Invasive Mosquito Research Articles

Genetic divergence between populations of feral and domestic forms of a mosquito disease vector assessed by transcriptomics.

Culex pipiens, an invasive mosquito and vector of West Nile virus in the US, has two morphologically indistinguishable forms that differ dramatically in behavior and physiology. Cx. pipiens form pipiens is primarily a bird-feeding temperate mosquito, while the sub-tropical Cx. pipiens form molestus thrives in sewers and feeds on mammals. Because the feral form can diapause during the cold winters but the domestic form cannot, the two Cx. pipiens forms are allopatric in northern Europe and, although viable, hybrids are rare. Cx. pipiens form molestus has spread across all inhabited continents and hybrids of the two forms are common in the US. Here we elucidate the genes and gene families with the greatest divergence rates between these phenotypically diverged mosquito populations, and discuss them in light of their potential biological and ecological effects. After generating and assembling novel transcriptome data for each population, we performed pairwise tests for nonsynonymous divergence (Ka) of homologous coding sequences and examined gene ontology terms that were statistically over-represented in those sequences with the greatest divergence rates. We identified genes involved in digestion (serine endopeptidases), innate immunity (fibrinogens and α-macroglobulins), hemostasis (D7 salivary proteins), olfaction (odorant binding proteins) and chitin binding (peritrophic matrix proteins). By examining molecular divergence between closely related yet phenotypically divergent forms of the same species, our results provide insights into the identity of rapidly-evolving genes between incipient species. Additionally, we found that families of signal transducers, ATP synthases and transcription regulators remained identical at the amino acid level, thus constituting conserved components of the Cx. pipiens proteome. We provide a reference with which to gauge the divergence reported in this analysis by performing a comparison of transcriptome sequences from conspecific (yet allopatric) populations of another member of the Cx. pipiens complex, Cx. quinquefasciatus.

Avian malaria parasites in the last supper: identifying encounters between parasites and the invasive Asian mosquito tiger and native mosquito species in Italy.

BackgroundThe invasive Asian tiger mosquito Aedes albopictus has dramatically expanded its distribution range, being catalogued as one of the world’s 100 worst invasive alien species. As vectors of pathogens, Ae. albopictus may create novel epidemiological scenarios in the invaded areas.MethodsHere, the frequency of encounters of Ae. albopictus with the avian malaria parasite Plasmodium and the related Haemoproteus was studied in an area with established populations in northeastern Italy and compared with those from four native mosquito species, Anopheles maculipennis s.l., Culex hortensis, Culex pipiens, and Ochlerotatus caspius. The abdomens of mosquitoes with a recent blood meal were used to identify both the blood meal source and the parasites harboured.ResultsAedes albopictus had a clear antropophilic behaviour while An. maculipennis and Oc. caspius fed mainly on non-human mammals. Birds were the most common hosts of Cx. pipiens and reptiles of Cx. hortensis. Parasites were isolated from three mosquito species, with Cx. pipiens (30%) showing the highest parasite prevalence followed by Cx. hortensis (9%) and Ae. albopictus (5%).ConclusionsThese results are the first identifying the avian malaria parasites harboured by mosquitoes in Italy and represent the first evidence supporting that, although Ae. albopictus could be involved in the transmission of avian malaria parasites, the risk of avian malaria parasite spread by this invasive mosquito in Europe would be minimal.

Geographic expansion of the invasive mosquito Aedes albopictus across Panama--implications for control of dengue and Chikungunya viruses.

