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Panigrahi A.,Central Institute of Brackishwater Aquaculture
Journal of Applied Aquaculture | Year: 2012

An epidemiological investigation was done in brackish water culture systems in three coastal districts of West Bengal. A total of 198 farms were randomly surveyed with a structured questionnaire. The data showed that there was a significant difference in outbreak of white spot disease (WSD) (p < 0.01), shell-associated problems (p < 0.01), and gill-associated problems (p < 0.05) among the culture systems. Among all systems, stunted and uneven growth and white fecal disease (only in shrimp monoculture) were the dominant emerging disorders. WSD remained the most prevalent disease. Some farms tested (polymerase chain reaction [PCR]) positive for WSD, but the animals were apparently healthy. Chlorination, use of PCR screening, application of immunostimulants, and strict bio-security measures play major roles in containing disease outbreaks. © 2012 Copyright Taylor and Francis Group, LLC.


Kathiravan V.,Central Institute of Brackishwater Aquaculture | Krishnani K.K.,National School of Management
World Journal of Microbiology and Biotechnology | Year: 2014

In the present work, novel heterotrophic nitrifying and aerobic denitrifying bacteria have been isolated from greenwater system of coastal aquaculture. Based on the 16S rRNA gene, FAME analysis and biochemical test, the isolates have been identified as Pseudomonas aeruginosa and Achromobacter sp. These have been named as P. aeruginosa strain DBT1BNH3 and Achromobacter sp. strain DBTN3. Denitrifying functional genes such as nitrite reductase (nirS), nitric oxide reductase (qnorB) and nitrous oxide reductase (nosZ) genes have been identified. These strains found to have a 27 kb plasmid coding for nirS and nosZ. The possibility of horizontal transfer of plasmid among Pseudomonadaceae and Alcaligenaceae families in coastal aquaculture has been explored. Further, we have studied combined nitrification and oxygen tolerant denitrification potential in the same isolates. © 2013 Springer Science+Business Media Dordrecht.


Velusamy K.,Central Institute of Brackishwater Aquaculture | Krishnani K.K.,National School of Management
Applied Biochemistry and Biotechnology | Year: 2013

In this work, herbivorous fish Mugil cephalus has been cultured to secrete protein rich green slime, which helps nitrifying and oxygen tolerant denitrifying bacteria to grow and colonize. Four strains representing Alcaligenaceae family have been isolated from greenwater system and characterized using biochemical test, fatty acid methyl ester (GCFAME) analysis, 16S rRNA and functional gene approaches. They were tested for an ability to nitrify ammonia and nitrite aerobically. Two strains showed notable nitrification activity, when grown in a mineral salts medium containing ammonium sulfate and potassium nitrite. Functional gene analysis confirmed the presence of nitrous oxide reductase (nosZ) gene showing that they have an oxygen-tolerant denitrification system. It has been proposed that Alcaligenes faecalis strains heterotrophically nitrify ammonia into nitrite via formation of hydroxyl amine, which is oxidized to nitrous oxide using oxygen or nitrite as electron acceptor. These results provide a possible advantage of having nitrification and denitrification capabilities in the same organism, which plays an important role in biological wastewater system. © Springer Science+Business Media New York 2013.


Alavandi S.V.,Central Institute of Brackishwater Aquaculture | Poornima M.,Central Institute of Brackishwater Aquaculture
Indian Journal of Virology | Year: 2012

Viruses are abundant biological entities on earth and the emergence of viral pathogens has become a serious threat to aquaculture and fisheries worldwide. However, our response to viral pathogens has been largely reactive, in the sense that a new pathogen is usually not discovered until it has already reached epidemic proportions. Current diagnostic methods such as PCR, immunological assays and pan-viral microarrays are limited in their ability to identify novel viruses. In this context, the knowledge on the diversity of viruses in healthy and disease situations becomes important for understanding their role on the health of animals in aquaculture species. Viral metagenomics, which involves viral purification and shotgun sequencing, has proven to be useful for understanding viral diversity and describing novel viruses in new diseases and has been recognized as an important tool for discovering novel viruses in human and veterinary medicine. With the advancements in sequencing technology and development of bioinformatics tools for nucleic acid sequence assembly and annotation, information on novel viruses and diversity of viruses in marine ecosystems has been rapidly expanding through viral metagenomics. Novel circoviruses and RNA viruses in Tampa bay pink shrimp, annelovirus in sea lion, picornavirus in ringed seals and several new viruses of marine animals have been recently described using viral metagenomics and this tool has been also recently used in describing viral diversity in aquaculture ponds. Further, a large amount of information has been generated on the diversity of viruses in the marine environment using viral metagenomics during the last decade. There exists a great potential with viral metagenomics for discovering novel viruses in asymptomatic marine candidate animals of aquaculture/mariculture, some of which may assume pathogenic status under high density culture and stress. Additionally, viral metagenomics can help our understanding of viruses present in aquaculture/mariculture settings and routine pathogen surveillance programmes. © 2012 Indian Virological Society.


