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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.

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).

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 of Ae. 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 env...

4 ABSTRACT. 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.

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 posi- tives (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 influ- ence the RAMP testing efficacies. We discuss the potential issues and solutions that could prevent the high rate of false positives.

In the midwestern United States, the summertime rise in infection rate by the West Nile virus is associated with a seasonal shift in the abundance of two mosquito populations, Culex restuans and Culex pipiens. This seasonal shift usually precedes the time of the peak infection rate in mosquitoes by 2–3 weeks and generally occurs earlier in the summer with above normal temperatures and later in the summer with below-normal temperatures. Two empirical models were developed to predict this seasonal shift in mosquito species, or the “crossover,” and have been run operationally since 2004 by the Midwestern Regional Climate Center located at the Illinois State Water Survey. These models are based on daily temperature data and have been verified by use of a unique dataset of daily records of mosquito species abundance collected by the Illinois Natural History Survey. An unfortunate characteristic of the original temperature models was that the crossover date often was reached with little or no lead time. In 2009, the models were modified to incorporate National Weather Service (NWS) model output statistics (MOS) 10-day temperature forecasts. This paper evaluates the effectiveness of these models to predict the crossover date and thus the period of increased risk of West Nile virus in the Midwest. For the 8-yr period from 2002 to 2009, 6 yr had at least one model predicting the crossover within one week of the actual crossover date, and for 7 yr at least one of the model predictions was within 2 weeks of the actual crossover date. Incorporation of MOS temperature forecasts for a 10-day period, although not substantially changing the predicted crossover date, greatly improved the forecast lead time by about 9 days. From a disease management point of view, this improvement in advanced notice is significant. In 2009, there was an unprecedented early crossover date and a failed forecast. The poor forecast was likely caused by an unusually early summer prolonged and intense heat wave, followed immediately by a record cold July.

The chronic effects of exposure of Culex restuans (Theobald) and Aedes albopictus (Skuse) (Diptera: Culicidae) to low concentrations of malathion were examined by exposing larvae of the two species to four malathion doses at 20, 25, and 30 degrees C and maintaining the resulting adults at 25 degrees C. For both species, a significant temperature by malathion interaction on survival was found. Greater temperatures at the highest malathion dosage had significantly lower survivorship than in other treatments, but this effect was not observed at 20 degrees C. These results suggest that low temperature may buffer against some of the negative effects of malathion. For both species, temperature but not malathion had significant effects on female developmental time to adulthood and adult longevity. Temperature also affected adult female size for Ae. albopictus but not Cx. restuans. Ae. albopictus females developed faster as temperature increased, lived longer when larvae were maintained at 30 degrees C than at 20 degrees C, and were larger when larvae were maintained at 25 degrees C than at 20 or 30 degrees C. Cx. restuans females developed faster at 25 and 30 degrees C than at 20 degrees C and lived longer at 25 degrees C than at 20 or 30 degrees C. The estimated finite rate of increase (lambda') for Cx. restuans was significantly lower at 20 degrees C than at 25 and 30 degrees C, whereas that of Ae. albopictus was significantly influenced by an interaction between temperature and malathion with significantly lower lambda' at 20 degrees C than at 25 and 30 degrees C for all malathion treatments except 0.014 mg liter(-1). Understanding how pesticides interact with abiotic environmental conditions will contribute to management decisions about vector control practices.

One of the unique characteristics of West Nile virus (WNV) in North America is the large number of bird species for which the virus can be fatal. WNV mortality has been documented through experimental infections of captive birds and necropsies of free-ranging birds. Investigations of WNV-related mortality in wild birds often focus on species with dramatic population declines (e.g., American Crow, Corvus brachyrhynchos); however, few studies have addressed WNV-related mortality in species not exhibiting marked population declines since the arrival of WNV. We conducted a mark-recapture study of 204 Northern Cardinals (Cardinalis cardinalis) in an area with endemic WNV activity to estimate WNV-related mortality. Previous research has shown that once a bird is infected and recovers from WNV it develops antibodies making it resistant to future infection. Assuming that mortality risks from non-WNV causes were the same for individuals with (had been exposed to WNV) and without antibodies (had not been exposed to WNV), we compared the survival rates of birds with and without WNV antibodies to estimate the impact of WNV on wild birds. An information theoretic approach was used, and the apparent survival was found to be 34.6% lower for individuals without antibodies during the period when WNV was most active (July-September). However, the apparent survival rate was 9.0% higher for individuals without antibodies over the rest of the year. These differences in apparent survival suggest that WNV increases mortality during the WNV season and that chronic effects of WNV infection may also be contributing to mortality. Although WNV appears to have increased mortality rates within the population, population trend data do not indicate declines, suggesting that some cardinal populations can compensate for WNV-related mortality.