The Asian tiger mosquito, Aedes (Stegomyia) albopictus, is an invasive species that has expanded its territory to over 40% of the earth’s terrestrial landmass in the last 30 years [1]. Ae. albopictus is an efficient vector of all serotypes of dengue, a disease that has increased in frequency over the past 30 years in the Americas [2], where it represents an annual cost of 2,100,000,000 USD per year [3]. This mosquito is also an efficient vector of the three genotypes of Chikungunya virus, a worldwide emerging pathogen that causes fever, fatigue, and joint swelling in humans. Since 2006, Chikungunya outbreaks have been increasingly recorded outside the virus’s native range in tropical Africa, perhaps because of a mutation in the virus’s envelope gene, which increases the replication and dissemination capacity of the virus in Ae. albopictus [4]. During the second quarter of 2014, Chikungunya has been detected throughout much of the Americas, with major outbreaks occurring in several Caribbean nations, and local transmission confirmed or suspected in the United States, Panama, Venezuela, Peru, and Chile, creating an imminent threat for humans throughout the Americas, who have no prior exposure to this infection [5]. The first cases of Chikungunya disease in Panama were reported in May 2014, occurring in nonresidents who most likely picked up the virus in their Caribbean countries of origin. On 23 July 2014, Panama’s health authority reported autochthonous transmission of Chikungunya virus. Coincidentally, the earliest cases involved patients located in Juan Diaz, an urban area on the eastern outskirts of Panama City, where the first specimen of invasive Ae. albopictus was collected in 2002. Ae. albopictus has expanded across much of Panama since that time, yet to date, no information exists about the degree of expansion or about the factors contributing to the geographic expansion of this important mosquito vector across Panama. Here, we map the temporal expansion of Ae. albopictus, use species distribution models to determine the ecological and nonecological factors associated with its expansion, and comment on the implications for vector and disease control programs in Panama and elsewhere in the American tropics.

Approaches to passive mosquito surveillance in the EU.

The recent emergence in Europe of invasive mosquitoes and mosquito-borne disease associated with both invasive and native mosquito species has prompted intensified mosquito vector research in most European countries. Central to the efforts are mosquito monitoring and surveillance activities in order to assess the current species occurrence, distribution and, when possible, abundance, in order to permit the early detection of invasive species and the spread of competent vectors. As active mosquito collection, e.g. by trapping adults, dipping preimaginal developmental stages or ovitrapping, is usually cost-, time- and labour-intensive and can cover only small parts of a country, passive data collection approaches are gradually being integrated into monitoring programmes. Thus, scientists in several EU member states have recently initiated programmes for mosquito data collection and analysis that make use of sources other than targeted mosquito collection. While some of them extract mosquito distribution data from zoological databases established in other contexts, community-based approaches built upon the recognition, reporting, collection and submission of mosquito specimens by citizens are becoming more and more popular and increasingly support scientific research. Based on such reports and submissions, new populations, extended or new distribution areas and temporal activity patterns of invasive and native mosquito species were found. In all cases, extensive media work and communication with the participating individuals or groups was fundamental for success. The presented projects demonstrate that passive approaches are powerful tools to survey the mosquito fauna in order to supplement active mosquito surveillance strategies and render them more focused. Their ability to continuously produce biological data permits the early recognition of changes in the mosquito fauna that may have an impact on biting nuisance and the risk of pathogen transmission associated with mosquitoes. International coordination to explore synergies and increase efficiency of passive surveillance programmes across borders needs to be established.

Human‐aided and natural dispersal drive gene flow across the range of an invasive mosquito

Human-aided transport is responsible for many contemporary species introductions, yet the contribution of human-aided transport to dispersal within non-native regions is less clear. Understanding dispersal dynamics for invasive species can streamline mitigation efforts by targeting routes that contribute disproportionally to spread. Because of its limited natural dispersal ability, rapid spread of the Asian tiger mosquito (Aedes albopictus) has been attributed to human-aided transport, but until now, the relative roles of human-aided and natural movement have not been rigorously evaluated. Here, we use landscape genetics and information-theoretic model selection to evaluate 52 models representing 9240 pairwise dispersal paths among sites across the US range for Ae.albopictus and show that recent gene flow reflects a combination of natural and human-aided dispersal. Highways and water availability facilitate dispersal at a broad spatial scale, but gene flow is hindered by forests at the current distributional limit (range edge) and by agriculture among sites within the mosquito's native climatic niche (range core). Our results show that highways are important to genetic structure between range-edge and range-core pairs, suggesting a role for human-aided mosquito transport to the range edge. In contrast, natural dispersal is dominant at smaller spatial scales, reflecting a shifting dominance to natural movement two decades after introduction. These conclusions highlight the importance of (i) early intervention for species introductions, particularly those with readily dispersed dormant stages and short generation times, and (ii) strict monitoring of commercial shipments for transported immature stages of Ae.albopictus, particularly towards the northern edge of the US range.

