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 arbovirus vector Culex torrentium is more prevalent than Culex pipiens in northern and central Europe.

Two species of arbovirus vector, Culex torrentium and Culex pipiens (Diptera: Culicidae), occur in several European countries, but difficulties in their accurate identification and discrimination have hampered both detailed and large-scale distribution and abundance studies. Using a molecular identification method, we identified to species 2559 larvae of Cx. pipiens/torrentium collected from 138 sites in 13 European countries ranging from Scandinavia to the Mediterranean coast. In addition, samples of 1712 males of Cx. pipiens/torrentium collected at several sites in the Czech Republic were identified to species based on the morphology of their hypopygia. We found that the two species occur together in large areas of Europe, and that Cx. torrentium dominates in northern Europe and Cx. pipiens dominates south of the Alps. The transition in dominance occurs in central Europe, where both species are roughly equally common. There was a strong correlation between the length of the growing season at different sites and occurrences of the two species. As the growing season increases, the proportion and detection of Cx. torrentium decrease, whereas those of Cx. pipiens increase. The present findings have important consequences for the interpretation of the results of studies on major enzootic and link-vectors of mosquito-borne bird-associated viruses (i.e. Sindbis, West Nile and Usutu viruses), especially in central Europe and Scandinavia.

Prevalence of Coxiella burnetii in ticks after a large outbreak of Q fever.

Q fever has emerged as an important human and veterinary public health problem in the Netherlands with major outbreaks in three consecutive years. Goat farms are probably the prime source from which Coxiella burnetii have spread throughout the environment, infecting people living in the vicinity. Coxiella burnetii infection not only spilled over from animal husbandry to humans but could also have spread to neighbouring wildlife and pets forming novel reservoirs and consequently posing another and lingering threat to humans, companion animals and livestock. In these cases, transmission routes other than airborne spread of contaminated aerosols may become significant. Therefore, the role of ticks in the transmission of Coxiella burnetii in the current situation was investigated. A total of 1891 questing Ixodes ricinus ticks and 1086 ticks feeding on pets, wildlife and livestock were tested by a recently developed multiplex Q-PCR. All ticks were negative, except for a few ticks feeding on a herd of recently vaccinated sheep. Coxiella-positive ticks were not detected after resampling this particular herd three months later. Based on these data we conclude that the current risk of acquiring Q fever from questing ticks in the Netherlands is negligible. However, for future risk assessments, it might be relevant to sample more ticks in the vicinity of previously C. burnetii infected goat farms and to assess whether C. burnetii can be transmitted transovarially and transstadially in I. ricinus ticks.

Age-grading the biting midge Culicoides sonorensis using near-infrared spectroscopy.

Age-grading of insects is important in the control and monitoring of both insect populations and vector-borne diseases. Microscopy and morphological techniques exist to age-grade most blood-feeding flies, but these techniques are laborious, often destructive to the insects, and slow. Near-infrared spectroscopy (NIRS) can be automated and is a non-destructive technique for age-grading. We applied NIRS techniques to age-grade females of the biting midge, Culicoides sonorensis Wirth & Jones (Diptera: Ceratopogonidae), the vector of bluetongue and other arboviruses in North America. Female flies of five known age cohorts (1, 3, 6, 9 and 12 days post-emergence) from three laboratory colonies were used. The data indicate that NIRS can be used to differentiate age groups of C. sonorensis.

First report of a North American invasive mosquito species Ochlerotatus atropalpus (Coquillett) in the Netherlands, 2009.

In late August and early September 2009, numerous larvae, pupae, and actively flying adult specimens of Ochlerotatus atropalpus were discovered in the Province of Brabant, southern Netherlands, during surveillance activities for Aedes albopictus at two trading companies that import used tires. No Ae. albopictus were found. Both companies mainly import used tires from countries in Europe, but also from North America. Oc. atropalpus is endemic to North America and has so far only been found outside of its endemic range in Europe, namely France and Italy, where it was subsequently eradicated. A preliminary modelling study shows that the weather conditions in the Netherlands are unlikely to prevent establishment of Oc. atropalpus. This species has so far only been shown to serve as a vector for virus transmission under laboratory conditions. Studies on potential human and veterinary health risks, as well as possible control strategies are currently ongoing.

The hidden passenger of lucky bamboo: do imported Aedes albopictus mosquitoes cause dengue virus transmission in the Netherlands?

Since 2005, Aedes albopictus mosquitoes have been detected in companies in the Netherlands that import ornamental plants from China. To assess the risks of dengue transmission, a study was carried out in 48 persons who are professionally exposed to these mosquitoes. No evidence for non-travel-related flavivirus infections was found.

Accidental importation of the mosquito Aedes albopictus into the Netherlands: a survey of mosquito distribution and the presence of dengue virus.

In the summer of 2005, the Asian tiger mosquito, Aedes albopictus (Skuse) (Diptera: Culicidae) was found for the first time in the Netherlands. It was collected on the premises of several horticultural companies that import the ornamental plant Dracaena sanderiana (Sparagalus: Dracaenaceae [Agavaceae]), known as Lucky bamboo, from southern China, an area endemic for this mosquito species and for arboviruses transmitted by this vector. Here we report the results of a 1-year survey of the distribution and vector status of Ae. albopictus in Lucky bamboo nurseries in the Netherlands (July 2006-June 2007). As it had been established previously that the presence of this species was linked to the import of Lucky bamboo, the survey was conducted only on sites owned by relevant import companies. In total, 569 adult Ae. albopictus were collected with mosquito traps from 15 of the 17 (88%) glasshouses used by Lucky bamboo importers, none of which were found to be infected with dengue virus. On two occasions there was evidence that Ae. albopictus had escaped from the glasshouses, but, overall, there was no evidence that a population had become established in the greenhouses or elsewhere.