Competitive interactions with Aedes albopictus alter the nutrient content of Aedes aegypti.

Competition is often cited as a central force that affects the distribution and performance of organisms. Ecological stoichiometry is the balance of elements within animal bodies that can be affected by resource acquisition and processing, as well as by intra- or interspecific interactions. Though relatively underexplored for mosquitoes, stoichiometry may provide a wealth of information linking ecological interactions to body nutrient content, and potentially on to pathogen transmission. Detritus, which often varies in nutrient content, forms the base of the food web within the small aquatic habitats occupied by larval mosquitoes, and detrital nutrient content can alter mosquito growth, survival, and population growth. The invasive mosquitoes Aedes albopictus (Diptera: Culicidae) and Aedes aegypti (Diptera: Culicidae) interact as larvae in aquatic systems, often altering their adult populations. Herein, we investigated how different detritus combinations as well as how intra- and interspecific densities of Ae. albopictus and Ae. aegypti would affect coexistence; we also measured how nutrient composition (carbon and nitrogen) and stoichiometry (C:N) of adults would vary with those interactions. Ae. albopictus survival, population growth, and stoichiometry were not affected by intra- or interspecific competition; nutrient values did vary with detritus ratios. However, Ae. aegypti nutrient content and stoichiometry and survival were negatively affected within the lowest nutrient environments in the presence of Ae. albopictus, but in the highest nutrient environments, both species showed high survival rates and population growth. This is the first study to show that adult mosquito body nutrients can be altered by interspecific interactions, and as nutrient content in adults has been linked to pathogen transmission, it provides a novel role of competition in affecting disease dynamics.

Non-target effects of methoprene and larvicidal surface films on invertebrate predators of mosquito larvae.

Mosquito larvicides are used across a variety of aquatic habitats, although when applied they likely affect other aquatic organisms. The removal or impairment of top insect predators via larvicides could be beneficial to mosquitoes by allowing their populations to rebound once pesticide levels dissipate. Our goal was to determine if two larvicide types, growth regulators (IGRs) and surface films (SFs), harm non-target aquatic insect communities, and if these chemicals influence the ability of predatory aquatic insects to regulate mosquitoes. We surveyed aquatic sites before and after IGR and SF-application, then compared changes in insect community structure. Evenness was lower in SF treated habitats, and when analyzing prey/controphic taxa only, evenness and diversity changed in untreated reference areas, suggesting that differences measured were due to other environmental factors, not larvicide presence. A field experiment was then conducted by exposing specific predatory aquatic insects to varying doses of IGRs and SFs and then placing them in mesocosms containing mosquito larvae. Surface films were directly lethal to adult dytiscids at recommended and high concentrations. Although we found no significant differences in mosquito emergence among all treatment levels, there was a trend of negative controls (no predator mesocosms) and SF-treated predators allowing the most mosquitoes to emerge compared to positive controls (predators not exposed to larvicides) and IGR-treated predators. Thus, these larvicides may have minimal effects on mosquito larvae predators, but the direct effects of surface films on insects that interact with the water's surface require further investigation.

Plant pollen as a resource affecting the development and survival of the mosquitoes Anopheles quadrimaculatus and Culex quinquefasciatus (Diptera: Culicidae).

