Variation in Freshwater Insect Osmoregulatory Traits: A Comparative Approach.

AbstractFreshwater salinity regimes vary naturally and are changing in response to anthropogenic activities. Few insect species tolerate saline waters, and biodiversity losses are associated with increasing salinity in freshwater. We used radiotracers (22Na, 35SO4, and 45Ca) to examine ion uptake rates across concentration gradients in mayflies (Ephemeroptera), caddis flies (Trichoptera), and mosquitoes (Diptera) and made observations for some traits in seven other taxa representing mayflies, stone flies (Plecoptera), true flies (Diptera), and true bugs (Hemiptera). We further assessed the permeability of the cuticle to 3H2O influx and 22Na efflux when faced with deionized water in these same taxa. We hypothesized a relationship between uptake rates and reported saline tolerances, but our data did not support this hypothesis, likely because acclimatory responses were not part of this experimental approach. However, we found several common physiological traits across the taxa studied, including (i) ionic uptake rates that were always positively correlated with dissolved concentrations, (ii) generally low Ca uptake rates relative to other freshwater taxa, (iii) greater Na loss than Na uptake in dilute conditions, (iv) ion uptake that was more variable in ion-rich conditions than in dilute conditions, and (v) 3H2O influx that occurs quickly (but this rapidly exchangeable pool of body water accounts for a surprisingly small percentage of the water content of species tested). There remains much to learn about the physiology of these important organisms in the face of changing salinity regimes worldwide.

Rickettsia africae infections in sub-Saharan Africa: A systematic literature review of epidemiological studies and summary of case reports.

Rickettsia africae is a tick-borne bacteria known to cause African tick bite fever (ATBF). While the disease was first described more than 100 years ago, knowledge of transmission risk factors and disease burden remain poorly described. To better understand the burden of R. africae, this article reviewed and summarized the published literature related to ATBF epidemiology and clinical management. Using a systematic approach, consistent with the PRISMA guidelines, we identified more than 100 eligible articles, including 65 epidemiological studies and 41 case reports. Most reports described R. africae in ticks and livestock, while human studies were less common. Human disease case reports were exclusively among returning travellers from non-endemic areas, which limits our disease knowledge among at-risk populations: people living in endemic regions. Substantial efforts to elucidate the ATBF risk factors and clinical manifestations among local populations are needed to develop effective preventative strategies and facilitate appropriate and timely diagnosis.

Don't go chasing waterfalls: the phenology of Amblyomma americanum and increased collection of medically relevant ticks further away from hiking trails in the Piedmont of North Carolina, USA.

Hiking is a popular recreational activity in North Carolina that may expose people to ticks and tick-borne pathogens. However, there is a lack of knowledge on how ticks are distributed on and near trails. Our study tested the hypothesis that ticks are more likely to be encountered when moving further away from trails by measuring differences in relative abundance at various distances. We tested 4 distances (middle of trail, edge of trail, 5 m, 20 m), as well as an on-trail and off-trail grouping. We collected significantly more ticks and ticks were more likely to be collected at our 20 m and 5 m sampling distances rather than directly on or adjacent to trails, and significantly more were collected during off-trail collections than on-trails. When looking only at Amblyomma americanum, post hoc comparisons revealed that significantly more juvenile stages were collected at 5 m and 20 m distances, but not for adults. Our monthly sampling also allowed us to describe the phenology of A. americanum in North Carolina, which is consistent with the phenology of this species in the southeastern United States with adults peaking May-Jun, nymphs Jun-Jul, and larvae in Jul-Aug. These results generally demonstrate the importance of utilizing established trails when hiking to decrease tick-borne disease risk and should be communicated to the public as a recommendation for reducing tick-encounter risk.

Microbiome of Invasive Tick Species Haemaphysalis Longicornis in North Carolina, USA.