Economic evaluation of an area-wide integrated pest management program to control the Asian tiger mosquito in New Jersey.

Aedes albopictus is the most invasive mosquito in the world, an important disease vector, and a biting nuisance that limits outdoor activities. Area-wide integrated pest management (AW-IPM) is the recommended control strategy. We conducted an economic evaluation of the AW-IPM project in Mercer and Monmouth Counties, New Jersey with a controlled design (AW-IPM vs. control) from 2009 through 2011. The study analyzed financial documents and staff time for AW-IPM and surveyed an average of 415 randomly chosen households in AW-IPM and control areas each fall from 2008 through 2011. Hours lost from yard and porch activities were calculated as differences between actual and potential hours of these activities in an average summer week if there had been no mosquito concerns. Net estimated benefits of AW-IPM were based on cross-over and difference-in-difference analyses. Reductions in hours lost were valued based on respondents' willingness to pay for a hypothetical extra hour free of mosquitoes spent on yard or porch activities and literature on valuation of a quality adjusted life year (QALY). The incremental cost of AW-IPM per adult was $41.18 per year. Number of hours lost due to mosquitoes in AW-IPM areas between the base year (2008) and the intervention years (2009-2011) declined by 3.30 hours per summer week in AW-IPM areas compared to control areas. Survey respondents valued this improvement at $27.37 per adult per summer week. Over the 13-week summer, an average adult resident gained 42.96 hours of yard and porch time, worth $355.82. The net benefit over the summer was $314.63. With an average of 0.0027 QALYs gained per adult per year, AW-IPM was cost effective at $15,300 per QALY gained. The benefit-cost ratio from hours gained was 8.64, indicating that each $1 spent on AW-IPM gave adults additional porch and yard time worth over $8.

Size Matters: Purple Pitcher Plant (Sarracenia purpurea) Pitcher Size Influences the Presence of Aedes albopictus Larvae

A vector of multiple arboviruses, the invasive Asian tiger mosquito Aedes albopictus was recently found to develop inside the pitchers of the purple pitcher plant Sarracenia purpurea in a forested environment. In this study larvae and pupae of Ae. albopictus were found in S. purpurea pitchers in a peridomestic habitat in central North Carolina. Evidence of direct oviposition on the inner sides of the pitchers was obtained by dissecting pitchers and examining the insides for eggs, which were then raised to adults. To determine if pitcher size and origin (same or different plant) affected the presence of immature Ae. albopictus, these variables were measured and compared against the presence/absence of larvae and/or pupae through a logistic regression analysis. Statistical evidence showed pitchers with larger openings were much more likely to contain larvae and/or pupae, and pitchers from the same plant were not significantly more often colonized than pitchers on other plants. These data, in combination with the presence of dead adult females in pitchers, suggest the morphology of some pitchers may prevent successful oviposition by gravid females.

Updated distribution of Aedes albopictus (Diptera: Culicidae) in Spain: new findings in the mainland Spanish Levante, 2013

In 2004, Aedes (Stegomyia) albopictus (Skuse, 1894) was observed for the first time in Catalonia, northeastern Spain. A decade later, it has spread throughout the eastern Mediterranean region of the country and the Balearic Islands. Framed within a national surveillance project, we present the results of monitoring in 2013 in the autonomous communities of the mainland Levante. The current study reveals a remarkable increase in the spread of the invasive mosquito in relation to results from 2012; the species was present and well-established in 48 municipalities, most of which were along the Mediterranean coastline from the Valencian Community to the Region of Murcia.

