Identification and DNA barcoding of Tabanid flies in the pasture area of sheep at Mannavanur, Palani Hills, Tamil Nadu, India

The present study was carried out to identify the tabanid flies creating annoyance to sheep in the grazing area of SRRC, Mannavanur by means of morphological keys and DNA barcoding targeting mitochondrial cytochrome c oxidase subunit I (COI) gene. Two different kinds of tabanid flies were caught from the pasture area by graziers during the months of May & June 2021. With the help of Dept. of Agricultural Entomology, Centre for Plant Protection studies, Tamil Nadu Agricultural University, Coimbatore, Tamil Nadu, India and Zoological Survey of India, Kolkata, West Bengal, India, it was identified that Haematopota nathani (Cleg fly) and Tabanus subcinerascens (Horse fly) were the two dipteran tabanid flies causing annoyance and aching bite in sheep and shepherds. The total genomic DNA isolated from the flies were subjected to COI gene based PCR assay using the universal primer set and the resultant PCR amplified DNA fragments were cloned into E.coli based vector. The confirmed recombinant plasmids were subjected for gene sequencing protocol through outsourcing. Upon the BLAST analysis, the nucleotide sequences obtained from the flies identified by TNAU and ZSI, were having the highest identity with COI gene of Haematopota sp and Tabanus sp., respectively. The obtained nucleotide sequences were analysed using the standard bioinformatics tools. Small number of fly specimens and the analysis based on the partial nucleotide sequences of COI gene are the limitations. It was concluded that the documentation of H. nathani and T. subcinerascens as well as barcoding of both tabanid flies from Palani hills, Tamil Nadu, India, based on mitochondrial COI DNA marker had been carried out for the first time.