Ticks are one of the most important vectors of human and animal disease worldwide. In addition to pathogens, ticks carry a diverse microbiota of symbiotic and commensal microorganisms. In this study, we used next-generation sequencing (NGS) to survey the microbiomes of Haemaphysalis longicornis (Acari: Ixodidae) at different life stages collected from field populations in North Carolina (NC), USA. Sequence analyses were performed using QIIME2 with the DADA2 plugin and taxonomic assignments using the Greengenes database. Following quality filtering and rarefaction, the bacterial DNA sequences were assigned to 4795 amplicon sequence variants (ASVs) in 105 ticks. A core microbiome of H. longicornis was conserved across all ticks analyzed, and included bacterial taxa: Coxiella, Sphingomonas, Staphylococcus, Acinetobacter, Pseudomonas, Sphingomonadaceae, Actinomycetales, and Sphingobium. Less abundant bacterial taxa, including Rickettsia and Aeromonas, were also identified in some ticks. We discovered some ASVs that are associated with human and animal infections among the identified bacteria. Alpha diversity metrics revealed significant differences in bacterial diversity between life stages. Beta diversity metrics also revealed that bacterial communities across the three life stages were significantly different, suggesting dramatic changes in the microbiome as ticks mature. Based on these results, additional investigation is necessary to determine the significance of the Haemaphysalis longicornis microbiome for animal and human health.

Phylogenomics reveals the history of host use in mosquitoes.

Mosquitoes have profoundly affected human history and continue to threaten human health through the transmission of a diverse array of pathogens. The phylogeny of mosquitoes has remained poorly characterized due to difficulty in taxonomic sampling and limited availability of genomic data beyond the most important vector species. Here, we used phylogenomic analysis of 709 single copy ortholog groups from 256 mosquito species to produce a strongly supported phylogeny that resolves the position of the major disease vector species and the major mosquito lineages. Our analyses support an origin of mosquitoes in the early Triassic (217 MYA [highest posterior density region: 188-250 MYA]), considerably older than previous estimates. Moreover, we utilize an extensive database of host associations for mosquitoes to show that mosquitoes have shifted to feeding upon the blood of mammals numerous times, and that mosquito diversification and host-use patterns within major lineages appear to coincide in earth history both with major continental drift events and with the diversification of vertebrate classes.

Exploring the effects of caffeine on Aedes albopictus (Diptera: Culicidae) survival and fecundity.

Investigating new avenues of mosquito control is an important area of entomological research. Examining the effects of various compounds on mosquito biology contributes to the foundation of knowledge from which novel control methods can be built. Caffeine, in particular, is a commonly consumed compound that has not been thoroughly studied for its potential in disrupting the natural life cycle of mosquitoes. In this exploratory study, we analyzed caffeine's effect on the blood-feeding behavior, survival, and fecundity of Aedes albopictus Skuse (Diptera: Culicidae) mosquitoes. Two outcomes, blood-feeding behavior and fecundity, were analyzed in the first experiment in which mosquitoes were exposed to caffeine doses ranging from 0.2 to 2.4 mg/ml. We found a negative linear relationship between dose and fecundity, but no significant impact on blood-feeding behavior. Adjustments were made to the experimental design in which mosquitoes were exposed to doses ranging from 2.5 to 20 mg/ml. From this experiment, we found that caffeine negatively affected blood-feeding behavior, survival, and fecundity especially at higher concentrations. These results suggest that caffeine could be a potential target for future mosquito control research.

Life-history stage and the population genetics of the tiger mosquito Aedes albopictus at a fine spatial scale.

As a widespread vector of disease with an expanding range, the mosquito Aedes albopictus Skuse (Diptera: Culicidae) is a high priority for research and management. A. albopictus has a complex life history with aquatic egg, larval and pupal stages, and a terrestrial adult stage. This requires targeted management strategies for each life stage, coordinated across time and space. Population genetics can aid in A. albopictus control by evaluating patterns of genetic diversity and dispersal. However, how life stage impacts population genetic characteristics is unknown. We examined whether patterns of A. albopictus genetic diversity and differentiation changed with life stage at a spatial scale relevant to management efforts. We first conducted a literature review of field-caught A. albopictus population genetic papers and identified 101 peer-reviewed publications, none of which compared results between life stages. Our study uniquely examines population genomic patterns of egg and adult A. albopictus at five sites in Wake County, North Carolina, USA, using 8425 single nucleotide polymorphisms. We found that the level of genetic diversity and connectivity between sites varied between adults and eggs. This warrants further study and is critical for research aimed at informing local management.

Prevalence of Knock-Down Resistance F1534S Mutations in Aedes albopictus (Skuse) (Diptera: Culicidae) in North Carolina.

