Study of Cytochrome B Gene Variations in Avian Fauna of Sargodha

Pakistan is home to approximately 765 bird species, but molecular-level studies for species identification remain scarce. The Sargodha region, with its agricultural land, canal systems, and hilly terrain, provides ideal habitats for various bird species. A total of 34 species belonging to 29 different families have been morphologically identified in this area. However, morphological methods alone are often insufficient for accurate species identification, which is crucial for conservation, taxonomy, and ecological research. This study aimed to identify avian species using molecular tools, specifically the cytochrome b gene, which is widely used for phylogenetic and evolutionary studies due to its conserved nature within species and variation among species. Tissue samples from eight bird species (totaling 24 samples) were collected from various locations in Sargodha through local hunters. DNA was extracted using the Russell and Sambrook method, and cytochrome b was amplified using PCR. The PCR products were sequenced at Macrogen Laboratory. Sequence analysis was performed using BioEdit, and phylogenetic relationships were assessed using MEGA6, employing methods such as Maximum Likelihood (ML), Maximum Parsimony (MP), Neighbor Joining (NJ), Minimum Evolution (ME), and UPGMA. All tree-building methods consistently revealed the phylogenetic relationships between and within species.The results confirmed significant genetic diversity among the bird species of Sargodha. Homogeneity and heterogeneity across sequences were also identified and recorded in tabular format. The study demonstrated that cytochrome b is a reliable genetic marker for species differentiation and phylogenetic analysis. Additionally, the organic DNA extraction method proved to be efficient and economical. Overall, this research highlights the effectiveness of molecular tools in avian identification and underlines the rich avian biodiversity present in the Sargodha region, emphasizing the importance of integrating genetic methods into regional biodiversity assessments.