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Several invasive mosquito species that are nuisances or of medical and veterinary importance have been introduced into the Southeastern region of the USA, posing a threat to other species and the local ecosystems and/or increasing the risk of pathogen transmission to people, livestock, and domestic pets. Prompt and effective monitoring and control of invasive species is essential to prevent them from spreading and causing harmful effects. However, the capacity for invasive mosquito species surveillance is highly variable among mosquito control programs in the Southeast, depending on a combination of factors such as regional geography and climate, access to resources, and the ability to interact with other programs. To facilitate the development of invasive mosquito surveillance in the region, we, the Mosquito BEACONS (Biodiversity Enhancement and Control of Non-native Species) working group, conducted a survey on the capacities of various public health agencies and pest control agencies engaged in mosquito surveillance and control in seven Southeastern states (Alabama, Florida, Georgia, Louisiana, Mississippi, North Carolina, and South Carolina). Ninety control programs completed the survey, representing an overall response rate of 25.8%. We report key findings from our survey, emphasizing the training and resource needs, and discuss their implications for future invasive mosquito surveillance and control capacity building. By increasing communication and collaboration opportunities (e.g., real-time sharing of collection records, coordinated multistate programs), the establishment of Mosquito BEACONS and the implementation of this survey can accelerate knowledge transfer and improve decision support capacity in response to or in preparation for invasive mosquito surveillance and can establish infrastructure that can be used to inform programs around the world.

Recurring outbreaks of mosquito-borne diseases, like dengue, in the Pacific region represent a major biosecurity risk to neighboring continents through potential introductions of disease-causing pathogens. Aedes mosquitoes, highly prevalent in this region, are extremely invasive and the predominant vectors of multiple viruses including causing dengue, chikungunya, and Zika. Due to the absence of vaccines for most of these diseases, Aedes control remains a high priority for public health. Currently, international organizations put their efforts into improving mosquito surveillance programs in the Pacific region. Also, a novel biocontrol method using Wolbachia has been tried in the Pacific region to control Aedes mosquito populations. A comprehensive understanding of mosquito biology is needed to assess the risk that mosquitoes might be introduced to neighboring islands in the region and how this might impact arboviral virus transmission. As such, we present a comprehensive review of arboviral disease outbreak records as well as Aedes mosquito biology research findings relevant to the Pacific region collected from both non-scientific and scientific sources.

Over 20 years since its introduction, the West Nile virus (WNV) continues to be the leading cause of arboviral disease in the USA. In Panama City Beach (Bay County, FL), WNV transmission is monitored using sentinel chickens and testing mosquito pools for presence of viral RNA. In the current work, we monitored WNV transmission from 2014 to 2020 through weekly serology sampling of sentinel chickens; mosquito populations through biweekly mosquito collections by suction traps (1 m and 9 m) and weekly gravid trap collections; and mosquito infection rates using a reverse transcriptase–polymerase chain reaction (RT-PCR) assay. Samples were sent to the Bureau of Public Health Laboratories (Tampa, FL) for testing presence/absence of WNV via RT-PCR assay. Our results indicated that canopy surveillance could augment ground collections, providing greater proportions of Culex mosquitoes with less bycatch compared with ground collections. Serology indicated 94 seroconversions to WNV in the study area from 2014 to 2020. The most active year was 2016, which accounted for 32% (n = 30) of all seroconversions reported during the study period. We detected 20 WNV-positive mosquito pools from Culex quinquefasciatus during 2014–17; mosquito infection rates ranged from 2.02 to 23.81 per thousand (95% CI). Climate data indicated anomalously high precipitation in 2014–19 preceding WNV transmission. Data analyzed herein indicate utility in year-round continuous and diversified surveillance methodologies. This information is needed to properly calibrate future models that could assist with predicting transmission events of WNV in Panama City Beach, FL.

The monarch butterfly, Danaus plexippus (Lepidoptera: Nymphalidae), is threatened by substantial loss of habitat, extreme weather events linked to global climate change, and nontarget impacts of broad-spectrum insecticides. To investigate the impact of chronic ingestion of pyrethroids on monarchs, wild-type Florida D. plexippus were reared on milkweed (Asclepias curassavica) that was exposed to ultra-low volume applications of DeltaGard by a truck-mounted fogger, at distances of 25 and 50 m. We observed significant negative impacts on monarchs reared on milkweed at 25 m from the DeltaGard spray route, including significant decreases in survival, and significantly longer development times, compared with untreated controls. Larvae reared on host plants closest to the truck spray route were 3 times more likely to experience a mortality event than the control cohort in trial 1 and 6 times in trial 2. Survival of monarch caterpillars reared on milkweed sprayed at 50 m was not significantly different from controls. For monarchs that survived to adulthood, we did not observe statistically significant differences among cohorts for variables measured. These data demonstrate that ultra-low volume treatments of pyrethroids can result in significant mortality in monarchs, but that the effects diminish with distance from the spray route.

