Standardized Laboratory Feeding of Larval Aedes japonicus japonicus (Diptera: Culicidae).

The Asian bush mosquito (Aedes japonicus japonicus, Theobald 1901) is an invasive culicid species which originates in Asia but is nowadays present in northern America and Europe. It is a competent vector for several human disease pathogens. In addition to the public health threat, this invasive species may also be an ecological threat for native container-breeding mosquitoes which share a similar larval habitat. Therefore, it is of importance to gain knowledge on ecological and eco-toxicological features of the Asian bush mosquito. However, optimal laboratory feeding conditions have not yet been established. Standardized feeding methods will be needed in assessing the impact of insecticides or competitional strength of this species. To fill this gap, we performed experiments on food quality and quantity for Ae. j. japonicus larvae. We found out that the commercial fish food TetraMin (Tetra, Melle, Germany) in a dose of 10 mg per larva is the most suitable food tested. We also suggest a protocol with a feeding sequence of seven portions for all larval stages of this species.

The potential invasion into North America and Europe by non-native mosquito, Aedes koreicus (Diptera: Culicidae).

Aedes koreicus (Edward, 1917) (Diptera: Culicidae), a mosquito species native to East Asia, has spread to parts of Europe and Central Asia since 2008. The species shares ecological characteristics with Aedes japonicus (Theobald, 1901) (Diptera: Culicidae), which has already successfully invaded and established in North America and Europe. Given these similarities, it is plausible that Ae. koreicus may also invade North America in the future. However, the invasion of Ae. koreicus may be masked or have delayed detection due to their similar morphologies with Ae. japonicus. This study highlights the potential risks of invasion of Ae. koreicus into North America, especially in the northeastern United States, and for further expansion in Europe. We used the maximum entropy model to identify areas with a high likelihood of presence in North America and Europe using comprehensive occurrence records from East Asia, Central Asia, and Europe. We have identified 15 additional countries in Europe and 7 states in the United States that will likely have suitable environments for Ae. koreicus. Additionally, we reviewed the morphological characteristics of Ae. koreicus and Ae. japonicus and provided morphological keys to distinguish the 2 species. Morphological results contradicting previous studies suggested that finding the origin by morphological comparison between Ae. koreicus populations may need re-evaluation. The information presented here will be useful for researchers and public health professionals in high-risk areas to be informed about morphological characteristics to distinguish Ae. koreicus from similar-looking Ae. japonicus. These tools will allow more careful monitoring of the potential introduction of this highly invasive species.

First Nationwide Monitoring Program for the Detection of Potentially Invasive Mosquito Species in Austria.

In Austria, only fragmented information on the occurrence of alien and potentially invasive mosquito species exists. The aim of this study is a nationwide overview on the situation of those mosquitoes in Austria. Using a nationwide uniform protocol for the first time, mosquito eggs were sampled with ovitraps at 45 locations in Austria at weekly intervals from May to October 2020. The sampled eggs were counted and the species were identified by genetic analysis. The Asian tiger mosquito Aedes albopictus was found at two sites, once in Tyrol, where this species has been reported before, and for the first time in the province of Lower Austria, at a motorway rest stop. The Asian bush mosquito Aedes japonicus was widespread in Austria. It was found in all provinces and was the most abundant species in the ovitraps by far. Aedes japonicus was more abundant in the South than in the North and more eggs were found in habitats with artificial surfaces than in (semi-) natural areas. Further, the number of Ae. japonicus eggs increased with higher ambient temperature and decreased with higher wind speed. The results of this study will contribute to a better estimation of the risk of mosquito-borne disease in Austria and will be a useful baseline for a future documentation of changes in the distribution of those species.

Diapause characterisation and seasonality of Aedes japonicus japonicus (Diptera, Culicidae) in the northeast of France.

