of organic (organic film) and growth and the recruitment and settlement of macro- inorganic particles onto the surface, followed by the organisms (Chambers et al. 2006) which results in attachment of pioneer microorganisms, the growth and biofouling. Thus, marine biofouling represents a major reproduction of primary colonizers, and then the economic drawback in the maritime industries, since maturation of the biofilm matrix (Dang and Lovell the microbial slime films and the large numbers of 2000). The composition of the biofilm depends on barnacles, mussels, and tunicates which accumulate on the ions, glycoproteins, and humic and fulvic acids ships’ hulls increase drag forces and surface corrosion, Downloaded by [Nat Institute of Ocean Technology] at 03:15 20 July 2015 available in the liquid phase. Marine bacterial biofilm thereby causing additional fuel and maintenance costs formation might be a survival strategy as it provides (Zambon et al. 1984; Pereira et al. 2002). microorganisms with increased access to nutrients, Methods commonly employed to prevent biofilm protection against toxins and antibiotics, maintenance formation include chemical treatment of the water of extracellular enzyme activities, and shelter from column by biocides or coating the surfaces with predation. The forces that promote the adsorption and antifouling paints. As these methods invariably lead conditioning of the surface include electrostatic inter- to pollution, environmental friendly methods are actions and Van der Waal’s forces (Vigeant et al. desirable. After the ban on the use of tin and copper 2002). based antifouling paints, subsequent studies largely In seawater, the microbial population produces concentrated on a few novel approaches (Zambon a primary biofilm on surfaces which is generally et al. 1984; Beveridge et al. 1997) such as natural thought to be a prerequisite for the attachment and product based non metallic and ecofriendly coatings *Corresponding author. Email: [email protected] Published online 27 October 2010 ISSN 0892-7014 print/ISSN 1029-2454 online Ó 2010 Taylor & Francis DOI: 10.1080/08927014.2010.530347 http://www.informaworld.com 894 D. Inbakandan et al. (Kristensen et al. 2008), surface modification appro- PCR products were analyzed by agarose gel electro- aches such as engineered topographies (Magin et al. phoresis, purified with High Pure PCR Product 2010), foul release polymer based coatings (Chaudhury Purification Kit (Roche, Germany), and sequenced et al. 2005) and nanotechnological approaches (Gladis using an ABI PRISM 3100 Genetic Analyzer (Applied et al. 2010). But for any kind of study to prevent Biosystems, USA). biofilm formation, knowledge of the marine bacterial composition of the target biofilm layer would be of considerable importance. In the present study, the Sequence analysis and phylogenetic analyses authors report on the culturable marine bacterial The sequencing was performed using primer 518r isolates that occupy the surfaces or the hulls of ships (50-GTA TTA CCG CGG CTG CTG-30) and 338f and trace their possible origins. (50-ACT CCT ACG GGA GGC AGC-30) (Kwon et al. 2002) and sequences of the 16S rDNA between 362 and 484 bp (average 451 bp) were submitted to the Materials and methods Advanced BLAST search program of the National Biofilm collection and culture condition Center for Biotechnology Information (NCBI) to The biofilms were collected from the air–seawater determine whether they aligned with closely related interface at the bottom of the hull of a fishing vessel organisms. The related sequences were preliminarily berthed for dry docking at Ennore harbor (138150 aligned
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