Intraspecific genetic variation in the lymphatic filariasis vector Mansonia dives (Diptera: Culicidae) in Thailand: Hidden species or genetically divergent populations?

Mansonia dives is recognized as a vector for brugian filariasis in Thailand. A recent study analyzing the cytochrome c oxidase subunit I (COI) gene revealed two distinct clades within the Ma. dives population in Thailand. This study aimed to examine the genetic diversity and structure of Ma. dives using the COI gene and the internal transcribed spacer 2 (ITS2) region to determine the presence of distinct species or genetically divergent populations. We analyzed 60 COI and 60 ITS2 sequences from Ma. dives populations in Narathiwat, Ranong, Tak, and Trat. The results showed a nucleotide diversity of 0.019 and a haplotype diversity of 0.979 for the COI gene, while the ITS2 region displayed a nucleotide diversity of 0.005 and a haplotype diversity of 0.545. Phylogenetic and haplotype network analyses of the COI gene identified two genetic lineages: one confined to Trat and another encompassing the other sites. However, species delimitation methods suggested that these genetic differences were insufficient to classify the lineages as distinct species. In contrast, the ITS2 analysis indicated a uniform genetic pattern across all populations. We conducted neutrality tests and mismatch distribution to examine the demographic history. For the COI gene, Tajima's D was slightly positive and non-significant (0.014), while Fu's Fs was negative (-9.750), indicating a potential expansion phase. Conversely, for the ITS2 region, Tajima's D and Fu's Fs were positive and non-significant, suggesting that the population might be in equilibrium or undergoing contraction. Moreover, the mismatch distribution analysis for the ITS2 region was inconclusive. The apparent discrepancies between these markers indicate the presence of genetically divergent populations, rather than distinct species.