Tick and mosquito management is important to public health protection. At the same time, growing concerns about declines of pollinator species raise the question of whether vector control practices might affect pollinator populations. We report the results of a task force of the North American Pollinator Protection Campaign (NAPPC) that examined potential effects of vector management practices on pollinators, and how these programs could be adjusted to minimize negative effects on pollinating species. The main types of vector control practices that might affect pollinators are landscape manipulation, biocontrol, and pesticide applications. Some current practices already minimize effects of vector control on pollinators (e.g., short-lived pesticides and application-targeting technologies). Nontarget effects can be further diminished by taking pollinator protection into account in the planning stages of vector management programs. Effects of vector control on pollinator species often depend on specific local conditions (e.g., proximity of locations with abundant vectors to concentrations of floral resources), so planning is most effective when it includes collaborations of local vector management professionals with local experts on pollinators. Interventions can then be designed to avoid pollinators (e.g., targeting applications to avoid blooming times and pollinator nesting habitats), while still optimizing public health protection. Research on efficient targeting of interventions, and on effects on pollinators of emerging technologies, will help mitigate potential deleterious effects on pollinators in future management programs. In particular, models that can predict effects of integrated pest management on vector-borne pathogen transmission, along with effects on pollinator populations, would be useful for collaborative decision-making.

Hard ticks (Acari: Ixodidae) are blood-sucking ectoparasites characterized by the extended period of their attachment to their host. To access their bloodmeal, ticks secrete saliva containing a range of molecules that target the host's inflammation, immune system, and hemostatic components. Some of these molecules reportedly possess antiangiogenic and antitumor properties. The present study describes our investigation, the first of its kind, of the antiangiogenic and antitumoral effects of the Hyalomma dromedarii Koch, 1844 (Acari: Ixodidae), salivary gland extract (SGE), which inhibited the adhesion and migration of Human Umbilical Vein Endothelial Cells (HUVECs) in a dose-dependent manner, as well as angiogenesis in the Chick Chorioallantoic Membrane model. Interestingly, H. dromedarii SGE exerted an antiproliferative effect on U87 glioblastoma cells and inhibited their adhesion and migration to fibrinogen. These results open up new possibilities for characterizing and developing new molecules involved in the key steps of tumor progression.

Bed bugs (Cimex lectularius L.) are now endemic in most major cities, but information regarding their basic biology is still largely based on research done over four decades ago. We investigated the effects of starvation, mating, sperm storage, and female and male age on egg production and hatch. Egg production cycles varied with the number of bloodmeals that females received. Once-mated females fed every 5 d had constant egg production for ∼75 d followed by a monotonic decline to near zero. Percentage egg hatch was high and constant, but declined after ∼30 d to near zero. To determine whether the age of the female, male, or sperm affected these patterns, we mated newly eclosed females to 60-d-old virgin males, 60-d-old mated males, or newly eclosed males. Females produced the most eggs when mated to young males, followed by old mated males, and then old virgin males; percentage hatch followed a similar pattern, suggesting that sperm stored within males for long was deficient. To examine effects of sperm stored within females, we mated newly eclosed females, starved them for 30 or 60 d, then fed them every 5 d. The 60-d starved group produced fewer eggs than the 30-d starved group, and both produced fewer eggs than young females mated to old or young males. Longer periods of sperm storage within females caused lower corresponding percentage hatch. These findings indicate egg production and hatch are governed by complex interactions among female and male age, frequency of feeding and mating, and sperm condition.

Sarcosaprophagous flesh flies are insect species frequently found on corpses, and their developmental patterns can be used as reliable indicators for estimating minimum postmortem interval (PMI_min). In this study, the Boettcherisca peregrina Robineau-Desvoidy was reared at seven constant temperatures ranging from 16 to 34 °C to investigate the indexes of development duration and larval body length. Using these results, we generated three development models, including isomorphen diagram, isomegalen diagram, and thermal summation model. Regression analysis was conducted to obtain simulation equations of the variation in larval body length with time after larviposition, and variation in time after larviposition with body length. The developmental durations of B. peregrina from larviposition to adult eclosion under 16, 19, 22, 25, 28, 31 and 34 °C were 1,064.7 ± 34.8, 756.0 ± 19.0, 559.6 ± 5.5, 414.3 ± 3.9, 315.0 ± 2.0, 278.0 ± 4.0, and 258.0 ± 3.5 h, respectively. The developmental threshold temperature of B. peregrina was 10.87 ± 0.49 °C, and the thermal summation constant was 5,809.7 ± 291.4 degree days. The results of this study provide basic data for the use of B. peregrina for estimating PMI_min.

Each biome has its own fauna and intrinsic local conditions that determine the succession patterns of insects on carcasses. For this reason, regional studies are very important to forensics. This study deals with the flies that visit carcasses of Rattus rattus (L., 1758) in the Atlantic Forest remnant of Floresta da Tijuca, Rio de Janeiro, Brazil. We compare the diversity and relative abundance of fly families and species throughout the four seasons of the year and test for correlations between certain environmental variables and fly richness. In each of the four seasons, we exposed the carcasses of three rats. The carcasses were put in traps that were installed on the ground, separated by 100 m. The flies were collected from the carcasses on a daily basis, and were taken to the laboratory and kept in a climate chamber (28 °C day and 26 °C night). A total of 5,537 flies were captured, identified into 12 families: Calliphoridae (4,884 specimens), followed by Sarcophagidae, Micropezidae, Fannidae, Muscidae, Phoridae, and Neridae. Canonical correspondence and redundancy analyses of the environmental variables indicated no relationship between the seasons and fly abundance. Carcass decomposition lasted longer in the winter (12 d) than in the summer (8 d). The Margalef richness index indicated higher richness in the spring and winter, and lower richness in the summer. The Shannon–Wiener index and Simpson dominance indicated greater diversity in the spring. Diptera colonized all stages of decomposition. The families Calliphoridae and Sarcophagidae were more prevalent during the Fermentation stage. Our data have important implications for forensic entomology.

