VectAbundance: a spatio-temporal database of Aedes mosquitoes observations.

Modelling approaches play a crucial role in supporting local public health agencies by estimating and forecasting vector abundance and seasonality. However, the reliability of these models is contingent on the availability of standardized, high-quality data. Addressing this need, our study focuses on collecting and harmonizing egg count observations of the mosquito Aedes albopictus, obtained through ovitraps in monitoring and surveillance efforts across Albania, France, Italy, and Switzerland from 2010 to 2022. We processed the raw observations to obtain a continuous time series of ovitraps observations allowing for an extensive geographical and temporal coverage of Ae. albopictus population dynamics. The resulting post-processed observations are stored in the open-access database VectAbundance.This initiative addresses the critical need for accessible, high-quality data, enhancing the reliability of modelling efforts and bolstering public health preparedness.

Random mutations induced by a sub-sterilizing dose of gamma ray on Aedes albopictus male pupae and transmission to progeny.

The application of the Sterile Insect Technique (SIT) to mosquito control is based on the systematic release of large numbers of adult males that have been previously sterilized by irradiation. Ionizing radiation doses inducing full sterility also cause somatic damages that reduce the capacity of the treated males to compete with wild males. The optimal dose inducing high levels of male sterility and minimal impact on competitiveness can be assessed by establishing a dose-response curve. Sub-sterile males are, to a variable degree, still fertile and might be able to transmit to the progeny and following generation(s) sub-lethal random mutations resulting from irradiation. To investigate this, we treated Ae. albopictus male pupae with a sub-sterilizing (2-4 % of egg hatching) dose of gamma rays and explored expressed mutated genes in treated males and their progeny using RNA-seq. Single nucleotide polymorphisms (SNPs) were called using two independent pipelines. Only SNPs common to both pipelines (less than 5 % of the total SNPs predicted) were considered reliable and were annotated to genes. Over 600 genes with mutations likely induced by irradiation were found in the treated Ae. albopictus males. A part of the genes found mutated in irradiated males were also found in (and therefore probably passed on to) males of the F1 and F2 progeny, indicating that genetic variations induced by irradiation may be transmitted along generations. The mutated genes in irradiated males did not seem to significantly affect biological processes, except in one case (i.e., oxidative phosphorylation). Only in four cases (i.e., oxidative phosphorylation, UDP-glucose metabolic process, proton transmembrane transport and riboflavin metabolism) we found biological processes to be significantly affected by mutated genes that were likely transmitted to the male progeny. Our results suggest that random mutations induced by a sub-sterilizing dose of gamma ray in Ae. albopictus male pupae and transmitted to the male progeny of the irradiated mosquitoes do not affect biological processes potentially harmful, from a public-health point of view.

First Detection of West Nile Virus Lineage 2 in Mosquitoes in Switzerland, 2022.

West Nile virus (WNV) is one of the most widespread flaviviruses in the world, and in recent years, it has been frequently present in many Mediterranean and Eastern European countries. A combination of different conditions, such as a favourable climate and higher seasonal average temperatures, probably allowed its introduction and spread to new territories. In Switzerland, autochthonous cases of WNV have never been reported, and the virus was not detected in mosquito vectors until 2022, despite an entomological surveillance in place in Canton Ticino, southern Switzerland, since 2010. In 2022, 12 sites were monitored from July to October, using BOX gravid mosquito traps coupled with honey-baited FTA cards. For the first time, we could detect the presence of WNV in FTA cards and mosquitoes in 8 out of the 12 sampling sites monitored, indicating an unexpectedly widespread circulation of the virus throughout the territory. Positive findings were recorded from the beginning of August until mid-October 2022, and whole genome sequencing analysis identified a lineage 2 virus closely related to strains circulating in Northern Italy. The entomological surveillance has proved useful in identifying viral circulation in advance of possible cases of WNV infection in humans or horses.

Optical recognition of the eggs of four Aedine mosquito species (Aedes albopictus, Aedes geniculatus, Aedes japonicus, and Aedes koreicus).

The continuous expansion of exotic Aedine mosquito species potential vectors of pathogens into new areas is a public health concern. In continental Europe, the surveillance of these mosquitoes is hindered by the simultaneous presence of three main invasive species (i.e., Aedes albopictus, Ae. japonicus, and Ae. koreicus). Standard low-cost surveillance methods (i.e., the deployment of oviposition traps and count of eggs under stereoscopic microscope) fail to distinguish the eggs of the different species. Identification of eggs by molecular methods is costly and time consuming and prevents measuring the density of invasive species and detecting early new invaders. Here we tested whether certain species could be identified by the patterns on the exochorionic membrane of their eggs. In a first step, we examined Aedine eggs of the three mentioned invasive and one indigenous (i.e., Ae. geniculatus) species with a high-resolution stereomicroscope and we identified each egg by MALDI-TOF mass spectrometry. In a second step, we submitted images of the eggs to 60 entomology experts and non-experts and tested their ability to distinguish among the species after an initial short training. The results obtained were consistent. Participants did not encounter difficulties in determining Ae. albopictus and Ae. geniculatus, while they had more difficulties in distinguishing Ae. japonicus from Ae. koreicus. In general, the quality of the exochorion seemed to play a more important role than the expertise level of the rater. The feasibility to differentiate Ae. albopictus from the other two invasive species is a significant achievement, as this is currently the most problematic species at the level of public health in Europe. Due to the presence of multiple invasive species that might prevent the correct quantification of mosquito population densities using standard surveillance methods and due to Ae. aegypti threat, it is recommended to optically determine also other species.

