Fish Karyome: A karyological information network database of Indian Fishes

‘Fish Karyome’, a database on karyological information of Indian fishes have been developed that serves as central source for karyotype data about Indian fishes compiled from the published literature. Fish Karyome has been intended to serve as a liaison tool for the researchers and contains karyological information about 171 out of 2438 finfish species reported in India and is publically available via World Wide Web. The database provides information on chromosome number, morphology, sex chromosomes, karyotype formula and cytogenetic markers etc. Additionally, it also provides the phenotypic information that includes species name, its classification, and locality of sample collection, common name, local name, sex, geographical distribution, and IUCN Red list status. Besides, fish and karyotype images, references for 171 finfish species have been included in the database. Fish Karyome has been developed using SQL Server 2008, a relational database management system, Microsoft's ASP.NET-2008 and Macromedia's FLASH Technology under Windows 7 operating environment. The system also enables users to input new information and images into the database, search and view the information and images of interest using various search options. Fish Karyome has wide range of applications in species characterization and identification, sex determination, chromosomal mapping, karyo-evolution and systematics of fishes

EXPLORING FISH MISCELLANY FROM BIODIVERSITY HOTSPOT REGION OF NORTHEAST INDIA THROUGH CHROMOSOME PROFILING

Sjeveroistočne države Indije obiluju ekstenzivnim ekoklimatskim uvjetima te sadrže veliki dio indijske endemske flore i faune. Ova regija obiluje ribljom raznolikošću s mnoštvom vrsta akvarijskih riba, uključujući zebricu. Među potencijalnim ukrasnim ribama, Ctenops nobilis označena je kao gotovo ugrožena, a Danio dangila uključena je na popis najmanje zabrinutosti IUCN-a. Ovdje je poduzeto citogenetsko profiliranje kako bi se osigurali komplementarni podaci za preciznu identifikaciju vrsta i proučavanje genetskog nasljeđa, organizacije genoma i evolucije vrste. Citogenetskim analizama utvrđen je diploidni broj kromosoma 44 i 50 s formulom kariotipa 8m+20sm+10st+6t (FN=72) odnosno 20m+24sm+6st (FN=94) u C. nobilis i D. dangila. Bojanje nukleolarnih regija otkrilo je prisutnost Ag-NORs, CMA3 mjesta i signala gena 18S rDNA na jednom paru subtelocentričnog kromosoma i jednom paru submetacentričnog kromosoma u C. nobilis i D. dangila. Ovi biomarkeri važan su izvor za molekularnu taksonomiju, evolucijske/filogenetske studije i genetiku očuvanja C. nobilis i D. dangila.North Eastern states of India are blessed with extensive ecoclimatic conditions and much of India’s endemic flora and fauna. This region is rich in fish diversity with plenty of species of ornamental importance, including model species zebrafish. Among the potential ornamental fishes, Ctenops nobilis is designated as Near Threatened and Danio dangila is included in the Least Concern list by IUCN. Cytogenetic profiling was undertaken here to provide complementary data for precise species identification and to study genetic inheritance, genome organization and evolution of the species. The cytogenetic analyses revealed diploid chromosome numbers 44 and 50 with karyotype formula 8m+20sm+10st+6t (FN=72) and 20m+24sm+6st (FN=94), respectively, in C. nobilis and D. dangila. The staining of nucleolar organizer regions revealed the presence of Ag-NORs, CMA3 sites and 18S rDNA probe signals on one pair sub-telocentric chromosome and one pair sub-metacentric chromosome in C. nobilis and D. dangila, respectively. These biomarkers are an important resource for molecular taxonomy, evolutionary/phylogenetic studies and conservation genetics of C. nobilis and D. dangila

DEVELOPMENT OF CAUDAL FIN CELL LINE FROM HILL TROUT Barilius bendelisis (HAMILTON, 1807) FOR CYTOTOXICITY AND TRANSFECTION STUDIES