The invasive mosquito Aedes japonicus japonicus (Theobald, 1901) settled in 2013 in the Alsace region, in the northeast of France. In this temperate area, some mosquito species use diapause to survive cold winter temperatures and thereby foster settlement and dispersal. This study reports diapause and its seasonality in a field population of Ae. japonicus in the northeast of France. For two years, eggs were collected from May to the beginning of November. They were most abundant in summer and became sparse in late October. Diapause eggs were determined by the presence of a fully developed embryo in unhatched eggs after repeated immersions. Our study showed effective diapause of Ae. japonicus in this part of France. At the start of the egg-laying period (week 20), we found up to 10% of eggs under diapause, and this rate reached 100% in October. The 50% cut-off of diapause incidence was determined by the end of summer, leading to an average calculated maternal critical photoperiod of 13 h 23 min. Interestingly, diapause was shown to occur in part of the eggs even at the earliest period of the two seasons, i.e. in May of each year. Even though we observed that the size of eggs was positively correlated with diapause incidence, morphology cannot be used as the unique predictive indicator of diapause status due to overlapping measurements between diapausing and non-diapausing eggs. This study provides new knowledge on diapause characterisation and invasive traits of Ae. japonicus.

TITLE: Caractérisation de la diapause et saisonnalité d'Aedes japonicus japonicus (Diptera, Culicidae) dans le nord-est de la France. ABSTRACT: Aedes japonicus japonicus (Theobald, 1901), un moustique invasif, s'est installé en 2013, en Alsace, dans le nord-est de la France. Dans une telle région tempérée, certaines espèces de moustiques ont recours à la diapause pour survivre aux températures froides de l'hiver, ce qui favorise donc l'établissement et la dispersion de ces espèces. Cette étude montre le phénomène de diapause et la saisonnalité d'une population sauvage d'Ae. japonicus dans le nord-est de la France. Pendant deux ans, les œufs ont été collectés de mai à début novembre. Ils étaient plus abondants durant l'été et plus rarement présents en octobre. Les œufs diapausants ont été déterminés par la présence d'un embryon entièrement développé dans les oeufs non éclos après des immersions répétées. Notre étude a mis en évidence une diapause effective d'Ae. japonicus dans le nord-est de la France. Au début de la période de ponte (semaine 20), jusqu'à 10 % des œufs a été trouvé à l'état de diapause et ce taux atteint 100 % en octobre. Un taux de 50 % de diapause a été atteint à la fin de l'été, amenant à une photopériode critique maternelle calculée de 13 heures 23 minutes en moyenne. De façon intéressante, il a été montré que la diapause se produisait chez une partie des œufs, et ce même dès le début des deux saisons, en mai de chaque année. Bien que nous ayons observé que les mesures des œufs étaient positivement corrélées avec le taux de diapause, la morphologie des œufs seule ne peut pas être utilisée comme unique indicateur de prédiction du statut de diapause, à cause d'un chevauchement des mesures entre les œufs diapausants et non diapausants. Cette étude offre de nouvelles connaissances sur les caractéristiques de la diapause et sur les traits d'invasion d'Ae. japonicus.

Emergence of the invasive Asian bush mosquito, Aedes (Finlaya) japonicus japonicus, in an urban area, Romania.

BACKGROUND: A study conducted at the International Airport of Cluj-Napoca, Romania, with the aim of investigating the presence/absence of invasive Aedes mosquito species resulted in finding Aedes japonicus japonicus (Theobald 1901) eggs in one of the ovitraps placed on site. METHODS: The study was carried out between 30 June and 29 September 2020. On 24 August, 26 eggs were collected and later hatched at the University of Agricultural Sciences and Veterinary Medicine of Cluj-Napoca's insectary. On 15 October another adult female Ae. japonicus was caught entering a building in the center of the city, about 7 km from the first sampling spot. RESULTS: The mosquitoes were identified morphologically and confirmed by molecular analysis, based on the genetic analysis of the mitochondrial gene cytochrome c oxidase subunit 1 (COI). CONCLUSION: This is the first report of the species in Romania, highlighting the need for surveillance and implemented control methods. However, in Romania to our knowledge only Aedes albopictus has been established; further studies are required to learn about this new invasive species' status in Romania.

At the tip of an iceberg: citizen science and active surveillance collaborating to broaden the known distribution of Aedes japonicus in Spain.