In Slovenia, two invasive mosquito species are present, Aedes albopictus (Skuse, 1895) (Diptera: Culicidae) and Aedes japonicus (Theobald, 1901) (Diptera: Culicidae). In this study, we examined their actual distribution and suitable habitats for new colonizations. Data from survey of species presence in 2013 and 2015, bioclimatic variables and altitude were used for the construction of predictive maps. We produced various models in Maxent software and tested two bioclimatic variable sets, WorldClim and CHELSA. For the variable selection of A. albopictus modeling we used statistical and expert knowledge-based approach, whereas for A. j. japonicus we used only a statistically based approach. The best performing models for both species were chosen according to AIC score-based evaluation. In 2 yr of sampling, A. albopictus was largely confined to the western half of Slovenia, whereas A. j. japonicus spread significantly and can be considered as an established species in a large part of the country. Comparison of models with WorldClim and CHELSA variables for both species showed models with CHELSA variables as a better tool for prediction. Finally, we validated the models performance in predicting distribution of species according to collected field data. Our study confirms that both species are co-occurring and are sympatric in a large part of the country area. The tested models could be used for future prevention of invasive mosquitoes spreading in other countries with similar bioclimatic conditions.

Aedes albopictus (Skuse) (Diptera: Culicidae) is distributed widely and is common in much of Japan. In Japan, female adults begin to bite in between April and June, except in the southern subtropics where the mosquito has no dormant period. It is difficult to estimate the first Ae. albopictus biting day because it varies annually depending on the location. Over several years, we surveyed the mosquitoes at different locations that covered a range of warmer to cooler areas of Japan. We found an association between the timing of first biting day by Ae. albopictus and spring temperature. In spring months, the strongest correlation was found with mean April temperatures, followed by March. Based on these data, it may, therefore, be possible to apply a simple formula to predict the timing of the first biting day at various geographical locations in Japan. Forecasting maps were created using a simple prediction formula. We found that the first biting day for Ae. albopictus changed depending on early spring temperatures for each year.There was an approximate 20-d difference in first biting day between years with warmer and cooler springs. This prediction model will provide useful insight for planning and practice of Ae. albopictus control programs, targeting larvae and adults, in temperate regions globally.

Phlebotomus caucasicus Marzinovsky and Phlebotomus mongolensis Sinton are morphologically similar sand fly species. Finding a reliable, fast, and simple method to differentiate these two sand flies is important in understanding their role in the transmission of Leishmania parasite. In our study, 20 specimens of male P. caucasicus, 4 specimens of male P. mongolensis, and 16 specimens of female of both species (Caucasicus group) were examined by polymerase chain reaction (PCR). The result shows identical patterns with a visible fragment of about 500 bp in size. In restriction fragment length polymorphism (RFLP), we observed identical patterns with TasI used as the restriction enzyme. After alignment with sequences of the ITS2 partial gene in GenBank, a perfect match was obtained for the P. mongolensis, but not for P. caucasicus whose sequence was not present in the GenBank. Based on the results of our study, the RFLP-PCR method with nucleotide gene fragment ITS2 was a rapid and reliable method for differentiating these sand fly species.

The insect integument, formed by the cuticle and the underlying epidermis, is essential for insect fitness, regulation of lipid biosynthesis and storage, insect growth and feeding, together with development progress. Its participation in insecticide resistance has also been outlined. Triatoma infestans Klug (Hemiptera: Reduviidae) is one of the major vectors of Chagas disease in South America; however, genomic data are scarce. In this study, we performed a transcriptome analysis of the nymph integument in order to identify which genes are expressed and their putative role. Using the 454 GS-FLX sequencing platform, we obtained approximately 144,620 reads from the integument tissue. These reads were assembled into 6,495 isotigs and 8,504 singletons. Based on BLAST similarity searches, about 8,000 transcripts were annotated with known genes, conserved domains, and/or Gene Ontology terms. The most abundant transcripts corresponded to transcription factors and nucleic acid metabolism, membrane receptors, cell signaling, and proteins related to cytoskeleton, transport, and cell energy processes, among others. More than 10% of the transcripts-encoded proteins putatively involved in the metabolism of fatty acids and related components (fatty acid synthases, elongases, desaturases, fatty alcohol reductases), structural integument proteins, and the insecticide detoxification system (among them, cytochrome P450s, esterases, and glutathione transferases). Real-time qPCR assays were used to investigate their putative participation in the resistance mechanism. This preliminary study is the first transcriptome analysis of a triatomine integument, and together with prior biochemical information, will help further understandthe role of the integument in a wide array of mechanisms.

Simulium (Gomphostilbia) laosense sp. nov. is described based on adults, pupae, and mature larvae from Laos. This new species is placed in the Simulium batoense species-group of the subgenus Gomphostilbia Enderlein. It is characterized by the pupal gill with eight filaments arranged as 3 + 3 + 2 from dorsal to ventral, of which an inner filament of the ventral pair is slightly longer than its counter filament, and is 1.7–1.8 times as long as filaments of the middle triplet. Taxonomic notes are provided to distinguish this new species from Simulium (G.) johorense Takaoka, Sofian-Azirun & Ya’cob from Peninsular Malaysia and four other related species. The phylogenetic position of this new species in the S. batoense species-group is also presented based on the mitochondrial COI gene. This new species represents the second species known from Laos.

Simulium (Gomphostilbia) fukudae sp. nov. is described based on females, males, pupae, and larvae from Thailand. This new species is placed in the Simulium batoense species-group of Simulium (Gomphostilbia). It is characterized by the female frons almost bare, male paramere covered with minute setae, pupal gill composed of six slender filaments, pupal terminal hooks cone-shaped, and cocoon with an anterodorsal projection. Taxonomic notes are given to separate this new species from the most similar species, Simulium (G.) sextuplum Takaoka & Davies and Simulium (G.) paukatense Takaoka, both from Malaysia.

Thaumetopoea pityocampa (Denis & Schiffermüller) (Lepidoptera: Notodontidae) is harmful to conifer trees because of defoliation and to public health because of the release of urticating setae from the caterpillars. Contact with setae by humans and domestic animals induces dermatitis, usually localized to the exposed areas. Recent studies demonstrated the presence of a complex urticating mechanism where proteins present in the setae may play a role as activators of immune responses. Yet, limited information is available at present about the proteins occurring in the setae of T. pityocampa. Using a refined method for protein extraction from the setae, and a combination of liquid chromatography tandem-mass spectrometry (LC-MS/MS), de novo assembly of transcriptomic data, and sequence similarity searches, an extensive data set of 353 proteins was obtained. These were further categorized by molecular function, biological process, and cellular location. All the 353 proteins identified were found to match through BLAST search with at least one Lepidoptera sequence available in databases. We found the previously known allergens Tha p 1 and Tha p 2 described from T. pityocampa, as well as enzymes involved in chitin biosynthesis, one of the principal components of the setae, and serine proteases that were responsible for inflammatory and allergic reactions in other urticating Lepidoptera. This new proteomic database may allow for a better understanding of the complexity of allergenic reactions due to T. pityocampa and to other Lepidoptera sharing similar defense systems.

