54 research outputs found

    Development and characterization of a fibroblastic-like cell line from caudal fin of the red-line torpedo, Puntius denisonii (Day) (Teleostei: Cyprinidae)

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    A fibroblastic-like cell line was established from the ornamental ¢sh, red-line torpedo (Puntius denisonii). The red-line torpedo fin (RTF) cell line is being maintained in Leibovitz’s L-15 mediumsupplementedwith 10% fetal bovine serum (FBS) for over 1year at 28 1C on a continuous basis in normal atmosphere. The growth rate of RTF cells increased as the FBS proportion increased from 5% to 20% at 28 1C with optimum growth at the concentrations of 10% FBS. The morphology of RTF cell was predominantly fibroblastic like. Propagation of these cell lines was serum dependent, with a low plating efficiency (o15%). Karyotyping analysis of RTF cells at the 25th passage indicated that the modal chromosome number was 2n550. The cell line was cryopreserved in liquid nitrogen at �196 1C and could be recovered from storage after 6 months with good cell viability. Polymerase chain reaction amplification of a fragment of two mitochondrial genes, 16S rRNA and CO1, con¢rmed the identity of these cell lines with those reported from this animal species, confirming that the cell lines originated from P. denisonii. The bacterial extracellular products from Vibrio cholerae MTCC3904 and Aeromonas hydrophila were found to be toxic to RTF. The cell lines were not susceptible to viral nervous necrosis virus, a marine ¢sh virus

    Outbreak of Parasitic Dinoflagellate Piscinoodinium sp. Infection in an Endangered Fish from India: Arulius Barb (Dawkinsia arulius)

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    Freshwater velvet disease is caused by the dinoflagellate parasite, Piscinoodinium sp. This parasite has been reported in tropical and subtropical fishes, and it can cause devastating losses. Moreover, Piscinoodinium sp. is identified as one of the least studied finfish parasites, and the available molecular information about this parasite is meager. Recently, Piscinoodinium sp. was responsible for the 100% cumulative mortality of the captive-bred F1 generation of Arulius barb (Dawkinsia arulius), an endangered freshwater fish native to India. The trophont stages of the parasite were observed in the skin and gills of the affected fish. The total DNA was extracted from the trophonts collected from the affected Arulius barb and the partial nucleotide sequence of the rDNA complex region (2334 bp) was amplified using PCR. The amplified PCR product exhibited a high sequence identity (97.61%) with Piscinoodinium sp. In the phylogenetic analysis of the SSU rDNA, Piscinoodinium sp. emerged as a separate clade from other dinoflagellate species. This is the first report of the infection of Piscinoodinium sp. in Arulius barb and the molecular information generated from this study can serve as a baseline to study the diversity of the parasite in India. Furthermore, the impact of this parasite among wild fish stock is not known, and this parasite needs further research focus to generate more molecular information and to understand the host–pathogen interaction

    Renaissance in Fisheries: Outlook and Strategies - Book of Abstracts 9th Indian Fisheries Forum, December 19-23, 2011, Chennai, India

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    The Asian Fisheries Society – Indian Branch (AFSIB) since its inception in 1986, has been providing a platform for discussion at the national level on issues related to research, development, education and policies by organizing Indian Fisheries Forum (IFF) every three years in different parts of the country. The 9th Indian Fisheries Forum (9th iff) will be hosted by the Central Marine Fisheries Research Institute (CMFRI), at Chennai during 19-23 December 2011. The main theme of the 9th iff is “Renaissance in Fisheries: Outlook & Strategies”. It would have a comprehensive look for the fisheries and aquaculture sectors, for achieving greater synergy among the stakeholders and planning strategies for capture fisheries and aquafarming to build higher levels of sustainability and profitability. The forum would also address the issues of impact of climate change and its mitigation, resource constraint and species diversification for the expansion of fish production activity; and encourage young scientists to undertake need-based and resource specific research. An international symposium sponsored by the Bay of Bengal Large Marine Ecosystem (BoBLME) is scheduled to be held during the forum on 21 December, 2011 with theme: Bay of Bengal–Ecosystem Approach to Fisheries Management