Estimating the Impact of Consecutive Blood Meals on Vector Competence of Aedes albopictus for Chikungunya Virus.

The continuous expansion of Aedes albopictus in Europe and the increases in autochthonous arboviruses transmissions in the region urge a better understanding of the virus transmission dynamic. Recent work described enhanced chikungunya virus (CHIKV) dissemination in Aedes aegypti mosquitoes exposed to a virus-free blood meal three days after their infection with CHIKV. Our study investigated the impact of a second blood meal on the vector competence of Ae. albopictus from southern Switzerland infected with CHIKV. Seven-day-old Ae. albopictus females were exposed to CHIKV-spiked blood and incubated at constant (27 °C) and fluctuating (14-28 °C) temperatures. Four days post-infection (dpi), some of these females were re-fed with a non-infectious blood meal. Virus infectivity, dissemination, transmission rate, and efficiency were investigated at seven and ten dpi. No enhanced dissemination rate was observed among females fed a second time; however, re-fed females have shown higher transmission efficiency than those fed only once after seven days post-infection and incubated under a fluctuating temperature regime. Vector competence for CHIKV was confirmed in Ae. albopictus from southern Switzerland. We did not observe an increase in dissemination rates among mosquitoes fed a second time (second blood meal), regardless of the temperature regime.

The effects of microclimatic winter conditions in urban areas on the risk of establishment for Aedes albopictus.

The tiger mosquito, Aedes albopictus, has adjusted well to urban environments by adopting artificial water containers as oviposition sites. Its spread in temperate regions is favoured by the deposition of cold-tolerant diapausing eggs that survive winter temperatures to a certain degree. The probability of establishment in new geographical areas is estimated using predictive models usually based on meteorological data measured at coarse resolution. Here, we investigated if we could obtain more precise and realistic risk scenarios for the spread of Ae. albopictus when considering the winter microclimatic conditions of catch basins, one of the major sites of oviposition and egg overwintering in temperate urban areas. We monitored winter microclimatic conditions of catch basins in four Swiss cities and developed a regression model to predict the average microclimatic temperatures of catch basins, based on available meteorological parameters, accounting for the observed differences between cities. We then used the microclimatic model to correct the predictions of our previously developed risk model for the prediction of Ae. albopictus establishment. Comparison of the predictive model's results based on local climate data and microclimate data indicated that the risk of establishment for Ae. albopictus in temperate urban areas increases when microhabitat temperatures are considered.

Geographic distribution of the V1016G knockdown resistance mutation in Aedes albopictus: a warning bell for Europe.

BACKGROUND: Colonization of large part of Europe by the Asian tiger mosquito Aedes albopictus is causing autochthonous transmission of chikungunya and dengue exotic arboviruses. While pyrethroids are recommended only to reduce/limit transmission, they are widely implemented to reduce biting nuisance and to control agricultural pests, increasing the risk of insurgence of resistance mechanisms. Worryingly, pyrethroid resistance (with mortality < 70%) was recently reported in Ae. albopictus populations from Italy and Spain and associated with the V1016G point mutation in the voltage-sensitive sodium channel gene conferring knockdown resistance (kdr). Genotyping pyrethroid resistance-associated kdr mutations in field mosquito samples represents a powerful approach to detect early signs of resistance without the need for carrying out phenotypic bioassays which require availability of live mosquitoes, dedicated facilities and appropriate expertise. METHODS: Here we report results on the PCR-genotyping of the V1016G mutation in 2530 Ae. albopictus specimens from 69 sampling sites in 19 European countries. RESULTS: The mutation was identified in 12 sites from nine countries (with allele frequencies ranging from 1 to 8%), mostly distributed in two geographical clusters. The western cluster includes Mediterranean coastal sites from Italy, France and Malta as well as single sites from both Spain and Switzerland. The eastern cluster includes sites on both sides of the Black Sea in Bulgaria, Turkey and Georgia as well as one site from Romania. These results are consistent with genomic data showing high connectivity and close genetic relationship among West European populations and a major barrier to gene flow between West European and Balkan populations. CONCLUSIONS: The results of this first effort to map kdr mutations in Ae. albopictus on a continental scale show a widespread presence of the V1016G allele in Europe, although at lower frequencies than those previously reported from Italy. This represents a wake-up call for mosquito surveillance programs in Europe to include PCR-genotyping of pyrethroid resistance alleles, as well as phenotypic resistance assessments, in their routine activities.