Aedes aegypti (L.) and Ae. albopictus (Skuse) are important arbovirus vectors in the United States, and the recent emergence of Zika virus disease as a public health concern in the Americas has reinforced a need for tools to rapidly distinguish between these species in collections made by vector control agencies. We developed a duplex real-time PCR assay that detects both species and does not cross-amplify in any of the other seven Aedes species tested. The lower limit of detection for our assay is equivalent to ~0.03 of a first-instar larva in a 60-μl sample (0.016 ng of DNA per real-time PCR reaction). The assay was sensitive and specific in mixtures of both species that reflected up to a 2,000-fold difference in DNA concentration. In addition, we developed a simple protocol to extract DNA from sonicated first-instar larvae, and used that DNA to test the assay. Because it uses real-time PCR, the assay saves time by not requiring a separate visualization step. This assay can reduce the time needed for vector control agencies to make species identifications, and thus inform decisions about surveillance and control.

The global expansion of Ae. albopictus from its native range in Southeast Asia has been implicated in the recent emergence of dengue endemicity in Malaysia. Genetic variability studies of Ae. albopictus are currently lacking in the Malaysian setting, yet are crucial to enhancing the existing vector control strategies. The study was conducted to establish the genetic variability of maternally inherited mitochondrial DNA encoding for cytochrome oxidase subunit 1 (CO1) gene in Ae. albopictus. Twelve localities were selected in the Subang Jaya district based on temporal indices utilizing 120 mosquito samples. Genetic polymorphism and phylogenetic analysis were conducted to unveil the genetic variability and geographic origins of Ae. albopictus. The haplotype network was mapped to determine the genealogical relationship of sequences among groups of population in the Asian region. Comparison of Malaysian CO1 sequences with sequences derived from five Asian countries revealed genetically distinct Ae. albopictus populations. Phylogenetic analysis revealed that all sequences from other Asian countries descended from the same genetic lineage as the Malaysian sequences. Noteworthy, our study highlights the discovery of 20 novel haplotypes within the Malaysian population which to date had not been reported. These findings could help determine the genetic variation of this invasive species, which in turn could possibly improve the current dengue vector surveillance strategies, locally and regionally.

The continuing worldwide increase in urbanization can potentially have a major impact on the epidemiology of vector-borne diseases, as anthropogenic changes to the environment are known to favor a few species of mosquitoes that can thrive in urban environments. Culex quinquefasciatus Say (Diptera: Culicidae) is found extensively in urban habitats, where it lives in degraded, polluted areas and is often the only species of mosquito capable to surviving under such conditions. Traditional mosquito control strategies no longer have the desired effect due to the several factors such as insecticide resistance, abundance of breeding sites, lack of proper sewage and sanitation, and absence of natural predator, leading Cx. quinquefasciatus populations to increase its numbers in cities. In this study, five Cx. quinquefasciatus populations were analyzed using 12 microsatellite markers to investigate whether the dynamics of these populations are being modulated by urbanization and how they are structured in the city of São Paulo, Brazil. Our results indicate that only one of the study populations (the population from Anhanguera Park) exhibited evidence of expansion.The populations from Ibirapuera Park and Piqueri Park, the most urbanized regions of the areas studied, did not show signs of expansion. A better understanding of the mechanisms involved in the dispersal of Cx. quinquefasciatus and its colonization of new areas, as well as the species' demographic patterns and how these are associated with urbanization, particularly in areas undergoing a rural-to-urban transformation, such as Anhanguera Park, is of great importance for mosquito control.

The impact of the rapid expansion of rubber plantations in South-East Asia on mosquito populations is uncertain. We compared the abundance and diversity of adult mosquitoes using human-baited traps in four typical rural habitats in northern Lao PDR: secondary forests, immature rubber plantations, mature rubber plantations, and villages. Generalized estimating equations were used to explore differences in mosquito abundance between habitats, and Simpson's diversity index was used to measure species diversity. Over nine months, 24,927 female mosquitoes were collected, including 51 species newly recorded in Lao PDR. A list of the 114 mosquito species identified is included. More mosquitoes, including vector species, were collected in the secondary forest than immature rubber plantations (rainy season, odds ratio [OR] 0.33, 95% confidence interval [CI] 0.31–0.36; dry season, 0.46, 95% CI 0.41–0.51), mature rubber plantations (rainy season, OR 0.25, 95% CI 0.23–0.27; dry season, OR 0.25, 95% CI 0.22–0.28), and villages (rainy season, OR 0.13, 95% CI 0.12–0.14; dry season, 0.20, 95% CI 0.18–0.23). All habitats showed high species diversity (Simpson's indexes between 0.82–0.86) with vectors of dengue, Japanese encephalitis (JE), lymphatic filariasis, and malaria. In the secondary forests and rubber plantations, Aedes albopictus (Skuse), a dengue vector, was the dominant mosquito species, while in the villages, Culex vishnui (Theobald), a JE vector, was most common. This study has increased the overall knowledge of mosquito fauna in Lao PDR. The high abundance of Ae. albopictus in natural and man-made forests warrants concern, with vector control measures currently only implemented in cities and villages.

The mosquitoes Aedes (Stegomyia) aegypti (L.)(Diptera:Culicidae) and Ae. (Stegomyia) albopictus (Skuse) (Diptera:Culicidae) transmit dengue, chikungunya, and Zika viruses and represent a growing public health threat in parts of the United States where they are established.To complement existing mosquito presence records based on discontinuous, non-systematic surveillance efforts, we developed county-scale environmental suitability maps for both species using maximum entropy modeling to fit climatic variables to county presence records from 1960–2016 in the contiguous United States. The predictive models for Ae. aegypti and Ae. albopictus had an overall accuracy of 0.84 and 0.85, respectively. Cumulative growing degree days (GDDs) during the winter months, an indicator of overall warmth, was the most important predictive variable for both species and was positively associated with environmental suitability. The number (percentage) of counties classified as environmentally suitable, based on models with 90 or 99% sensitivity, ranged from 1,443 (46%) to 2,209 (71%) for Ae. aegypti and from 1,726 (55%) to 2,329 (75%) for Ae. albopictus. Increasing model sensitivity results in more counties classified as suitable, at least for summer survival, from which there are no mosquito records. We anticipate that Ae. aegypti and Ae. albopictus will be found more commonly in counties classified as suitable based on the lower 90% sensitivity threshold compared with the higher 99% threshold. Counties predicted suitable with 90% sensitivity should therefore be a top priority for expanded mosquito surveillance efforts while still keeping in mind that Ae. aegypti and Ae. albopictus may be introduced, via accidental transport of eggs or immatures, and potentially proliferate during the warmest part of the year anywhere within the geographic areas delineated by the 99% sensitivity model.

