Molecular detection of vector-borne pathogens from mosquitoes collected in two zoological gardens in Germany.

In Germany, knowledge of disease agents transmitted by arthropods in zoological gardens is scarce. In the framework of ecological studies, mosquitoes were therefore collected in German zoological gardens and examined for mosquito-borne pathogen DNA and RNA. In total, 3840 mosquitoes were screened for filarial nematodes and three groups of viruses (orthobunyaviruses, flaviviruses, alphaviruses) while 405 mosquitoes were tested for avian malaria parasites. In addition to the filarial nematode species Dirofilaria repens (n = 1) and Setaria tundra (n = 8), Sindbis virus (n = 1) and the haemosporidian genera Haemoproteus (n = 8), Leucocytozoon (n = 10) and Plasmodium (n = 1) were demonstrated. Identified pathogens have the potential to cause disease in zoo and wild animals, but some of them also in humans. Positive mosquitoes were collected most often in July, indicating the highest infection risk during this month. Most of the pathogens were found in mosquito specimens of the Culex pipiens complex, suggesting that its members possibly act as the most important vectors in the surveyed zoos, although the mere demonstration of pathogen DNA/RNA in a homogenised complete mosquito is not finally indicative for a vector role. Outcomes of the study are not only significant for arthropod management in zoological gardens, but also for the general understanding of the occurrence and spread of mosquito-borne disease agents.

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.

Mosquito species composition and phenology (Diptera, Culicidae) in two German zoological gardens imply different risks of mosquito-borne pathogen transmission.

Due to their large diversity of potential blood hosts, breeding habitats, and resting sites, zoological gardens represent highly interesting places to study mosquito ecology. In order to better assess the risk of mosquito-borne disease-agent transmission in zoos, potential vector species must be known, as well as the communities in which they occur. For this reason, species composition and dynamics were examined in 2016 in two zoological gardens in Germany. Using different methods for mosquito sampling, a total of 2,257 specimens belonging to 20 taxa were collected. Species spectra depended on the collection method but generally differed between the two zoos, while species compositions and relative abundances varied seasonally in both of them. As both sampled zoos were located in the same climatic region and potential breeding sites within the zoos were similar, the differences in mosquito compositions are attributed to immigration of specimens from surrounding landscapes, although the different sizes of the zoos and the different blood host populations available probably also have an impact. Based on the differences in species composition and the various biological characteristics of the species, the risk of certain pathogens to be transmitted must also be expected to differ between the zoos.

The Nuisance Mosquito Anopheles plumbeus (Stephens, 1828) in Germany-A Questionnaire Survey May Help Support Surveillance and Control.

The mosquito species Anopheles plumbeus is an aggressive biter and a potential vector of malaria parasites and West Nile virus. It occurs naturally at low population densities, as its larval development is adapted to the specific water qualities found in tree holes. However, probably owing to environmental changes, it has recently been observed in several European countries to use increasingly often artificial breeding habitats that may lead to mass development and severe annoyance to humans living close by. The perception of mosquito nuisance, however, is very subjective, and breeding habitats are not always known, thus impeding targeted surveillance and control. To relate nuisance by An. plumbeus to specific environmental conditions, a questionnaire survey was carried out addressing persons who had submitted specimens of this particular mosquito species to the German citizen science project "Mueckenatlas", an instrument of passive mosquito surveillance. The questionnaire was intended to find out whether a nuisance situation linked to An. plumbeus had existed, whether mosquito breeding habitats could be identified and whether control measures had been conducted. Despite some efforts, the participants who claimed to suffer from an An. plumbeus nuisance problem had rarely identified the source of the mosquitoes. Once control measures had been performed on abandoned manure pits, however, the nuisance problem disappeared or mosquito abundance was at least significantly reduced. Nevertheless, no significant effect of abandoned manure pits on the probability of an An. plumbeus nuisance could be demonstrated in a multivariate logistic regression model testing various variables. Instead, a significant positive effect of a disused farm nearby was found. The reason is probably that manure pits as the most frequent source of An. plumbeus mass development are often located on disused farms, without most people's knowledge about their existence. Disused farms are therefore appropriate candidates to consider when it comes to public health issues connected to An. plumbeus such as surveillance of mass development and implementation of control measures.

Anopheles plumbeus (Diptera: Culicidae) in Germany: updated geographic distribution and public health impact of a nuisance and vector mosquito.

OBJECTIVES: The aim of this study was to map the current spatial distribution of Anopheles plumbeus in Germany, a potential vector of malaria parasites and West Nile virus. Reports of mass occurrence and nuisance connected with artificial breeding site usage by this species were analysed. METHODS: Distribution data were collected from 2011 to 2014 mainly through trapping and submissions of adult mosquito specimens to a citizen science project. In the framework of the latter, additional information was gathered on recent nuisance incidents caused by An. plumbeus, including a longitudinal analysis of mosquito occurrence and the impact of management measures at a nuisance site in south-western Germany. RESULTS: Based on the most comprehensive set of collection data obtained during the last decades, An. plumbeus is shown to be widely distributed over Germany. The data also indicate a continuing extension of the breeding site repertoire of the species from natural to artificial habitats that facilitate mass development. Increasing incidents of persistent nuisance suggest that this mosquito species is rarely diagnosed correctly and managed adequately. CONCLUSIONS: As An. plumbeus is both a serious nuisance pest and a potential vector species, awareness of this species and the public health problems linked to it should be raised among pest managers and public health personnel.