Bacterial Communities and Midgut Microbiota Associated with Mosquito Populations from Waste Tires in East-Central Illinois.

Mosquito-microbe interactions tend to influence larval nutrition, immunity, and development, as well as fitness and vectorial capacity of adults. Understanding the role of different bacterial species not only improves our knowledge of the physiological and ecological consequences of these interactions, but also provides the basis for developing novel strategies for controlling mosquito-borne diseases. We used culture-dependent and culture-independent techniques to characterize the bacterial composition and abundance in water and midgut samples of larval and adult females of Aedes japonicus (Theobald), Aedes triseriatus (Say), and Culex restuans (Theobald) collected from waste tires at two wooded study sites in Urbana, IL. The phylum-specific real-time quantitative polymerase chain reaction assay revealed a higher proportion of Actinobacteria and a lower proportion of gamma-Proteobacteria and Bacteroidetes in water samples and larval midguts compared to adult female midguts. Only 15 of the 57 bacterial species isolated in this study occurred in both study sites. The number of bacterial species was highest in water samples (28 species from Trelease Woods; 25 species from South Farms), intermediate in larval midguts (13 species from Ae. japonicus; 12 species from Ae. triseriatus; 8 species from Cx. restuans), and lowest in adult female midguts (2 species from Ae. japonicus; 3 species from Ae. triseriatus). These findings suggest that the composition and richness of bacterial communities varies both between habitats and among mosquito species and that the reduction in bacteria diversity during metamorphosis is more evident among bacteria detected using the culture-dependent method.

The importance of oxidases in the tolerance of deciduous leaf infusions by Aedes (Stegomyia) aegypti and Aedes (Stegomyia) albopictus (Diptera: Culicidae).

Aedes (Stegomyia) aegypti (L.) and Aedes (Stegomyia) albopictus (Skuse) larvae rely on oxidases to reduce toxicity of water soluble toxins from some senescent tree leaf infusions. The mortality of third instar Ae. aegypti larvae in live oak and pin oak leaf infusions increased significantly in the presence of piperonyl butoxide (PBO), a broad inhibitor of cytochrome P450s (CYPs). In contrast, PBO treatment did not increase mortality in water controls or infusions of northern red oak or sugar maple leaf infusions for Ae. aegypti larvae. A similar pattern was observed for Ae. albopictus larvae, that is, an increase in mortality when CYPs were inhibited in live oak leaf infusions and no increase in sugar maple leaf infusions or water controls. However, the fresh live oak leaf infusion (5 d old) was the most toxic infusion to Ae. aegypti, but appeared less toxic to Ae. albopictus than the older infusions. A direct comparison of survival between the two Aedes species revealed Ae. aegypti exhibited a greater mortality than Ae. albopictus in PBO-treated live oak leaf infusions. These findings suggest that toxic components of some leaf litter in larval habitats may impose cryptic energy costs (detoxification).

Pesticide-induced release from competition among competing Aedes aegypti and Aedes albopictus (Diptera: Culicidae).

Competitive interactions between mosquitoes Aedes aegypti (L.) and Aedes albopictus (Skuse) may depend on environmental conditions. Pesticides may alleviate density-dependent competition for limited food, and a differential species response to sublethal concentrations may modify interspecific competition. We tested the hypothesis that exposure to malathion alters interspecific resource competition between these two species. In the absence of malathion, Ae. aegypti survivorship and per capita rate of population change were negatively affected by increasing densities of Ae. albopictus. However, the asymmetrical negative effect ofAe. albopictus on Ae. aegypti was eliminated in the presence of malathion. In addition, the presence of malathion resulted in shorter development time compared with the controls. The relative importance of pesticide-mediated coexistence in nature has not been evaluated, so its role in mediating coexistence is unclear; however, these findings underscore the potential of environmental concentrations of malathion, and perhaps other pesticides to facilitate coexistence between species.

West Nile virus infection rates and avian serology in east-central Illinois.

