Molecular Identification of Tilapia Using Mitochondrial COI DNA Barcoding from Major Freshwater Reservoirs of Akola District, Maharashtra, India

Abstract

Abstract Accurate identification of tilapia species is essential for fisheries management, biodiversity conservation and invasive fish population monitoring. Oreochromis species can be difficult to differentiate morphologically because of phenotypic plasticity and overlapping morphometric and meristic traits. The present study employed a mitochondrial cytochrome c oxidase subunit I (COI) DNA barcoding technique to confirm the identity of representative tilapia specimens collected from important freshwater reservoirs in Akola District, Maharashtra, India. The COI gene was amplified using universal fish primers and bidirectional Sanger sequencing after genomic DNA was extracted using the phenol-chloroform method. MEGA version 11 was used to analyse the modified sequences, and BLAST was used to compare them to verified reference sequences from the NCBI GenBank database. The presence of both species in the reservoirs under investigation was confirmed by sequence analysis, which showed 100% similarity between sample FTIL001 and Oreochromis mossambicus and sample FTIL002 with Oreochromis niloticus. Beyond traditional morphological analysis, the produced COI barcode sequences offered trustworthy molecular evidence for species identification. The current investigation validates molecular identification of O. niloticus as well as O. mossambicus from the main freshwater reservoirs in Akola District and offers baseline data for the evaluation of fisheries resources, documentation of biodiversity, and long-term surveillance of invasive tilapia populations in central India.