Prevalence of microplastics in commonly consumed fish species of the river Old Brahmaputra, Bangladesh.

Microplastics (MPs) are pervasive in aquatic environments, but inland waterbodies (rivers and floodplains) have received much less attention. The present study assesses the incidence of MPs in the gastrointestinal tracts of five commercially important edible fish species-two column feeders (n = 30) and three benthivores (n = 45) from upstream, midstream, and downstream of the Old Brahmaputra river in north-central Bangladesh. MPs were detected in 58.93% of fish, with the highest level in freshwater eel, Mastacembelus armatus (10.31 ± 0.75/fish). Fibers (49.03%) and pellets (28.02%) were the most frequent MPs. Nearly 72% MPs were smaller than 1 mm, and 50.97% were black. FTIR analysis showed 59% polyethelene (PE), followed by polyamide (40%) and unidentified (1%). MP ingestion was linked to fish size and weight, and a high incidence was recorded in the downstream river. Two omnivorous benthic fish ingest more MPs than others. The results corroborate the presence of MPs in the inland river and fish fauna and augment our understanding of heterogeneous MP uptake by fish.

A high-quality chromosome-level genome assembly of rohu carp, Labeo rohita, and its utilization in SNP-based exploration of gene flow and sex determination.

Labeo rohita (rohu) is a carp important to aquaculture in South Asia, with a production volume close to Atlantic salmon. While genetic improvements to rohu are ongoing, the genomic methods commonly used in other aquaculture improvement programs have historically been precluded in rohu, partially due to the lack of a high-quality reference genome. Here we present a high-quality de novo genome produced using a combination of next-generation sequencing technologies, resulting in a 946 Mb genome consisting of 25 chromosomes and 2,844 unplaced scaffolds. Notably, while approximately half the size of the existing genome sequence, our genome represents 97.9% of the genome size newly estimated here using flow cytometry. Sequencing from 120 individuals was used in conjunction with this genome to predict the population structure, diversity, and divergence in three major rivers (Jamuna, Padma, and Halda), in addition to infer a likely sex determination mechism in rohu. These results demonstrate the utility of the new rohu genome in modernizing some aspects of rohu genetic improvement programs.

Presence of microplastics in two common dried marine fish species from Bangladesh.

We examined microplastics (MP) in two commercially important dried fish, Bombay duck (Harpadon nehereus) and ribbon fish (Trichiurus lepturus), collected from two sites on the Bay of Bengal (Cox's Bazar and Kuakata). The number of MP found in dried Bombay duck and ribbon fish from Kuakata was significantly higher (41.33 g-1 and 46.00 g-1, respectively) than the MP present in samples collected from Cox's Bazar (28.54 g-1 and 34.17 g-1, respectively). Fibers were the most common type of MP identified in all samples (41-64%), followed by fragments (22-34%), microbeads (9-16%), films (3-4%), foams (1-4%), and pellets (0-2%). ATR-FTIR analysis revealed three different types of MP polymer - polyethylene (35-45%), polystyrene (20-30%) and polyamide (30-45%) in the dried fish samples. The study confirms the presence of high MP loads in dried fish from the Bay of Bengal, with high potential of trophic transfer of MP to the human body.

Diallel Cross Application and Histomolecular Characterization: An Attempt to Develop Reference Stock of Labeo ariza.

The objective of the present study was to evaluate the growth performance and genetic variation in diallel crosses of Ariza labeo (Labeo ariza) originating from three geographically separated rivers (Atrai, Jamuna and Kangsha) in Bangladesh. Intra (G1K♀K♂, G2J♀J♂, and G3A♀A♂) and inter (G4K♀A♂, G5K♀J♂, G6A♀K♂, G7A♀J♂, G8J♀K♂, and G9J♀A♂) stocks were produced following diallel cross (sex ratio-1:1 and n = 48; 16 from each river). Reproductive and growth performance, muscle cellularity and genetic variation following genotyping of eight microsatellite markers (Lr1, Lr2, Lr3, Lr22, Lr24, Lr27, Lr28 and Lr29) and analysis of all crossbreeds was performed. The fertilization (95% ± 2.11%), hatching (88% ± 1.03%), and survival rates (82% ± 1.88%) of G4K♀A♂ were higher compared to other groups. With respect to length and weight gains (2.67 ± 0.4 cm and 3.39 ± 0.2 g), SGR (3.23% ± 0.20%), and heterosis (8.87% and 24.74%) G4K♀A♂ was the superior group. A higher number of hyperplastic muscle fibers, mean number of alleles (2.75) and mean observed heterozygosity (0.417) from G4K♀A♂ could be interpreted to mean that G4K♀A♂ comprise better performance efficiency compared to others and are considered for continuing the L. ariza stock improvement program.