We compared climatic distribution ranges between Aedes albopictus (Skuse) (Diptera: Culicidae) and the five wild (nondomesticated) species of Albopictus Subgroup of Scutellaris Group of Aedes (Stegomyia) in southern Asia. Distribution sites of the wild species concentrate in seasonal forest and savannah climate zones in India, Indochina, and southern China. The distribution of Ae. albopictus is broader than the wild species under 1) tropical rain-forest climate, 2) steppe and temperate savannah climate, and 3) continental climate with large seasonal temperature variation (hot summer and cold winter) at temperate lowlands (northernmost sites 40°N in Ae. albopictus vs 32°N in the wild species). However, the distribution of Ae. albopictus is more limited at tropical and subtropical highlands where the climate is cool but less continental (small seasonal variation, mild summer, and winter). We discuss a possibility that the broader climate ranges of Ae. albopictus are ecological or eco-evolutionary consequences of adaptation to human habitats. We also propose a general scenario for the origin, dispersal, and adaptation of Ae. albopictus in Asia as a hypothesis for future research.

The potential of integrating the mycoinsecticide, Metarhizium anisopliae (Met.), into house fly control programs is tremendous. However, the interaction between the fungus and insecticide, when applied at poultry farms, remains poorly understood. This study investigated the interaction between M. anisopliae and two selected insecticides, cyromazine and ChCy (a mixture of chlorpyrifos and cypemethrin), with three objectives: to assess the compatibility of M. anisopliae and the insecticides by measuring fungal vegetative growth and conidia production in the presence of insecticides; to evaluate the effect of M. anisopliae on these insecticides by analyzing insecticidal residue using ultra performance liquid chromatography; and to study the synergistic effects of M. anisopliae and the insecticides by applying sublethal concentrations of insecticides with M. anisopliae to house fly larvae. Metarhizium anisopliae was more tolerant to ChCy than to cyromazine, as M. anisopliae showed significantly more growth when grown with this insecticide. The M. anisopliae þ ChCy combination resulted in significantly less chlorpyrifos residues compared to the ChCy plate, and 62–72% house fly larva mortality occurred when M. anisopliae and sublethal concentrations of ChCy were combined, implicating synergistic effects of the fungus with low concentrations of ChCy. Integrating M. anisopliae with compatible chemical at right concentration is crucial for poultry farm house fly control programs.

The brown dog tick, Rhipicephalus sanguineus (Latrielle) sensu lato, is an important ectoparasite of dogs and occasionally humans, capable of transmitting several pathogens, such as Rickettsia and Ehrlichia, which are of veterinary and medical importance. The brown dog tick is distributed worldwide and has an affinity for human habitations in much of its range. In some populations, lack of integrated pest management plans and overuse of pyrethroid pesticides and other sodium channel inhibitors has resulted in high levels of resistance to permethrin. Recently, a highly conserved region of the R. sanguineus sodium channel was sequenced, indicating that a single nucleotide polymorphism of thymine to cytosine on domain III segment VI of the sodium channel could confer resistance. A molecular assay targeting a point mutation in the sodium channel was developed and optimized to separate ticks expressing permethrin resistance from those from a susceptible colony. Thereafter, multiple field-collected phenotypically permethrin-resistant populations were evaluated using this molecular assay to determine genotype. As confirmed by DNA sequencing, a point mutation was present at a high rate in phenotypically resistant tick populations that was not present in the susceptible strain. These data suggest an additional permethrin resistance mechanism to metabolic resistance, which has been reported for this tick species, and confirm its association with phenotypic resistance. The results of this study further emphasize the need to preserve acaricide chemistry through rotation of active ingredients used to control ectoparasites.

The brown dog tick, Rhipicephalus sanguineus sensu lato (Latreille), is a cosmopolitan ectoparasite and vector of pathogens that kill humans and animals. Pyrethroids represent a class of synthetic acaricides that have been used intensely to try to control the brown dog tick and mitigate the risk of tick-borne disease transmission. However, acaricide resistance is an emerging problem in the management of the brown dog tick. Understanding the mechanism of resistance to acaricides, including pyrethroids, is important to adapt brown dog tick control strategies. The main objective of this study was to determine if target-site mutations associated with pyrethroid resistance in other pests could be associated with phenotypic resistance detected in a brown dog tick population from Florida. We amplified segment 6 of the domain III of the voltage-sensitive sodium channel protein, using cDNAs synthesized from pyrethroid-susceptible and pyrethroid-resistant tick strains. A single nucleotide point mutation (SNP) identified in a highly conserved region of domain III S6 in the resistant ticks resulted in an amino acid change from phenylalanine to leucine. This mutation is characteristic of resistance phenotypes in other tick species, and is the first report of this mutation in R. sanguineus. Molecular assays based on this knowledge could be developed to diagnose the risk for pyrethroid resistance, and to inform decisions on integrated brown dog tick management practices.

Puerto Rico (PR) has a long history of vector-borne disease and insecticide-resistant Aedes aegypti (L.). Defining contributing mechanisms behind phenotypic resistance is critical for effective vector control intervention. However, previous studies from PR have each focused on only one mechanism of pyrethroid resistance. This study examines the contribution of P450-mediated enzymatic detoxification and sodium channel target site changes to the overall resistance phenotype of Ae. aegypti collected from San Juan, PR, in 2012. Screening of a panel of toxicants found broad resistance relative to the lab susceptible Orlando (ORL1952) strain. We identified significant resistance to representative Type I, Type II, and nonester pyrethroids, a sodium channel blocker, and a sodium channel blocking inhibitor, all of which interact with the sodium channel. Testing of fipronil, a chloride channel agonist, also showed low but significant levels of resistance. In contrast, the PR and ORL1952 strains were equally susceptible to chlorfenapyr, which has been suggested as an alternative public health insecticide. Molecular characterization of the strain indicated that two common sodium channel mutations were fixed in the population. Topical bioassay with piperonyl butoxide (PBO) indicated cytochrome P450-mediated detoxification accounts for approximately half of the resistance profile. Transcript expression screening of cytochrome P450s and glutathione-S-transferases identified the presence of overexpressed transcripts. This study of Puerto Rican Ae. aegypti with significant contributions from both genetic changes and enzymatic detoxification highlights the necessity of monitoring for resistance but also defining the multiple resistance mechanisms to inform effective mosquito control.

