Asian Bush Mosquito Research Articles

Thermal experiments with the Asian bush mosquito (Aedes japonicus japonicus) (Diptera: Culicidae) and implications for its distribution in Germany

BackgroundAs ectothermic animals, temperature influences insects in almost every aspect. The potential disease spreading Asian bush mosquito (Aedes japonicus japonicus) is native to temperate East Asia but invasive in several parts of the world. We report on the previously poorly understood temperature-dependence of its life history under laboratory conditions to understand invasion processes and to model temperature niches.ResultsTo evaluate winter survival, eggs were exposed between 1 day and 14 days to low temperatures (5 °C, 0 °C, -5 °C and -9 °C). Hatching success was drastically decreased after exposure to 0 °C and -5 °C, and the minimal hatching success of 0% was reached at -9 °C after two days. We then exposed larvae to 14 temperatures and assessed their life trait parameters. Larval survival to adulthood was only possible between 10 °C and 31 °C. Based on this, we modelled the optimal (25 °C), minimal (7 °C) and maximal (31 °C) temperature for cumulative female survival. The time to adult emergence ranges from 12 days to 58 days depending on temperature. We used an age-at-emergence-temperature model to calculate the number of potential generations per year for the Asian bush mosquito in Germany with an average of 4.72 potential generations. At lower temperatures, individuals grew larger than at higher temperatures with female R1 length ranging from 3.04 ± 0.1 mm at 31 °C to 4.26 ± 0.2 mm at 15 °C.ConclusionsReduced egg hatch after exposure to sub-zero temperatures prohibits the establishment of the Asian bush mosquito in large parts of Germany. Larval overwintering is not possible at temperature ≤ 5 °C. The many potential generations displayed per year may contribute to the species’ invasion success. This study on the thermal ecology of the Asian bush mosquito adds to our knowledge on the temperature dependence of the species and data could be incorporated in epidemiological and population dynamic modelling.

Density Dependence, Landscape, and Weather Impacts on Aquatic Aedes japonicus japonicus (Diptera: Culicidae) Abundance Along an Urban Altitudinal Gradient.

The Asian Bush Mosquito, Aedes (Finlaya) japonicus japonicus (Theobald) is an important globally invasive mosquito species. In comparison with other major invasive mosquitoes, relatively little is known about Ae. j. japonicus population dynamics in the field. Here, we present results from a 54-biweek long study of Ae. j. japonicus abundance in ovitraps set across the altitudinal gradient of Mt. Konpira, Nagasaki, Japan. Spatially, we found that Ae. j. japonicus fourth instar larvae (Aj4il) were more abundant at the base and top of Mt. Konpira and in ovitraps with more platykurtic water temperature (WT) distributions. In contrast, we found that temporally Aj4il were more abundant when ovitrap WT was more leptokurtic with 2 weeks of lag, and with high relative humidity SD with 2 months of lag. We also found that Aj4il were unlikely present when ovitrap WT was below 12.41°C. Parameter estimates for the Ricker model suggested that Ae. j. japonicus population growth was under density-dependence regulation, with a stable population dynamics whose fluctuations were associated with changes in ovitrap WT kurtosis and demographic stochasticity. Our results suggest that Aj4il abundance is more sensitive to temperature changes in kurtosis than mean values, potentially limiting the predictive ability of Ae. j. japonicus niche models based on the increase of average temperatures with global warming, and suggesting this mosquito species has a relatively coarse-grained response to temperature changes.

Occurrence and Spread of the Invasive Asian Bush Mosquito Aedes japonicus japonicus (Diptera: Culicidae) in West and North Germany since Detection in 2012 and 2013, Respectively.

The invasive Asian bush mosquito Aedes japonicus japonicus was first recognised as established in Germany in 2008. In addition to the first known and quickly expanding population in the southwestern part of the country, three separate populations were discovered in West, North and southeastern Germany in 2012, 2013 and 2015, respectively, by means of the ‘Mueckenatlas’, a German instrument of passive mosquito surveillance. Since the first findings of mosquito specimens in West and North Germany, these regions were checked annually for continuing colonisation and spread of the species. Both affected areas were covered by a virtual 10x10km2 grid pattern in the cells of which cemeteries were screened for immature stages of the mosquito. The cells were considered populated as soon as larvae or pupae were detected, whereas they were classified as negative when no mosquito stages were found in the cemeteries of at least three different towns or villages. Presence was also recorded when Ae. j. japonicus adults were submitted to the ‘Mueckenatlas’ from the respective cell or when there was evidence of local occurrence in localities other than cemeteries. Based on this approach, a significant expansion of the populated area was documented in West Germany since the first detection of Ae. j. japonicus in 2012 (increase in positive grid cells by more than 400%), while the North German population appears not to be expanding so far (reduction of positive grid cells by ca. 30% since 2013). As Ae. j. japonicus finds suitable climatic and ecological conditions in Germany, the differential expansion of the two populations might be attributed to the West German population being older and thus more firmly established than the closely related but younger North German population that might still be in its founder phase. However, geographic spread of all German populations in the future is anticipated. Continuous surveillance is recommended, as Ae. j. japonicus is a competent vector of several pathogens in the laboratory.