Stanična linija, nazvana BBdF-1, uspostavljena iz repne peraje Barilius bendelisis, presađena je na više od 52 „oblika“(prolaza) a održavala se u medijima L-15 koji sadrže 20% FBS. Porijeklo stanične linije potvrđeno je pomoću pojačanja djelomičnog područja 16S i COI mitohondrijskih genskih sekvenci. Utvrđeno je da je optimalna temperatura za rast stanične linije BBdF-1 28°C. Kariotipizacijom se utvrdio diploidni broj kromosoma kao 50. Stanice su pokazale snažno vezanje za citokeratinski marker i, prema tome, utvrđeno je da sliče epitelu. Jaka zelena fluorescencija primijećena je u stanicama BBdF-1 transficiranim vektorom phrGFP-II-N, što ukazuje da je prikladna za upotrebu u ispitivanjima ekspresije i manipulacije gena. Izvršena je i uspješna procjena citotoksičnosti dvaju teških metala, žive i kroma. Stanična linija može poslužiti kao koristan izvorni materijal za ranu provjeru toksičnosti pesticida/zagađivača i ekspresije gena.A cell line named BBdF-1, established from the caudal fin of hill stream fish Barilius bendelisis, has been subcultured for more than 52 passages and is being maintained in L-15 media containing 20% FBS. Species origin of the cell line was confirmed using amplification of partial region of 16S and COI mitochondrial gene sequences. The optimum temperature for growth of BBdF-1 cell line was found to be 28°C. Karyotyping revealed diploid chromosome number as 50. Cells exhibited strong binding for cytokeratin marker and thus were found to be epithelial-like. Strong green fluorescence was observed in BBdF-1 cells transfected with phrGFP-II-N vector, indicating its suitability for utilization in gene expression and manipulation studies. Successful assessment of cytotoxicity of two heavy metals, viz. mercury and chromium, was performed. The cell line can serve as a useful resource material for early toxicity screening of pesticides/pollutant and gene expression

Toxicity of the Herbicide Atrazine: Effects on Lipid Peroxidation and Activities of Antioxidant Enzymes in the Freshwater Fish Channa Punctatus (Bloch)

The present study was undertaken to evaluate the toxicity and effects of a commercial formulation of the herbicide atrazine (Rasayanzine) on lipid peroxidation and antioxidant enzyme system in the freshwater air breathing fish Channa punctatus. The 12, 24, 48, 72 and 96 h LC50 of atrazine, calculated by probit analysis, were determined to be 77.091, 64.053, 49.100, 44.412 and 42.381 mg·L−1, respectively, in a semi static system with significant difference (p < 0.05) in LC10–90 values obtained for different times of exposure. In addition to concentration and time dependent decrease in mortality rate, stress signs in the form of behavioral changes were also observed in response to the test chemical. In fish exposed for 15 days to different sublethal concentrations of the herbicide (1/4 LC50 = ∼10.600 mg·L−1, 1/8 LC50 = ∼5.300 mg·L−1 and 1/10 LC50 = ∼4.238 mg·L−1) induction of oxidative stress in the liver was evidence by increased lipid peroxidation levels. The antioxidants superoxide dismutase (SOD), catalase (CAT) and glutathione reductase (GR) responded positively in a concentration dependent pattern, thus, suggesting the use of these antioxidants as potential biomarkers of toxicity associated with contaminations exposure in freshwater fishes

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Not AvailableCharacterization of the major (18S) and minor (5S) ribosomal RNA genes were carried out in three commercially important Indian major carp (IMC) species, viz. Catla catla, Labeo rohita and Cirrhinus mrigala along with their physical localizations using dual colour fluorescence in situ hybridization. The diploid chromosome number in the above carps was confirmed to be 50 with inter-species karyo-morphological variations. The 18S rDNA signals were observed on 3 pair of chromosomes in C. catla and L. rohita, and two pairs in C. mrigala. The 5S rDNA signal was found on single pair of chromosome in all the species with variation in their position on chromosomes. The sequencing of 18S rDNA generated 1804, 1805 and 1805 bp long fragments, respectively, in C. catla, L. rohita and C. mrigala with more than 98% sequence identity among them. Similarly, sequencing of 5S rDNA generated 191 bp long fragments in the three species with 100% identity in coding region and 23.2% overall variability in non-transcribed spacer region. Thus, these molecular markers could be used as species-specific markers for taxonomic identification and might help in understanding the genetic diversity, genome organization and karyotype evolution of these species.Department of Biotechnology, the Ministry of Science and Technology, Government of Indi

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Not AvailableChromosome preparations were obtained from kidney tissues of freshly dead fish Labeo rohita and Cirrhinus mrigala, using in vitro colchicine treatment. The chromosomal spreads were of good quality, suitable for karyotyping and chromosome banding studies. The present technique is highly suitable for cytogenetic studies in wild stock of fish species for which there is difficulty in getting live fish and also in those species which die immediately in captivity. This is the first report on use of tissues from dead fish for studying metaphase chromosomes.Not Availabl