Risk-Based Mapping Tools for Surveillance and Control of the Invasive Mosquito Aedes albopictus in Switzerland.

BACKGROUND: In Switzerland, Aedes albopictus is well established in Ticino, south of the Alps, where surveillance and control are implemented. The mosquito has also been observed in Swiss cities north of the Alps. Decision-making tools are urgently needed by the local authorities in order to optimize surveillance and control. METHODS: A regularized logistic regression was used to link the long-term dataset of Ae. albopictus occurrence in Ticino with socioenvironmental predictors. The probability of establishment of Ae. albopictus was extrapolated to Switzerland and more finely to the cities of Basel and Zurich. RESULTS: The model performed well, with an AUC of 0.86. Ten socio-environmental predictors were selected as informative, including the road-based distance in minutes of travel by car from the nearest cell established in the previous year. The risk maps showed high suitability for Ae. albopictus establishment in the Central Plateau, the area of Basel, and the lower Rhone Valley in the Canton of Valais. CONCLUSIONS: The areas identified as suitable for Ae. albopictus establishment are consistent with the actual current findings of tiger mosquito. Our approach provides a useful tool to prompt authorities' intervention in the areas where there is higher risk of introduction and establishment of Ae. albopictus.

AIMSurv: First pan-European harmonized surveillance of Aedes invasive mosquito species of relevance for human vector-borne diseases.

Human and animal vector-borne diseases, particularly mosquito-borne diseases, are emerging or re-emerging worldwide. Six Aedes invasive mosquito (AIM) species were introduced to Europe since the 1970s: Aedes aegypti, Ae. albopictus, Ae. japonicus, Ae. koreicus, Ae. atropalpus and Ae. triseriatus. Here, we report the results of AIMSurv2020, the first pan-European surveillance effort for AIMs. Implemented by 42 volunteer teams from 24 countries. And presented in the form of a dataset named "AIMSurv Aedes Invasive Mosquito species harmonized surveillance in Europe. AIM-COST Action. Project ID: CA17108". AIMSurv2020 harmonizes field surveillance methodologies for sampling different AIMs life stages, frequency and minimum length of sampling period, and data reporting. Data include minimum requirements for sample types and recommended requirements for those teams with more resources. Data are published as a Darwin Core archive in the Global Biodiversity Information Facility- Spain, comprising a core file with 19,130 records (EventID) and an occurrences file with 19,743 records (OccurrenceID). AIM species recorded in AIMSurv2020 were Ae. albopictus, Ae. japonicus and Ae. koreicus, as well as native mosquito species.

Effectiveness of integrated Aedes albopictus management in southern Switzerland.

BACKGROUND: The exotic invasive tiger mosquito, Aedes albopictus, appeared in southern Switzerland in 2003. The spread of the mosquito has been surveyed constantly since then, and an integrated vector management (IVM) has been implemented to control its numbers. The control measures focus on the aquatic phase of the mosquito with removal of breeding sites and applications of larvicides in public areas whereas private areas are reached through extensive public information campaigns. Here, we evaluated the efficacy of the IVM. METHODS: Since all the municipalities with Ae. albopictus in southern Switzerland are currently implementing the IVM, Italian municipalities just across the Swiss-Italian border, where Ae. albopictus is present but no coordinated intervention programme is in place, served as control. Ovitraps and adult female traps were used to measure mosquito abundance in 2019. Generalised mixed-effects models were used to model the numbers of Ae. albopictus eggs and adult females collected. These numbers of Ae. albopictus eggs were compared to the numbers of eggs collected in 2012 and 2013 in a previous assessment of the IVM, using a hurdle model. RESULTS: Mean numbers of Ae. albopictus eggs and adult females in 2019 were consistently higher in the municipalities not following an IVM programme. In these municipalities, there were about four times (3.8) more eggs than in the municipalities implementing an IVM programme. Also, the numbers of eggs and adult females increased steadily from the beginning of the Ae. albopictus reproductive season, reaching a peak in August. In contrast, the increase in numbers of Ae. albopictus was much more contained in the municipalities implementing an IVM programme, without reaching an evident peak. Comparison with data from 2012 and 2013 indicates that the gap between intervention and non-intervention areas may have almost doubled in the past 6 years. CONCLUSIONS: The results of the survey strongly support the efficacy of the IVM programme implemented in southern Switzerland compared to municipalities without defined control measures. With the constant implementation of an IVM, it appears possible to contain the numbers of Ae. albopictus at a manageable level, reducing the nuisance for the human population and the risk of arbovirus epidemics.