The mosquito microbiota affects many aspects of mosquito biology including development and reproduction. It also strongly impacts interactions between the mosquito host and pathogens that cause important disease in humans, such as dengue and malaria. Critically, the mosquito microbiota is highly diverse and can vary in composition in response to multiple environmental variables, but these effects are not always consistent. Understanding how the environment shapes mosquito microbial diversity is a critical step in elucidating the ubiquity of key host-microbe-pathogen interactions in nature. To that end, we examined the role of time of collection, collection location and host species on mosquito microbial diversity by repeating collections at two-month intervals on a trapping grid spanning three distinct biomes. We then used 16S rRNA sequencing to compare the microbiomes ofAedes taeniorhynchus,Anopheles crucians, andCulex nigripalpusmosquitoes from those collections. We saw that mosquito diversity was strongly affected by both time and collection location. We also observed that microbial richness and diversity increased from March to May, and thatAn. cruciansandCx. nigripalpushad greater microbial diversity thanAe. taeniorhynchus. However, we also observed that collection location had no impact on microbial diversity except for significantly lower bacterial richness observed in mosquitoes collected from the mangrove wetlands. Our results highlight that collection time, collection location, and mosquito species each affect aspects of mosquito microbial diversity, but their importance is context dependent. We also demonstrate that these variables have differing impacts on mosquito diversity and mosquito microbial diversity. Our findings suggest that the environment likely plays an important but variable role in influencing the composition of the mosquito microbiota.

Mosquito control agencies monitor mosquito diversity and abundance through a variety of trap types. Although various long-term ecological data sets exist, little work has been done to address the sampling effort required to capture mosquito community diversity by trap type and few spatiotemporal distributions of vector species have been described. Here, we describe the seasonal distributions of vector species of importance, assess trapping effort needed to capture the diversity of the mosquito community, and use a partial redundancy analysis to identify trap bias from four commonly deployed adult mosquito traps in Volusia County, Florida. Collections were made with American Biophysics Corporation (ABC) light traps, Biogents Sentinel (BGS) traps, chicken coop exit traps, and gravid traps. We collected a total of 238,301 adult female mosquitoes belonging to 11 genera and 36 species, 12 of which we deemed to be vector species of epidemiological importance. We found that ABC traps not only yielded the greatest abundance and diversity but also captured several nonvector species. BGS and gravid traps yielded the highest proportions of vector species; exit traps recorded the lowest abundances and species richness. Wintertime abundances of several species demonstrated a need for year-round surveillance in the study area; partial redundancy analysis revealed that trap type explained a significant proportion of the variance in our data set, with certain vector species associated with specific trap types. Increased awareness regarding the amount of trapping effort needed to detect vector species diversity will help to optimize efforts in the field, leading to more effective resource allocation.Graphical AbstractProportion of vector species of epidemiological importance from four commonly deployed adult mosquito traps in Volusia County, Florida, from 1 January 2015 to 31 December 2018.

A population of Aedes (Stegomyia) albopictus (Skuse) (Diptera: Culicidae), a vector of chikungunya, dengue, yellow fever, and Zika and West Nile viruses, has been detected in Windsor, Ontario, Canada from 2016 onwards. Here, we describe its seasonal distribution, as well as the various aquatic habitats from which this species was collected and its larval co-habitation. We collected immatures from tires, treeholes, extruded polystyrene foam containers, discarded plastic cups, old recycling bins and oviposition traps. Aedes albopictus larvae were collected with Aedes japonicus (Theobald), Anopheles punctipennis (Say), Culex pipiens Linnaeus, Ochlerotatus hendersoni (Cockerell), Ochlerotatus triseriatus (Say) and Orthopodomyia signifera (Coquillett). Adult female and male specimens were collected from Biogents sentinel traps (Biogents AG, Regensburg, Germany), as well as Centers for Disease Control and Prevention miniature light traps (CDC, Atlanta, GA, U.S.A.), and also as they alighted on the investigators. Peak adult collections occurred in September during epidemiological week 37. We also collected Aedes (Stegomyia) aegypti (Linnaeus), a new record for Canada, in 2016 and from two new collection sites in 2017. The 2017 collections were 3.5 km north and 19.4 km south of the index site. The present study adds to the increasing number of studies reporting range expansions of these mosquito species.

