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DNA barcoding based on the molecular techniques has become a highly promising method for identification of species from non-invasive samples. This study presents the preliminary findings on generating robust DNA barcodes of Accipitridae family raptors of Tamilnadu using their molted feathers. Molted feathers of five different birds collected were subjected to extraction of DNA and sequencing of mitochondrial COI gene, widely accepted as DNA barcode region with approximately 650 bp in length. BLAST analysis of the COI gene was performed to identify the five molted feathers to the species level. Phylogenetic relationship of all the five samples with other 7 vulture sequences from NCBI were analysed using MEGA software based on the Neighbour Joining (NJ) method and the interspecies distances were calculated based on the Kimura 2 parameter model. The dataset comprising the DNA barcode of five species were identified as four raptors included in the family Accipitridae. The species level identification revealed the raptors were Gyps bengalensis, Haliastur indus, Elanus careleus, Milvus migrans and Hierrococcyx varius of Cuculidae family. Phylogenetic relationship determined based on the phylogenetic tree represented raptors in a clustered manner and Hierrococcyx varius which belongs to the different family was branched into a different group. Further, the interspecies distances based on Kimura 2 parameter model also supported the results. The average interspecies sequence divergence between the 12 species was 0.13 (+0.01). The present study had exhibited that DNA barcoding had discriminating power to delineate boundaries among the closely related bird species even using molted feathers as sample. The data generated are of high importance to the law enforcement agencies in effective identification of bird species where physical observation is challenging. The similar approach can be utilized for generating DNA barcodes for other endangered birds with critical ecosystem roles.
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