BACKGROUND: Active surveillance aimed at the early detection of invasive mosquito species is usually focused on seaports and airports as points of entry, and along road networks as dispersion paths. In a number of cases, however, the first detections of colonizing populations are made by citizens, either because the species has already moved beyond the implemented active surveillance sites or because there is no surveillance in place. This was the case of the first detection in 2018 of the Asian bush mosquito, Aedes japonicus, in Asturias (northern Spain) by the citizen science platform Mosquito Alert. METHODS: The collaboration between Mosquito Alert, the Ministry of Health, local authorities and academic researchers resulted in a multi-source surveillance combining active field sampling with broader temporal and spatial citizen-sourced data, resulting in a more flexible and efficient surveillance strategy. RESULTS: Between 2018 and 2020, the joint efforts of administrative bodies, academic teams and citizen-sourced data led to the discovery of this species in northern regions of Spain such as Cantabria and the Basque Country. This raised the estimated area of occurrence of Ae. japonicus from < 900 km2 in 2018 to > 7000 km2 in 2020. CONCLUSIONS: This population cluster is geographically isolated from any other population in Europe, which raises questions about its origin, path of introduction and dispersal means, while also highlighting the need to enhance surveillance systems by closely combining crowd-sourced surveillance with public health and mosquito control agencies' efforts, from local to continental scales. This multi-actor approach for surveillance (either passive and active) shows high potential efficiency in the surveillance of other invasive mosquito species, and specifically the major vector Aedes aegypti which is already present in some parts of Europe.

An unwanted companion reaches the country: the first record of the alien mosquito Aedes japonicus japonicus (Theobald, 1901) in Slovakia.

BACKGROUND: Invasive mosquitoes of the genus Aedes are quickly spreading around the world. The presence of these alien species is concerning for both their impact on the native biodiversity and their high vector competence. The surveillance of Aedes invasive mosquito (AIM) species is one of the most important steps in vector-borne disease control and prevention. METHODS: In 2020, the monitoring of AIM species was conducted in five areas (Bratislava, Zvolen, Banská Bystrica, Presov, Kosice) of Slovakia. The sites were located at points of entry (border crossings with Austria and Hungary) and in the urban and rural zones of cities and their surroundings. Ovitraps were used at the majority of sites as a standard method of monitoring. The collected specimens were identified morphologically, with subsequent molecular identification by conventional PCR (cox1) and Sanger sequencing. The phylogenetic relatedness of the obtained sequences was inferred by the maximum likelihood (ML) method. The nucleotide heterogeneity of the Slovak sequences was analysed by the index of disparity. RESULTS: A bush mosquito, Aedes japonicus japonicus, was found and confirmed by molecular methods in three geographically distant areas of Slovakia-Bratislava, Zvolen and Presov. The presence of AIM species is also likely in Kosice; however, the material was not subjected to molecular identification. The nucleotide sequences of some Slovak strains confirm their significant heterogeneity. They were placed in several clusters on the ML phylogenetic tree. Moreover, Ae. j. japonicus was discovered in regions of Slovakia that are not close to a point of entry, where the mosquitoes could find favourable habitats in dendrothelms in city parks or forests. CONCLUSION: Despite being a first record of the Ae. j. japonicus in Slovakia, our study indicates that the established populations already exist across the country, underlining the urgent need for intensified surveillance of AIM species as well as mosquito-borne pathogens.

Global occurrence data improve potential distribution models for Aedes japonicus japonicus in non-native regions.

