Species composition, geographical distribution and seasonal abundance of the Anopheles maculipennis complex along the Upper Rhine, Germany.

Between May and September 2016, mosquitoes were collected on a biweekly basis at 55 locations with CO-baited encephalitis vector surveillance traps along the Upper Rhine, Germany, to evaluate the species composition, geographical distribution and abundance of the Anopheles maculipennis complex, some members of this complex being considered vectors of historical malaria in Germany. A total of 2115 Anopheles maculipennis complex specimens were collected during the season, of which a sample of 1252 individuals was determined to species level by amplification of species-specific internal transcribed spacer 2 (ITS2) sequences. A total of 856 individuals of Anopheles daciae (68.37%), 394 Anopheles messeae (31.47%) and 2 Anopheles maculipennis s.s. (0.16%) were recorded. The number and proportion of A. daciae was remarkably higher in the northern meandering zone of the Upper Rhine (843 specimens, 79.90%), than in the more canalised southern furcation zone where A. messeae with 183 collected specimens represented 92.89% of 197 classified individuals. The average number of collected A. maculipennis s.l. individuals per trapping site was 38.45, equalling 0.64% of the total mosquito collection. Despite an increase in imported malaria cases, this comparatively low abundance of A. maculipennis s.l. may indicate a low risk of endemic malaria transmission by members of the A. maculipennis complex today. The proportionally dominance of A. daciae suggests that this species could be suspected the main historical vector of malaria in the Upper Rhine region. Sequence analyses of the ITS2 fragment revealed intraindividual polymorphisms within 3 of 5 diagnostic nucleotides in all specimens of A. daciae, raising the question if additional loci should be considered, to gain further insight into the taxonomical relation to A. messeae.