We assessed the structure and latitudinal selection that might result in sensitivities to critical day-lengths that trigger diapause between Culex pipiens populations distributed along North-South and East-West axes in eastern North America. Strong population structure between Cx. p. pipiens and Cx. p. quinquefasciatus existed. Among Cx. p. pipiens, a 100-km increase in the latitudinal change resulted in an increased square root of F(ST) by 0.002. A 100-km increase in the longitudinal change caused an increased square root of F(ST) by 0.035. A lack of latitudinal influence on the structure between Cx. p. pipiens populations suggests a uniform signal using the 12 microsatellite markers, which might increase the risk of West Nile virus (WNV) transmission toward northern areas because of longer breeding season, extend host-seeking period, and larger population size. Northern Cx. p. pipiens may have undergone additional generations before diapause is triggered, magnifying population size when WNV amplification is peaking.

The identification of the members of the Culex pipiens L. complex in arbovirus surveillance programs relies heavily on the use of morphology. In this work, we studied Cx. pipiens complex male mosquitoes collected from nine different locations, from northern, southern, and the hybrid zone sites in North America; Cairo, Egypt; and Nairobi, Kenya. Specimens were identified using DV/D ratio and also using amplification of the acetylcholinesterase (ACE.2) gene by both conventional and real-time polymerase chain reaction, and examination of the CQ11 locus. Consistent with previous findings, the morphological and molecular identifications did not always agree, particularly in regions of extensive introgression. There was an increased frequency of hybrid forms in late summer and early fall in Champaign Co., IL, that is north of the previously described Cx. pipiens complex hybrid zone. This represents an expansion of the North American hybrid zone. The biological and epidemiological relevance of the high degree of introgression and the late season increase in the proportion of intermediate forms is discussed.

In North America, West Nile and St. Louis encephalitis viruses have been detected in a wide range of vector species, but the majority of isolations continue to be from pools of mixed mosquitoes in the Culex subgenus Culex. Unfortunately, the morphologic identification of these important disease vectors is often difficult, particularly in regions of sympatry. We developed a sensitive real-time TaqMan polymerase chain reaction assay that allows reliable identification of Culex mosquitoes including Culex pipiens pipiens, Cx. p. quinquefasciatus, Cx. restuans, Cx. salinarius, Cx. nigripalpus, and Cx. tarsalis. Primers and fluorogenic probes specific to each species were designed based on sequences of the acetylcholinesterase gene (Ace2). Both immature and adult mosquitoes were successfully identified as individuals and as mixed species pools. This identification technique provides the basis for a rapid, sensitive, and high-throughput method for expounding the species-specific contribution of vectors to various phases of arbovirus transmission.

The potential role of many urban passerine birds in the transmission of West Nile virus (WNV) is well-documented by studies on host competency, seroprevalence in wild birds, and identification of vector blood meal source. In contrast, the impact of bird behavior on transmission dynamics is largely unexplored. Bird roosting (perching) behavior may be a critical component regulating WNV transmission because of the crepuscular/nocturnal feeding behavior of Culex mosquitoes, the primary vectors of WNV. We used radio telemetry to determine the roosting behavior of American crows (Corvus brachyrhynchos) and northern cardinals (Cardinalus cardinalus). On average, healthy crows moved slightly shorter distances between roosts than viremic crows, 1,038.3 meters versus 1,255.5 meters, while cardinals only moved 54.7 meters. Given the average movements of crows and cardinals between roosts, crows, which are viremic for five days, could spread the virus throughout a mean ± SE area of 20.84 ± 0.79 km 2 , while viremic cardinals would, on average, only spread the virus over a mean ± SE area of 0.03 ± 0.01 km 2 . Because the crow population in Illinois is decreasing at a rate of 11.5% per year and up to 35.6% per year in certain locations, crows are becoming scarce in some areas, thus reducing their role as wild bird sentinels. We suggest that if crows are important in dispersing WNV, large decreases in their abundance will shift transmission cycles to a more focal nature because of the differences in roosting behavior of crows compared with other urban birds, such as cardinals.

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.

After a severe outbreak of West Nile virus (WNV) in Cook County, Illinois, in 2002, detections of WNV in mosquitoes were frequent across the state in the following years despite small numbers of human cases. We conducted a spatio-temporal analysis of Culex (subgenus Culex) mosquitoes collected in 2004 in three mosquito abatement districts (MAD) in Cook County by calculating monthly estimates of mosquito density, prevalence of infected mosquitoes, and exposure intensity, which in turn is a product of mosquito density and infection rates. Mosquito infections were detected early at three sites in late May and were widely detected throughout the three MADs in the summer with infection rates as high as 13 per 1000 in August. Exposure intensities were higher at sites adjacent to the Des Plaines River, especially in August and September. The aggregated pattern of WNV transmission along the river might be related to the existence of substantial forest preserves and wetlands that might produce ecological conditions favorable for mosquito proliferation and interactions between mosquitoes and birds.

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.

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.