BACKGROUND: There is great interest in modelling the distribution of invasive species, particularly from the point of view of management. However, distribution modelling for invasive species using ecological niche models (ENMs) involves multiple challenges. Owing to the short time span since the introduction or arrival of a non-indigenous species and the associated dispersal limitations, applying regular ENMs at an early stage of the invasion process may result in an underestimation of the potential niche in the new ranges. This topic is dealt with here using the example of Aedes japonicus japonicus, a vector competent mosquito species for a number of diseases. RESULTS: We found high niche unfilling for the species' non-native range niches in Europe and North America compared with the native range niche, which can be explained by the early stage of the invasion process. Comparing four different ENMs based on: (i) the European and (ii) the North American non-native range occurrence data, (iii) (derived) native range occurrence data, and (iv) all available occurrence data together, we found large differences in the projected climatic suitability, with the global data model projecting larger areas with climatic suitability. CONCLUSION: ENM in biological invasions can be challenging, especially when distribution data are only poorly available. We suggest one possible way to project climatic suitability for Aedes j. japonicus despite poor data availability for the non-native ranges and missing occurrences from the native range. We discuss aspects of the lack of information and the associated implications for modelling. © 2020 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Oviposition of Aedes japonicus japonicus (Diptera: Culicidae) and associated native species in relation to season, temperature and land use in western Germany.

BACKGROUND: Aedes japonicus japonicus, first detected in Europe in 2000 and considered established in Germany 10 years later, is of medical importance due to its opportunistic biting behaviour and its potential to transmit pathogenic viruses. Its seasonal phenology, temperature and land use preference related to oviposition in newly colonised regions remain unclear, especially in the context of co-occurring native mosquito species. METHODS: Focussing on regions in Germany known to be infested by Ae. japonicus japonicus, we installed ovitraps in different landscapes and their transition zones and recorded the oviposition activity of mosquitoes in relation to season, temperature and land use (arable land, forest, settlement) in two field seasons (May-August 2017, April-November 2018). RESULTS: Ae. japonicus japonicus eggs and larvae were encountered in 2017 from June to August and in 2018 from May to November, with a markedly high abundance from June to September in rural transition zones between forest and settlement, limited to water temperatures below 30 °C. Of the three native mosquito taxa using the ovitraps, the most frequent was Culex pipiens s.l., whose offspring was found in high numbers from June to August at water temperatures of up to 35 °C. The third recorded species, Anopheles plumbeus, rarely occurred in ovitraps positioned in settlements and on arable land, but was often associated with Ae. japonicus japonicus. The least frequent species, Aedes geniculatus, was mostly found in ovitraps located in the forest. CONCLUSIONS: The transition zone between forest and settlement was demonstrated to be the preferred oviposition habitat of Ae. japonicus japonicus, where it was also the most frequent container-inhabiting mosquito species in this study. Compared to native taxa, Ae. japonicus japonicus showed an extended seasonal activity period, presumably due to tolerance of colder water temperatures. Higher water temperatures and arable land represent distribution barriers to this species. The frequently co-occurring native species An. plumbeus might be useful as an indicator for potentially suitable oviposition habitats of Ae. japonicus japonicus in hitherto uncolonised regions. The results contribute to a better understanding of mosquito ecology and provide a basis for more targeted monitoring, distribution modelling and risk management of mosquitoes.

First detection of Aedes japonicus in Spain: an unexpected finding triggered by citizen science.

BACKGROUND: Aedes japonicus is an invasive vector mosquito from Southeast Asia which has been spreading across central Europe since the year 2000. Unlike the Asian Tiger mosquito (Aedes albopictus) present in Spain since 2004, there has been no record of Ae. japonicus in the country until now. RESULTS: Here, we report the first detection of Ae. japonicus in Spain, at its southernmost location in Europe. This finding was triggered by the citizen science platform Mosquito Alert. In June 2018, a citizen sent a report via the Mosquito Alert app from the municipality of Siero in the Asturias region (NW Spain) containing pictures of a female mosquito compatible with Ae. japonicus. Further information was requested from the participant, who subsequently provided several larvae and adults that could be classified as Ae. japonicus. In July, a field mission confirmed its presence at the original site and in several locations up to 9 km away, suggesting a long-time establishment. The strong media impact in Asturias derived from the discovery raised local participation in the Mosquito Alert project, resulting in further evidence from surrounding areas. CONCLUSIONS: Whilst in the laboratory Ae. japonicus is a competent vector for several mosquito-borne pathogens, to date only West Nile virus is a concern based on field evidence. Nonetheless, this virus has yet not been detected in Asturias so the vectorial risk is currently considered low. The opportunity and effectiveness of combining citizen-sourced data to traditional surveillance methods are discussed.