Mosquito larvae often subsist on inputs of terrestrial-derived resources, including leaves and dead insects. However, seasonal inputs of plant pollen is an underexplored resource for many species. We compared the effects of three levels (low, medium, high) of two pollen types (corn, pine) on development, mass, and survival in Anopheles quadrimaculatus (Say) and Culex quinquefasciatus (Say) (Diptera: Culcidae). We also examined the nutrient content of adults (%nitrogen, %carbon, C:N) and stable isotopes (δ15N, δ13C). Culex quinquefasciatus had the highest survival rates when grown on high and medium pine pollen compared with low pine. Survival of Culex quinquefasciatus was generally higher compared to that of An. quadrimaculatus on any level of pine, with the latter species having higher survival in high corn. Nutrient content for An. quadrimaculatus did not vary significantly in either pollen type or amount but were more enriched in δ15N in corn pollen relative to pine pollen. For Cx. quinquefasciatus, %N decreased and C:N ratio increased across low to high amounts of corn. Adults raised in corn had generally more δ13C compared to pine pollen. No developmental differences across diets were observed for either species, however both sexes of Cx. quinquefasciatus were generally larger when grown in high pine and medium and high corn pollen compared with other treatments. The poor performance of An. quadrimaculatus on corn pollen was unexpected, however, we show a benefit of corn pollen to Cx. quinquefasciatus with implications for West Nile virus transmission in the United States, especially around agricultural areas where corn is grown.

Ontogenetic Changes in Nutrients and Stoichiometry in the Invasive Mosquito, Aedes albopictus (Diptera: Culicidae).

A variety of physiological, morphological, and behavioral changes occur throughout the life cycle of mosquitoes, which can be correlated with a shift from the aquatic to terrestrial environment. Aedes albopictus Skuse is an abundant invasive species from Asia that was introduced into the Americas in the 1980's and is responsible for transmitting several important human disease-causing pathogens. How physiological and anatomical changes within each instar and throughout the developmental stages are related to changes in carbon (C) and nitrogen (N) levels are an unexplored area of mosquito ecology. We hypothesized that these changes as well as stoichiometry (C:N) would vary with instar stage and larval diet. Cohorts of larvae were grown in three different diets: animal only (crickets), plant only (red maple leaves), and a mixture containing both types. Larval instars (1st-4th), pupae, and adults were raised in each diet and were separately analyzed for nutrient content (%C, %N) and stoichiometry (C:N). Significant changes in nutrient values occurred across the life cycle, with C:N values being lower in early instars versus adults or pupae, especially in animal only or mixed diets; few differences were detected in %C or %N across ontogeny. This knowledge may lead to a better understanding of mosquito ecology and pathogen transmission.

Mosquito Surveillance for West Nile Virus.

Identifying the mosquitoes responsible for transmitting human disease-causing pathogens is of critical importance for effective control of mosquito-borne outbreaks. West Nile virus is often transferred by adult female mosquitoes in the genus Culex, which deposit eggs in a variety of aquatic habitats throughout the world. Herein we describe several methodological approaches to monitor these species in nature, as well as offering details for data collection and analysis.

Robust network stability of mosquitoes and human pathogens of medical importance.

BACKGROUND: The exact number of mosquito species relevant to human health is unknown, posing challenges in understanding the scope and breadth of vector-pathogen relationships, and how resilient mosquito vector-pathogen networks are to targeted eradication of vectors. METHODS: We performed an extensive literature survey to determine the associations between mosquito species and their associated pathogens of human medical importance. For each vector-pathogen association, we then determined the strength of the associations (i.e., natural infection, lab infection, lab dissemination, lab transmission, known vector). A network analysis was used to identify relationships among all pathogens and vectors. Finally, we examined how elimination of either random or targeted species affected the extinction of pathogens. RESULTS: We found that 88 of 3578 mosquito species (2.5%) are known vectors for 78 human disease-causing pathogens; however, an additional 243 species (6.8%) were identified as potential or likely vectors, bringing the total of all mosquitos implicated in human disease to 331 (9.3%). Network analysis revealed that known vectors and pathogens were compartmentalized, with the removal of six vectors being enough to break the network (i.e., cause a pathogen to have no vector). However, the presence of potential or likely vectors greatly increased redundancies in the network, requiring more than 41 vectors to be eliminated before breaking the network. CONCLUSION: Although < 10% of mosquitoes are involved in transmitting pathogens that cause human disease, our findings point to inherent robustness in global mosquito vector-pathogen networks.

Mapping Yellow fever epidemics as a potential indicator of the historical range of Aedes aegypti in the United States.