Understanding the geographic role of different species of mosquito vectors and vertebrate hosts in West Nile virus (WNV) transmission cycles can facilitate the development and implementation of targeted surveillance and control measures. This study examined the relationship between WNV-antibody rates in birds and mosquito infection rates and bloodfeeding patterns in east-central Illinois. The earliest detection of WNV-RNA by reverse transcription-polymerase chain reaction TaqMan was from Culex restuans; however, amplification typically coincided with an increase in abundance of Cx. pipiens. Trap type influenced annual estimates of infection rates in Culex species, as well as estimation of blood meal source. Bird species with the highest WNV-antibody rates (i.e., Mourning Doves [Zenaida macroura], Northern Cardinals [Cardinalis cardinalis], American Robins [Turdus migratorius], and House Sparrows [Passer domesticus]) were also the common species found in Culex blood meals. Although antibody rates were not directly proportional to estimated avian abundance, the apparent availability of mammal species did influence proportion of mammal to bird blood meals. Antibody prevalence in the American Robin was lower than expected based on the strong attraction of Culex to American Robins for blood meals. Age-related differences in serology were evident, antibody rates increased in older groups of robins and sparrows, whereas 1st-year hatch and older adults of Mourning Doves and Northern Cardinals had equally high rates of antibody-positive serum samples. The vector and host interactions observed in east-central Illinois (Champaign County), an urban area surrounded by agriculture, are compared to studies in the densely population areas of southern Cook County.

First record of Aedes japonicus japonicus in Illinois: defining its spatial distribution and associated mosquito species.

Since the initial discovery of Aedes japonicus japonicus in 1998 from sites in the northeastern United States (New York, Connecticut, and New Jersey), this invasive mosquito species now permeates local faunas throughout eastern North America and continues to expand its distribution westward. In July and August of 2006, the 1st reported specimens of Ae. japonicus were detected in east central Illinois at 2 woodlots in southern Urbana. Sampling efforts were concentrated around the site of the original detection. Dry ice-baited light traps and gravid traps caught 26,421 mosquitoes, with 5 and 6 Ae. japonicus recovered from each trap type, respectively. Although numerous larvae were found in 1 woodlot, they were restricted to a single container, a child's plastic swimming pool. Routine weekly mosquito sampling for West Nile virus (WNV) surveillance in Urbana and the adjacent city of Champaign (gravid and dry ice-baited traps placed in agricultural, industrial, residential, and urban parks) failed to detect Ae. japonicus outside of the 2 woodlots. Despite previous reports of Ae. japonicus from the adjacent states of Indiana and Missouri in 2004 and 2005, the extremely localized spatial distribution and low abundance of this species strongly suggests a recent introduction. If Ae. japonicus follows previous trends in the United States and Canada, within 1-3 years after it becomes established, its abundance, frequency of detection, and spatial distribution will significantly expand. Aedes japonicus has some characteristics of a bridge vector for West Nile virus because of its vector competency, ability to feed on mammals and birds, and potential abundance near sites of known WNV transmission in Illinois.

Modeling the impact of variable climatic factors on the crossover of Culex restauns and Culex pipiens (Diptera: culicidae), vectors of West Nile virus in Illinois.

The aim of this study was to model the impact of temperature on the timing of the seasonal shift in relative proportion of Culex restuans Theobald and Culex pipiens L. in Illinois. The temporal pattern of West Nile virus (WNV) and St. Louis encephalitis virus transmission in the midwest exhibits a late summer to early fall peak in activity, which parallels the temporal increase in the abundance of Cx. pipiens. The daily number of egg rafts oviposited by each species has been monitored at multiple surveillance sites in Urbana-Champaign in central Illinois for more than 13 years. The time when the two Culex species are in equal abundance (crossover) varies considerably from year to year. Our investigation of several thermal measures indicated that this variation was related in large part to climatic conditions with warmer (cooler) temperatures correlated to earlier (later) crossover dates. Models based on degree days and the number of days in which the daily maximum temperature exceeded an upper temperature threshold explained more than 60% of the variance in crossover dates. In contrast, models based on the number of days in which the daily minimum temperature exceeded a lower temperature threshold explained no more than 52% of the variance. An evaluation of these models demonstrated that they provide relatively simple and accurate estimates of crossover date from daily temperature data, a necessary component for developing an overall climatic index for the risk of WNV transmission in Illinois.