Although pesticide resistance is common in insect vectors of human diseases, the evolution of resistance might be delayed if management practices are adopted that limit selection of resistance alleles. Outbreaks of dengue fever have occurred in Queensland, Australia, since the late 1800s, leading to ongoing attempts to control the mosquito vector, Aedes aegypti (L.). Since the 1990s, pyrethroid insecticides have been used for this purpose, but have been applied in a strategic manner with a variety of delivery methods including indoor residual spraying, lethal ovitraps, and use of insect growth regulators as larvicides. Separate selection experiments on mosquitoes from Queensland using Type I and Type II pyrethroids did not produce resistant lines of Ae. aegypti, and bioassays of field material from Queensland showed only weak tolerance in comparison with a susceptible line. There was no evidence of knockdown resistance (kdr) mutations in Ae. aegypti from Queensland, in stark contrast to the situation in nearby southeast Asia. We suspect that careful management of pyrethroid insecticide use combined with surveillance and interception of exotic incursions has helped to maintain pyrethroid (and particularly kdr-based) susceptibility in Ae. aegypti in Australia.

Mosquitoes are a nuisance and potentially transmit pathogens causing numerous diseases worldwide. Homeowners and others may hire private companies to alleviate mosquito-related issues. Here, two pyrethroids (Suspend Polyzone [deltamethrin] and Bifen Insecticide/Termiticide [bifenthrin]) were evaluated on properties in North Carolina for 23 wk from 18 May through 19 October 2015. Properties were treated using backpack mist blowers every 21 d. At 17 fixed sampling locations, Centers for Disease Control and Prevention carbon dioxide-baited traps were deployed overnight once/week for the duration of the experiment. Oviposition traps were deployed weekly at the same locations. Differences were observed in mosquito abundance between neighborhoods, treatments, and weeks and differences varied between species. Mosquito abundance was generally significantly higher in traps placed on control properties (no insecticide) compared to traps placed on treatment properties. Bifenthrin and deltamethrin showed differences from each other in efficacy, but this varied between neighborhoods and species. Future studies could test the efficacy of barrier sprays at different application frequencies and/or in conjunction with weather monitoring. Coupled with regular mosquito surveillance and using integrated pest management principles, barrier sprays can be an effective tool for suppression of mosquito populations.

Aedes aegypti (L.) (Diptera: Culicidae) are known to flourish in a variety of natural and residential habitats and are competent vectors of at least 22 different arboviruses, including dengue, chikungunya, and Zika. Their global distribution, anthropophilic nature, and vector competency make them species of interest for control. A novel durable dual-action lethal ovitrap (DDALO) with combined larviciding and adulticiding effects, as well as a slow-release polymer (isobutyl methacrylate), was designed to target Ae. aegypti.The use of the DDALO resulted in high adult mosquito mortality (~95–100%) in no-choice laboratory cage studies targeting gravid females and successfully prevented all deposited eggs from hatching. Aging of the traps caused some loss in activity over time, but they still caused adult mortality (~50%) and continued to prevent successful hatching of eggs for 6 mo. Oviposition preference studies resulted in ~4.5 times as many larvae developing in untreated DDALOs compared with the other containers combined. Small-cage multigenerational studies resulted in significantly lower populations of adult mosquitoes in cages containing treated DDALOs after 4 wk. Successful laboratory studies show that the DDALO appears to be a promising tool that could be used for controlling wild vector populations of Ae. aegypti in combination with other mosquito control practices.

The main malaria vectors in sub-Saharan Africa, the Anopheles gambiae (Giles; Diptera: Culicidae), normally breed in clean water sources. However, evidence suggests an on-going adaptation of Anopheline species to polluted breeding habitats in urban settings. This study aimed at understanding the adaptation to breeding in water bodies with different qualities, in five selected mosquito breeding sites in urban Accra, Ghana. The study sites were also evaluated for the WHO water-quality parameters as a measure of pollution, and insecticide residues. Field mosquitoes were evaluated for five genes; CYP6P3, CYP4H19, CYP4H24, GSTD1-4, and ABCC11—associated with insecticide detoxification—using quantitative RT-PCR, as well as Mono-oxygenase, Alpha Esterase, Glutathione S-transferase, and insensitive acetylcholinesterase (AChE) using biochemical enzyme assays. The lab-reared, insecticide susceptible An. gambiae Kisumu strain was bred in the most polluted water source for 10 generations and evaluated for the same genes and enzymes. The results revealed that the fold expression of the genes was higher in the larvae compared with the adults. The results also suggest that detoxification enzymes could be involved in the adaptation of An. gambiae to polluted breeding sites. Correlation analysis revealed a highly positive significant correlation between calcium levels and all five genes (P < 0.05). Stepwise linear regression to understand which of the variables predicted the expression of the genes revealed that sulphate was responsible for ABCC11 and CYP4H24, alkalinity for GSTD1-4, and calcium for CYP4H19 and CYP6P3. The detailed genetic basis of this adaptation need to be further investigated. A further understanding of this adaptation may provide outlooks for controlling malaria and other disease vectors adapted to polluted breeding water sources.

Essential oils are potential alternatives to synthetic insecticides because they have low mammalian toxicity, degrade rapidly in the environment, and possess complex mixtures of bioactive constituents with multi-modal activity against the target insect populations.Twenty-one essential oils were initially screened for their toxicity against Aedes aegypti (L.) larvae and three out of the seven most toxic essential oils (Manuka, oregano, and clove bud essential oils) were examined for their chemical composition and combined toxicity against Ae. aegypti larvae. Manuka essential oil interacted synergistically with oregano essential oil and antagonistically with clove bud essential oil. GC–MS analysis revealed the presence of 21 components in Manuka essential oil and three components each in oregano and clove bud essential oils. Eugenol (84.9%) and eugenol acetate (9.6%) were the principal constituents in clove bud essential oil while carvacrol (75.8%) and m-isopropyltoluene (15.5%) were the major constituents in oregano essential oil. The major constituents in Manuka essential oil were calamenene (20%) and 3-dodecyl-furandione (11.4%). Manuka essential oil interacted synergistically with eugenol acetate and antagonistically with eugenol, suggesting that eugenol was a major contributor to the antagonistic interaction between Manuka and clove bud essential oils. In addition, Manuka interacted synergistically with carvacrol suggesting its contribution to the synergistic interaction between Manuka and oregano essential oils. These findings provide novel insights that can be used to develop new and safer alternatives to synthetic insecticides.

