Molecular characterization of scarab beetles (Coleoptera: Scarabaeidae) from the Malakand division, Pakistan

Dung beetles (Coleoptera: Scarabaeidae) play a pivotal role in agroecosystem functioning by promoting nutrient recycling, enhancing soil aeration, and suppressing dung-breeding pests. Despite their ecological significance, molecular data on regional scarab beetle fauna in Pakistan remain limited. This study provides a comprehensive molecular characterization of scarab beetles from Malakand division, Khyber Pakhtunkhwa, Pakistan, integrating traditional morphological identification with mitochondrial Cytochrome c Oxidase subunit I (COI) DNA barcoding. Specimens representing eight species, Onitis falcatus, O. philemon, Digitonthophagus gazella, Onthophagus nuchicornis, Copris lunaris, Catharsius molossus, and Oniticellus cinctus, were collected using standardized dung-baited pitfall traps and manual sampling. COI sequences were generated, submitted to GenBank, and analyzed using the Maximum Likelihood method under the Kimura 2-Parameter model with 1,000 bootstrap replicates. Phylogenetic analyses showed strong concordance between morphological and molecular identifications, with all species resolved as well-supported monophyletic clades. Intraspecific genetic divergence was generally low, whereas interspecific divergence was substantial, confirming clear taxonomic boundaries. Some taxa exhibited evidence of geographic structuring, suggesting the influence of regional isolation on genetic differentiation. This study establishes a reliable COI barcode reference library for scarab beetles of Malakand division and highlights the importance of molecular tools in biodiversity assessment, conservation planning, and sustainable agroecosystem management.