BACKGROUND: Yellow fever (YF) plagued the United States from the 1690s until 1905, resulting in thousands of deaths. Within the US, Aedes aegypti is the only YF vector and almost no data exists for the location of this species prior to the early 1900s. OBJECTIVES: To determine the historical range of Ae. aegypti we examined the occurrence of YF epidemics across time and space. We hypothesized that historically Ae. aegypti was driven by human population density, like its contemporary range suggests. METHODS: To test this hypothesis, we compiled a list of YF cases in the US, human population density, location, and the number of people infected. This data was mapped using ArcGIS and was analyzed using linear regression models to determine the relationship among variables. FINDINGS: The historic range was generally south of 40º latitude, from Texas in the west to Florida in the east, with concentrations along major waterways like the Mississippi River. Infected individuals and human population density were strongly correlated across the whole dataset as well as by decade. MAIN CONCLUSIONS: Although other factors likely affected the range of Ae. aegypti, we found that human population density was related to the number of people infected with historic YF infections.

Mosquitoes (Diptera: Culicidae) on the islands of Puerto Rico and Vieques, U.S.A.

We conducted an island-wide survey of the Caribbean islands Puerto Rico and Vieques, U.S.A. during late 2018 and early 2019 to document the current richness of mosquitoes (Diptera: Culicidae). We used a combination of larval surveys and adult trapping using baited CDC light traps and BG-Sentinel traps across 41 of the 78 municipalities. We collected 9 genera, 12 subgenera, and 31 species, which when combined with past studies yields 44 species on the islands. We also note species occurrences across habitat types and elevations from around the islands. One new record, Aedes (Ochlerotatus) obturbator Dyar and Knab, is noted. However we found no evidence of the presence of Aedes albopictus (Skuse), an invasive found throughout the Caribbean, or Aedes (Fredwardsius) vittatus (Bigot), an exotic species recently reported in the Dominican Republic and Cuba. Habitat associations and information regarding the medical importance of species are also included. Given that the islands often experience outbreaks of several arboviruses, obtaining a complete picture of the species present is of high importance.

Correction: Quantifying species traits related to oviposition behavior and offspring survival in two important disease vectors.

[This corrects the article DOI: 10.1371/journal.pone.0239636.].

Temporal and Spatial Blood Feeding Patterns of Urban Mosquitoes in the San Juan Metropolitan Area, Puerto Rico.

Urban ecosystems are a patchwork of habitats that host a broad diversity of animal species. Insects comprise a large portion of urban biodiversity which includes many pest species, including those that transmit pathogens. Mosquitoes (Diptera: Culicidae) inhabit urban environments and rely on sympatric vertebrate species to complete their life cycles, and in this process transmit pathogens to animals and humans. Given that mosquitoes feed upon vertebrates, they can also act as efficient samplers that facilitate detection of vertebrate species that utilize urban ecosystems. In this study, we analyzed DNA extracted from mosquito blood meals collected temporally in multiple neighborhoods of the San Juan Metropolitan Area, Puerto Rico to evaluate the presence of vertebrate fauna. DNA was collected from 604 individual mosquitoes that represented two common urban species, Culex quinquefasciatus (n = 586) and Aedes aegypti (n = 18). Culex quinquefasciatus fed on 17 avian taxa (81.2% of blood meals), seven mammalian taxa (17.9%), and one reptilian taxon (0.85%). Domestic chickens dominated these blood meals both temporally and spatially, and no statistically significant shift from birds to mammals was detected. Aedes aegypti blood meals were from a less diverse group, with two avian taxa (11.1%) and three mammalian taxa (88.9%) identified. The blood meals we identified provided a snapshot of the vertebrate community in the San Juan Metropolitan Area and have potential implications for vector-borne pathogen transmission.

Transmission Potential of Zika Virus by Aedes aegypti (Diptera: Culicidae) and Ae. mediovittatus (Diptera: Culicidae) Populations From Puerto Rico.