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Not AvailableThe freshwater snakehead fishes are members of the family Channidae and order Perciform, which are native to parts of Africa and Asia with widespread distributions. The present study was aimed to assess the genetic variability and molecular phylogeny among 22 species of genus Channa, viz. C. argus, C. asiatica, C. aurantimaculata, C. bankanensis, C. barca, C. bleheri, C. burmanica, C. diplogramma, C. gachua, C. lucius, C. maculata, C. marulius, C. melasoma, C. micropeltis, C. orientalis, C. ornatipinnis, C. panaw, C. pleurophthlama, C. pulchra, C. punctata, C. stewartii and C. striata, using partial 513 bp long consensus sequence of cytochrome-c oxidase subunit I mitochondrial DNA retrieved from NCBI (March, 2018). The study showed 281 haplotypes with 267 variable sites (i.e., 33 singletons and 234 parsimoniously informative) in nucleotide positions. Most of the sequences were A + T rich, except in C. argus which is G + C rich (50.25%). The NJ and MP trees showed similar topology with discrete branches, each as a distinct species of the Channa with slight differences in bootstrap values. Maximum K2P distance was found between the C. ornatipinnis and the C. melasoma. The maximum numbers of polymorphic loci were present in C. gachua.Not Availabl

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Not AvailableSeveral options have been proposed for eradication of germ cells (GCs) in mammals such as treatment with cytotoxic drugs, irradiation, cold ischemia and hyperthermic treatment. Some of these methods have been also tried in fish but conditions for complete sterilisation of gonads have not been established. Here, we report the production of sterile adult common carp Cyprinus carpio in 10 weeks by the heat and chemical treatments. The cytotoxic drug busulfan (40 mg/kg) was intraperitoneally injected into the animals at 2-week intervals (5 doses in total), and they were maintained in water at 38 °C between Weeks 1 and 10. The effectiveness of the treatments was assessed using gonadal index, histology, and vasa gene expression. At the end of Week 10, very severe gonadal degeneration was observed in fish treated with the heat–chemical combination, and 100% of male and female fish were devoid of endogenous GCs. The average levels of vasa transcript were 0.01 ± 0.005 and 0.02 ± 0.016 for males and females, respectively. By contrast, high temperature alone caused minor gonadal degeneration and the gene transcript were 0.59 ± 0.131 for male and 0.62 ± 0.13 for female. In Week 20, after the recovery period of 10 weeks at 25 °C, the gonadal germ cell did not recover from the sterile condition in any of the sampled individuals. The change in colouration of gonads was an additional useful index of the degree of gonadal sterility.Not Availabl

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Not AvailableDose dependent genotoxic effect of malathion pesticide on Channa punctatus was evaluated by micronuclei test assay. Single cell gel electrophoresis (comet assay) was also standardised by using Channa punctatus. The results confirm that both assays are sensitive to detect genotoxicity in fishes even at very low level of pollutants in aquatic medium. Thus these assays can be used as genotoxic bioassays for monitoring pollutants in aquatic medium.Not Availabl

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Not AvailableA wide range of diploid number of chromosomes and the body size of Channa congeners are useful combination of characters for studying the factors controlling the body size. In this study, the karyological information was superimposed on the evolutionary tree generated by 16S rRNA mitochondrial gene sequences. Here, the metaphase chromosome complements stained with Giemsa, AgNO3 and CMA3 were prepared from six snakehead murrel fish species collected from northeast India. The diploid chromosome numbers and the fundamental arms of C. aurantimaculata (2n = 52, NF = 98), C. gachua (2n = 56, NF = 84), C. marulius (2n = 44, NF = 58), C. orientalis (2n = 52, NF = 74), C. punctata (2n = 32, NF = 60) and C. striata (2n = 40, NF = 48) were calculated by the analysis of metaphase chromosome complements. Both methods of nucleolar organizer region (NOR) localization, silver nitrate and chromomycin A3, revealed NOR pairs of 1, 2, 3, 1, 4 and 3 in C. aurantimaculata, C. gachua, C. marulius, C. orientalis, C. punctata and C. striata, respectively. The subject species showed primitive type of asymmetrical chromosomes, except the C. punctata. The variation in 2n for C. orientalis (2n = 52, 78) and C. gachua (2n = 52, 78, 104) of a complete haploid set indicates the possibility of either ploidy change in C. orientalis and C. gachua, if we consider 2n = 52 or the Robertsonian rearrangements in different populations of these two species. The chromosome evolution tree was constructed on 16S rRNA ML-phylogenetic tree using ChromEvol 1.3. The analysis of chromosome evolution explained the loss or gain of chromosome, duplications or semiduplications mechanism. For time scaling the chromosome evolution, the node age of available 16S rRNA gene of Channa species were estimated, which was also used for estimating the time when chromosomal changes occurred in context of geological time-scaleDepartment of Biotechnology, Government of India, New Delh