Modelling seasonal dynamics, population stability, and pest control in Aedes japonicus japonicus (Diptera: Culicidae).

BACKGROUND: The invasive temperate mosquito Aedes japonicus japonicus is a potential vector for various infectious diseases and therefore a target of vector control measures. Even though established in Germany, it is unclear whether the species has already reached its full distribution potential. The possible range of the species, its annual population dynamics, the success of vector control measures and future expansions due to climate change still remain poorly understood. While numerous studies on occurrence have been conducted, they used mainly presence data from relatively few locations. In contrast, we used experimental life history data to model the dynamics of a continuous stage-structured population to infer potential seasonal densities and ask whether stable populations are likely to establish over a period of more than one year. In addition, we used climate change models to infer future ranges. Finally, we evaluated the effectiveness of various stage-specific vector control measures. RESULTS: Aedes j. japonicus has already established stable populations in the southwest and west of Germany. Our models predict a spread of Ae. j. japonicus beyond the currently observed range, but likely not much further eastwards under current climatic conditions. Climate change models, however, will expand this range substantially and higher annual densities can be expected. Applying vector control measures to oviposition, survival of eggs, larvae or adults showed that application of adulticides for 30 days between late spring and early autumn, while ambient temperatures are above 9 °C, can reduce population density by 75%. Continuous application of larvicide showed similar results in population reduction. Most importantly, we showed that with the consequent application of a mixed strategy, it should be possible to significantly reduce or even extinguish existing populations with reasonable effort. CONCLUSION: Our study provides valuable insights into the mechanisms concerning the establishment of stable populations in invasive species. In order to minimise the hazard to public health, we recommend vector control measures to be applied in 'high risk areas' which are predicted to allow establishment of stable populations to establish.

The Asian bush mosquito Aedes japonicus japonicus (Diptera: Culicidae) in Europe, 17 years after its first detection, with a focus on monitoring methods.

After the first detection of the Asian bush mosquito Aedes japonicus japonicus in the year 2000 in France, its invasive nature was revealed in 2008 in Switzerland and Germany. In the following years, accumulating reports have shown that Ae. j. japonicus succeeded in establishing in several European countries. Surveillance efforts suggest that there are currently four populations in Europe, with the largest one, formed by the recent fusion of several smaller populations, ranging from West Germany, with extensions to Luxembourg and French Alsace, southwards to Switzerland and continuing westwards through Liechtenstein to western Austria. This paper summarises the present distribution of Ae. j. japonicus in Europe, based on published literature and hitherto unpublished findings by the authors, and critically reviews the monitoring strategies applied. A proposal for a more standardised monitoring approach is provided, aiming at the harmonisation of future data collections for improving the comparability between studies and the suitability of collected data for further research purposes, e.g. predictive modelling approaches.

First Record of Aedes japonicus japonicus in Oklahoma, 2017.

In spring 2017, mosquito larvae were collected from 25 sites across eastern Oklahoma as part of a Zika virus vector surveillance effort. Aedes japonicus japonicus larvae were collected from horse troughs at 2 sites in Ottawa County, OK. Identification was made using 1 larva stored in 70% ethanol and 3 adult females reared from the larvae. Another invasive mosquito species, Culex coronator, was detected at 2 different sites, adding 2 additional counties to the 9 where the species had been previously reported. The presence of these invasive species in Oklahoma may have an impact on future regional arbovirus concerns.

Mosquito Species (Diptera: Culicidae) Persistence and Synchrony Across an Urban Altitudinal Gradient.