High levels of genetic variability and differentiation in hilsa shad, Tenualosa ilisha (Clupeidae, Clupeiformes) populations revealed by PCR-RFLP analysis of the mitochondrial DNA D-loop region.

The hilsa shad, Tenualosa ilisha (Clupeidae, Clupeiformes) is an important anadromous clupeid species from the Western division of the Indo-Pacific region. It constitutes the largest single fishable species in Bangladesh. Information on genetic variability and population structure is very important for both management and conservation purposes. Past reports on the population structure of T. ilisha involving morphometric, allozyme and RAPD analyses are contradictory. We examined genetic variability and divergence in two riverine (the Jamuna and the Meghna), two estuarine (Kuakata and Sundarbans) and one marine (Cox's Bazar) populations of T. ilisha by applying PCR-RFLP analysis of the mtDNA D-loop region. The amplified PCR products were restricted with four restriction enzymes namely, XbaI, EcoRI, EcoRV, and HaeIII. High levels of haplotype and gene diversity within and significant differentiations among, populations of T. ilisha were observed in this study. Significant F(ST) values indicated differentiation among the river, estuary and marine populations. The UPGMA dendrogram based on genetic distance resulted in two major clusters, although, these were subsequently divided into three, corresponding to the riverine, estuarine and marine populations. The study underlines the usefulness of RFLP of mtDNA D-loop region as molecular markers, and detected at least two differentiated populations of T. ilisha in Bangladesh waters.

First draft genome assembly and identification of SNPs from hilsa shad ( Tenualosa ilisha) of the Bay of Bengal.

Background: Hilsa shad ( Tenualosa ilisha), a widely distributed migratory fish, contributes substantially to the economy of Bangladesh. The harvest of hilsa from inland waters has been fluctuating due to anthropological and climate change-induced degradation of the riverine habitats.  The whole genome sequence of this valuable fish could provide genomic tools for sustainable harvest, conservation and productivity cycle maintenance. Here, we report the first draft genome of  T. ilisha from the Bay of Bengal, the largest reservoir of the migratory fish. Methods: A live specimen of T. ilisha was collected from the Bay of Bengal. The whole genome sequencing was performed by the Illumina HiSeqX platform (2 × 150 paired end configuration). We assembled the short reads using SOAPdenovo2 genome assembler and predicted protein coding genes by AUGUSTUS. The completeness of the T. ilisha genome assembly was evaluated by BUSCO (Benchmarking Universal Single Copy Orthologs). We identified single nucleotide polymorphisms (SNPs) by calling them directly from unassembled sequence reads using discoSnp++. Results: We assembled the draft genome of 710.28 Mb having an N50 scaffold length of 64157 bp and GC content of 42.95%. A total of 37,450 protein coding genes were predicted of which 29,339 (78.34%) were annotated with other vertebrate genomes. We also identified 792,939 isolated SNPs with transversion:transition ratio of 1:1.8. The BUSCO evaluation showed 78.1% completeness of this genome. Conclusions: The genomic data generated in this study could be used as a reference to identify genes associated with physiological and ecological adaptations, population connectivity, and migration behaviour of this biologically and economically important anadromous fish species of the Clupeidae family.

Accounting for Contribution of Trade Openness and Foreign Direct Investment in Life Expectancy: The Long-Run and Short-Run Analysis in Pakistan.

This paper examines the impact of trade openness and foreign direct investment (FDI) on life expectancy using time series data over the period of 1972-2013. We have applied structural break unit root as well as cointegration tests to examine integrating properties of the variables and cointegration among the variables. The causal linkage between the variables has been tested by applying the VECM Granger causality. The empirical evidence confirms the presence of cointegration amid the variables. Moreover, trade openness and FDI increase population health measured by life expectancy in the long-run. Furthermore, the analysis suggests that trade openness and FDI cause life expectancy in the short-run. These findings have several policy implications to improve life expectancy for the people of Pakistan in particular and other developing countries in general.

Population genetic characterization and family reconstruction in brood bank collections of the Indian major carp Labeo rohita (Cyprinidae:Cypriniformes).

