DNA identification of species of the Anopheles maculipennis complex and first record of An. daciae in Belgium.

N Smitz,W Dekoninck,S Gombeer,T Backeljau,M De Meyer,J De Witte,A Schneider,I Deblauwe,I Verlé,K Meganck,R Müller,W Van Bortel,K De Wolf,A Vanslembrouck,A Gheysen,A Vanderheyden

doi:10.1111/mve.12519

Abstract

The present study aimed at identifying the members of the Anopheles maculipennis complex (Diptera: Culicidae) occurring in Belgium. Therefore, the second internal transcribed spacer of nuclear ribosomal DNA (ITS2) and the mitochondrial cytochrome oxidase subunit I (COI) loci were sequenced in 175 and 111 specimens, respectively, collected between 2007 and 2019. In parallel, the suitability of two species‐diagnostic PCR‐RFLP assays was tested. The identified specimens included: An. maculipennis s.s. (N = 105), An. daciae (N = 62), An. atroparvus (N = 6) and An. messeae (N = 2). Each species was characterized by unique ITS2 haplotypes, whereas COI only supported the monophyly of An. atroparvus, a historical malaria vector in Belgium. Species identification results were further supported by unique PCR‐RFLP banding patterns. We report for the first time An. daciae in Belgium, where it was found to co‐occur with An. maculipennis s.s. The latter was the most prevalent in the collection studied (60%) and appears to have the widest distribution in Belgium. As in other studies, An. daciae and An. messeae appeared the most closely related species, up to the point that their species status remains debatable, while their ecological differences, including vector competences, need further study.

Highlights

IntroductionIn the Paleartic region, ten cryptic species of the Anopheles maculipennis subgroup (Maculipennis complex) are presently recognized (Harbach, 2013), with An. messeae Falleroni, 1926 being the most widely distributed (Becker et al, 2020)
In the Paleartic region, ten cryptic species of the Anopheles maculipennis subgroup (Maculipennis complex) are presently recognized (Harbach, 2013), with An. messeae Falleroni, 1926 being the most widely distributed (Becker et al, 2020).Seven of these cryptic species are distributed throughout continental Europe, viz. An. atroparvus van Thiel, 1927, An. daciae Linton, Nicolescu & Harbach, 2004, An. labranchiae Falleroni, 1926, An. maculipennis s.s
Anopheles atroparvus specimens were all collected at the locations Kallo (N = 5) and Vrasene (N = 1) (Fig. 1) using the Mosquito Magnet and the BG-Sentinel traps over two successive years

Summary

Introduction

In the Paleartic region, ten cryptic species of the Anopheles maculipennis subgroup (Maculipennis complex) are presently recognized (Harbach, 2013), with An. messeae Falleroni, 1926 being the most widely distributed (Becker et al, 2020) Seven of these cryptic species are distributed throughout continental Europe, viz. An. atroparvus van Thiel, 1927, An. daciae Linton, Nicolescu & Harbach, 2004, An. labranchiae Falleroni, 1926, An. maculipennis s.s. Meigen, 1818, An. melanoon Hackett, 1934, An. messeae and An. sacharovi Favre, 1903. It is important to monitor the distribution of Anopheles species across Europe and investigate their link with human travel and climate change. This can only be achieved if Maculipennis species are correctly identified

Methods

Results

Conclusion