Patterns of mosquito spatial persistence and temporal presence, as well as synchrony, i.e., the degree of concerted fluctuations in abundance, have been scarcely studied at finely grained spatial scales and over altitudinal gradients. Here, we present a spatial persistence, temporal presence, and synchrony analysis of four common mosquito species across the altitudinal gradient of Mt. Konpira in Nagasaki, Japan. We found that Aedes albopictus (Skuse) was more frequently found at the mountain base. In contrast, Aedes japonicus (Theobald) and Aedes flavopictus Yamada were common higher in the mountain, while Armigeres subalbatus (Coquillet) was uniformly present across the mountain, yet less frequently than the other species during the studied period. Our analysis showed that these spatial heterogeneities were associated with differences in landscape and microclimatic elements of Mt. Konpira. Temporally we found that presence across sampling locations was mainly synchronous across the four species and positively associated with rainfall and temperature. With the exception of Ae albopictus, where no significant synchrony was observed, mosquito species mainly showed flat synchrony profiles in Mt. Konpira when looking at the geographic (2-D) distance between their sampling locations. By contrast, when synchrony was studied across altitude, it was observed that Ae. flavopictus tracked the temperature synchrony pattern, decreasing its synchrony with the separation in altitude between sampling locations. Finally, our results suggest that differences in mosquito species persistence, temporal presence, and synchrony might be useful to understand the entomological risk of vector-borne disease transmission in urban landscapes.

Aedes albopictus and Aedes japonicus - two invasive mosquito species with different temperature niches in Europe.

BACKGROUND: Aedes albopictus and Ae. japonicus are two of the most widespread invasive mosquito species that have recently become established in western Europe. Both species are associated with the transmission of a number of serious diseases and are projected to continue their spread in Europe. METHODS: In the present study, we modelled the habitat suitability for both species under current and future climatic conditions by means of an Ensemble forecasting approach. We additionally compared the modelled MAXENT niches of Ae. albopictus and Ae. japonicus regarding temperature and precipitation requirements. RESULTS: Both species were modelled to find suitable habitat conditions in distinct areas within Europe: Ae. albopictus within the Mediterranean regions in southern Europe, Ae. japonicus within the more temperate regions of central Europe. Only in few regions, suitable habitat conditions were projected to overlap for both species. Whereas Ae. albopictus is projected to be generally promoted by climate change in Europe, the area modelled to be climatically suitable for Ae. japonicus is projected to decrease under climate change. This projection of range reduction under climate change relies on the assumption that Ae. japonicus is not able to adapt to warmer climatic conditions. The modelled MAXENT temperature niches of Ae. japonicus were found to be narrower with an optimum at lower temperatures compared to the niches of Ae. albopictus. CONCLUSIONS: Species distribution models identifying areas with high habitat suitability can help improving monitoring programmes for invasive species currently in place. However, as mosquito species are known to be able to adapt to new environmental conditions within the invasion range quickly, niche evolution of invasive mosquito species should be closely followed upon in future studies.

Spread of Aedes japonicus japonicus (Theobald, 1901) in Austria, 2011-2015, and first records of the subspecies for Hungary, 2012, and the principality of Liechtenstein, 2015.

BACKGROUND: The Asian bush mosquito, Aedes (Hulecoeteomyia) japonicus japonicus (Theobald, 1901) (Diptera: Culicidae), was first identified in Austria in August 2011 in the federal state of Styria at the border to Slovenia. METHODS: Between 2011 and 2015 the spread of Ae. j. japonicus was monitored in southern, eastern and western Austrian provinces as well as in neighbouring countries by checking natural and man-made container habitats for the aquatic stages. The search concentrated around the most recent occurrence of Ae. j. japonicus and extended up to several kilometres until the subspecies could not be found anymore. RESULTS: Between May and July 2012 the distribution area of Ae. j. japonicus was found to be extended westwards into Carinthia, and eastwards towards the federal state of Burgenland. In August 2012, the subspecies was found in Hungary, representing the first record of an invasive mosquito species in this country. In 2013 its expansion was confirmed at several sites in Austria. Additionally, between April and July 2015, the subspecies was detected in all districts of the westernmost Austrian state Vorarlberg reaching the alpine Montafon valley at the end of October 2015, at all three examined sites in southern Bavaria bordering Vorarlberg, and in the adjacent Principality of Liechtenstein, for which it also represents the first record of an invasive mosquito species. One remarkable finding of the subspecies was located close to the city of Kufstein in the lower Inn valley of the Tyrol in September 2015, which is an isolated occurrence without spatial connection to any known established population. CONCLUSIONS: Our findings demonstrate the ongoing spread of Ae. j. japonicus towards all directions within Austria and beyond. Together with the absence of supposed natural barriers, e.g. high mountain chains, at the borders of the current subspecies' distribution area in south-eastern Austria, these findings suggest a further spread to the Austrian capital Vienna and the Hungarian tourist region of Lake Balaton within the upcoming few years. The observed intrusions in western Austria represent most probably extensions of the population established and spreading in eastern Switzerland and southern Germany. The putative role of the subspecies in pathogen transmission together with its rapid spread observed argues for the implementation of comprehensive nation-wide surveillance and response preparedness.

