An Evaluation of Characters for the Separation of Two Culex Species (Diptera: Culicidae) Based on Material From the Upper Midwest

Mosquitoes (Diptera: Culicidae) in the Culex pipiens complex play a key role in the transmission and therefore epidemiology of a number of human and animal pathogens globally. These mosquitoes, and sympatric species of the genus Culex Linnaeus that are not within the Cx. pipiens complex, are often considered ‘impossible’ to distinguish by morphology in the adult female stage. In the United States, this is particularly true for Culex pipiens s.l. and Culex restuans Theobald, both of which are competent vectors of West Nile virus, but likely play different roles in the transmission cycle. Therefore, we undertook an in-depth morphological evaluation of matched larval exuviae and adult specimens that revealed five useful morphological characters that are informative to distinguish Cx. pipiens s.l. from Cx. restuans in the adult stage. Herein, we provide a comprehensive review of the literature on these species of interest, and four additional, morphologically similar, Culex species, and a proposed key to adult female specimens.

Ecology of West Nile Virus in the Danube Delta, Romania: Phylogeography, Xenosurveillance and Mosquito Host-Feeding Patterns

The ecology of West Nile virus (WNV) in the Danube Delta Biosphere Reserve (Romania) was investigated by combining studies on the virus genetics, phylogeography, xenosurveillance and host-feeding patterns of mosquitoes. Between 2014 and 2016, 655,667 unfed and 3842 engorged mosquito females were collected from four sampling sites. Blood-fed mosquitoes were negative for WNV-RNA, but two pools of unfed Culex pipiens s.l./torrentium collected in 2014 were tested positive. Our results suggest that Romania experienced at least two separate WNV lineage 2 introductions: from Africa into Danube Delta and from Greece into south-eastern Romania in the 1990s and early 2000s, respectively. The genetic diversity of WNV in Romania is primarily shaped by in situ evolution. WNV-specific antibodies were detected for 19 blood-meals from dogs and horses, but not from birds or humans. The hosts of mosquitoes were dominated by non-human mammals (19 species), followed by human and birds (23 species). Thereby, the catholic host-feeding pattern of Culex pipiens s.l./torrentium with a relatively high proportion of birds indicates the species’ importance as a potential bridge vector. The low virus prevalence in combination with WNV-specific antibodies indicate continuous, but low activity of WNV in the Danube Delta during the study period.

Detection of Usutu, Sindbis, and Batai Viruses in Mosquitoes (Diptera: Culicidae) Collected in Germany, 2011–2016

Due to the emergence of non-endemic mosquito vectors and the recent outbreaks of mosquito-borne diseases, mosquito-borne pathogens are considered an increasing risk to public and animal health in Europe. To obtain a status quo regarding mosquito-borne viruses and their vectors in Germany, 97,648 mosquitoes collected from 2011 to 2016 throughout the country were screened for arboviruses. Mosquitoes were identified to species, pooled in groups of up to 50 individuals according to sampling location and date, and screened with different PCR assays for Flavi-, Alpha- and Orthobunyavirus RNA. Two pools tested positive for Usutu virus-RNA, two for Sindbis virus-RNA, and 24 for Batai virus-RNA. The pools consisted of Culex pipiens s.l., Culex modestus, Culex torrentium, Culiseta sp., Aedes vexans, Anopheles daciae, and Anopheles messeae mosquitoes and could be assigned to nine different collection sites, with seven of them located in northeastern Germany. Phylogenetic analyses of the viral RNA sequences showed relationships with strains of the viruses previously demonstrated in Germany. These findings confirm continuing mosquito-borne zoonotic arbovirus circulation even though only a rather small percentage of the screened samples tested positive. With respect to sampling sites and periods, virus circulation seems to be particularly intense in floodplains and after flooding events when mosquitoes develop in excessive numbers and where they have numerous avian hosts available to feed on.