Changes in seroprevalence of West Nile virus across Illinois in free-ranging birds from 2001 through 2004.

Of the 5,236 birds sampled for antibodies to West Nile virus (WNV) in Illinois from 2001 through 2004, 348 (6.6%) birds were seropositive. Our multiple year surveillance identified several avian species that had particularly high percentages of seropositive individuals. The importance of these species in the enzootic and/or epizootic transmission of WNV is discussed relative to their regional abundance and literature on host competency. The species with the highest exposure rates to WNV differed both temporally and regionally. In general, birds that bred or were born in Illinois were more likely to have antibodies than transient birds. There was also a significant difference in the seroprevalence between adults (12.1%) and juveniles (5.5%), indicating that the acquired antibody response from previous years is a critical concern when interpreting seroprevalence rates in wild-caught birds. The most common hosts for St. Louis encephalitis virus were also the most common hosts for WNV, which strongly supports the role of similar vectors for both flaviviruses. Avian species with high WNV seroprevalence rates tended to be those that bred throughout the year, have open cup nests, and live in close proximity to humans.

A comparison of two West Nile virus detection assays (TaqMan reverse transcriptase polymerase chain reaction and VecTest antigen assay) during three consecutive outbreaks in northern Illinois.

Mosquitoes identified as female Culex (Culex) species, primarily mixtures or uniform batches of Culex pipiens and Culex restuans, were collected daily from gravid traps by 2 mosquito abatement districts (MADs) in Cook County, Illinois. From 2002 through 2004, batches (pools) of mosquitoes were tested by the MADs for West Nile virus (WNV) by using VecTest WNV antigen assays and the same samples were retested, usually within 1-2 wk, for WNV RNA by the TaqMan reverse transcriptase polymerase chain reaction (RT-PCR). There were 952 TaqMan-positive pools out of 3,953 pools over the 3 years, and about one half of that number were VecTest-positive. The difference between the 2 detection assays varied between and within years. The VecTest assays detected about 57% and 69% of the TaqMan RT-PCR-positive pools from Des Plaines Valley MAD and Northwest MAD in 2002, but only about 40% and 46% in 2003, and 36% and 55% in 2004, respectively. Based on a subset of the 2004 data, a linear relationship was found between VecTest detection of WNV and TaqMan cycle threshold between 18 and 28 cycles. A temporal decrease in the difference between the 2 assays was observed in 2003 and 2004, which we conjecture is due, at least partially, to a seasonal decline in the proportion of recently infected mosquitoes. This trend was not observed in 2002 because infection rates indicated a high likelihood of more than 1 infected mosquito per pool at the peak of transmission. Unlike a previous study, the 95% confidence intervals of infection rates based on the 2 detection methods did not always overlap. The highest infection rates occurred in 2002 when mean monthly temperatures were above average.

Evaluation of selected modifications to CO2 and infusion-baited mosquito traps in Urbana, Illinois.

The efficacy of granular media CO2 sachets used in conjunction with Center for Disease Control and Prevention (CDC) miniature light traps with light bulbs removed and modifications of bucket size and color to infusion-baited gravid traps was assessed from June through August of 2004 in Urbana, IL. The sachet packets caught significantly more mosquitoes and mosquito species compared with traps containing no CO2 bait; however, they also caught significantly fewer mosquitoes and fewer mosquito species compared with traps baited with dry ice. Pairing CO2-generating sachets with various dosages of octenol did not significantly increase the numbers of mosquitoes or mosquito species compared with traps baited only with sachets. When comparing modifications in the bucket size of gravid traps, the larger bucket (1,645-cm2 surface area holding about 5.3 liters of infusion) caught significantly more mosquitoes compared with the smaller bucket (759-cm2 surface area holding about 3.0 liters of infusion). Bucket color (green, blue, or black) did not affect the total number of mosquitoes caught.

