Global genetic diversity and phylogenetic analysis of Pseudolynchia canariensis based on the COI gene with first molecular detection of Wolbachia (wsp and 16S rRNA) from Iran.

Pseudolynchia canariensis (Diptera: Hippoboscidae) is a hematophagous ectoparasite of pigeons with recognized veterinary and epidemiological importance due to its association with avian hemoparasites and bacterial microorganisms. Despite its wide geographic distribution, molecular data remain limited in several regions, including Iran, and Wolbachia infection has not previously been documented in Iranian populations of this species. The present study aimed to molecularly confirm P. canariensis from southern Iran, evaluate global genetic diversity based on mitochondrial COI sequences, and screen for Wolbachia infection. A total of 100 adult flies were collected from five districts of Shiraz, Iran, and identified using morphological characters and COI barcoding. Phylogenetic relationships were reconstructed using Maximum Likelihood inference, and global diversity indices were calculated from a comparative COI dataset comprising 45 sequences trimmed to a 261 bp overlapping region. Genetic diversity analysis revealed moderate haplotype diversity (Hd = 0.736) and low nucleotide diversity (π = 0.00773) at the global level, with no statistically significant deviation from neutrality. The overall mean evolutionary divergence was low (K2P = 0.01 substitutions per site), indicating limited mitochondrial differentiation among global populations. Wolbachia infection was detected in pooled samples from two districts using both wsp and 16S rRNA markers. Phylogenetic analyses of both markers consistently placed Iranian Wolbachia sequences within supergroup B. These findings represent the first molecular evidence of Wolbachia infection in P. canariensis from Iran and provide baseline genetic data supporting future investigations into host-symbiont interactions and the potential epidemiological role of this ectoparasitic fly.