Detection of Invasive Mosquito Vectors Using Environmental DNA (eDNA) from Water Samples.

Repeated introductions and spread of invasive mosquito species (IMS) have been recorded on a large scale these last decades worldwide. In this context, members of the mosquito genus Aedes can present serious risks to public health as they have or may develop vector competence for various viral diseases. While the Tiger mosquito (Aedes albopictus) is a well-known vector for e.g. dengue and chikungunya viruses, the Asian bush mosquito (Ae. j. japonicus) and Ae. koreicus have shown vector competence in the field and the laboratory for a number of viruses including dengue, West Nile fever and Japanese encephalitis. Early detection and identification is therefore crucial for successful eradication or control strategies. Traditional specific identification and monitoring of different and/or cryptic life stages of the invasive Aedes species based on morphological grounds may lead to misidentifications, and are problematic when extensive surveillance is needed. In this study, we developed, tested and applied an environmental DNA (eDNA) approach for the detection of three IMS, based on water samples collected in the field in several European countries. We compared real-time quantitative PCR (qPCR) assays specific for these three species and an eDNA metabarcoding approach with traditional sampling, and discussed the advantages and limitations of these methods. Detection probabilities for eDNA-based approaches were in most of the specific comparisons higher than for traditional survey and the results were congruent between both molecular methods, confirming the reliability and efficiency of alternative eDNA-based techniques for the early and unambiguous detection and surveillance of invasive mosquito vectors. The ease of water sampling procedures in the eDNA approach tested here allows the development of large-scale monitoring and surveillance programs of IMS, especially using citizen science projects.

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.

BackgroundThe 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.MethodsBetween 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.ResultsBetween 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.ConclusionsOur 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.

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

BackgroundIn 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.FindingsIn 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.ConclusionsAe. 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.

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.

BackgroundThe 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.MethodsThe 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 km2 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.ResultsBased on the grid cells, an area of approximately 900 km2 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.ConclusionAs 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.Electronic supplementary materialThe online version of this article (doi:10.1186/s13071-016-1447-z) contains supplementary material, which is available to authorized users.

Field evaluation of baited traps for surveillance of Aedes japonicus japonicus in Switzerland.

The efficacy of Centers for Disease Control (CDC) miniature light traps and ovitraps was tested in the outskirts of the city of Zurich in Switzerland for their use in the surveillance of Aedes (Hulecoeteomyia) japonicus japonicus (Theobald) (Diptera: Culicidae), the invasive Asian bush mosquito. Sets of single CDC traps were run overnight (n = 18) in three different environments (forest, suburban and urban) in 3 × 3 Latin square experimental designs. Traps were baited with: (a) carbon dioxide (CO2 ); (b) CO2 plus light, or (c) CO2 plus lure blend [Combi FRC 3003 (iGu® )]. At the same locations, mosquito eggs were collected weekly using standard ovitraps baited with different infusions (oak, hay or tap water) and equipped with different oviposition substrates (a block of extruded polystyrene, a germination paper strip or a wooden stick). Data were analysed using Poisson and negative binomial general linear models. The use of light (P < 0.001) or lure (P < 0.001) significantly increased the attractiveness of CDC traps baited with CO2 . Oak and hay infusions did not increase the attractiveness of ovitraps compared with standing tap water (P > 0.05), and extruded polystyrene blocks were preferred as an oviposition substrate over wooden sticks (P < 0.05) and seed germination paper (P < 0.05). Carbon dioxide-baited CDC miniature light traps complemented with light or iGu® lure and ovitraps containing standing tap water and polystyrene oviposition blocks can be considered as efficient and simple tools for use in Ae. j. japonicus surveillance programmes.

Aedes japonicus japonicus (Diptera: Culicidae) arrives at the most easterly point in North America

AbstractAedes japonicus japonicus (Theobald) (Diptera: Culicidae), the Asian bush mosquito, is a keen biter linked to the transmission to humans of a variety of diseases. It has moved significantly from its historical Asian distribution, with its arrival in North America first noted in 1998 in New York and New Jersey, United States of America. Here we report the presence of A. j. japonicus within our collections of mosquitoes in the capital city of the easternmost province in Canada: St. John’s, Newfoundland and Labrador, in 2013. This observation provides further evidence of this mosquito’s ability to significantly expand its geographic range, potentially affecting connectivity between subpopulations globally.