A thorough understanding of the local mosquito fauna is required to develop effective mosquito abatement programs and assess risk of arbovirus transmission in an area. Although many mosquito control districts routinely survey adult populations of mosquitoes, few studies address trap bias, attractant bias, and the sampling effort needed to fully describe mosquito community diversity. Quantifying and visualizing differences in mosquito community composition and abundance, collected in adjacent mosquito control districts that use different attractants, provides a first step toward understanding and communicating cross-district arbovirus risk in continuous geographic areas. We obtained female mosquito collection data from CDC (Centres for Disease Control and Prevention) suction light traps from St. Johns County (baited with octenol) and Duval County (baited with dry ice) in Florida, USA, presenting a unique opportunity to summarize and compare collections across 2 attractants in adjacent Florida mosquito control programs. In the current work, we describe the seasonal distribution of mosquito species, highlight proportions of vector species of importance, quantify and assess diversity, and summarize the variation explained by attractant type using partial redundancy analysis. Numerous actual and potential vector species of importance were abundant throughout the sampling period that included Aedes atlanticus (Dyar & Knab), Anopheles crucians Weidemann, Culex erraticus (Dyar & Knab), and Culex nigripalpus Theobald (all Diptera: Culicidae). Dry ice collections yielded the greatest diversity of species with the least trapping effort. Traps baited with octenol yielded the greatest number of mosquitoes with a greater proportion of vector species. The results of the partial redundancy analysis revealed that attractant explained a significant proportion of the variance in the data set. But a significant linear trend also was present indicating that additional spatially structured variables were responsible for a large proportion of the variance in the data. It is necessary to know the differences between mosquito species and trap numbers when varying collection methods and attractants are used to assess arbovirus transmission risk across mosquito control district administrative boundaries. This information can be used to provide more robust and comprehensive surveillance information capable of identifying new challenges to public health safety. Resumen Se requiere un conocimiento profundo de la fauna local de mosquitos para desarrollar programas efectivos de eliminacion de mosquitos y evaluar el riesgo de transmision de arbovirus en un area. Aunque muchos distritos de control de mosquitos realizan sondeos de forma rutinaria a poblaciones adultas de mosquitos, pocos estudios abordan el sesgo de trampas, el sesgo de atrayentes y el esfuerzo de muestreo necesario para describir completamente la diversidad de la comunidad de mosquitos. La cuantificacion y visualizacion de las diferencias en la composicion y abundancia de las comunidades de mosquitos, recopiladas en distritos de control de mosquitos adyacentes que utilizan diferentes atrayentes, proporciona un primer paso para comprender y comunicar el riesgo de arbovirus entre distritos en areas geograficas continuas. Obtuvimos datos de recoleccion de mosquitos hembra de trampas de luz de succion del CDC del condado de St. Johns (cebado con octenol) y el condado de Duval (cebado con hielo seco) en la Florida, EE. UU., lo que presenta una oportunidad unica para resumir y comparar colecciones de 2 atrayentes en la Florida en programas adyacentes para el control de mosquitos. En el trabajo actual, describimos la distribucion estacional de las especies de mosquitos, resaltamos las proporciones de las especies de vectores de importancia, cuantificamos y evaluamos la diversidad y resumimos la variacion explicada por el tipo de atrayente mediante un analisis de redundancia parcial. Numerosas especies de vectores reales y potenciales de importancia fueron abundantes durante el periodo de muestreo que incluyeron Aedes atlanticus (Dyar & Knab), Anopheles crucians Weidemann, Culex erraticus (Dyar & Knab), y Culex nigripalpus Theobald (todos Diptera: Culicidae). Las recolecciones de hielo seco produjeron la mayor diversidad de especies con el menor esfuerzo de captura. Las trampas cebadas con octenol produjeron el mayor numero de mosquitos con una mayor proporcion de especies de vectores. Los resultados del analisis de redundancia parcial revelaron que el atrayente explicaba una proporcion significativa de la varianza en el conjunto de datos. Pero tambien estuvo presente una tendencia lineal significativa que indica que las variables estructuradas espacialmente adicionales fueron responsables de una gran proporcion de la varianza en los datos. Es necesario conocer las diferencias entre las especies de mosquitos, el numero de trampas al variar los metodos de recoleccion y los atrayentes al evaluar el riesgo de transmision de arbovirus a traves de las divisiones administrativos de los distritos con control de mosquitos. Esta informacion se puede utilizar para proporcionar un dato de vigilancia mas solido y completo capaz de identificar nuevos desafios para la seguridad de la salud publica. Key Words: surveillance; vector ecology; rarefaction, redundancy analysis; Culicidae View this article in BioOne