Concentrations of malathion and permethrin typical in droplets generated from ultra-low-volume and low-volume applications used to control mosquito populations were evaluated for efficacy against multiple-aged Culex quinquefasciatus Say (Diptera: Culicidae), using a topical bioassay. Although insecticide droplets will impinge on many exoskeletal body regions and a range of ages of mosquitoes in a population, traditional mosquito topical bioassays focus pesticide application to the mesothoracic pleural or dorsal regions across an average mosquito age (e.g., 3–7 d). Our results document nonuniform insecticide sensitivity across body regions at ages not previously assessed in mosquitoes (teneral and 14-d old). We expect our findings to influence the topical bioassay process, illustrating the difference in mosquito body regions and ages that ultimately may explain insecticide effectiveness wherever droplets impinge upon the mosquito body during field control applications.

Numerous advantages over the standard incandescent lamp favor the use of light-emitting diodes (LEDs) as an alternative and inexpensive light source for sampling medically important insects in surveillance studies. Previously published studies examined the response of mosquitoes to different wavelengths, but data on anopheline mosquito LED attraction are limited. Center for Disease Control and Prevention-type light traps were modified by replacing the standard incandescent lamp with 5-mm LEDs, one emitting at 520 nm (green) and the other at 470 nm (blue). To test the influence of moon luminosity on LED catches, the experiments were conducted during the four lunar phases during each month of the study period. A total of 1,845 specimens representing eight anopheline species were collected. Anopheles (Nyssorhynchus) evansae (35.2%) was the most frequently collected, followed by An. (Nys.) triannulatus (21.9%), An. (Nys.) goeldii (12.9%), and An. (Nys.) argyritarsis (11.5%). The green LED was the most attractive light source, accounting for 43.3% of the individuals collected, followed by the blue (31.8%) and control (24.9%) lights. The LED traps were significantly more attractive than the control, independent of the lunar phase. Light trapping of anopheline mosquitoes was more efficient when the standard incandescent lamp was replaced with LEDs, regardless of the moon phase. The efficiency of LEDs improves light trapping results, and it is suggested that the use of LEDs as an attractant for anopheline mosquitoes should be taken into consideration when sampling anopheline mosquitoes.

The detection and identification of natural infections in sand flies by Leishmania protozoan species in endemic areas is a key factor in assessing the risk of leishmaniasis and in designing prevention and control measures for this infectious disease. In this study, we analyzed the Leishmania DNA using nuclear ribosomal internal transcript spacer (ITS) sequences. Parasite DNA was extracted from naturally infected, blood-fed sand flies collected in nine localities considered leishmaniasis-endemic foci in Ecuador.

The species of parasites identified in sand flies were Leishmania major-like, Leishmania naiffi, Leishmania mexicana, Leishmania lainsoni, and “Leishmania sp. siamensis”. Sand fly specimens of Brumptomyia leopoldoi, Mycropigomyia cayennensis, Nyssomyia yuilli yuilli, Nyssomyia trapidoi, Pressatia triacantha, Pressatia dysponeta, Psychodopygus carrerai carrerai, Psychodopygus panamensis, and Trichophoromyia ubiquitalis were found positive for Leishmania parasite. These findings contribute to a better understanding of the epidemiology and transmission dynamics of the disease in high-risk areas of Ecuador.

Flies are known to be mechanical vectors of bacterial, viral, and parasitic diseases. Although flies are known to transmit disease, the effects of cleaning behavior have not been well studied. This study quantified the cleaning effectiveness and behavior of three fly species: Sarcophaga bullata, Musca domestica L., and Drosophila virilis. Flies were transferred to plates of Escherichia coli or Pseudomonas aeruginosa and allowed to walk on the bacteria for a total of 5 min. After the flies were contaminated, they were either immediately collected to quantify bacteria or were placed onto sterile plates to clean for 5 or 10 min. After cleaning, flies were placed into tubes with 1 ml of sterile 0.85% saline and were gently shaken for 1 min to remove bacteria. A serial dilution was made and 50-μl spot titers were plated. Cleaning behavior was also monitored and scored for a period of 5 min. Results demonstrate a bacterial reduction for both bacteria on all three fly species. Sarcophaga bullata and D. virilis both showed a significant reduction of both bacteria within 10 min, whereas M. domestica only showed a significant reduction in P. aeruginosa. Cleaning behavior increased significantly in flies that were exposed to bacteria compared to flies that were not exposed to bacteria. This study is important, as it demonstrates that fly cleaning could affect mechanical transmission of disease, and additional studies should look at flies' abilities to remove other types of microorganisms.

Little is known about tick-borne rickettsial pathogens in Belize, Central America. We tested ixodid ticks for the presence of Rickettsia species in three of the six northern and western Belizean districts. Ticks were collected from domestic animals and tick drags over vegetation in 23 different villages in November 2014, February 2015, and May 2015. A total of 2,506 collected ticks were identified to the following species: Dermacentor nitens Neumann (46.69%), Rhipicephalus sanguineus (Latreille) (19.55%), Rhipicephalus microplus (Canestrini) (19.47%), Amblyomma cajennense complex (9.74%), Amblyomma maculatum Koch (3.47%), Amblyomma ovale Koch (0.68%), Ixodes nr affinis (0.16%), Amblyomma nr maculatum (0.12%), and Amblyomma nr oblongoguttatum (0.12%). Ticks were pooled according to species, life stage (larva, nymph, or adult), and location (n = 509) for DNA extraction and screened for genus Rickettsia by quantitative real-time polymerase chain reaction (qPCR). All 42 positive pools were found to be positive for spotted fever group (SFG) Rickettsia in pools of A. cajennense complex (n = 33), A. maculatum (n = 4), A. nr maculatum (n = 1), A. ovale (n = 1), R. sanguineus (n = 1), and I. nr affinis (n = 2). Rickettsia amblyommatis was identified from A. cajennense complex and A. nr maculatum. Rickettsia parkeri was found in A. maculatum, and Rickettsia sp. endosymbiont was detected in I. nr affinis. The presence of infected ticks suggests a risk of tick-borne rickettsioses to humans and animals in Belize. This knowledge can contribute to an effective tick management and disease control program benefiting residents and travelers.