Recurrence of local transmission of Zika virus in Puerto Rico is a major public health risk to the United States, where mosquitoes Aedes aegypti (Linnaeus) and Aedes mediovittatus (Coquillett) are abundant. To determine the extent to which Ae. mediovittatus are capable of transmitting Zika virus and the influence of viremia, we evaluated infection and transmission in Ae. mediovittatus and Ae. aegypti from Puerto Rico using serial dilutions of infectious blood. Higher doses of infectious blood resulted in greater infection rates in both mosquitoes. Aedes aegypti females were up to twice as susceptible to infection than Ae. mediovittatus, indicating a more effective midgut infection barrier in the latter mosquito species. Aedes aegypti exhibited higher disseminated infection (40-95%) than Ae. mediovittatus (<5%), suggesting a substantial midgut escape barrier in Ae. mediovittatus. For Ae. aegypti, transmission rates were low over a range of doses of Zika virus ingested, suggesting substantial salivary gland barriers.

No Evidence That Salt Water Ingestion Kills Adult Mosquitoes (Diptera: Culicidae).

Various products and insecticides are available that purport to reduce wild populations of adult mosquitoes. Recently, several manufacturers and general public comments on the internet have promoted devices that claim that ingestion of salt will significantly reduce populations of wild mosquitoes to near zero; there are no known scientific efficacy data that support these claims. We tested the survival of nine mosquito species of pest and public health importance across four adult diets: Water Only, Sugar Water Only (8.00%), Salt Water Only (1.03%), and Sugar + Salt Water. Species included the following: Aedes aegypti (L.), Aedes albopictus (Skuse), Aedes dorsalis (Meigen), Aedes notoscriptus (Skuse), Aedes vigilax (Skuse), Anopheles quadrimaculatus (Say), Culex pipiens (L.), Culex quinquefasciatus (Say), and Culex tarsalis (Coquillett). Male and female mosquitoes were placed in cages and allowed to feed on liquid diets under controlled environmental conditions for 1 wk. For seven of the nine species, adult survival was significantly higher in the presence (Sugar Water, Sugar + Salt Water) versus the absence (Water Only, Salt Only) of sugar, with no indication that salt had any effect on survival. Anopheles quadrimaculatus showed intermediate survival in Sugar + Salt to either Sugar Only or no sugar diets, whereas Aedes dorsalis showed low survival in Salt Only versus other diets. Based on our data and coupled with the fact that mosquitoes have physiological and behavioral adaptations that allow them to avoid or process excess salt (as found in blood meals), we conclude that there is no scientific foundation for salt-based control methods of mosquitoes.

Quantifying species traits related to oviposition behavior and offspring survival in two important disease vectors.

Animals with complex life cycles have traits related to oviposition and juvenile survival that can respond to environmental factors in similar or dissimilar ways. We examined the preference-performance hypothesis (PPH), which states that females lacking parental care select juvenile habitats that maximize fitness, for two ubiquitous mosquito species, Aedes albopictus and Culex quinquefasciatus. Specifically, we examined if environmental factors known to affect larval abundance patterns in the field played a role in the PPH for these species. We first identified important environmental factors from a field survey that predicted larvae across different spatial scales. We then performed two experiments, the first testing the independent responses of oviposition and larval survival to these environmental factors, followed by a combined experiment where initial oviposition decisions were allowed to affect larval life history measures. We used path analysis for this last experiment to determine important links among factors in explaining egg numbers, larval mass, development time, and survival. For separate trials, Aedes albopictus displayed congruence between oviposition and larval survival, however C. quinquefasciatus did not. For the combined experiment path analysis suggested neither species completely fit predictions of the PPH, with density dependent effects of initial egg number on juvenile performance in A. albopictus. For these species the consequences of female oviposition choices on larval performance do not appear to fit expectations of the PPH.

Do socioeconomic factors drive Aedes mosquito vectors and their arboviral diseases? A systematic review of dengue, chikungunya, yellow fever, and Zika Virus.