Chikungunya in the United States

Chikungunya is characterized by severe and often enduring febrile arthralgia, which is rarely fatal. The clinical symptoms include febrile poly-arthralgia, and small joints are frequently affected. The general symptomatology is most often headache, asthenia, myalgia and chills, combined with joint swelling, rash, buccal bleeding and mouth ulcer; chronic arthralgia is a frequent complication of acute disease. The capability of Aedes albopictus to effectively transmit the Asian genotype of the chikungunya virus that was recently found in the Caribbean could facilitate local transmission and spread into the United States towards areas where this invasive mosquito is widespread. Locally acquired infections have been confirmed in the Caribbean, where North American travelers frequently visit. The three major destinations of passengers departing the regions of the Caribbean are the US, France, and the Netherlands Antilles. Seven months after the disease was detected in the Caribbean Islands, the first autochthonous transmission of infection was reported on July 17, 2014 in Florida in a male patient who had not recently visited any foreign country. The arrival of Chikungunya in the US underscores the risks posed by this and other exotic pathogens.

Distribution and genetic structure of Aedes japonicus japonicus populations (Diptera: Culicidae) in Germany.

In recent years, the number of imported cases of arthropod-borne diseases in Europe, such as dengue fever, has increased steadily, as did the emergence and distribution of invasive insect vectors. Consequently, the risk of disease spreading into previously unaffected regions through invasive mosquitoes is also increasing. One example of an invasive mosquito is Aedes japonicus japonicus (A. j. japonicus), which spread from its original habitat in Japan to North America and Europe. This species has been shown to act as a vector for Japanese encephalitis and West Nile viruses. In Europe, A. j. japonicus has been detected in Switzerland, Belgium, Slovenia, and Germany, where it has become a resident species. Here, we describe the recent spread and genetic structure of A. j. japonicus populations in Germany. By monitoring the species in Baden-Württemberg in 2011 and 2012, we observed a considerable enlargement of the infested area from 54 municipalities in 2011 to 124 municipalities in 2012. To elucidate the colonization of Europe by A. j. japonicus, seven microsatellite loci were studied in 106 individuals sampled in Germany and Switzerland in 2012. The same markers were genotyped in 31 North American and 26 Japanese specimens. Population genetic analyses indicated that A. j. japonicus in Baden-Württemberg and North Rhine-Westphalia represented two genetically distinct populations with FST-values of 0.073-0.152, suggesting that they originated from two independent introduction events in the past. These results are of particular interest in light of vectorial variability for the transmission of viruses and other pathogens in Europe.

Chikungunya Virus: A Major Emerging Threat

Chikungunya Virus: A Major Emerging Threat

Insecticide resistance status of United States populations of Aedes albopictus and mechanisms involved.

Aedes albopictus (Skuse) is an invasive mosquito that has become an important vector of chikungunya and dengue viruses. Immature Ae. albopictus thrive in backyard household containers that require treatment with larvicides and when adult populations reach pest levels or disease transmission is ongoing, adulticiding is often required. To assess the feasibility of control of USA populations, we tested the susceptibility of Ae. albopictus to chemicals representing the main insecticide classes with different modes of action: organochlorines, organophosphates, carbamates, pyrethroids, insect growth regulators (IGR), naturalytes, and biolarvicides. We characterized a susceptible reference strain of Ae. albopictus, ATM95, and tested the susceptibility of eight USA populations to five adulticides and six larvicides. We found that USA populations are broadly susceptible to currently available larvicides and adulticides. Unexpectedly, however, we found significant resistance to dichlorodiphenyltrichloroethane (DDT) in two Florida populations and in a New Jersey population. We also found resistance to malathion, an organophosphate, in Florida and New Jersey and reduced susceptibility to the IGRs pyriproxyfen and methoprene. All populations tested were fully susceptible to pyrethroids. Biochemical assays revealed a significant up-regulation of GSTs in DDT-resistant populations in both larval and adult stages. Also, β-esterases were up-regulated in the populations with suspected resistance to malathion. Of note, we identified a previously unknown amino acid polymorphism (Phe → Leu) in domain III of the VGSC, in a location known to be associated with pyrethroid resistance in another container-inhabiting mosquito, Aedes aegypti L. The observed DDT resistance in populations from Florida may indicate multiple introductions of this species into the USA, possibly from tropical populations. In addition, the mechanisms underlying DDT resistance often result in pyrethroid resistance, which would undermine a remaining tool for the control of Ae. albopictus. Continued monitoring of the insecticide resistance status of this species is imperative.

