UTTAR PRADESH JOURNAL OF ZOOLOGY

This study assessed the molecular identity and phylogenetic placement of Garra mullya and Parambassis ranga from the Ujani Reservoir, Maharashtra, using mitochondrial cytochrome c oxidase subunit I (COI) sequences. Fish specimens were collected during 2022–2023, and fin or muscle tissues were preserved in 95% ethanol. Genomic DNA was extracted, the COI region was amplified using universal primers, and purified amplicons were subjected to bidirectional sequencing. Sequence identities were evaluated against the NCBI nucleotide database, and phylogenetic relationships were examined using comparative GenBank records. The Garra mullya sequences generated in the study clustered with records from Maharashtra, indicating close regional affinity and limited divergence within the sampled lineage. Sequences from other Indian regions formed adjacent but distinguishable subclusters, suggesting geographic structuring across river basins. Parambassis ranga showed greater phylogenetic subdivision, with one sequence associated with northeastern Indian records and two sequences grouped with Maharashtra records. These patterns indicate geographically structured lineages within the available dataset. However, the observed clustering should be interpreted cautiously because the number of locally generated sequences was limited and the analysis relied on a single mitochondrial marker. The study provides additional COI reference sequences for both species from the Ujani Reservoir and supports the use of molecular evidence alongside morphology, broader geographic sampling, and multilocus analyses for future taxonomic and conservation assessments.