J. K. Jena

Complete mitochondrial genome sequence of Catla catla and its phylogenetic consideration

Complete nucleotide sequence of mitochondrial genome (mitogenome) of the Catla catla (Ostariophysi: Cypriniformes: Cyprinidae) was determined in the present study. Its length is 16,594 bp and contains 13 protein coding genes, 22 transfer RNAs, two ribosomal RNAs and one non-coding control region. Most of the genes were encoded on the H-strand, while the ND6 and eight tRNA (Gln, Ala, Asn, Cys, Tyr, Ser (UCN), Glu and Pro) genes were encoded on the L-strand. The reading frames of two pair of genes overlapped: ATPase 8 with 6 and ND4L with ND4 by seven nucleotides each. The main non-coding region was 929 bp, with three conserved sequence blocks (CSB-I, CSB-II, and CSB-III) and an unusual simple sequence repeat, (TA)7. Phylogenetic analyses based on complete mitochondrial genome sequences were in favor of the traditional taxonomy of family Cyprinidae. In conclusion present mitogenome of Catla catla adds more information to our understanding of diversity and evolution of mitogenome in fishes.

Complete mitochondrial genome of Labeo rohita.

The complete mitochondrial genome of Labeo rohita, an important cultivable fish, was determined for the first time. The genome is 16,611 bp in length and consists of 13 protein-coding genes, 22 tRNA genes, 2 rRNA genes, and one control region. The gene organisation and its order were similar to other vertebrates. The overall base composition on heavy strand was as follows A: 32.5%, G: 15.2%, C: 27.7%, T: 24.47%, and the A+T content 56.9%. The control region contains a microsatellite, (TA)12, a putative termination-associated sequence and three conserved sequence blocks. This mitogenome sequence data would play an important role in population genetics and phylogenetics of Indian major carps.

Mitochondrial ATPase 6/8 genes to infer the population genetic structure of silver pomfret fish Pampus argenteus along the Indian waters

Abstract Silver pomfret, Pampus argenteus is an economically important seafood species. The fishery resource of pomfret in Indian waters shows a dwindling catch since the last few years and the pomfrets caught were mostly undersized which calls for immediate attempts for management of resources. An accurate definition of population structure is important for management of this species. The genetic stock structure of P. argenteus distributed along Indian coast was identified using analysis of 842 bp of complete ATPase 6/8 genes of mitochondrial DNA. Altogether, 83 silver pomfret (P. argenteus) collected from 4 locations along Indian coast (Gujarat, Kerala, Tamil Nadu and West Bengal) were sequenced. Twenty four haplotypes were identified among 83 individuals with haplotype diversity (0.87) and nucleotide diversity (0.0025). The significant pair-wise FST and AMOVA values, between samples from West Bengal (east coast) and other locations along the west coast (Gujarat and Kerala) indicated the occurrence of distinct population structure in silver pomfret along the coast.

Molecular phylogenetics of three species of the genus Rastrelliger using mitochondrial DNA markers

Abstract In the present study three species of mackerel, Rastrelliger present in Indian waters were taken for genetic identification using cytochrome c oxidase subunit I and 16s rRNA sequences. Accurate identification of these species is important for fishery management as its morphological characters are very similar. In this study, the sequences of COI and 16S rRNA were determined from 19 individuals of three Rastrelliger species, Rastrelliger kanagurta, Rastrelliger brachysoma and Rastrelliger faughni from Andamans and Indian mainland to study the phylogenetic relationship. The intraspecies and interspecies genetic distance ranged from 0.000 to 0.002 and 0.007 to 0.015 respectively based on 16S rRNA sequences. Using COI data analysis, the intraspecies genetic distance ranged from 0.000 to 0.012, while it varied from 0.039 to 0.086 for interspecies. The present study clearly demarcates three species of mackerel based on the mitochondrial genetic sequences and also showed a non-descriptive genetic distance of R. kanagurta from mainland and Andaman Islands.

Complete sequence and characterization of mitochondrial genome in great snakehead, Channa marulius (Hamilton, 1822)

Abstract The complete nucleotide sequence of mitogenome of the great snakehead, Channa marulius (Channidae), was determined and found to be 16,569 base pairs in length. The content and arrangement of different genes on the mitogenome was found similar to other typical teleosts. The overall base composition of the L-strand was found to be T (19.1%), C (31.5%), A (34.8%) and G (14.6%). The control region was 915 nt long and without any repetitive region. The mitogenome sequence data would be useful for studying phylogenetic relationship of C. marulius with other perciform species.