Comparison of the vector potential of different mosquito species for the transmission of heartworm, Dirofilaria immitis, in rural and urban areas in and surrounding Stillwater, Oklahoma, U.S.A.

Dirofilaria immitis Leidy (Spirurida: Onchocercidae), or heartworm, is a mosquito-borne nematode that causes a fatal disease in carnivores. Although infection is preventable through prophylactic drugs, compliance and the spectre of resistance suggest vector control is a viable alternative. There were two main objectives in this study: (a) to evaluate the relationships between landscape and social factors and the number and species of heartworm-positive mosquitoes, with a specific focus on the importance of the invasive Asian tiger mosquito, Aedes albopictus (Skuse) (Stegomyia albopicta) (Diptera: Culicidae), and (b) to test the hypothesis that dog heartworm is more prevalent in suburban than in rural areas. To achieve these objectives, mosquitoes were collected from May to November 2010 at 16 rural and 16 urban locations in Payne County, Oklahoma, U.S.A. using three trapping methods that utilized, respectively, resting boxes, carbon dioxide traps and BG Sentinel traps. Urban areas showed greater numbers of Ae. albopictus and a higher overall likelihood of infection with D. immitis. Because many species of mosquito are responsible for heartworm transmission, current prophylactic treatment remains the best method of controlling this parasite.

Book Reviews Kay, Brian H. and Richard C. Russell [Eds.] 2013. Mosquito Eradication. The Story of KillingCampto, CSIRO Publishing, Collingwood, VIC. Australia, pp., paperback. ISBN 9781486300570, $69.95 (AU).

Kay, Brian H. and Richard C. Russell [Eds.] 2013. Mosquito Eradication. The Story of Killing. Campto, CSIRO Publishing, Collingwood, VIC. Australia. xxii + 256 pp., paperback. ISBN 9781486300570, $69.95 (AU). This multi-authored volume chronicles stages and strategic planning of a remarkable achievement, the unprecedented eradication of an invasive salt marsh mosquito pest and potential disease vector. The book is a collaboration between Australian experts, including the editors, who are doyens of mosquito biology and control Down Under, and New Zealand scientists, government workers, and response teams whose timely engagement was responsible for the eventual purging of the Australian southern salt marsh mosquito Aedes camptorhynchus from their country. The cooperation between these neighbors is appropriate and not unexpected, given their shared concerns about invasive species and ‘biosecurity’, plus the fact that the unwanted Campto was inadvertently introduced into NZ from Australia. View this article in BioOne

Distribution of two invasive mosquito species in Slovenia in 2013

In the recent years there has been growing interest in the establishment and spread of invasive mosquito species in Europe. Apart from the most dispersed, Aedes albopictus, there are five other alien aedine mosquito species spreading in Europe. Their spreading represents a considerable threat for public health as they can serve as vectors of mosquito-borne diseases. A survey in whole Slovenia was performed in 2013 to determine the distribution of invasive mosquitoes. The study was based on the search for invasive mosquito larvae in artificial water containers at the cemeteries, around human dwellings and in used tires at vulcanizing companies. Additionally, a call for citizens to report any nuisance from unusual mosquitoes was published in local media. With the study we have revealed the presence of two invasive mosquito species in Slovenia, Ae. albopictus and Ae. japonicus. Altogether we caught 494 larvae of Ae. albopictus and 1318 larvae of Ae. japonicus. The results show that Ae. albopictus is present mostly in Southwestern and central part of the country, with some isolated locations in other parts of Slovenia. Nevertheless, Ae. japonicus was found in a large part of the investigated area. This species was previously collected from a small area near the Austrian border in 2011, and in only two years it has colonized the majority of Northeastern part of Slovenia. In Slovenia no systematic mosquito surveillance or control is currently done. There are only some minor projects, which are carried out by local institutions at a regional level. Ae. albopictus and Ae. japonicus are spreading fast and they both present high nuisance for the humans, as well as a public health risk since transmitting vector borne diseases. Therefore an effective national monitoring program of invasive mosquito species is highly needed. With well planned, long term strategy we will be able to detect and control their occurrence, monitor their pathogens and prevent the establishment of new foci of mosquito-borne diseases.