Knock-down resistance (kdr) mutations in the voltage-gated sodium channel gene of Aedes species mosquitoes are biomarkers for resistance to pyrethroid insecticides. In the United States, few studies have reported kdr mutations among Aedes albopictus (Skuse) (Diptera: Culicidae) populations. In this study, we sought to compare the presence of kdr alleles among Ae. albopictus mosquitoes collected from Fort Bragg and Wake County, North Carolina. We collected 538 Ae. albopictus mosquitoes, including 156 from 4 sites at Fort Bragg, North Carolina and 382 from 15 sites in Wake County, North Carolina to compare the prevalence of kdr mutations. Of those successfully sequenced, we identified 12 (3.0%) mosquitoes with kdr mutations, all of which were attributed to variants at position 1534 within domain 3. All mutations were found in mosquitoes collected at Wake County sites; no mutations were identified in collections from Fort Bragg. There was a focus of mutations observed at the Wake County sites with approximately 92% (11 of 12) of the mosquitoes with the mutation coming from one site, where kdr mutations represented 24.4% (11 of 45) of all mosquitoes collected. We observed highly focal resistance in a suburban area of Raleigh, which may be attributable to peri-domestic mosquito control activities that involve area dispersal of pyrethroid insecticides. More robust surveillance is needed to monitor the emergence and spread of resistance.

Effectiveness of autocidal gravid trapping and chemical control in altering abundance and age structure of Aedes albopictus.

BACKGROUND: Aedes albopictus is a nuisance pest mosquito of public health importance commonly managed with adulticides and larvicides. We investigated whether adding Gravid Aedes Traps (GATs), Autocidal Gravid Ovitraps (AGOs) or In2Care traps would extend the effectiveness of chemical control methods in Wake County, North Carolina, USA, by combining barrier sprays and larval habitat management (LHM) with each trap type at suburban households. We compared these three treatment groups to untreated controls and to backyards treated only with barrier sprays and LHM. Once a week, for ten weeks, we collected adult mosquitoes at each house using lure-baited surveillance traps and dissected a portion of Ae. albopictus females to determine parity. RESULTS: Barrier sprays and LHM alone or combined with any supplemental autocidal ovitrap significantly reduced female Ae. albopictus through Week 3 post-treatment. GATs significantly extended chemical control effectiveness for the duration of the study. Compared to the untreated control, the AGO and GAT treatment groups had significant overall female Ae. albopictus reductions of 74% and 80.4%, respectively, with populations aging significantly slower at houses treated with AGOs. CONCLUSION: This household-level study, though limited in size, observed significant reductions in nuisance Ae. albopictus when combining AGOs and GATs with chemical controls for an eight-week period. Delayed population aging in AGO-treated yards suggests that traps also could mitigate disease transmission risk. Future studies should test these control methods at the neighborhood level to evaluate large-scale effectiveness as well as assess the effect of autocidal ovitraps without chemical intervention. © 2022 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Invasive Plants as Foci of Mosquito-Borne Pathogens: Red Cedar in the Southern Great Plains of the USA.

West Nile virus (WNV) is the most significant mosquito-borne disease affecting humans in the United States. Eastern redcedar (ERC) is a native encroaching plant in the southern Great Plains that greatly alters abiotic conditions and bird and mosquito populations. This study tested the hypotheses that mosquito communities and their likelihood of WNV infection differ between ERC and other habitats in the southern Great Plains of the United States. We found support for our first hypothesis, with significantly more Culex tarsalis and Culex erraticus in ERC than deciduous and grass habitats. Mosquito communities in Central Oklahoma were more diverse (21 species) than western Oklahoma (11 species) but this difference was not associated with vegetation. Our second hypothesis was also supported, with significantly more WNV-infected Culex from ERC in both regions, as was our third hypothesis, with significantly more Culex tarsalis and Culex pipiens collected in ERC than other habitats in urban areas. The connection of mosquito-borne disease with invasive plants suggests that land management initiatives can affect human health and should be considered in light of public health impact. Evidence from other vector-borne disease suggests invasive plants, both in the Great Plains and globally, may facilitate the transmission of vector-borne pathogens.

Effects of host extinction and vector preferences on vector-borne disease risk in phylogenetically structured host-hector communities.

