Mosquitoes of the Anopheles maculipennis group (Diptera: Culicidae) in Romania, with the discovery and formal recognition of a new species based on molecular and morphological evidence

Abstract

Mosquitoes of the Anopheles maculipennis group were collected in five districts of Romania (Constant,a, Giurgiu, Ilfov, Mehedint,i and Suceava) between March 2000 and June 2003. Two hundred and ninety-seven specimens were identified by molecular methods. Nuclear rDNA ITS2 sequences of 178 specimens were compared with GenBank sequences for nine known Palaearctic species of the group, and 119 specimens were identified using an ITS2 PCR-RFLP assay developed during the study. Five genetically distinct species of the group were identified: A. atroparvus van Thiel, A. maculipennis Meigen, A. melanoon Hackett and A. messeae Falleroni and a previously unrecognized species. The new species, herein formally described and named A. daciae sp. n., was collected in the Black Sea coastal region and plains adjacent to the Danube River in southern Romania. Anopheles daciae is most similar to and sympatric with A. messeae. It is contrasted with A. messeae and characterized on the basis of unique nuclear ITS2 and mitochondrial COI DNA sequences and morphological characters of the eggs. The larval, pupal and adults stages of the two species were also compared, but no reliable characters were found to distinguish them. It seems likely that A. daciae is more widespread in eastern Europe and the Balkan States, and could be responsible for malaria transmission in these regions that is currently attributed to A. messeae. Anopheles melanoon is reported from Romania for the first time.

The ITS2 ribosomal DNA of Anopheles beklemishevi and further remarks on the phylogenetic relationships within the Anopheles maculipennis group of species (Diptera: Culicidae)

“…U60410)=outgroup) phylogenetic relationships. He added two more sibling species to the palaearctic branch of An maculipennis group found in Iran and in Romania: An persiensis (Sedaghat et al 2003) and An daciae (Nicolescu et al 2004). Moreover, An beklemishevi is not grouped together with the other palaearctic siblings in the 'maculipennis subgroup' but in the 'quadrimaculatus subgroup'.…”

Molecular approach for identification of mosquito species (Diptera: Culicidae) in Province of Alessandria, Piedmont, Italy

“…50 bp is almost not visible in lane 33-1 on Fig. 1, but other visible fragments are specific only for A. sacharovi (Nicolescu et al, 2004 atroparvus can suck blood all year round, but do not lay eggs in winter. Therefore, a correct identification of the vectors is essential for the development of control strategies.…”

“…sacharovi, however A. messeae, A. melanoon, A. atroparvus, and A. labranchia have a fragment of similar length (Marinucci et al, 1999). Recently, a complex of PCR-RFLP methods was developed to identify some cryptic species in the Anopheles maculipennis complex (Nicolescu et al, 2004;Ejov et al, 2008).…”

Re-Emergence of Malaria and Dengue in Europe

“…Moreover a new species of the An. maculipennis complex, named Anopheles daciae, was recently first described in Romania (Nicolescu et al, 2004). It seems likely that An.…”

“…Table 2. Anopheles species with endemic presence in Europe and indication of historical data about its vectorial role (Ramsdale & Snow, 2000;Schaffner et al, 2001;Beck et al, 2003;Nicolescu et al, 2004;Linton et al, 2005;Becker et al, 2010;European Mosquito Taxonomists [MOTAX], 2010).…”