Modeling of the putative distribution of the arbovirus vector Ochlerotatus japonicus japonicus (Diptera: Culicidae) in Germany.

Today, international travel and global freight transportation are increasing and have a direct influence on the introduction and establishment of non-native mosquito species as well as on the spread of arthropod (mosquito)-borne diseases inside Europe. One of the mosquito species that has become invasive in many areas is the Asian rock pool or bush mosquito Ochlerotatus japonicus japonicus (synonyms: Aedes japonicus japonicus or Hulecoeteomyia japonica japonica). This species was detected in Germany in 2008 for the first time. Until today, three different Oc. j. japonicus populations have been documented. Laboratory studies have shown that Oc. j. japonicus can act as a vector for a variety of disease agents. Thus, the knowledge on its current distribution is essential for different measurements. In the present study, ecological niche models were used to estimate the potential distribution of Oc. j. japonicus in Germany. The aim was to detect areas within Germany that could potentially function as habitats for this species. According to our model, areas in western, southern, and central Germany offer suitable conditions for the mosquito and may therefore be at risk for an invasion of the species. We strongly suggest that those areas should be monitored more intensively in the future. For this purpose, it would also be essential to search for possible dispersal routes as well as for natural barriers.

Mosquitoes as vectors for exotic pathogens in Germany

As a result of intensified globalization of international trade and of substantial travel activities, mosquito-borne exotic pathogens are becoming an increasing threat for Europe. In Germany some 50 different mosquito species are known, several of which have vector competence for pathogens. During the last few years a number of zoonotic arboviruses that are pathogenic for humans have been isolated from mosquitoes in Germany including Usutu, Sindbis and Batai viruses. In addition, filarial worms, such as Dirofilaria repens have been repeatedly detected in mosquitoes from the federal state of Brandenburg. Other pathogens, in particular West Nile virus, are expected to emerge sooner or later in Germany as the virus is already circulating in neighboring countries, e.g. France, Austria and the Czech Republic. In upcoming years the risk for arbovirus transmission might increase in Germany due to increased occurrence of new so-called "invasive" mosquito species, such as the Asian bush mosquito Ochlerotatus japonicus or the Asian tiger mosquito Aedes albopictus. These invasive species are characterized by high vector competence for a broad range of pathogens and a preference for human blood meals. For risk assessment, a number of mosquito and pathogen surveillance projects have been initiated in Germany during the last few years; however, mosquito control strategies and plans of action have to be developed and put into place to allow early and efficient action against possible vector-borne epidemics.

Rapid protein profiling for arthropod vector identification

Identification of insect vectors is primarily carried out using morphological features, which can be a time-consuming and difficult task. PCR-based approaches have been developed for the identification of a number of vector species. We have recently established matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF MS), which has come of age for the high throughput identification of microorganisms, to identify imagos and larvae of biting midges (Culicoides spp.), and the technique has proven its suitability to accurately identify field-collected biting midges on a large scale. We have extended our insect vector reference database to also include culicid mosquito and phlebotomine species. Currently, biomarker mass sets are determined for the imagos of 15 phlebotomines (9 Phlebotomus spp., 3 Lutzomyia spp., 3 Sergentomyia spp.), as well as for both imagos and larvae of 38 Central European culicid species, including all established and presumptive Aedine invasive species such as the Asian tiger mosquito Aedes albopictus or the Asian bush mosquito Ae. japonicus. In addition, biomarker mass sets of eggs were determined for indigenous and invasive container-inhabiting Aedine mosquitoes (Ae. aegypti, Ae. albopictus, Ae. atropalpus, Ae. cretinus, Ae. geniculatus, Ae. japonicus, Ae. koreicus, Ae. phoeniciae, Ae. triseriatus) allowing to identify eggs collected in oviposition traps in the framework of surveillance programs. Eggs can be identified by MALDI-TOF MS either singly or in pools (demonstrated for pools of 10 eggs, but possibly also for larger ones) for those species of which at least 3 eggs are present in the sample. We are continuously expanding our database, e.g. to also include the remaining mosquito species whose eggs can be encountered in surveillance programs of container-inhabiting mosquitoes in Europe, i.e. species usually breeding in tree holes or rock pools in southern Europe (Ae. berlandi, Ae. echinus, Ae. gilcolladoi, Ae. mariae, Ae. pulcritarsis, Ae. zammitii, and Orthopodomyia pulcripalpis). Taken together, protein profiling by MALDI-TOF MS is a quick and inexpensive tool to accurately identify adult and immature stages of insect vectors collected in the field, and the technique has the potential to become the method of choice for a centralized, robust and high throughput screening of insect vector populations in connection with surveillance programs. The simple sample preparation can be done in peripheral laboratories and the slides be sent to the measuring laboratory. The future application of the method will include the accomplishment of the measurement with a mass spectrometry device anywhere and the identification via our online platform.

