Evaluating perceptions of risk in mosquito experts and identifying undocumented pathways for the introduction of invasive mosquito species into Europe.

In several reported cases of the entry of invasive mosquito species (IMSs) into Europe, the introduction was associated with a specific pathway of introduction or dispersal. The identification of potential pathways for the introduction of IMSs and evaluations of the importance of the different pathways are key to designing proper surveillance strategies to promptly detect and control introductions in non-infested areas. The main goals of the present study were to identify other, previously undocumented, pathways of introduction into Europe, and to identify mosquito experts' perceptions regarding control measures against IMS introductions via different documented pathways. At the European Mosquito Control Association (EMCA) conference in Montenegro in March 2017, a questionnaire was distributed among meeting participants to collect expert data. Results showed that ground transportation (by cars, trucks, etc.), passive natural dispersal and the shipping of used tyres are perceived as the most likely pathways. Introduction via aircraft did not appear to be well known and was not perceived as probable. This study shows that there were no pathways unknown to European experts that could lead to cryptic introductions into the experts' countries. Furthermore, the findings demonstrated that the perceived efficacy of surveillance and control is key to overcoming the constraints experienced and to supporting the implementation of actions against introductions.

National Mosquito (Diptera: Culicidae) Survey in The Netherlands 2010-2013.

From 2010 onwards, a nationwide mosquito monitoring scheme has been conducted in The Netherlands with the aim of gaining crucial information about mosquito (Diptera: Culicidae) species composition, geographical distributions, biodiversity, and habitat preferences. The results of this study are based on 778 randomly sampled mosquito locations. These are divided into three main habitat types: urban, rural-agricultural, and natural areas. Twenty-seven mosquito species were found: 26 indigenous and 1 exotic, Aedes japonicus japonicus (Theobald, 1901). The preliminary results are presented here, with details of their species distribution and seasonality. Monitoring the temporal and spatial distribution of mosquitoes is an essential step in the risk analysis of emerging mosquito-borne diseases.

Real-time PCR Tests in Dutch Exotic Mosquito Surveys; Implementation of Aedes aegypti and Aedes albopictus Identification Tests, and the Development of Tests for the Identification of Aedes atropalpus and Aedes japonicus japonicus (Diptera: Culicidae).

Since 2009, The Netherlands Food and Consumer Product Safety Authority carries out surveys focusing on, amongst others, the presence of invasive mosquito species (IMS). Special attention is given to exotic container-breeding Aedes species Aedes aegypti (L.), Aedes albopictus (Skuse), Aedes atropalpus (Coquillett), and Aedes japonicus japonicus (Theobald). This study describes the implementation of real-time PCR tests described by Hill et al. (2008) for the identification of Ae. aegypti and Ae. albopictus, and the development of two novel real-time PCR tests for the identification of Ae. atropalpus and Ae. j. japonicus. Initial test showed that optimization of elements of the Ae. aegypti and Ae. albopictus tests was needed. Method validation tests were performed to determine if the implemented and newly developed tests are fit for routine diagnostics. Performance criteria of analytical sensitivity, analytical specificity, selectivity, repeatability, and reproducibility were determined. In addition, experiments were performed to determine the influence of environmental conditions on the usability of DNA extracted from mosquito specimens trapped in BG-Sentinel traps. The real-time PCR tests were demonstrated to be sensitive, specific, repeatable, reproducible, and are less prone to false negative results compared to partial cytochrome c oxidase I gene sequencing owing to the DNA fragmentation caused by environmental influences.

No evidence for the persistence of Schmallenberg virus in overwintering mosquitoes.

In 2011, Schmallenberg virus (SBV), a novel member of the Simbu serogroup, genus Orthobunyavirus, was identified as the causative agent of a disease in ruminants in Europe. Based on the current knowledge on arthropods involved in the transmission of Simbu group viruses, a role of both midges and mosquitoes in the SBV transmission cycle cannot be excluded beforehand. The persistence of SBV in mosquitoes overwintering at SBV-affected farms in the Netherlands was investigated. No evidence for the presence of SBV in 868 hibernating mosquitoes (Culex, Anopheles, and Culiseta spp., collected from January to March 2012) was found. This suggests that mosquitoes do not play an important role, if any, in the persistence of SBV during the winter months in northwestern Europe.