A passive surveillance program monitored ticks submitted by the public in Iowa from 1990–2013. Submitted ticks were identified to species and life stage, and Ixodes scapularis Say nymphs and adults were tested for the presence of Borrelia burgdorferi. An average of 2.6 of Iowa's 99 counties submitted first reports of I. scapularis per year over the surveillance period, indicating expansion of this tick species across the state. The proportion of vector ticks infected by B. burgdorferi increased over time between 1998 and 2013. In 2013, 23.5% of nymphal and adult I. scapularis were infected with B. burgdorferi, the highest proportion of any year. Active surveillance was performed at selected sites from 2007–2009. Ixodes scapularis nymphs collected at these sites were tested for the presence of B. burgdorferi, Anaplasma phagocytophilum, and spotted fever group Rickettsia spp. (likely representing Rickettsia buchneri). Nymphs tested were 17.3% positive for B. burgdorferi, 28.9% for A. phagocytophilum, and 67.3% for Rickettsia spp. The results of these surveillance programs indicate an increasing risk of disease transmission by I. scapularis in Iowa.

A total of 6,255 ticks belonging to three genera and six species (Haemaphysalis flava Neumann, Haemaphysalis longicornis Neumann, Haemaphysalis phasiana Saito, Ixodes nipponensis Kitaoka & Saito, Ixodes persulcatus Schulze, and Amblyomma testudinarium Koch) collected from May–August, 2013, at four southwestern provinces in the Republic of Korea (ROK) were submitted to the Armed Forces Research Institute of Medical Sciences and assayed for selected tick-borne pathogens. One pool each of H. flava and H. phasiana was positive by PCR and sequencing for a Francisella-like endosymbiont, while all pools were negative for Francisella tularensis, the causative agent of tularemia.

Rickettsia parkeri is an emerging human pathogen transmitted by Amblyomma ticks in predominately tropical and subtropical regions of the western hemisphere. In 2014 and 2015, one confirmed case and one probable case of R. parkeri rickettsiosis were reported from the Pajarita Wilderness Area, a semi-arid mountainous region in southern Arizona. To examine more closely the potential public health risk of R. parkeri in this region, a study was initiated to investigate the pervasiveness of Amblyomma maculatum Koch group ticks in mountainous areas of southern Arizona and to ascertain the infection frequencies of R. parkeri in these ticks. During July 2016, a total of 182 adult ticks were collected and evaluated from the Pajarita Wilderness Area in Santa Cruz County and two additional sites in Cochise and Santa Cruz counties in southern Arizona. DNA of R. parkeri was detected in a total of 44 (24%) of these ticks. DNA of “Candidatus Rickettsia andeanae” and Rickettsia rhipicephali was detected in three (2%) and one (0.5%) of the samples, respectively. These observations corroborate previous collection records and indicate that established populations of A. maculatum group ticks exist in multiple foci in southern Arizona. The high frequency of R. parkeri in these tick populations suggests a public health risk as well as the need to increase education of R. parkeri rickettsiosis for those residing, working in, or visiting this area.

Knowledge of the blood-feeding patterns exhibited by arthropod vectors is essential for understanding the complex dynamics of vector-borne disease transmission. Some species of mosquitoes belonging to the genus Culex have been implicated as having major roles in the transmission of arboviruses such as West Nile virus, Saint Louis encephalitis virus, and Western equine encephalitis virus. Although the host-feeding patterns for many of these Culex species are well studied, the host-feeding patterns of Culex stigmatosoma Dyar are relatively poorly studied, even though this species is suspected to be an important maintenance vector for West Nile virus and other arboviruses. In the current study, bloodmeals from 976 blood-engorged Cx. stigmatosoma, collected from 30 sites in southern California from 2009–2012, were processed for vertebrate host identification by nucleotide sequencing following polymerase chain reaction to amplify portions of the cytochrome oxidase I and cytochrome b genes of vertebrate animals. Vertebrate DNA was amplified, sequenced, and identified from a total of 647 Cx. stigmatosoma bloodmeals, revealing that 98.6% of bloodmeals were from birds, 1.2% from three mammal species, and a single bloodmeal was from a reptile species. In total, 40 different host species were identified. The greatest number of bloodmeals identified was from domestic chicken (Gallus gallus domesticus L.) (38% of bloodmeals), house sparrow (Passer domesticus L.) (23%), house finch (Haemorhous mexicanus Müller) (17%), American crow (Corvus brachyrhynchos L.) (4%), and mourning dove (Zenaida macroura L.) (3%). However, chicken bloodmeals were identified almost entirely from a single site where mosquito collection devices were placed in the near vicinity of confined domestic chickens. The strongly ornithophilic feeding behavior shown in this study for Cx. stigmatosoma supports the hypothesis that this mosquito species may be an important maintenance (or endemic) vector for arboviruses that circulate among susceptible birds.

Anopheles arabiensis (Patton; Diptera: Culicidae) is a major malaria vector in the southern African region. In South Africa, effective control of this species using indoor-based interventions is reduced owing to its tendency to rest outdoors. As South Africa moves towards malaria elimination there is a need for complementary vector control strategies. One of the methods under consideration is the use of the sterile insect technique (SIT). Key to the successful implementation of an SIT programme is prior knowledge of the size and spatial distribution of the target population. Understanding mosquito population dynamics for both males and females is critical for efficient programme implementation. It is thus necessary to use outdoor-based population monitoring tools capable of sampling both sexes of the target population. In this project mosquito surveillance and evaluation of tools capable of collecting both genders were carried out at Mamfene in northern KwaZulu-Natal Province, South Africa, during the period January 2014 to December 2015. Outdoor- and indoor-resting Anopheles mosquitoes were sampled in three sections of Mamfene over the 2-yr sampling period using modified plastic buckets, clay pots and window exit traps. Morphological and molecular techniques were used for species identifications of all samples. Wild-caught adult females were tested for Plasmodium falciparum (Welch; Haemosporida: Plasmodiidae) infectivity. Out of 1,705 mosquitoes collected, 1,259 (73.8%) and 255 (15%) were identified as members of either the Anopheles gambiae complex or Anopheles funestus group respectively. An. arabiensis was the most abundant species contributing 78.8% of identified specimens. Mosquito density was highest in summer and lowest during winter. Clay pots yielded 16.3 mosquitoes per trap compared to 10.5 for modified plastic buckets over the 2-yr sampling period. P. falciparum infection rates for An. arabiensis were 0.7% and 0.5% for 2014 and 2015, respectively. Logistic regression analysis showed an association between An. arabiensis catches with Section and season of collection but not with sex and collection methods. These data confirmed the presence of a perennial An. arabiensis population at Mamfene and constitute the first records of P. falciparum infective An. arabiensis from South Africa, confirming this species as a major vector in the malaria endemic provinces of the country.