The further spread of Aedes japonicus japonicus (Diptera, Culicidae) towards northern Germany

After its first detection in 2008 in the south German federal state of Baden-Wuerttemberg, another distinct population of the invasive Asian bush mosquito Aedes japonicus japonicus was unexpectedly found in western Germany in 2012. Range expansion had already been observed for the southern German population and was anticipated for the western German one. Here, we report on a third, apparently independent and even more northerly German colonization area of Aedes j. japonicus in southern Lower Saxony and northeastern North Rhine-Westphalia, which was discovered in spring 2013. In a snapshot study, intended to determine the presence or absence of Aedes j. japonicus in an area close to Hanover, the capital of the northern German federal state of Lower Saxony, where a specimen had been collected in late 2012, central water basins of cemeteries were checked for pre-imaginal mosquito stages at the beginning of the mosquito season 2013. Almost 20% of the inspected cemeteries were found positive (25 out of 129), with many of them being located in towns and villages close to the motorways A2 and A7. Being of Far Eastern origin, the Asian bush mosquito is well adapted to moderate climates and appears to be further expanding its distribution area in Central Europe. As it is a proven laboratory vector of several mosquito-borne disease agents, its present and future distribution areas should be carefully monitored.

A new focus of Aedes japonicus japonicus (Theobald, 1901) (Diptera, Culicidae) distribution in Western Germany: rapid spread or a further introduction event?

BackgroundThe Asian bush mosquito, Aedes japonicus japonicus, a potential vector of several viruses, was first detected in Germany in 2008 on the Swiss-German border. In the following years, this invasive species apparently succeeded in establishing populations in southern Germany and in spreading northwards. In 2011, its distribution area already covered large areas of the federal state of Baden-Wurttemberg, and its northernmost German collection point was reported to be close to Stuttgart. Several independent submissions to our laboratories of Ae. j. japonicus specimens in July 2012, originating from the same area in the federal state of North Rhine-Westphalia, western Germany, prompted us to carry out an immediate surveillance in this region in the expectation of finding a further distribution focus of Ae. j. japonicus in Germany.MethodsAfter inspecting the places of residence of the collectors of the submitted mosquito specimens, all kinds of water containers in 123 cemeteries in surrounding towns and villages were checked for mosquito developmental stages. These were collected and kept to produce adults for morphological species identification. One specimen per collection site was identified genetically by COI sequence analysis.ResultsAedes j. japonicus adults and immature stages were found in 36 towns/villages that were checked (29%) over an area of approximately 2,000 km2 in southern North Rhine-Westphalia and northern Rhineland Palatinate. The species could not be demonstrated further south when monitoring towards the northernmost previous collection sites in southern Germany. It therefore remains to be elucidated whether the species has entered western Germany from the south, from Belgium in the west where it has been demonstrated to occur locally since 2002, or through a new introduction.ConclusionsAedes j. japonicus is obviously much more widely distributed in Germany than previously thought. It appears to be well adapted, to have a strong expansion tendency and to replace indigenous mosquito species. Thus, a further spread is anticipated and elimination seems hardly possible anymore. The vector potency of the species should be reason enough to thoroughly monitor its future development in Germany.

FINDING NEEDLES IN THE HAYSTACK: SINGLE COPY MICROSATELLITE LOCI FOR AEDES JAPONICUS (DIPTERA: CULICIDAE)

First identified in three North American states in 1998, Aedes japonicus japonicus, the Asian bush mosquito, has since spread to 21 states, plus Ontario in Canada, northern France, and Belgium. Analyses of the introduction and expansion of this potentially deadly disease vector will be radically improved by including powerful genetic markers like microsatellites. Useful microsatellite loci have, however, been difficult to identify for mosquitoes in the genus Aedes because of the high amount of repetitive DNA in these species. We isolated single-copy DNA from Ae. j. japonicus and then used a standard enrichment method to identify regions containing microsatellites. Here we describe seven polymorphic microsatellite loci that were tested in American populations of Ae. j. japonicus. These loci were also found to be polymorphic in two other of the four Ae. japonicus subspecies and in Aedes koreicus.