Kiruthika J.,Central Institute of Brackishwater Aquaculture | Rajesh S.,Central Institute of Brackishwater Aquaculture | Ponniah A.G.,Central Institute of Brackishwater Aquaculture | Shekhar M.S.,Central Institute of Brackishwater Aquaculture
Developmental and Comparative Immunology | Year: 2013

Acyl-CoA binding protein (ACBP), a protein present ubiquitously in wide range of organisms play significant role in transport of acyl groups for macromolecular biosynthesis involved in various functional and regulatory processes. In crustaceans, ACBP has functional role in growth, reproduction and temperature tolerance. In the present study, two suppression subtractive hybridization (SSH) cDNA libraries were performed using gut tissues of shrimp Penaeus monodon exposed to low (3ppt) and high (55ppt) salinity stress conditions. SSH library resulted in identification of differentially expressed genes that belonged to various functional classes such as the nucleic acid regulation and replication, defence proteins, allergen protein, signal transduction pathways, apoptosis, energy and metabolism, cell cycle regulation and hypothetical proteins. ACBP was identified as one of the differentially expressed gene in both the SSH libraries of shrimp P. monodon subjected to low and high salinity stress. The full-length cDNA of P. monodon ACBP gene was isolated and the sequence revealed 273. bp open reading frame encoding 90 amino acids with molecular mass of 10. kDa and pI 6.8. The ORF showed presence of four phosphorylation sites, with absence of signal peptide sequence and glycosylation sites. The deduced amino acid sequence of ACBP exhibited high sequence identity (92%) with ACBP class of protein identified from Fenneropenaeus chinensis. Real time PCR analysis of shrimps subjected to 3ppt salinity conditions after 2. weeks revealed an increase in expression of ACBP transcripts, in the gut (28.08-folds), gills (11.71-folds) and in the muscle tissues (1.70-folds). Whereas, shrimps exposed to 55ppt salinity conditions after 2. weeks exhibited increased ACBP transcript levels in the gut (11.95-folds), gills (1.052-folds) and muscle tissues (7.35-folds). The significant increase in expression levels of ACBP in various tissues of shrimps suggests a functional role of this gene in salinity stress tolerance and adaptation. © 2013 Elsevier Ltd.


Shekhar M.S.,Central Institute of Brackishwater Aquaculture | Ponniah A.G.,Central Institute of Brackishwater Aquaculture
Journal of Fish Diseases | Year: 2015

Viral disease outbreaks are a major concern impeding the development of the shrimp aquaculture industry. The viral disease due to white spot syndrome virus (WSSV) observed in early 1990s still continues unabated affecting the shrimp farms and cause huge economic loss to the shrimp aquaculture industry. In the absence of effective therapeutics to control WSSV, it is important to understand viral pathogenesis and shrimp response to WSSV at the molecular level. Identification and molecular characterization of WSSV proteins and receptors may facilitate in designing and development of novel therapeutics and antiviral drugs that may inhibit viral replication. Investigations into host-pathogen interactions might give new insights to viral infectivity, tissue tropism and defence mechanism elicited in response to WSSV infection. However, due to the limited information on WSSV gene function and host immune response, the signalling pathways which are associated in shrimp pathogen interaction have also not been elucidated completely. In the present review, the focus is on those shrimp proteins and receptors that are potentially involved in virus infection or in the defence mechanism against WSSV. In addition, the major signalling pathways involved in the innate immune response and the role of apoptosis in host-pathogen interaction is discussed. © 2014 John Wiley & Sons Ltd.


Rajesh S.,Central Institute of Brackishwater Aquaculture | Kiruthika J.,Central Institute of Brackishwater Aquaculture | Ponniah A.G.,Central Institute of Brackishwater Aquaculture | Shekhar M.S.,Central Institute of Brackishwater Aquaculture
Fish and Shellfish Immunology | Year: 2012

O-methyltransferase (OMT), a protein present ubiquitously in wide range of organisms plays significant role in methylation of small macro molecules for various functional and regulatory purposes. In crustaceans, OMT has functional role in growth, reproduction, ovarian development and molting. In the present study, suppression subtractive hybridization (SSH) performed using gill tissues of low (3ppt) and high (55ppt) salinity stressed shrimp Penaeus monodon resulted in identification of differentially expressed genes involved in signal transduction pathways, metabolism, defense proteins, DNA repair and synthesis, apoptosis, cell cycle regulation along with unknown and hypothetical proteins. Catechol-O-methyltransferase (COMT) a type of OMT was identified by SSH as one of the differentially expressed genes of shrimp P. monodon subjected to low and high salinity stress. The full length cDNA of COMT was cloned from the gills of P. monodon which consisted an open reading frame of 666 bp, encoding 221 amino acids. The ORF revealed one each of N-glycosylation and O-glycosylation sites and nine phosphorylation sites. The deduced amino acid sequence of COMT exhibited high sequence identity (92%) with COMT class of protein from Fenneropenaeus chinensis. Real time PCR analysis of the shrimp samples exposed to low salinity conditions at 3ppt revealed significant increase in expression of COMT transcripts in the guts at 24 h, 48 h, 1 week and 2 weeks, gills at 24 h and in the muscle tissues at 48 h, with maximum expression of the COMT levels by 5 fold in guts (1 week), 1 fold in gills (24 h) and 1.5 fold in muscle (48 h) respectively. The increased expression level of COMT at different time intervals in different tissues suggests a possible role of this gene in salinity stress tolerance in shrimps under low salinity conditions. © 2012 Elsevier Ltd.