The Effectiveness of Asian Bush Mosquito (Aedes japonicus japonicus) Control Actions in Colonised Peri-urban Areas in the Netherlands.

The Asian bush mosquito (Aedes japonicus japonicus (Theobald)) is an invasive mosquito species in Europe. In 2012, it was for the first time detected in the Netherlands, in the municipality of Lelystad. After further research, thousands of specimens were found in the surrounding peri-urban areas of the city. A targeted mosquito control campaign began in 2015 with the objective of reducing populations in locations with the highest concentrations of Ae. japonicus breeding sites: allotment garden complexes. Mosquito control consisted of source reduction combined with application of the larvicide Vectomax in breeding sites. At eight complexes, mosquito control effectiveness has been systematically measured by sampling larvae from breeding sites. Six measurements were performed between 2015 and 2016. Results show that the effectiveness of mosquito control actions was similar in all treated allotment gardens and resulted in a significant reduction in Ae. japonicus larval abundance. Rain barrels at the allotments represent the most frequent breeding site in Lelystad, but every water filled artificial container is a potential breeding site for the species. Ae. japonicus was not found in the samples taken in other allotment gardens in the province of Flevoland; however, the collection methodology used proven to be effective in detecting this species when it has newly colonized surrounding areas. Targeted mosquito control actions at the breeding sites are crucial for successful reduction of populations of an invasive mosquito species, and systematic measurements of the effectiveness, is in this case, the base to understand the dynamics of Ae. japonicus populations after mosquito control.

The first detected airline introductions of yellow fever mosquitoes (Aedes aegypti) to Europe, at Schiphol International airport, the Netherlands.

BACKGROUND: Air-borne introduction of exotic mosquitoes to Schiphol airport in the Netherlands has been considered plausible based upon findings of mosquitoes in aircraft cabins during 2008, 2010 and 2011. Beginning in 2013, surveillance efforts at Schiphol had focused on promptly detecting accidental introductions at the airport facilities in order to quickly react and avoid temporary proliferation or establishment of mosquito populations, identify the origin of the introductions, and avoid potential transmission of vector-borne diseases. METHODS: BG-Mosquitaire mosquito traps were set at the most likely locations for arrival of the invasive Aedes mosquitoes as part of the mosquito monitoring program at Schiphol airport. Samples were collected bi-weekly. Upon detection of exotic specimens, information about the origin of the flights arriving to the particular location at the airport where specimens were captured was requested from airport authorities. The GIS tool Intersect was then used to identify airports of origin common to positive trapping locations during the specific trapping period. Captured Aedes aegypti mosquitoes were subsequently genotyped at 12 highly polymorphic microsatellite markers and compared to a reference database of 79 populations around the world to further narrow down their location of origin. RESULTS: In 2016, six adult yellow fever mosquitoes were captured indoors and outdoors at the airport of Schiphol in the Netherlands confirming, for the first time, air-borne transport of this mosquito vector species into Europe. Mosquitoes were captured during three time periods: June, September and October. Containers carried by aircrafts are considered the most likely pathway for this introduction. GIS analysis and genetic assignment tests on these mosquitoes point to North America or the Middle East as possible origins, but the small sample size prevents us from reliably identifying the geographic origin of this introduction. CONCLUSIONS: The arrival of Ae. aegypti mosquitoes to Schiphol airport from flights arriving from overseas, demonstrates the potential risk of international flights to public health as carriers of arthropod vectors of disease. The results strongly suggest that disinsection of containers and their storage compartments inside the aircrafts could contribute to preventing future introductions of mosquito vectors. Invasive mosquito species introduced by aircrafts from overseas could become seasonally established during the warmer months in Europe, or permanently in certain climatically suitable areas for the species, with major consequences for human health.