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.

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.

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.

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.

A Statewide Survey for Container-Breeding Mosquitoes in Mississippi.

Container-breeding mosquitoes are important in public health due to outbreaks of Zika, chikungunya, and dengue viruses. This paper documents the distribution of container-breeding mosquito species in Mississippi, with special emphasis on the genus Aedes. Five sites in each of the 82 Mississippi counties were sampled monthly between May 1 and August 31, 2016, and 50,109 mosquitoes in 14 species were collected. The most prevalent and widely distributed species found was Ae. albopictus, being found in all 82 counties, especially during July. A recent invasive, Ae. japonicus, seems to be spreading rapidly in Mississippi since first being discovered in the state in 2011. The most abundant Culex species collected were Cx. quinquefasciatus (found statewide), Cx. salinarius (almost exclusively in the southern portion of the state), and Cx. restuans (mostly central and southern Mississippi). Another relatively recent invasive species, Cx. coronator, was found in 20 counties, predominantly in the southern one-third of the state during late summer. Co-occurrence data of mosquito species found in the artificial containers were also documented and analyzed. Lastly, even though we sampled extensively in 410 sites across Mississippi, no larval Ae. aegypti were found. These data represent the first modern statewide survey of container species in Mississippi, and as such, allows for better public health readiness for emerging diseases and design of more effective vector control programs.

Interspecific Interactions Between Adult Aedes albopictus and Culex quinquefasciatus (Diptera: Culicidae).

Single and mixed species densities of adult Aedes albopictus and Culex quinquefasciatus were manipulated to determine if different combinations affected their egg laying preference or mortality rates. Oviposition was measured in environments that contained containers of different surface areas (small cups vs. larger bowls), and the number of eggs (Aedes) and egg rafts or larvae (Culex) deposited by each species was examined with respect to intra- and interspecific density treatment levels. Mixed species densities did not have an effect on survivorship, but single species densities did affect longevity, with higher densities leading to shorter life spans in Cx. quinquefasciatus: Cx. quinquefasciatus lived longer than Ae. albopictus overall. There was no significant effect of density combinations on oviposition patterns in either species, but Cx. quinquefasciatus laid more rafts in bowls compared with cups. There is little evidence of adult interactions between these species; however, future experimental work is necessary to more fully characterize the possible effects of adult interactions on these species.

Mosquito Larvae in Tires from Mississippi, United States: The Efficacy of Abiotic and Biotic Parameters in Predicting Spatial and Temporal Patterns of Mosquito Populations and Communities.

Container systems, including discarded vehicle tires, which support populations of mosquitoes, have been of interest for understanding the variables that produce biting adults that serve as both nuisances and as public health threats. We sampled tires in six sites at three times in 2012 across the state of Mississippi to understand the biotic and abiotic variables responsible for explaining patterns of larvae of common species, species richness, and total abundance of mosquitoes. From 498 tires sampled, we collected >58,000 immatures representing 16 species, with the most common species including Aedes albopictus (Skuse), Culex quinquefasciatus (L.), Orthopodomyia signifera (Coquillett), Aedes triseriatus (Say), Toxorhynchites rutilus septentrionalis (Coquillett), and Culex territans (Walker) accounting for ∼97% of all larvae. We also documented 32 new county records for resident species and recent arrivals in the state, including Aedes japonicus japonicus (Theobald) and Culex coronator (Dyar & Knab). Cluster analysis, which was used to associate sites and time periods based on similar mosquito composition, did reveal patterns across the state; however, there also were more general patterns between species and genera and environmental factors. Broadly, Aedes was often associated with factors related to detritus, whereas Culex was frequently associated with habitat variables (e.g., tire size and water volume) and microorganisms. Some Culex did lack factors connecting variation in early and late instars, suggesting differences between environmental determinants of oviposition and survival. General patterns between the tire environment and mosquito larvae do appear to exist, especially at the generic level, and point to inherent differences between genera that may aid in predicting vector locations and populations.

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.

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.