There are several main vectors of scrub typhus in China, and Leptotrombidium rubellumWang et Liao, 1984 is one of them, which was previously considered to be restricted to the coastal regions of Changle to Xiamen, Fujian province of east China. Ecological investigation of chigger mites in recent years demonstrated the presence of L. rubellum also in Yunnan province. Chigger mites were collected from 34 counties, in which 127,460 larval mites were collected from 14,381 small mammal hosts. A total of 277 species belonging to 26 genera and three subfamilies were identified. A total of 705/127,460 (0.55%) L. rubellum were collected from eight counties. Leptotrombidium rubellum was collected mainly at low elevations in southern Yunnan. A total of 663/705 (94.04%) of L. rubellum were collected from Rattus flavipectus (Milne-Edwards, 1871) found in outdoor habitats with relatively high infestation prevalence and mean intensity. Although it was collected from several hosts, the primary host was Rattus tanezumi. This represents a new distribution record of L. rubellum for Yunnan province.

Gas stations often provide windshield wash basins (WWBs) that customers may use to clean their windshields. Motivated by casual observations, we conducted a survey of WWBs in and around Raleigh, NC, to determine whether these WWBs also serve as larval habitats for mosquitoes. We found that 27.7% (95% CI: 12.4–43.14%) of the 36 surveyed gas stations had mosquito larvae in their WWBs, and 22.4% (95% CI: 15.07–29.1%) of the 152 WWBs surveyed were positive for mosquito larvae. Two species were identified inhabiting these containers: Aedes albopictus (Skuse) and Culex quinquefasciatus Say. Aedes albopictus was associated with clear, unturbid water, whereas Cx. quinquefasciatus did not have any significant association with water characteristics. Pupae of both species were observed, suggesting these habitats could be sources of pest mosquitoes. Gas stations may be a convenient surveillance target for vector control specialists and may provide insight into human-aided mosquito dispersal.

Factors affecting the survival of Ixodes scapularis Say (Acari: Ixodidae) during diapause are poorly known. This is partially due to the difficulty involved in collecting ticks that are not actively questing. A possible method to overcome this issue involves the use of microcosms containing litter material and soil, but an effective method for tick recovery is required. This study tested three methods for the recovery of I. scapularis nymphs from soil microcosms during their active and inactive periods, as well as recovery of engorged larval I. scapularis. The first method was hand sorting for 120 min; the second was sorting for 30 min before placing the contents of the microcosm into a Berlese funnel for 72 h; and the third method was placing the microcosm contents into a Berlese funnel for 72 h with no prior hand sorting. Hand sorting alone and the combination of hand sorting plus the Berlese funnel were the most effective recovery methods for both active nymphs and those in diapause. Hand sorting alone was not an effective method for the recovery of engorged larvae and Berlese funnel extraction alone was not the most effective method for any of the I. scapularis physiological states tested. Overall, a combination of hand sorting and Berlese extraction was an effective recovery method for all physiological states and was 58.3% more time-effective compared against hand-sorting alone. This method will allow researchers to process microcosm samples effectively and efficiently, improving our ability to investigate the ecology of I. scapularis during their inactive periods.

Numerous researchers have observed a form of punctate corneal lesions causing leukomas (corneal opacities) in humans, domestic animals, and wild animals in different parts of the world.This condition has been reported under different names, including West Indian (or Caribbean) punctate keratopathy, West Indian dots, tropical punctate keratopathy, Rice's keratopathy, Florida keratopathy, and Florida spots. Many of these cases, appear to have a common cause, the stings of a small red ant, Wasmannia auropunctata (Hymenoptera: Formicidae), originally from the Neotropics, but spread to other parts of the world through human commerce. The purpose of this article is to link disparate literature on punctate or nummular corneal lesions published in medical, veterinary, wildlife, and entomology journals, because many researchers seem largely unaware of the literature from disciplines other than their own. Recognizing a common cause of this corneal condition is important to insure proper medical treatment and foster efforts to limit the spread and negative impact of W. auropunctata.

Accurate blood meal identification is critical to understand hematophagous vector-host relationships. This study describes a customizable Next-Generation Sequencing (NGS) approach to identify blood meals from Rhodnius pallescens (Hemiptera: Reduviidae) triatomines using multiple barcoded primers and existing software to pick operational taxonomic units and match sequences for blood meal identification. We precisely identified all positive control samples using this method and further examined 74 wild-caught R. pallescens samples. With this novel blood meal identification method, we detected 13 vertebrate species in the blood meals, as well as single and multiple blood meals in individual bugs. Our results demonstrate the reliability and descriptive uses of our method.

The citizen science project ‘Mueckenatlas' (mosquito atlas) was implemented in early 2012 to improve mosquito surveillance in Germany. Citizens are asked to support the spatiotemporal mapping of culicids by submitting mosquito specimens collected in their private surroundings. The Mueckenatlas has developed into an efficient tool for data collection with close to 30,000 mosquitoes submitted by the end of 2015. While the vast majority of submissions included native mosquito species, a small percentage represented invasive species. The discovery of Aedes albopictus (Skuse) (Diptera: Culicidae), Aedes japonicus japonicus (Theobald) (Diptera: Culicidae) and Aedes koreicus (Edwards) (Diptera: Culicidae) specimens via the Mueckenatlas project prompted targeted monitoring activities in the field which produced additional information on the distribution of these species in Germany. Among others, Mueckenatlas submissions led to the detection of three populations of Ae. j. japonicus in West, North and Southeast Germany in 2012, 2013, and 2015, respectively. As demonstrated by on-site monitoring, the origins of Ae. j. japonicus specimens submitted to the Mueckenatlas mirror the distribution areas of the four presently known German populations as found by active field sampling (the fourth population already reported prior to the launch of the Mueckenatlas). The data suggest that a citizen science project such as the Mueckenatlas may aid in detecting changes in the mosquito fauna and can therefore be used to guide the design of more targeted field surveillance activities.