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    Aquaculture of popular freshwater species, Nile tilapia (Oreochromis niloticus), accounts for around 71% of the total global tilapia production. Frequent use of antibiotics for treating bacterial infections in tilapia leads to the emergence of antimicrobial resistance. To mitigate the issue, proper evaluation methods and control strategies have to be implemented. This study was aimed to analyze the antimicrobial resistance of bacterial isolates from the infected Nile tilapia cultured in freshwater. The recovered isolates were identified as Pseudomonas entomophila, Edwardsiella tarda, Comamonas sp, Delftia tsuruhatensis, Aeromonas dhakensis, A. sobria, A. hydrophila, A. lacus, Plesiomonas shigelloides and Vogesella perlucida through phenotypic and genotypic analyses. Using Primer-E software, Shannon Wiener diversity index of the isolates was determined as H' (loge) = 2.58. Antibiotic susceptibility test of the recovered strains through disk diffusion using 47 antibiotics, showed an elevated resistance pattern for Aeromonas hydrophila, Pseudomonas entomophila and Comamonas with higher multiple antibiotic resistance indexes (MAR index > 0.3). The minimum inhibitory concentration of antibiotics was > 256 mcg/ml for most of the resistant isolates. Meanwhile, all the recovered isolates were susceptible to amikacin, aztreonam, kanamycin, cefalexin, cefotaxime, levofloxacin, norfloxacin, piperacillin, and polymyxin-B.Department of Science and Technology (SERB) for providing National Postdoctoral Research fellowship (PDF/2017/000378

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    Not AvailableCommon carp (Cyprinus carpio) is a widely cultivated freshwater fish for human consumption, while koi carp, is a farmed colored sub species of common carp used for ornamental purposes. Since 1998, both common carp and koi carp are severely affected by a viral disease called as Koi herpes virus disease (KHVD). This disease is caused by Koi herpes virus (KHV), also known as cyprinid herpes virus-3. The virus causes interstitial nephritis and gill necrosis in carps, so it is also termed as carp interstitial nephritis and gill necrosis virus. KHV is a double stranded icosahedral DNA virus belonging to family Alloherpesviridae, with a genome size of 295 kbp, larger than any member of Herpesviridae. The viral genome encodes 156 potential protein coding open reading frames. Each virion consists of forty structural proteins, which are classified as capsid (3), envelope (13), tegument (2) and unclassified (22) structural proteins. Diagnosis of KHVD is mainly based on detection of viral DNA by polymerase chain reaction amplification using specific primers or loop mediated isothermal amplification. Temperature dependent latent infection is unique to KHV; and carrier fish are often not detected, thereby possibly resulting in spread of this pathogen to newer areas. The disease is now known to occur in, or has been recorded from at least 26 different countries of the world. Fortunately, KHVD has not been reported from India or from Indian major carps. To monitor the disease status of the country, a total of 254 fish samples collected from different parts of India were screened by PCR for the presence of KHV. None of the tested samples were found to be positive for KHV. These results demonstrate that tested samples from different parts of India were apparently free from KHV. Preliminary risk assessment of KHV suggest that in the event of unrestricted importation of koi carps into our country, there is a higher probability of risk to aquaculture as compared to natural waters. So there is strong need to develop diagnostic capabilities and launch surveillance programmes for KHV in India.Department of Biotechnology (DBT), Government of Indi

