Molecular confirmation of the occurrence in Germany of Anopheles daciae (Diptera, Culicidae)

Mandy Kronefeld,Doreen Werner,Helge Kampen,Marko Dittmann,Dorothee Zielke

doi:10.1186/1756-3305-5-250

Abstract

BackgroundAnopheles daciae, a newly described member of the Maculipennis group, was recently reported from western, southern and eastern Europe. Before its recognition, it had commonly been listed under the name of An. messeae, due to its extreme morphological and genetic similarities. As the sibling species of the Maculipennis group are known to differ in their vector competences for malaria parasites and other pathogens, the occurrence of An. daciae in a given region might have an impact on the epidemiology of mosquito-borne diseases. Mosquito collections from different localities in Germany were therefore screened for An. daciae.MethodsAdult and immature Maculipennis group mosquitoes were collected between May 2011 and June 2012 at 23 different sites in eight federal states of Germany. A standard PCR assay was used to differentiate the previously known sibling species while the ITS2 rDNA of specimens preliminarily identified as An. messeae/daciae was sequenced and analysed for species-specific nucleotide differences.ResultsFour hundred and seventy-seven Anopheles specimens were successively identified to Maculipennis group level by morphology and to species level by DNA-based methods. Four species of the Maculipennis group were registered: An. messeae (n = 384), An. maculipennis (n = 82), An. daciae (n = 10) and An. atroparvus (n = 1). Anopheles daciae occurred at four sites in three federal states of Germany, three of the sites being located in north-eastern Germany (federal states of Brandenburg and Saxony) while one collection site was situated in the northern Upper Rhine Valley in the federal state of Hesse, south-western Germany.ConclusionsThe detection of An. daciae represents the first recognition of this species in Germany where it was found to occur in sympatry with An. messeae and An. maculipennis. As the collection sites were in both north-eastern and south-western parts of Germany, the species is probably even more widely distributed in Germany than demonstrated, albeit apparently with low population densities. Research is needed that confirms the species status of An. daciae and elucidates its vector competence as compared to An. messeae and the other species of the Maculipennis group, in order to optimize management of possible future outbreaks of diseases caused by pathogen transmission through Maculipennis group mosquitoes.

Highlights

Anopheles daciae, a newly described member of the Maculipennis group, was recently reported from western, southern and eastern Europe
The recognition of sibling species within the Maculipennis group of the culicid genus Anopheles in the early 20th century and of their different roles as vectors of malaria parasites was a historical milestone in malaria research [1,2]
Mosquitoes belonging to the Maculipennis group were further identified by a speciesspecific polymerase chain reaction (PCR) assay [8] performed on DNA extracted from whole single specimens using the DNeasy Blood & Tissue Kit (Qiagen, Germany) and the NucleoSpin RNA Virus Kit (Macherey-Nagel, Germany) according to the instruction manuals

Summary

Introduction

A newly described member of the Maculipennis group, was recently reported from western, southern and eastern Europe Before its recognition, it had commonly been listed under the name of An. messeae, due to its extreme morphological and genetic similarities. The recognition of sibling species within the Maculipennis group of the culicid genus Anopheles in the early 20th century and of their different roles as vectors of malaria parasites was a historical milestone in malaria research [1,2] It triggered in-depth research on the biology artemievi, a mosquito species described in 2005 [4], the Palaearctic members of the Maculipennis group, including An. atroparvus, An. labranchiae, An. maculipennis, An. melanoon, An. messeae, An. sacharovi, An. artemievi, An. martinius and An. persiensis, form the Maculipennis subgroup. Using that PCR assay, An. daciae is erroneously identified as An. messeae and remains unrecognized

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