Anthropogenic disturbance impacts the phylogenetic composition and diversity of ecological communities. While changes in diversity are known to dramatically change species interactions and alter disease dynamics, the effects of phylogenetic changes in host and vector communities on disease have been relatively poorly studied. Using a theoretical model, we investigated how phylogeny and extinction influence network structural characteristics relevant to disease transmission in disturbed environments. We modelled a multi-host, multi-vector community as a bipartite ecological network, where nodes represent host and vector species and edges represent connections among them through vector feeding, and we simulated vector preferences and threat status on host and parasite phylogenies. We then simulated loss of hosts, including phylogenetically clustered losses, to investigate how extinction influences network structure. We compared effects of phylogeny and extinction to those of host specificity, which we predicted to strongly increase network modularity and reduce disease prevalence. The simulations revealed that extinction often increased modularity, with higher modularity as species loss increased, although not as much as increasing host specificity did. These results suggest that extinction itself, all else being equal, may reduce disease prevalence in disturbed communities. However, in real communities, systematic patterns in species loss (e.g. favoring high competence species) or changes in abundance may counteract these effects. Unexpectedly, we found that effects of phylogenetic signal in host and vector traits were relatively weak, and only important when phylogenetic signal of host and vector traits were similar, or when these traits both varied.

Estimations of Fine-Scale Species Distributions of Aedes aegypti and Aedes albopictus (Diptera: Culicidae) in Eastern Florida.

Many species distribution maps indicate the ranges of Aedes aegypti (Linnaeus) and Aedes albopictus (Skuse) overlap in Florida despite the well-documented range reduction of Ae. aegypti. Within the last 30 yr, competitive displacement of Ae. aegypti by Ae. albopictus has resulted in partial spatial segregation of the two species, with Ae. aegypti persisting primarily in urban refugia. We modeled fine-scale distributions of both species, with the goal of capturing the outcome of interspecific competition across space by building habitat suitability maps. We empirically parameterized models by sampling 59 sites in south and central Florida over time and incorporated climatic, landscape, and human population data to identify predictors of habitat suitability for both species. Our results show human density, precipitation, and urban land cover drive Ae. aegypti habitat suitability, compared with exclusively climatic variables driving Ae. albopictus habitat suitability. Remotely sensed variables (macrohabitat) were more predictive than locally collected metrics (microhabitat), although recorded minimum daily temperature showed significant, inverse relationships with both species. We detected minor Aedes habitat segregation; some periurban areas that were highly suitable for Ae. albopictus were unsuitable for Ae. aegypti. Fine-scale empirical models like those presented here have the potential for precise risk assessment and the improvement of operational applications to control container-breeding Aedes mosquitoes.

Short-Term, Large-Area Survey of Container Aedes spp. (Diptera: Culicidae): Presence and Abundance is Associated with Fine-scale Landscape Factors in North Carolina, USA.

Container Aedes mosquitoes are responsible for the transmission of anthroponotic and zoonotic viruses to people. The surveillance and control of these mosquitoes is an important part of public health protection and prevention of mosquito-borne disease. In this study, we surveyed 327 sites over 2 weeks in late June and early July in 2017 in North Carolina, USA for the presence and abundance of Aedes spp. eggs in an effort to better target potential Ae. aegypti collections. We examined the ability of 2 types of landscape data, Light Detection And Ranging (LIDAR) and National Land Cover Database (NLCD) to explain the presence and abundance of eggs using principal component analysis to deal with collinearity, followed by generalized linear regression. We explained variation of both egg presence and abundance for Aedes albopictus (Skuse) and Aedes triseriatus (Say) using both NLCD and LIDAR data. However, the ability to make robust predictions was limited by variation in the data. Increased sampling time and better landscape data would likely improve the predictive ability of our models, as would a better understanding of oviposition behavior.

Assessing Insecticide Resistance in Adult Mosquitoes: Perspectives on Current Methods.

Mosquito insecticide resistance (IR) is a growing global issue that must be addressed to protect public health. Vector control programs (VCPs) should regularly monitor local mosquito populations for IR and plan control measures accordingly. In some cases, state/federal resources financially support this testing with expertise and/or training programs. Standardization of methods (eg, Centers for Disease Control and Prevention bottle bioassay, World Health Organization tube testing, dose-mortality bioassay) for monitoring IR must be prioritized. One solution is regional hubs of IR monitoring at the state or other level. Training programs on methodology and interpretation of results should be developed and routinely offered to local VCPs conducting IR testing in mosquitoes. Here, current methods for assessing mosquito IR are discussed and insights into a variety of questions from VCPs are considered. It is critical that methods for IR monitoring and data interpretation are standardized through routine training, with the goal of evidence-driven decision making to improve control of mosquitoes and mosquito-borne disease.