Newly discovered population of Aedes japonicus japonicus (Diptera: Culicidae) in Upper Bavaria, Germany, and Salzburg, Austria, is closely related to the Austrian/Slovenian bush mosquito population.

BACKGROUND: The German mosquito surveillance instrument 'Mueckenatlas' requests the general public to collect and submit mosquito specimens. Among these, increasing numbers of individuals of invasive species have been registered. Specimens of the Asian bush mosquito Aedes japonicus japonicus submitted from German Upper Bavaria, where this species had not previously been recorded, triggered regional monitoring in mid-2015. METHODS: The search for Ae. j. japonicus breeding sites and developmental stages concentrated on cemeteries in the municipality of origin of the submitted specimens and, subsequently, in the whole region. A virtual grid consisting of 10 × 10 km(2) cells in which up to three cemeteries were checked, was laid over the region. A cell was considered positive as soon as Ae. j. japonicus larvae were detected, and regarded negative when no larvae could be found in any of the cemeteries inspected. All cells surrounding a positive cell were screened accordingly. A subset of collected Aedes j. japonicus specimens was subjected to microsatellite and nad4 sequence analyses, and obtained data were compared to individuals from previously discovered European populations. RESULTS: Based on the grid cells, an area of approximately 900 km(2) was populated by Ae. j. japonicus in Upper Bavaria and neighbouring Austria. Genetic analyses of microsatellites and nad4 gene sequences generated one genotype out of two previously described for Europe and three haplotypes, one of which had previously been found in Europe only in Ae. j. japonicus samples from a population in East Austria and Slovenia. The genetic analysis suggests the new population is closely related to the Austrian/Slovenian population. CONCLUSION: As Ae. j. japonicus is well adapted to temperate climates, it has a strong tendency to expand and to colonise new territories in Central Europe, which is facilitated by human-mediated, passive transportation. The new population in Upper Bavaria/Austria is the seventh separate population described in Europe. According to our data, it originated from a previously detected population in eastern Austria/Slovenia and not from an introduction event from abroad. The dispersal and population dynamics of Ae. j. japonicus should be thoroughly surveyed, as this species is a potential vector of disease agents.

First record of the Asian bush mosquito, Aedes japonicus japonicus, in Italy: invasion from an established Austrian population.

BACKGROUND: In 2011 we identified the Asian bush mosquito, Aedes japonicus japonicus (Theobald, 1901) (Diptera: Culicidae) for the first time in northern Slovenia and in the bordering Austrian federal state of Styria. Between May and July 2012 the distribution area of Ae. j. japonicus was already found to be extended westwards into Carinthia and eastwards towards Burgenland and bordering Hungary. In August 2012 the species was first detected in a western province of Hungary. In subsequent years, follow-up field studies demonstrated an active spread westwards throughout Carinthia, reaching the border to northern Italy. FINDINGS: In July 2015 several aquatic-stage specimens of the species were discovered at three different sites in the Friuli Venezia Giulia region, north-eastern Italy. In September 2015, co-occurrence of Ae. j. japonicus and Aedes albopictus (Skuse, 1895) was observed in the same sample in that region. CONCLUSIONS: Ae. j. japonicus actively extended its geographic range from an established population in Carinthia (Austria) southwards to northern Italy by crossing Alpine ranges. Since Ae. albopictus and Aedes koreicus (Edwards, 1917) are already well established in northern Italy, it will be pivotal to monitor the consequences of a third invasive mosquito species trying to populate the same geographic region.