Satellite data of Landsat TM, IRS 1B LISS II, IRS 1C LISS III and IRS 1D LISS III were used to monitor aquaculture development and its spatial distribution in Cuddalore District, south-east coast of India. Analysis of satellite images using image processing software ERDAS Imagine 9.0 and Geographic Information System (GIS) software Arc GIS 9.0 indicated that the area under aquaculture was 6.91 ha in 1987, gradually increasing to 67.71 ha in 1994, and reaching a maximum of 380.19 ha in 1998. There was a little shrimp culture development (6.14 ha) from 1998 to 2005 because of risk of White Spot Syndrome Virus (WSSV), epizootics and constraints related to social and environmental concerns raised against shrimp farming. Shrimp farms initially started in the marsh areas close to creeks and then moved inland towards agricultural and fallow lands. Farms developed in clusters along the creeks without affecting mangrove reserve forests between 1988 and 2005 and about 177.02 ha agriculture land including 234.25 ha fallow lands were converted for aquaculture farming. Shrimp culture was abandoned in 109.47 ha and 15.13 ha of agricultural land previously converted to aquaculture farms, was reconverted for agriculture. The present study shows that remote sensing data and GIS are useful tools in assessing and monitoring the development of aquaculture for larger areas and suggests that national level planning using multi-spectral satellite data can be used to assess the existing land use and the potential for sustainable use of coastal resources.


Shekhar M.S.,Central Institute of Brackishwater Aquaculture | Kiruthika J.,Central Institute of Brackishwater Aquaculture | Ponniah A.G.,Central Institute of Brackishwater Aquaculture
Fish and Shellfish Immunology | Year: 2013

Four suppression subtractive hybridization (SSH) cDNA libraries were constructed to identify differentially expressed salinity stress responsive genes of black tiger shrimp, Penaeus monodon exposed to low (3ppt) salinity conditions. Forward and reverse SSH cDNA libraries were developed from the gill and gut tissues of shrimp and clones having inserts larger than 300bp were unidirectionally sequenced. Based on the sequence homology search, the identified genes were categorized for their putative functions related to a wide range of biological roles, such as nucleic acid regulation and replication, immune response, energy and metabolism, cell signaling, cellular process, cytoskeleton and membrane structure, stress and osmoregulation. Gene expression levels in response to low salinity conditions at 2 weeks post salinity stress of thirteen selected differentially expressed genes identified from SSH cDNA libraries (14-3-3 like protein, crustin, lysozyme, arginine kinase, Na+/K+-ATPase α-subunit, intracellular fatty acid binding protein, cathepsin B, anti-lipopolysaccharide factor, ferritin, ubiquitin conjugating enzyme E2, calreticulin, innexin 2 and heat shock protein 21) were analyzed by RT-PCR. The highest gene expression levels were observed for Na+/K+-ATPase α-subunit (34.28-folds) in gill tissues, intracellular fatty acid binding protein (13.30-folds) in gut tissues and innexin 2 (14.43-folds) in muscle tissues respectively. The differential and significant levels of gene expression indicate the functional role of these genes in shrimp salinity stress adaptive mechanisms. © 2013 Elsevier Ltd.


Krishnani K.K.,Central Institute of Brackishwater Aquaculture
Aquaculture | Year: 2010

Nucleic acid methods based on sequencing of clone libraries provide sequence and the phylogenetic information of an individual clone. In the present study, ammonia monooxygenase (amoA), nitrite-oxido-reductase(norB), nitrite reductase (nirS) and nitrous oxide reductase (nosZ) genes are chosen to detect nitrifiers and denitrifiers in coastal aquaculture using gene-specific primers. The abundance of these functional genes revealed the presence of nitrifying and denitrifying organisms in coastal aquaculture. Nitrifying and denitrifying communities were analyzed by parallel DNA extractions and clone library construction for amoA, norB, nirS and nosZ obtained from coastal aquaculture soil. Amino acids in parts of the amoA, norB, nirS and nosZ encoded proteins were aligned to know conserved amino acid residues. The amoA genes exhibited 81-82% identity to Nitrosomonas europaea, Nitrosococcus mobilis and Nitrosomonas eutropha which were also similar to particulate methane monooxygenase (pmoA) gene sequences. The norB genes are closely affiliated with Nitrobacter winogradskyi and other uncultured beta-proteobacteria. The nosZ genes exhibited 78-82% identity with Marinobacter sp. and other uncultured denitrifying gamma-proteobacteria. The present study could be useful for making bioremediation strategy for nitrogenous metabolites and understanding of nitrogen fluxes generated through these two functional groups in coastal shrimp aquaculture. © 2010 Elsevier B.V. All rights reserved.

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