Molecular appraoch for identification of unknown species by using Cytochrome b gene is an effective and reliable as compared with morphological based identification. For DNA barcoding universal molecular genes were used to identify the species. Cytochrome b is a specific gene used for identification purpose. DNA barcoding is a reliable and effective method compared to the different traditional morphological methods of specie identification. So,in the present study which was conducted to identify the species, a total of 50 fish samples were collected from five different sites. DNA was extracted by using the Phenol Chloroform method from muscle tissue. Five sequences were sequenced (one from each site), analyzed, and identified specific species as Pangasius pangasius. Identified sequences were variable in length from 369 bp (Site 1), 364 bp (Site 2), 364 bp (Site 3), 352 bp (Site 4), and 334 bp (Site 5). Identity matches on the NCBI database confirmed the specific specie as P. pangasius. A distancing tree was drawn to show maximum likelihood among the same and different species. Yet, in many cases fishes on diverse development stages are difficult to identify by morphological characters. DNA-based identification methods offer an analytically powerful addition or even an alternative tool for species identification and phylogenetic study. This work intends to provide an updated and extensive overview on the DNA based methods for fish species identification by using Cytochrome b gene as targeted markers for identification purpose.

Migratory birds globally face escalating anthropogenic threats, with crane species being particularly vulnerable. This study assessed human-perceived threats to cranes (Grus virgo & Grus grus) along Pakistan’s vital Indus Flyway using 400 stakeholder questionnaires across eight districts (2021–2024). We quantified perceived threat based on frequency (1 = Very Rare; 5 = Very Frequent) and severity (1 = Not Severe; 5 = Extremely Severe), revealing poaching (illegal killing) as the dominant threat (frequency = 4.9; severity = 4.8), followed by illegal wildlife trade (4.7; 4.5) and taming (4.6; 4.3). Spatial analysis showed strikingly higher perceived threats in southern Pakistan (Rajanpur: frequency = 4.88, severity = 4.82) versus central regions (Khushab: 3.76, 4.02; p < 0.001), with riverbanks identified as high-risk poaching zones (42 incidents). Cluster analysis also confirmed Rajanpur as a critical hotspot within three distinct threat tiers. Critically, analysis of socio-demographic drivers revealed threat type (frequency: F = 104.92, p < 0.001; severity: F = 153.64, p < 0.001) and poaching method (frequency: F = 10.14, p < 0.001; severity: F = 15.43, p < 0.001) as significant perception-shapers, while education, occupation, and crane species preference (frequency: F = 1.17, p = 0.310) exerted a non-significant influence. These results highlight that individual backgrounds minimally modulate threat perceptions. The study aligns with global evidence of uniform crane threats demanding the following urgent conservation action: region-specific enforcement (especially southern hotspots), community-led anti-poaching initiatives, and targeted awareness programs to shift high-threat communities toward crane-friendly coexistence practices.