Rapid protein profiling facilitates surveillance of invasive mosquito species

BackgroundInvasive aedine mosquito species have become a major issue in many parts of the world as most of them are recognised vectors or potentially involved in transmission of pathogens. Surveillance of these mosquitoes (e.g. Ae. aegypti, Yellow fever mosquito, Aedes albopictus, Asian tiger mosquito) is mainly done by collecting eggs using ovitraps and by identification of the larvae hatched in the laboratory. In order to replace this challenging and laborious procedure, we have evaluated matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF MS) for easy and rapid species identification.MethodsIndividual protein profiles were generated using five eggs each of nine aedine species (Ae. aegypti, Ae. albopictus, Ae. atropalpus, Ae. cretinus, Ae. geniculatus, Ae. japonicus, Ae. koreicus, Ae. phoeniciae, Ae. triseriatus) from various geographical origins, and species-specific biomarker mass sets could be generated. A blinded validation using our reference data base for automated egg identification was performed. In addition, pools of 10 aedine eggs (132 two-species and 18 three-species pools) in different ratios were evaluated.ResultsSpecific biomarker mass sets comprising 18 marker masses could be generated for eggs of nine container-inhabiting aedine species, including all the major invasive and indigenous species of Europe and North America. Two additional masses shared by all investigated aedine species are used as internal calibrators. Identification of single eggs was highly accurate (100% specificity, 98.75% sensitivity), and this method is also of value for the identification of species in pools of ten eggs. When mixing two or three species, all were identified in all pools in at least 2 or 1 of the 4 loaded replicates, respectively, if the “lesser abundant” species in the pool accounted for three or more eggs.ConclusionsMALDI-TOF MS, which is widely applied for routine identification of microorganisms in clinical microbiology laboratories, is also suited for robust, low-cost and high throughput identification of mosquito vectors in surveillance programmes. This tool can further be developed to include a wide spectrum of arthropods but also other Metazoa for which surveillance is required, and might become the method of choice for their centralised identification via online platforms.

Climatic suitability of Aedes albopictus in Europe referring to climate change projections: comparison of mechanistic and correlative niche modelling approaches

The Asian tiger mosquito, Aedes albopictus, is capable of transmitting a broad range of viruses to humans. Since its introduction at the end of the 20th century, it has become well established in large parts of southern Europe. As future expansion as a result of climate change can be expected, determining the current and projected future climatic suitability of this invasive mosquito in Europe is of interest. Several studies have tried to detect the potential habitats for this species, but differing data sources and modelling approaches must be considered when interpreting the findings. Here, various modelling methodologies are compared with special emphasis on model set-up and study design. Basic approaches and model algorithms for the projection of spatio-temporal trends within the 21st century differ substantially. Applied methods range from mechanistic models (e.g. overlay of climatic constraints based on geographic information systems or rather process-based approaches) to correlative niche models. We conclude that spatial characteristics such as introduction gateways and dispersal pathways need to be considered. Laboratory experiments addressing the climatic constraints of the mosquito are required for improved modelling results. However, the main source of uncertainty remains the insufficient knowledge about the species' ability to adapt to novel environments.

First case of laboratory-confirmed Zika virus infection imported into Europe, November 2013

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Sexual harassment and feeding inhibition between two invasive dengue vectors.

Two invasive mosquitoes, Aedes aegypti and Aedes albopictus, have been interacting during the course of a rapid range expansion by A. albopictus. We investigated the potential for interspecific feeding interference by male mosquitoes interacting with females within and between these species. A. aegypti feeding on both sugar and blood was suppressed when females of this species were exposed to A. albopictus males, but no change was observed when exposed to conspecifics. A. albopictus feeding was not affected by males of either species. The potential consequences of these behaviors are discussed within the context of other known interspecific effects, all of which appear to favor the displacement of A. aegypti by A. albopictus.

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.