Efficacy and Spatial Extent of Yard-Scale Control of Aedes (Stegomyia) albopictus (Diptera: Culicidae) Using Barrier Sprays and Larval Habitat Management.

The Asian tiger mosquito, Aedes (Stegomyia) albopictus (Skuse), is a peridomestic, container-ovipositing mosquito commonly found throughout the southeastern United States. In the United States, Ae. albopictus is typically considered a nuisance pest; however, it is capable of transmitting multiple pathogens. Ae. albopictus is an important pest species and the target of numerous mosquito control efforts in the United States. Here, we evaluate the effectiveness and spatial extent of Ae. albopictus population reduction using a bifenthrin (AI Bifen IT, 7.9%) barrier spray and larval habitat management (LHM) in a temperate, suburban setting. Sixteen pairs of adjoining neighbors were randomly assigned to treatment groups with one neighbor receiving a treatment and the other monitored for evidence of a spillover effect of the treatments. Ae. albopictus populations in both yards were monitored for 33 d, with treatments occurring on the eighth day. Barrier sprays, both alone and combined with LHM, resulted in a significant reduction in Ae. albopictus abundance posttreatment. While LHM alone did not result in a significant reduction over the entire posttreatment period, Ae. albopictus populations were observed to be in decline during this period. No treatments were observed to have any reduction in efficacy 25 d posttreatment, with treatments involving LHM having a significantly increased efficacy. Yards neighboring treated yards were also observed to have reduced population sizes posttreatment, but these differences were rarely significant. These results provide insights into the population dynamics of Ae. albopictus following two common treatments and will be useful for integrated pest management plans.

Mosquito diversity and dog heartworm prevalence in suburban areas.

BACKGROUND: Urbanization is occurring rapidly on a global scale and is altering mosquito communities, creating assemblages that are characteristically less diverse. Despite high rates of urbanization and ample examples of vector-borne diseases transmitted by multiple species, the effects of urbanization-driven mosquito diversity losses on disease transmission has not been well explored. We investigated this question using the dog heartworm, a filarial parasite vectored by numerous mosquito species. METHODS: We trapped host-seeking mosquitoes in undeveloped areas and neighborhoods of different ages in Wake County, North Carolina, USA, analyzing captured mosquitoes for heartworm DNA. We compared within-mosquito heartworm infection across land-use types by Kruskal-Wallis and likelihood ratio tests. Using zip code level data acquired from dogs in a local shelter, we performed linear regressions of within-host heartworm prevalence by within-mosquito heartworm prevalence as well as by three mosquito diversity measures. We also determined the best predictor of host-level prevalence among models including within-mosquito infection, mosquito diversity and abundance, and socioeconomic status as variables. RESULTS: Suburban areas had lower within-mosquito heartworm prevalence and lower likelihood of heartworm-positive mosquitoes than did undeveloped field sites, although no differences were seen between suburban and undeveloped wooded sites. No relationships were noted between within-mosquito and within-host heartworm prevalence. However, mosquito diversity metrics were positively correlated with host heartworm prevalence. Model selection revealed within-host prevalence was best predicted by a positive relationship with mosquito Shannon-Wiener diversity and a negative relationship with household income. CONCLUSIONS: Our results demonstrate that decreases in mosquito diversity due to urbanization alter vector-borne disease risk. With regard to dog heartworm disease, this loss of mosquito diversity is associated with decreased heartworm prevalence within both the vector and the host. Although the response is likely different for diseases transmitted by one or few species, mosquito diversity losses leading to decreased transmission could be generalizable to other pathogens with multiple vectors. This study contributes to better understanding of the effects of urbanization and the role of vector diversity in multi-vectored pathosystems.

Context-Dependent Accuracy of the BG-Counter Remote Mosquito Surveillance Device in North Carolina.