The founder stock of a captive breeding program is prone to changes in genetic structure due to inbreeding and genetic drift. Genetic characterization of the founder population using suitable molecular markers may help monitor periodic changes in the genetic structure in future. To develop benchmark information about the genetic structure we analyzed six microsatellite loci in the Brodbank collections of rohu (Labeo rohita) originated from three major rivers-the Jamuna, the Padma and the Halda. A total of 28 alleles were detected in 90 individuals with an average of 4.6 alleles per locus. The average observed heterozygosity ranged from 0.655 to 0.705 and the expected heterozygosity ranged from 0.702 to 0.725. The mean F IS values were 0.103, 0.106 and 0.018 for the Jamuna, Padma and Halda fishes respectively. The population pair-wise F ST values ranged from 0.0057 to 0.0278. Structure analysis grouped the fishes of the three rivers into two clusters. The numbers of half-sib families were 5, 5 and 4 and the numbers of full-sib families were 12, 10 and 18 for the Halda, Jamuna and the Padma samples respectively. Bottleneck was detected in all the river samples. We recommend to collect more fish from different locations of the major rivers to broaden the genetic variability of the founder stocks of the Brood bank.

Novel expression of importin alpha homologue in marine teleost, Pagrus major.

Importin alpha proteins are critical modulators of the classical nuclear protein import pathway. Although the physiological roles of importin alpha have been extensively studied in invertebrates and mammals, very little is known about their counterparts in lower vertebrates. In this study, to elucidate the roles of importin alpha in a teleost species, we isolated and characterized red seabream (Pagrus major) importin alpha cDNA derived from ovary and found changes in the mRNA levels of importin alpha in male and female red seabream during sexual maturation. The 1846-bp cDNA encodes a 520 amino acid protein that includes the importin beta-binding domain, a short acidic domain, and an armadillo (arm) repeat domain. Northern blot analysis and reverse transcription-polymerase chain reaction (RT-PCR) showed transcription of red seabream importin alpha in testis and ovary but not in the other tissues. The importin alpha mRNA levels in males increase in association with testicular development, whereas those in females remain high throughout sexual maturation. These findings suggest that red seabream ovary-derived importin alpha may be controlled in a tissue-specific manner and may perform unique functions in the gonad in addition to its involvement in nuclear transport.

Genetic structure of different populations of walking catfish (Clarias batrachus L.) in Bangladesh.

Information on genetic variation is essential for conservation and stock improvement programs. Seven dinucleotide microsatellite loci were analyzed to reveal genetic variability in three wild populations (Kella beel, Hakaluki haor, and Shobornokhali beel) and one hatchery population of the freshwater walking catfish, Clarias batrachus, in Bangladesh. Upon PCR amplification, the alleles were separated on polyacrylamide gel using a sequencing gel electrophoresis system and visualized by the silver-staining method. The loci were polymorphic (P95) in all the populations. Differences were observed in number and frequency of alleles as well as heterozygosity in the studied populations. Current gene diversity (He) was higher than expected under mutation-drift equilibrium, significantly in the Hakaluki haor and Shobornokhali beel populations, indicating a recent genetic bottleneck. Population differentiation (FST) values were significant (P<0.05) in all the population pairs. A relatively high level of gene flow and a low level of FST values were found between wild population pairs compared to hatchery-wild pairs. The unweighted pair group method with averages dendrogram based on genetic distance resulted in two major clusters: the hatchery population was alone in one cluster whereas the three wild populations made another cluster. The results reflect some degree of genetic variability in C. batrachus populations indicating potentialities for improving this species through a selective breeding program. The results revealed a recent bottleneck in some wild populations of C. batrachus. Protection of habitat may help increase the population size and lower the risk of vulnerability of the species in the future.

High levels of genetic variability and differentiation in hilsa shad, Tenualosa ilisha (Clupeidae, Clupeiformes) populations revealed by PCR-RFLP analysis of the mitochondrial DNA D-loop region

The hilsa shad, Tenualosa ilisha (Clupeidae, Clupeiformes) is an important anadromous clupeid species from the Western division of the Indo-Pacific region. It constitutes the largest single fishable species in Bangladesh. Information on genetic variability and population structure is very important for both management and conservation purposes. Past reports on the population structure of T. ilisha involving morphometric, allozyme and RAPD analyses are contradictory. We examined genetic variability and divergence in two riverine (the Jamuna and the Meghna), two estuarine (Kuakata and Sundarbans) and one marine (Cox's Bazar) populations of T. ilisha by applying PCR-RFLP analysis of the mtDNA D-loop region. The amplified PCR products were restricted with four restriction enzymes namely, XbaI, EcoRI, EcoRV, and HaeIII. High levels of haplotype and gene diversity within and significant differentiations among, populations of T. ilisha were observed in this study. Significant F ST values indicated differentiation among the river, estuary and marine populations. The UPGMA dendrogram based on genetic distance resulted in two major clusters, although, these were subsequently divided into three, corresponding to the riverine, estuarine and marine populations. The study underlines the usefulness of RFLP of mtDNA D-loop region as molecular markers, and detected at least two differentiated populations of T. ilisha in Bangladesh waters.