Identification of Labeo rohita, L. bata and L. gonius is sometimes problematic when usual morphological features are lost and it is difficult to differentiate them with traditional morphological features at their diverse developmental stages. PCR-sequencing provides an authentic alternative means of identification of individuals at species level. Three local carp fishes were collected and 16S rRNA genes were sequenced by sanger sequencing method after PCR amplification using universal primers. Obtained sequences were found accurate with blast search result which showed maximum range of similarity with the existing respective gene fragments present in GenBank database. Sequences were compared and multiple sequence alignment has revealed some polymorphic sites which can be used to differentiate these three species from one another. This study may provide valuable understanding to study their population in future. Dhaka Univ. J. Biol. Sci. 26(2): 167-174, 2017 (July)
Copy’s frog, commonly observed in Bangladesh, is ecologically important for evergreen forest. Sample was collected from Satchari National park, Hobigonj, Sylhet. Attempts were made to combine morphological and molecular techniques for identification and analysis of evolutionary relation of Copy’s frog with anuran species. Morphological identification of Hydrophylax sp. was conducted based on distinguishing characteristics of the body and different body measurement parameter. The species was identified morphologically as Hydrophylax leptoglossa based on the taxonomic key of finger toes and formula was 3>1>4>2. Simultaneously genomic DNA was isolated using CTAB extraction method. The 16S rRNA gene was amplified from the genomic DNA by polymerase chain reaction method using the pair of universal primers. Amplified product was a 308 bp fragment that generated AT/GC ratio of 1.08% once sequenced. In the present study comparative analysis of intraspecific and interspecific variation was conducted based on nucleotide sequence of 16S rRNA gene collected from GenBank. The intraspecific variation compares between Southeast Asia and Bangladesh was 1.29% for 16S rRNA gene. The interspecific genetic divergence was studied between H. leptoglossa and H. tytleri was observed 14.29%. The nucleotide sequence of the present study will be used as DNA barcode for that particular individual collected from Sylhet, Bangladesh. Furthermore, a molecular phylogenetic tree was constructed, where all the anuran species from a monophyletic group. Four species of Hydrophylax form monophyletic group and is consistent with relation based on morphology. Bangladesh J. Zool. 49(1): 105-115, 2021
Global amphibian populations are facing a novel threat, chytridiomycosis, caused by the fungus Batrachochytrium dendrobatidis (Bd), which is responsible for the severe decline of a number of species across several continents. Chytridiomycosis in Asia is a relatively recent discovery yet there have been no reports on Bd-presence in Bangladeshi amphibians. We conducted a preliminary study on 133 wild frogs from seven sites in Bangladesh between April and July 2018. Nested PCR analysis showed 20 samples (15.04%) and 50% of the tested taxa (9 species from 6 genera and 4 families) as Bd-positive. Eight of the nine species are discovered as newly infected hosts. Analysis of Bd-positive samples shows prevalence does not significantly vary among different land cover categories, although the occurrence is higher in forested areas.The prevalence rate is similar in high and low disturbed areas but the range of occurrence is statistically higher in low disturbance areas. Maximum entropy distribution modelling indicates high probabilities of Bd occurrence in hilly and forested areas in south-east and central-north Bangladesh. The Bd-specific ITS1-5.8S-ITS2 ribosomal gene sequence from the Bd-positive samples tested is completely identical. A neighbor-joining phylogenetic tree reveals that the identified strain shares a common ancestry with strains previously discovered in different Asian regions. Our results provide the first evidence of Bd-presence in Bangladeshi amphibians, inferring that diversity is at risk. The effects of environmental and climatic factors along with quantitative PCR analysis are required to determine the infection intensity and susceptibility of amphibians in the country.
Esterase isozyme of different tissues of Puntius sophore was analyzed using 7.5 % polyacrylamide gel electrophoresis (PAGE). Fish specimens were collected from local market. The tissues used were taken from different muscles, stomach, fore-, mid- and hind-gut, liver, eyes, gill, heart, kidney, fore-, mid- and hind- brain, eggs and ovary. Six different esterase bands were detected, named Est-1, Est-2, Est-3, Est-4, Est-5 and Est-6 and their relative mobility were 1.0, 0.84, 0.62, 0.33, 0.26 and 0.13 respectively, each of them representing a single allele. The highest esterase activity was found in liver, followed by gill, kidney, heart, brain, intestine, stomach, eye, reproductive organ and skeletal muscles as detected in the staining intensity. Staining intensity of Est-4 and Est-5 was higher and Est-6 was the least stained in all the tissues. Asiat. Soc. Bangladesh, Sci. 42(2): 201-208, December 2016
Heteropneustesfossilis, Clariasbatrachus and C. gariepinus are three major catfishes ofecological and economic importance. Identification of these fish species becomes aproblem when the usual external morphological features of the fish are lost or removed,such as in canned fish. Also, newly hatched fish larva is often difficult to identify. PCRsequencingprovides accurate alternative means of identification of individuals at specieslevel. So, 16S rRNA genes of three locally collected catfishes were sequenced after PCRamplification and compared with the same gene sequences available from othergeographical regions. Multiple sequence alignment of the 16S rRNA gene fragments ofthe catfish species has revealed polymorphic sites which can be used to differentiate thesethree species from one another and will provide valuable insight in choosing appropriaterestriction enzymes for PCR-RFLP based identification in future. Asiat. Soc. Bangladesh, Sci. 41(1): 51-58, June 2015
Attempts were made to combine morphological and molecular techniques for identification and analysis of evolutionary relationship of commercially important marine lizardfish (Bombay duck) from the Bay of Bengal. For molecular identification, mitochondrial DNA (mtDNA) was extracted using CTAB extraction protocol and 16S rRNA gene was amplified by PCR method. Morphological identification was supported by molecular data generated by 99% identity of BLAST search result of Harpadon nehereus. The nucleotide sequence of H. nehereus was 586 bp long and the G+C content was 51% and intraspecies polymorphic sites were 0.5%. According to interspecies polymorphic sites analysis, 4.9% polymorphism was observed when H. nehereus and H. microchir were compared. Phylogenetic tree was constructed and both the Harpadon spp. formed a monophyletic clade. This analysis will be used to study molecular diversity as well as conservation of this fish species. Bangladesh J. Zool. 47(1): 49-57, 2019
DNA barcoding has been proposed as a means of quick species identification using a short standardized segment of DNA. Two species (Eleotris fusca and Glossogobius giuris) from the family Gobiidae and Eleotridae were selected for DNA barcoding using samples collected from different regions of Bangladesh. Cytochrome Oxidase Subunit I (COI) gene was sequenced from two different gobi fishes and compared with two previously published similar sequences from the genera Eleotris and Glossogobius. Multiple sequence alignment and the molecular systematic study were performed. The DNA barcode technique identified the two species. The study provides a good example of how DNA barcoding can build upon its primary mission of species identification and use available data to integrate genetic variation investigated at the local scale into a global framework.
Work on two freshwater Gobi fish species such as Awaous guamensis and A. grammepomus was carried out on morphometric and meristic study for their morphological identification. Sequence data of mitochondrial cytochrome oxidase subunit I (COI) gene was used for this purpose. Around 620 bp sequences were obtained when universal primers were used. An alignment was performed between the sequences which showed some inter-specific variation. An entropy plot was generated for all the aligned positions which showed that the entropy rarely touches a scale of one, which is a sign of better alignment in the region. This work can be utilized for species identification, phylogenetic analysis, to study molecular diversity and evolution, and most importantly for the conservation of these fauna in near future. Dhaka Univ. J. Biol. Sci. 26(1): 83-90, 2017 (January)
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