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.

Concurrent effects of resource pulse amount, type, and frequency on community and population properties of consumers in detritus-based systems.

Episodic resource inputs (i.e., pulses) can affect food web properties and community dynamics, but detailed mechanistic understanding of such effects remain elusive. Natural aquatic microsystems (e.g., tree holes, human-made containers) are colonized by invertebrates that form complex food webs dependent on episodic and sometimes sizeable inputs of allochthonous detritus from adjacent terrestrial environments. We investigated how variation in pulse frequency, amount, and resource type interacted to affect richness, abundance, composition, and population sizes of colonizing invertebrates in water-filled tires and tree hole analogs in a forest habitat. Different container types were used to assess the generality of effects across two environmental contexts. Containers received large infrequent or small frequent pulses of animal or leaf detritus of different cumulative amounts distributed over the same period. Invertebrates were sampled in June and September when cumulative detritus input was equal for the two pulse frequencies. Pulse frequency and detritus type interacted to affect the responses of richness and abundance in both months; pulse frequency alone in June affected the relationship between richness and abundance. Richness and abundance were also greater with more detritus regardless of detritus type. One group, the filter feeders, were most important in driving the response of abundance and richness to pulses, especially in June. This work highlights the potential complex nature of responses of communities and populations to resource pulses and implicates the ability of certain groups to exploit pulses of detrital resources as a key to understanding community-level responses to pulses.

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.

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.

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.].

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.

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.

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.

Environmental correlates of abundances of mosquito species and stages in discarded vehicle tires.

Discarded vehicle tires are a common habitat for container mosquito larvae, although the environmental factors that may control their presence or abundance within a tire are largely unknown. We sampled discarded vehicle tires in six sites located within four counties of central Illinois during the spring and summer of 2006 to determine associations between a suite of environmental factors and community composition of container mosquitoes. Our goal was to find patterns of association between environmental factors and abundances of early and late instars. We hypothesized that environmental factors correlated with early instars would be indicative of oviposition cues, whereas environmental factors correlated with late instars would be those important for larval survival. We collected 13 species of mosquitoes, with six species (Culex restuans, Cx. pipiens, Aedes albopictus, Cx. salinarius, Ae. atropalpus, and Ae. triseriatus) accounting for r95% of all larvae. There were similar associations between congenerics and environmental factors, with Aedes associated with detritus type (fine detritus, leaves, seeds) and Culex associated with factors related to the surrounding habitat (human population density, canopy cover, tire size) or microorganisms (bacteria, protozoans). Although there was some consistency in factors that were important for early and late instar abundance, there were few significant associations between early and late instars for individual species. Lack of correspondence between factors that explain variation in early versus late instars, most notable for Culex, suggests a difference between environmental determinants of oviposition and survival within tires. Environmental factors associated with discarded tires are important for accurate predictions of mosquito occurrence at the generic level.

Stable Isotope Analysis Reveals Detrital Resource Base Sources of the Tree Hole Mosquito, Aedes triseriatus.

1. Detritus that forms the basis for mosquito production in tree hole ecosystems can vary in type and timing of input. We investigated the contributions of plant- and animal-derived detritus to the biomass of Aedes triseriatus (Say) pupae and adults by using stable isotope ((15)N and (13)C) techniques in lab experiments and field collections.2. Lab-reared mosquito isotope values reflected their detrital resource base, providing a clear distinction between mosquitoes reared on plant or animal detritus.3. Isotope values from field-collected pupae were intermediate between what would be expected if a single (either plant or animal) detrital source dominated the resource base. However, mosquito isotope values clustered most closely with plant-derived values, and a mixed feeding model analysis indicated tree floral parts contributed approximately 80% of mosquito biomass. The mixed model also indicated that animal detritus contributed approximately 30% of mosquito tissue nitrogen.4. Pupae collected later in the season generally had isotope values that were consistent with an increased contribution from animal detritus, suggesting this resource became more nutritionally important for mosquitoes as plant inputs declined over the summer.