Genetic diversity, phylogenetic and phylogeographic analysis of Anopheles culicifacies species complex using ITS2 and COI sequences

Anopheles culicifacies is the major vector of malaria in Sri Lanka and the Indian subcontinent which is characterized as a species complex with five sibling species provisionally designated as A, B, C, D and E. The current study was carried out to understand the phylogenetic and phylogeographic relationships between the sibling species of the species complex while observing their genetic diversity and genetic differentiation. Thirty-five ITS2 and seventy-seven COI sequences of An. culicifacies species complex reported from different geographical locations of Asia and China at the NCBI public database were used for the analysis. Bayesian likelihood trees were generated for the phylogenetic analysis. The divergence of the species complex was obtained from the Bayesian phylogeographic model in BEAST. There were two clades of the sibling species of An. culicifacies species complex as A, D and B, C and E in both phylogenetic and phylogeographic analysis using ITS2 sequences. Based on the highly divergent COI sequences and the high mutation rate of the mitochondrial genome, there were four and three clades in both phylogenetic and phylogeographic analysis using COI sequences. The diversification of An. culicifacies species complex was obtained as ranging from 20.25 to 24.12 Mya and 22.37 to 26.22 Mya based on ITS2 and COI phylogeographic analysis respectively. There was a recent diversification of the sibling species A and D than the sibling species B, C and E. Low haplotype diversity was observed in the sequences reported from Sri Lanka in both ITS2 and COI analysis that can be due to bottlenecks resulting from the intense malaria control efforts. A high genetic differentiation was achieved for some populations due to the large geographical distance. The high genetic diversity based on the five sibling species implies the possibility of maintaining a relatively high effective population size despite the vector control efforts.