As the threat of arboviral diseases continues to escalate worldwide, the question of, "What types of human communities are at the greatest risk of infection?" persists as a key gap in the existing knowledge of arboviral diseases transmission dynamics. Here, we comprehensively review the existing literature on the socioeconomic drivers of the most common Aedes mosquito-borne diseases and Aedes mosquito presence/abundance. We reviewed a total of 182 studies on dengue viruses (DENV), chikungunya virus (CHIKV), yellow fever virus (YFVV), Zika virus (ZIKV), and presence of Aedes mosquito vectors. In general, associations between socioeconomic conditions and both Aedes-borne diseases and Aedes mosquitoes are highly variable and often location-specific. Although 50% to 60% of studies found greater presence or prevalence of disease or vectors in areas with lower socioeconomic status, approximately half of the remaining studies found either positive or null associations. We discuss the possible causes of this lack of conclusiveness as well as the implications it holds for future research and prevention efforts.

Oviposition Responses and Potential Larval Control Methods of Aedes albopictus (Diptera: Culicidae) in Downspout Extensions.

Aedes albopictus (Skuse) is an important invasive species and vector of several important arboviruses across the globe. This species uses small water-holding cryptic containers as egg laying sites, which pose serious challenges to effective control of adult mosquito populations. Herein, we examined the response of gravid female Ae. albopictus to various features of common downspout extension tubes associated with human dwellings and the effectiveness of control efforts to eliminate larvae. Controlled field trials quantified oviposition in 1) extensions versus rubber bowls meant to mimic other container types, 2) among different shapes and materials of extensions, and 3) among different colors of extensions. We also investigated how flushing and use of Bti larvicides could control larvae. Females were more likely to lay eggs in flat plastic or metal extensions compared to rubber bowls. Eggs were also more plentiful in flat plastic extensions versus either corrugated or metal, and dark brown corrugated extensions had more eggs compared to tan or white. Flushing reduced nearly all larvae when the extensions were properly angled, and applications of Bti pellets or dunks were effective at killing most larvae. We show that dark extensions were preferred over other colors, and that larvae can be effectively removed with minimal effort. However, effective control will likely only come from better education of the public about proper installation of extensions.

Linking nutrient stoichiometry to Zika virus transmission in a mosquito.

Food quality and quantity serve as the basis for cycling of key chemical elements in trophic interactions; yet the role of nutrient stoichiometry in shaping host-pathogen interactions is under appreciated. Most of the emergent mosquito-borne viruses affecting human health are transmitted by mosquitoes that inhabit container systems during their immature stages, where allochthonous input of detritus serves as the basal nutrients. Quantity and type of detritus (animal and plant) were manipulated in microcosms containing newly hatched Aedes aegypti mosquito larvae. Adult mosquitoes derived from these microcosms were allowed to ingest Zika virus-infected blood and then tested for disseminated infection, transmission, and total nutrients (percent carbon, percent nitrogen, ratio of carbon to nitrogen). Treatments lacking high-quality animal (insect) detritus significantly delayed development. Survivorship to adulthood was closely associated with the amount of insect detritus present. Insect detritus was positively correlated with percent nitrogen, which affected Zika virus infection. Disseminated infection and transmission decreased with increasing insect detritus and percent nitrogen. We provide the first definitive evidence linking nutrient stoichiometry to arbovirus infection and transmission in a mosquito using a model system of invasive Ae. aegypti and emergent Zika virus.

Linking Water Quality to Aedes aegypti and Zika in Flood-Prone Neighborhoods.