There is a clear need for improved vector surveillance approaches that are affordable, labor efficient, and safer than traditional methods. The BG-Counter (Biogents USA, Moorefield, WV) is a device for remotely monitoring mosquito activity in combination with the BG-Sentinel (Biogents USA), a widely used trap for the collection of host-seeking mosquitoes. The BG-Counter uses a wireless connection to provide real-time counts of mosquitoes captured by the BG-Sentinel, allowing users to remotely monitor mosquito populations. This study tested the effectiveness of the BG-Counter in 5 North Carolina counties. A total of 96 trap-days resulted in the collection of >45,000 individual mosquitoes representing 35 species. Aedes albopictus was the most common species collected in all counties, except for New Hanover County where Culex nigripalpus was the most common. The mean daily accuracy ranged from 80.1% (New Hanover County) to 9.4% (Jackson County). There was a significant linear relationship between the actual number of mosquitoes collected and the device counts at all sites except Jackson County, the site with the lowest relative mosquito abundance compared with nontarget organisms. A linear regression of daily BG-Counter accuracy and the daily proportion of mosquitoes to the total number of arthropods collected revealed a significant positive linear relationship, supporting the premise that the BG-Counter is less effective when the relative abundance of mosquitoes is low. Mosquito surveillance programs using the BG-Counter should recognize its context-dependent accuracy and routinely evaluate the accuracy of the device based on local conditions.

Comparative Vector Efficiency of Two Prevalent Mosquito Species for Dog Heartworm in North Carolina.

The dog heartworm, Dirofilaria immitis (Leidy) (Spirurida: Onchocercidae), is a devastating parasite of domestic and wild canines vectored by a multitude of mosquito species. Although many species are implicated as vectors, not all contribute equally to disease transmission, with demonstrated variation in vector efficiency between and within species. We investigated the vector efficiency of mosquitoes derived from wild-caught North Carolina populations of two known heartworm vectors: a native species, Aedes triseriatus (Say) (Diptera: Culicidae), and an invasive species, Aedes albopictus (Skuse). We compared the parasite developmental times within the mosquito, mosquito longevity and fecundity, and the vector efficiency index between the two species. We found that the tested composite North Carolina population of Ae. triseriatus was an efficient vector of D. immitis under laboratory conditions, whereas the local composite population of Ae. albopictus was a competent but relatively poor vector. Compared with Ae. triseriatus, Ae. albopictus showed a longer time for parasite development, lower infection rates, and lower vector efficiency. Additionally, Ae. albopictus was the sole species to exhibit significant parasite-induced mortality. These results are in contrast to prior studies of populations of Ae. albopictus from locations outside of North Carolina, which have implicated the species as a highly competent heartworm vector. The variation seen for different strains of the same species emphasizes the heritable nature of D. immitis vector competence and highlights the need for local infection studies for accurate transmission risk assessment in a particular locale.

Patterns of genetic divergence among populations of Aedes aegypti L. (Diptera: Culicidae) in the southeastern USA.

BACKGROUND: The yellow fever mosquito, Aedes aegypti is a public health concern in the USA, especially in the wake of emergent diseases such as Zika and chikungunya. Aedes aegypti populations dwindled after the invasion of Aedes albopictus in the 1980s and many populations were extirpated. However, in some areas Ae. aegypti persisted in small populations and there are reports of recent resurgences of Ae. aegypti in Florida, Louisiana, Nevada and California. We assessed the population genetic structure of Ae. aegypti in Florida and Georgia, which has concomitant consequences related to mosquito dispersal, pesticide resistance and vectorial capacity. METHODS: We collected Ae. aegypti across Florida and in Georgia using ovitraps. We hatched the eggs and reared them to adults, and after sacrifice we extracted their DNA. We then probed each individual for variation in 6 microsatellite markers, which we used to address population genetic characteristics. RESULTS: We collected Ae. aegypti and genotyped seven Florida populations and one Georgia population using microsatellite markers. We found evidence of isolation by distance model of gene flow supported by driving distance among cities within Florida and two theoretic genetic clusters. CONCLUSIONS: Significant genetic structure between some populations with substantial gene flow between geographically distant cities suggests regional genetic structuring of Ae. aegypti in Florida. This study provides information on the genetic exchange between populations of Ae. aegypti in the southeastern USA and suggests potential routes of spread of this species.

First Record of Mansonia dyari From Saint Croix, United States Virgin Islands.

The first report of Mansonia dyari on Saint Croix, United States Virgin Islands (USVI), is confirmed. Adult and larval specimens were collected in 2018 and 2019 through adult surveillance and larval collections. Specimens were identified by microscopic methods, and a representative specimen was confirmed by DNA sequencing (mitochondrial cytochrome c oxidase subunit I). Morphological features are reviewed and compared with Mansonia flaveola, a species previously reported in the USVI. Notes are provided on the locations, collection methods, and mosquito associates found with Ma. dyari in the USVI.