The diagnostic utility of sequence-based assays for the molecular delimitation of the epidemiologically relevant Culex pipiens pipiens taxa (Diptera: Culicidae).

The northern house mosquito (Culex pipiens pipiens L.) is a vector of several important pathogens and comprises two epidemiologically distinct ecotypes (molestus Forskål and pipiens). The delimitation of its ecotypes is a crucial, yet controversial step in vector surveillance due to varying diagnostic values of different characters. Therefore, we reviewed the success of a diagnostic assay based on the mitochondrial cytochrome c oxidase subunit I locus (COI) by analyzing previously published sequences of molestus and pipiens sampled in different geographical areas. Next, by genotyping individuals from Northern Serbia at this locus, we additionally assessed whether genetic structure of urban and rural Cx. p. pipiens ecotypes corresponded to the admixture pattern. Finally, to account for the different susceptibility of genetic markers to introgression, we also analyzed genetic structuring based on the ribosomal internal transcribed spacer 2 (ITS2). No latitude-dependent differentiation of Cx. p. pipiens ecotypes was found at a global level, with the COI assay further failing to accurately identify molestus and pipiens ecotypes. Likewise, both individual- (BAPS) and population-based (analysis of molecular variance and FST estimates) methods showed no significant urban/rural genetic differentiation in Serbia, indicating unhindered gene flow between different Cx. p. pipiens habitat types. The findings challenge the previous instances of Cx. p. pipiens ecotype identification, while also spotlighting the vectorial capacity of their hybrid offspring.

Multi-character approach reveals a discordant pattern of phenotypic variation during ontogeny in Culex pipiens biotypes (Diptera: Culicidae).

Culex (Culex) pipiens s.l. (Diptera: Culicidae) comprises two distinct biotypes, pipiens ('rural') and molestus ('urban'), both of which are thought to have differing capacities due to different host preferences. To better understand West Nile encephalitis epidemiology and improve risk assessment, local distinction between these forms is essential. This study assesses phenotypic variation at larval and adult stages of 'urban' and 'rural' biotypes of the species by complementary use of meristic, univariate and multivariate traits analyzed by traditional and geometric morphometrics. Third- and fourth-instar larvae from a broad area of the city of Novi Sad (Serbia) were collected and reared in the laboratory. After adult eclosion, the sex of each larva was recorded based on the sex of the corresponding adult. Examination of the association between variations of larval traits revealed contrasting variations regarding pecten spines vs. siphonal size and siphonal shape in the 'rural' biotype. Siphons of larvae collected in marshes and forest ecosystems outside urban areas were found to be the largest, but possessed the smallest number of pecten spines. In addition, statistically significant female-biased sexual dimorphism was observed in siphonal size, wing size and wing shape. Finally, we propose that an integrative approach is essential in delimitation of Cx. pipiens s.l. biotypes, since their differentiation was not possible based solely on larval and adult traits. Our findings shed light on the phenotypic plasticity important for population persistence in the changing environment of these medically important taxa.

West Nile virus surveillance in mosquitoes, April to October 2013, Vojvodina province, Serbia: implications for the 2014 season.

After the West Nile virus (WNV) outbreak in 2012, we collected mosquito samples from Vojvodina province, Serbia, in 2013. We found high WNV infection rate in two species, Culex pipiens and Anopheles maculipennis. Phylogenetic analysis showed that Serbian WNV strains from 2013 were most closely related to Italian and Greek strains isolated in 2012 and 2010, respectively. Public health authorities should be aware of a potentially increased risk of WNV activity during the 2014 season.