Establishment and characterization of a continuous cell line from thymus of striped snakehead, Channa striatus (Bloch 1793)

The establishment and characterization of a continuous cell line from the thymus of air-breathing fish Channa striatus are described. The cell line, designated C. striatus thymus (CST), has been subcultured over 71 times and shows optimal growth at 28°C in Leibovitz’s-15 (L-15) medium supplemented with 20% fetal bovine serum. The CST cells exhibited low plating efficiency which improved with increase in seeding density. The karyotype analysis revealed that CST cells have a normal diploid karyotype with 2n = 40. Partial amplification and sequencing of two mitochondrial genes, viz. 16S ribosomal RNA (rRNA) and cytochrome oxidase I, confirmed that the cell line originated from C. striatus. CST cells were successfully transfected indicating their potential application for expression of recombinant proteins. In immunocytochemical staining, CST cells showed characteristics of epithelial cells. These cells were sensitive to extracellular products of Vibrio cholerae MTCC 3904 as well as to heavy metal mercuric chloride. The CST cell line would be a useful tool in functional genomic studies such as RNA interference and gene knockout as well as for cytotoxicity studies.

Mass mortality in ornamental fish, Cyprinus carpio koi caused by a bacterial pathogen, Proteus hauseri.

Moribund koi carp, Cyprinus carpio koi, from a farm with 50% cumulative mortality were sampled with the aim of isolating and detecting the causative agent. Three bacterial species viz., Citrobacter freundii (NSCF-1), Klebsiella pneumoniae (NSKP-1) and Proteus hauseri [genomospecies 3 of Proteus vulgaris Bio group 3] (NSPH-1) were isolated, identified and characterized on the basis of biochemical tests and sequencing of the 16S rDNA gene using universal bacterial primers. Challenge experiments with these isolates using healthy koi carp showed that P. hauseri induced identical clinical and pathological states within 3 d of intramuscular injection. The results suggest P. hauseri (NSPH-1) was the causative agent. In phylogenetic analysis, strain NSPH-1 formed a distinct cluster with other P. hauseri reference strains with ≥99% sequence similarity. P. hauseri isolates were found sensitive to Ampicillin, Cefalexin, Ciprofloxacin and Cefixime and resistant to Gentamycin, Oxytetracycline, Chloramphenicol, and Kanamycin. The affected fish recovered from the infection after ciprofloxacin treatment.

Invasion and impacts of alien fish species in the Ganga River, India

In this study, we document an increasing trend of catches of alien fish species from the Ganga River system. The changing fishery during 1980 to 2011 and the fish yield rate (kg km−1) are positively correlated with an invasion coefficient index (Ixi) of alien species within the river. The reproductive propagule pressure (PPP) of alien fishes (Cyprinus carpio, Oreochromis niloticus, Aristichthys nobilis, Ctenopharyngodon idella, Hypophthalmichthys molitrix and Clarias gariepinus) was determined and found to be 19.45×106 for C. carpio, 0.33×106 for O. niloticus and 0.82×106 for A. nobilis at confluences suggesting that these alien species may be migrating into new habitats. Further, these invaded species exhibited all reproductive stages, indicating their likely establishment within the river. The trophic spectrum of alien fishes spanned all levels and the gut repetitive index (GRI) indicated that that food items in most of the fishes were similar showing early trends of food-web alteration and biotic homogenization. The results of this study suggested a continuous decline in the catch of local fishes by wet weight, particularly Indian major carps (IMC) from 128.91 kg km−1 to 38.58 kg km−1 owing to increased catches of alien species from nil to 384.27 kg km−1. Invasion of alien species in the Ganga River represented one of many possible causes of the decline in river health and overall loss of native aquatic communities.

Population genetic structure of Macrobrachium rosenbergii (Palaemonidae) from Indian waters using mitochondrial ATPase 6/8 gene.

Abstract Macrobrachium rosenbergii, giant freshwater prawn, is one of the most commercially important crustaceans. In the present study, primers for ATPase 6/8 region of mt-DNA were designed and successfully amplified (827 bp) in the species. The nucleotide variation in ATPase 6/8 gene revealed the population structuring in natural populations of M. rosenbergii in Indian waters. A total of 35 haplotypes were observed in 93 individuals collected from different locations. Low nucleotide diversity and high haplotype diversity were noticed for the ATPase 6/8 gene. Significant pairwise FST and, haplotype network indicated occurrence of distinct populations. Observed mismatch distribution and Tajima’s D test suggested demographical stability of giant freshwater prawn. The genetic stock structure revealed in this study will be helpful for conservation and management of stocks of M. rosenbergii in Indian waters.

Genetic characterization of Silond catfish, Silonia silondia (Hamilton, 1822) inferred from two mitochondrial markers

Abstract Silonia silondia is a commercially important food fish. Samples collected through commercial catches from four rivers in India are described by sequence analysis of two molecular markers. Cytochrome b (1140 bp) and ATPase 6/8 (842 bp) genes were analyzed, which represented high level of genetic differentiation within populations of S. silondia. The sequence alignments of cytochrome b and ATPase 6/8 genes revealed 13 and 11 different haplotypes, respectively. The sequences of both the mitochondrial regions revealed high haplotype and low nucleotide diversities. The patterns of genetic diversity and haplotype networks clearly indicated two distinct mitochondrial lineages, however, haplotypes from both the lineages were not specifically assigned to any population. The results confirm the utility of molecular markers generating baseline information, useful for planning effective strategies for conservation, management and sustainability of Silond catfish fishery.