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    Research papers on the causes, effects, treatments, and prevention of diseases of marine and freshwater organisms, particularly fish and shellfishCommercially available culture media and supplements were tested for their potential to produce primary cell cultures from tissues of Indian mud crabs Scylla serrata. Eight commercially available culture media from Sigma-Aldrich (Leibovitz's L-15, Medium 199, Grace's Insect Medium, Minimal Essential Medium, Dulbecco's Modified Eagle Medium, TC-100 Insect Medium, IPL-41 Insect Medium, and Roswell Park Memorial Institute) were examined. Three different supplements (amino acid and sugar [AS], crab muscle extract [CME], and natural seawater [NSW]) were also examined. The hemocyte culture appeared to grow well for a maximum period of 21 d in 2 × L-15 medium supplemented with AS and 15% fetal bovine serum (FBS). Partial amplification and sequencing of the cytochrome oxidase subunit I (COI) gene confirmed that the primary hemocytes originated from Indian mud crabs. The effects of four metals on hemocyte viability were evaluated using the MTT assay. Of the four metals examined (arsenic, lead, cobalt, and nickel), cobalt and nickel were more toxic to the crab cells than the other metals. Both acridine orange/ethidium bromide and Hoechst staining showed the presence of apoptosis and necrosis in metal-treated groups, which suggests that metals in an aquatic environment induce death of the Indian mud crab's hemocytes. The hemocyte primary cell culture was also used to study the cytotoxicity effect of bacterial extracellular products from Vibrio harveyi and white spot syndrome virus. This study demonstrates that hemocyte primary cell culture can be used as a tool to study viral and bacterial pathogenesis and to assess the cytotoxicity of pollutants present in aquatic environments.Not Availabl

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    Describe novel and comprehensively tested methods which enhance human, animal, plant, bacterial or environmental virology and prions research and discovery.The lack of shrimp cell lines and difficulty in establishing shrimp cell culture systems, with an appropriate medium is a major concern in the aquaculture sector. The present study attempts to address this issue by developing an in vitro cell culture system from various tissues (hemocytes, heart, lymphoid tissue, hepatopancreas, gill, eye stalk, and muscle) of Penaeus vannamei (P.vannamei) using commercially available L-15 medium. The cell culture medium was formulated using five different media such as HBSCM-1, HBSCM-2, HBSCM-3, HBSCM-4, and HBSCM-5 containing L-proline and glucose with fetal bovine serum (FBS) supplements. Among the different media used, the HBSCM-5 medium with supplements showed good attachment and proliferation of cells with fibroblast-like, epithelioid, round, and adherent cell morphology in hemocyte culture. The same medium was further screened using different tissues to enhance the cell growth. The hemocytes, heart, and lymphoid tissue cells were passaged five times and maintained up to 20 days. Hepatopancreas and gill cells initially showed good morphological features and survived for more than ten days following subculture cells. Eye stalks and muscle cells perished within five days and did not show any unique morphology. The primary hemocyte cells were subjected to species identification, using cytochrome oxidase subunit I (COI) gene. To assess the primary hemocyte cell culture, cells were used for in vitro propagation of white spot syndrome virus (WSSV) and confirmed by the conventional polymerase chain reaction (PCR). Similarly, the primary cells were treated with bacterial extracellular products (ECPs) from Vibrio parahaemolyticus and Vibrio harveyi, to evaluate the cytotoxicity.Not Availabl

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    Not AvailableCell lines provide an important biological tool for carrying out investigations into physiology, virology, toxicology, carcinogenesis and transgenics. Teleost fish cell lines have been developed from a broad range of tissues such as ovary, fin, swim bladder, heart, spleen, liver, eye muscle, vertebrae, brain, skin. One hundred and twenty-four new fish cell lines from different fish species ranging from grouper to eel have been reported since the last review by Fryer and Lannan (J Tissue Culture Methods 16: 87–94, 1994). Among the cell lines listed, more than 60% were established from species from Asia, which contributes more than 80% of total fish production. This includes 59 cell lines from 19 freshwater, 54 from 22 marine and 11 from 3 brackish water fishes. Presently, about 283 cell lines have been established from finfish around the world. In addition to the listing and a scientific update on new cell lines, the importance of authentication, applications, cross-contamination and implications of overpassaged cell lines has also been discussed in this comprehensive review. The authors feel that the review will serve an updated database for beginners and established researchers in the field of fish cell line research and development.Department of Biotechnology (DBT), Government of Indi
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