Mosquito control districts in the United States are limited to two main classes of adulticides, pyrethroids and organophosphates, to control mosquitoes. Two adulticides used to control domestic mosquitoes are Fyfanon EW (malathion, organophosphate) and DeltaGard (deltamethrin, pyrethroid). While the effect of these pesticides on European honeybees (Apis mellifera L., Hymenoptera: Apidae) has been investigated, effects on native pollinators need additional research. The purpose of this study was to investigate the acute nontarget effects of these pesticides on Bombus impatiens Cresson (Hymenoptera: Apidae), a native North American bumble bee species, and compare these effects to wild and laboratory strains of mosquitoes (Aedes aegypti (L.) and Culex quinquefasciatus Say, Diptera: Culicidae) through field and laboratory assays. Bombus impatiens was found to be resistant to Fyfanon EW (x̅ = 6.7% mortality at 50-µg malathion per bottle) at levels that caused significant mortality to study mosquitoes (86.2 ≥ x̅ ≥ 100% mortality) in laboratory bottle bioassays. Comparatively, B. impatiens demonstrated greater mortality to DeltaGard (93.3%) at 2.5-µg deltamethrin/bottle than any mosquito colony assayed (14.1 ≥ x̅ ≥ 87.0% mortality). Only DeltaGard was tested in field applications. In the field, we observed acute effects of DeltaGard on mosquitoes and B. impatiens at 25- and 75-m distance from a truck-mounted ultra-low volume fogger, although treatment effects were not significant for B. impatiens. Additional wild-caught nontarget mortality to DeltaGard field trials was also evaluated. This study indicated that common mosquito control adulticides do cause nontarget mortality to B. impatiens but that impacts are variable depending on pesticide and further studies are needed.

The purpose of this study was to establish geospatial and seasonal distributions of West Nile virus vectors in southern Ontario, Canada using historical surveillance data from 2002 to 2014. We set out to produce mosquito abundance prediction surfaces for each of Ontario’s thirteen West Nile virus vectors. We also set out to determine whether elevation and proximity to conservation areas and provincial parks, wetlands, and population centres could be used to improve our model. Our results indicated that the data sets for Anopheles quadrimaculatus, Anopheles punctipennis, Anopheles walkeri, Culex salinarius, Culex tarsalis, Ochlerotatus stimulans, and Ochlerotatus triseriatus were not suitable for geospatial modelling because they are randomly distributed throughout Ontario. Spatial prediction surfaces were created for Aedes japonicus and proximity to wetlands, Aedes vexans and proximity to population centres, Culex pipiens/restuans and proximity to population centres, Ochlerotatus canadensis and elevation, and Ochlerotatus trivittatus and proximity to population centres using kriging. Seasonal distributions are presented for all thirteen species. We have identified both when and where vector species are most abundant in southern Ontario. These data have the potential to contribute to a more efficient and focused larvicide program and West Nile virus awareness campaigns.

Zika virus (ZIKV), belonging to the family Flaviviridae, genus Flavivirus, is an arthropod-borne virus that was first discovered from the Zika forest in Uganda in 1947. Recent outbreaks in South America have linked ZIKV to cases of microcephaly and Guillain-Barre syndrome in humans. With the increased interest in ZIKV, protocols must be established to facilitate proper research. Here we describe the laboratory techniques required to quantify, propagate, and store ZIVK. We also review the proper safety protocol for the handling of ZIKV, which is classified as a Biosafety Level 2 pathogen by the United States Centers for Disease Control and Prevention. © 2016 by John Wiley & Sons, Inc.

We provide an updated checklist of 67 endemic mosquito species known from Ontario, Canada. Nine endemic species are added to the checklist found in Darsie and Ward (2005): Aedes cantator, Ae. churchillensis, Ae. nigripes, Ae. pullatus, Anopheles perplexens, An. crucians, An. smaragdinus, Culex erraticus, and Cx. salinarius. Only 4 specimens of Ae. albopictus have been recorded in Ontario since 2001 despite concerted efforts to find this species; therefore, it is considered an “accidental” species and is excluded from the checklist.

Culex (Melanoconion) erraticus (Dyar and Knab) is now established in southern Ontario, Canada. This species was first discovered in 2002 during a province-wide adult mosquito surveillance program for West Nile virus. Using CO2-baited CDC miniature light traps, a few Cx. erraticus were collected from 2002 to 2011, but the total number increased during the 2012 and 2013 seasons. The number of Ontario Public Health Units with records for Cx. erraticus has also increased since 2002, demonstrating that the geographic distribution of this species is expanding northward. Cx. erraticus is a potential arboviral bridge vector for a number of pathogens and its establishment in Ontario should be considered a potential public health concern.