The ability of ecosystems to regulate water quality and flood events has been linked to health outcomes, including mosquito-borne illnesses. In the San Juan Bay Estuary watershed of Puerto Rico, habitat alterations and land-use development have disrupted watershed hydrology, exacerbating wastewater discharges and subjecting some neighborhoods to frequent flooding events. In 2016, the mosquito-borne illness Zika became a new cause for concern. We hypothesized that nutrient-enriched flood water could provide pulses of supplemental nutrients to local mosquito populations. We conducted a field study in six neighborhoods adjacent to the estuary to assess whether environmental variability of nutrient inputs could be linked to breeding habitat containers, Aedes aegypti larvae and adults, and the acquisition of Zika virus by adult mosquitoes. The most frequently flooded neighborhood had consistently higher levels of nitrogen in estuary water, leaf detritus, containers, and adult mosquitoes compared to other neighborhoods. Adult body nitrogen was significantly related to both nitrogen content of containers and leaf detritus from the local trapping area. Disseminated Zika concentration in adult Ae. aegypti tended to decrease as body carbon and nitrogen increased. Our study provides preliminary evidence that environmental variability in nutrient inputs can influence viral acquisition by mosquito vectors. This suggests that management actions to reduce flooding and improve water quality should go hand-in-hand with more traditional vector control methods, such as aerial spraying, to help control spread of vector-borne diseases.

The role of temperature in affecting carry-over effects and larval competition in the globally invasive mosquito Aedes albopictus.

BACKGROUND: Ectotherms, like mosquitoes, have evolved specific responses to variation in environmental conditions like temperature, and these responses could confer a fitness benefit or cost when carried-over to different life stages. However, effects of temperature on animals with complex life-cycles often only focus on part of their life-cycle, or only consider how single aspects of life-history may carry over to new stages. Herein we investigated how temperature affects intraspecific larval competition and carry-over effects from larval to adult stages in the widespread invasive Asian tiger mosquito Aedes albopictus. METHODS: For larval competition, larvae were reared at three densities (10, 20, and 40 individuals) across three source temperatures (21 °C, 27 °C and 34 °C). To test carry-over effects, adult survival was measured for individuals crossed with adult temperatures of 21 °C, 27 °C and 34 °C from the larval density of 20 individuals at each source temperature. Fecundity data also were obtained from mated females. RESULTS: For competition, there was a significant interaction between larval density and temperature, with the smallest females, who took the longest to develop, produced in the highest temperatures; density generally accentuated this effect. Regarding carry-over effects, adults exposed to higher temperatures lead to greater differences in fecundity and survival of adult populations. CONCLUSIONS: Temperature appears to affect life-history of developing larvae under competitive interactions and can also alter adult fitness as the disparity between larval rearing and adult habitat temperatures increases. This has importance for our understanding for how different life-history stages of Ae. albopictus and other vectors of disease may respond to changing climates.

Oviposition preference and offspring performance in container breeding mosquitoes: evaluating the effects of organic compounds and laboratory colonisation.

The preference-performance hypothesis predicts that organisms lacking parental care should oviposit in habitats that optimize offspring performance. We investigated preference-performance relationships for the Asian tiger mosquito (Aedes albopictus Skuse) and the southern house mosquito (Culex quinquefasciatus Say) (Diptera:Culicidae), two medically important container-breeding species, in response to an organic chemical blend mimicking decaying plant matter. Additionally, we evaluated the effects of long-term laboratory colonization of Cx. quinquefasciatus by using wild and laboratory strains.Oviposition bioassays were conducted by releasing gravid mosquitoes into field enclosures with automobile tires containing low and high concentrations of the chemical blend, and water controls. The offspring were then reared in water collected from the tires in which they were deposited.Aedes albopictus and wild Cx. quinquefasciatus laid more eggs in the chemical blend than water controls but did not differentiate between the low and high concentrations. Conversely, laboratory Cx. quinquefasciatus only preferred the high concentration to the low concentration. No statistical associations between oviposition preference and larval survival were found, as the chemical blend did not affect survivorship of either species.The oviposition preference for the chemical blend over water controls suggests that both species oviposit in the best available resource environment, but further studies are needed before conclusions regarding preference-performance relationships can be drawn.We found that long-term laboratory colonization affects oviposition behavior in Cx. quinquefasciatus, suggesting that behavioral studies on laboratory strains are not always applicable to wild populations.