Land use patterns and the risk of West Nile virus transmission in central Illinois.

Understanding how human land use patterns influence mosquito ecology and the risk of mosquito-borne pathogens is critical for the development of disease management strategies. We examined how different environments influenced mosquito species composition, abundance, and West Nile virus (WNV) infection rates in central Illinois. Using a combination of gravid traps and CDC light traps, adult mosquitoes were collected every other week from June 24 to September 16, 2012, in four major land use categories-row crops, prairies, forest fragments, and residential neighborhoods. The mosquitoes were identified to species morphologically, and pools of pure and mixed Culex mosquitoes (primarily Culex pipiens and Culex restuans) were tested for WNV-RNA by qRT-PCR. Mosquito species diversity was significantly higher in forest habitats compared to residential, agricultural, and prairie land use categories. All the four landscape types were equally important habitats for WNV vectors Cx. pipiens and Cx. restuans, contrary to previous findings that these species principally inhabit the residential areas. WNV-infected mosquito pools were observed in all land use types, and the infection rates overlapped among land use categories. Although our findings support the importance of residential habitats for WNV transmission to humans, they also establish that prairie, row crops, and wood lots are potentially important refuges for enzootic transmission. This is particularly important in urban ecosystems where these land use categories are small, interspersed fragments serving as potential refuge sites during periods of low rainfall.

Implications of spatial patterns of roosting and movements of American robins for West Nile virus transmission.

The arrival of West Nile virus (WNV) in North America has led to interest in the interaction between birds, the amplification hosts of WNV, and Culex mosquitoes, the primary WNV vectors. American robins (Turdus migratorius) are particularly important amplification hosts of WNV, and because the vector Culex mosquitoes are primarily nocturnal and feed on roosting birds, robin communal roosting behavior may play an important role in the transmission ecology of WNV. Using data from 43 radio-tracked individuals, we determined spatial and temporal patterns of robin roosting behavior, and how these patterns related to the distribution of WNV-infected mosquitoes. Use of the communal roost and fidelity to foraging areas was highly variable both within and among individual robins, and differed markedly from patterns documented in a previous study of robin roosting. Although there were clear seasonal patterns to both robin roosting and WNV occurrence, there was no significant relationship between communal roosting by robins and temporal or spatial patterns of WNV-positive mosquitoes. Our results suggest that, although robins may be important as WNV hosts, communal roosts are not necessarily important for WNV amplification. Other factors, including the availability and distribution of high-quality mosquito habitat and favorable weather for mosquito reproduction, may influence the importance of robin roosts for local WNV amplification and transmission.

Evaluation of a rapid analyte measurement platform for West Nile virus detection based on United States mosquito control programs.

The rapid analyte measurement platform (RAMP) system is an immunoassay test for West Nile virus (WNV) detection. Although reverse transcriptase-polymerase chain reaction (RT-PCR) methodology has been regarded as the gold standard for confirming WNV presence, usage of RAMP testing kits has increased in the past years. We collected RAMP test result data that were subsequently confirmed with RT-PCR methodology from mosquito control agencies to evaluate the efficacy of the RAMP testing. Results indicate that there are a high number of false positives (RAMP positive, RT-PCR negative) with RAMP testing. Correlation between RAMP unit values and RT-PCR cycle threshold values were varied depending on the primer/probe being compared. Comparison of RT-PCR results (on the same samples) between laboratories also indicates variation among the procedures and their potential to influence the RAMP testing efficacies. We discuss the potential issues and solutions that could prevent the high rate of false positives.