| Summary: | The blackwater fishes are frequently difficult to identify and differentiate within species due to the lack of well-defined characters for each species. The aim of this study is to use the cytochrome b (cyt b) gene to evaluate phylogenetic analysis on blackwater fishes in a peat swamp forest. A total of 23 blackwater fish were collected from Raja Musa Peat Swamp Forest in Bestari Jaya, Selangor. From the total number of individuals examined for the cyt b gene, eight species were identified, Helostoma temminkii, Hemibagrus capitulum, Mystus singaringan, Leiocassis poecilopterus, Leiocassis poecilopterus, Clarias meladerma, Clarias sp. and Belontia hasselti, with a range of similarity of 91.47 % to 99.64% in the NCBI (National Center for Biotechnology Information) database. This study also shows that the cyt b gene is still a conserved region and allows for molecular species identification. The genetic variation among blackwater fish species was determined by generating genetic distance with Molecular Evolutionary Genetics Analysis X (MEGA-X). The finding revealed that the nearest species is between Clarias sp and Clarias meladerma (5.2%) due to similar family, and the highest genetic distance (23.9%) was between Helostoma temminkii and Clarias sp due to distinct morphological characteristics. Based on the phylogenetic tree, two distinct clades were formed from the tree, with Myotis sp. acting as the basal of the tree, proving that this species is not a blackwater fish. This study concludes that the cyt b gene is suitable for molecular identification of blackwater fish species. © 2023 Author(s).
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