In 2002, the world’s largest outbreak of neuroinvasive West Nile virus (WNV) disease occurred. Illinois reported 21% of the total cases in the United States, the most among all states. The epidemiology of WNV in Illinois in 2002 was examined to determine factors associated with severe disease and death. A total of 884 cases were identified and there were 66 deaths. The overall attack rate of WNV infection was 7.1 per 100,000 population and this increased with age. The median ages of patients and patients who died were 56 and 78 years, respectively. Among patients who died, 91% were diagnosed with encephalitis and the case-fatality rate for patients with encephalitis was 18.6%. Patients more than 50 years old had a significantly higher risk of being reported with encephalitis (relative risk [RR] = 3.3, 95% confidence interval [CI] = 2.6–4.3%) and death (RR = 22.3, 95% CI = 5.5–90.4%). Clinicians evaluating elderly patients with WNV infection should assess patients closely for progression of disease.

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.

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).

As part of an on-going program to identify natural products from plants with mosquitocidal activity, two acetylenic compounds were isolated from Cryptotaenia canadensis, a native North American umbellifer frequently encountered in moist woodlands. Fresh foliage, roots, and fruits were extracted with a hot methanol and water mixture and then dried. The extract was partitioned into chloroform and water, and both phases were bioassayed against fourth instars of Culex pipiens at concentrations between 5 and 50 ppm. Only the organic phase was active. Gas–liquid chromatography revealed that the two main components of the organic phase were polyacetylenes. The compounds were isolated by vacuum chromatography on silica gel and identified as falcarinol and falcarindiol based on GC-MS-EI and GC-MS-CI fragmentation patterns, high-resolution MS, and 1H and 13C NMR spectrometric data. The dose–response curves with mosquito larvae were determined by probit analysis. The LC50 values were 3.5 ppm for falcarinol and 6.5 ppm for falcarindiol. The distribution of polyacetylenes varied among plant parts. Fruits contained an unknown compound not found in either the foliage or roots.

The olfactory response to volatile semiochemicals was determined for Diabrotica undecimpunctata howardi Barber, the spotted cucumber beetle, Diabrotica virgifera virgifera Le- Conte, the western corn rootworm, Diabrotica barberi Smith & Lawrence, the northern corn rootworm, and Diabrotica cristata (Harris), a nonpest species in a tallgrass prairie ecosystem and adjacent corn and cucurbit agroecosystems in Illinois. The results indicate that the divergence in response to plant volatiles by these Diabrotica species can be correlated with currently accepted phylogenetic groupings between the virgifera and Fucata groups, as well as within the virgifera group. For example, cinnamaldehyde is most attractive to D. u. howardi (fucata group), whereas 4-methoxycinnanlaldehyde is a specific attractant for D. v. virgiferal (virgifera group). The 2 closely related species D. barberi and D. cristata (virgifera group) were both attracted to eugenol, cinnamyl alcohol, and 4-methoxyphenethanol on one or more test dates. Although D. cristata is not normally found in cucurbit blossoms, adults were attracted to traps containing shredded blossoms of Cucurbita maxima Duchesne ex Poir. Furthermore, all 4 Diabrotica species responded to a multicomponent synthetic lure (a cucurbit blossom mimic), suggesting a commonality of response to cucurbit blossom aroma. A review of the literature on Diabroticite chemical ecology suggests cucurbit volatiles acted evolutionarily as synomones, providing a primitive means of pollination for cucurbits. For several Dillbrotica species, the olfactory response to these volatiles also may have facilitated the finding of noncucurbitaceous pollen sources.

Few studies have documented the developmental profile of both primary and secondary metabolites of plants; we set out to construct such a profile, in order to determine the extent to which primary and secondary metabolites covary. Primary and secondary chemistry have been documented for buds, female flowers and green fruits of wild parsnip Pastinaca sativa. A distinct qualitative shift in secondary chemistry occurs over the course of development. Mono- and sesquiterpenes were abundant in buds but absent from female flowers and green fruits. Furanocoumarins, which were found in all organs, were present at low concentration in buds, at intermediate concentrations in female flowers, and at highest concentrations in fruits. Among the primary metabolites, developmental shifts were, for the most part, quantitative. Soluble protein and fatty acid content declined with development. A qualitative change in fatty acid composition was observed in that linolenic acid inbuds was replaced by petroselinic acid in fruits. Overall variation in primary metabolites was rarely correlated with variation in secondary metabolites: of the 233 possible correlations, only 25 were significant.

The Report “Emerging vectors in the Culex pipiens complex” by D. M. Fonseca et al. (5 Mar., p. [1535][1]) advances sweeping extrapolations concerning mosquito blood-feeding behavior and West Nile virus (WNV) transmission. These findings led Fonseca to conclude ([1][2]) that a European

Olfactory synergism, where combinations of plant volatile kairomones are quantitatively more attractive to insects than the sum of attraction of the individual components, is an important but little-studied phenomenon in host plant selection and feeding and in pollination ecology. Diabroticite beetles (Coleoptera: Chrysomelidae) are strongly attracted toCucurbita blossoms, and 2- to 3-fold olfactory synergism has been demonstrated in four species by combinations of the key blossom volatiles, 1,2,4-trimethoxybenzene, indole, and (E)-cinnamaldehyde. This TIC mixture represents an optimizedCurcurbita blossom volatile kairomone mixture useful in monitoring Diabroticite populations and in studying their behavior and ecology. Indole, which exhibits a spectrum of attraction to these beetles ranging from moderate forDiabrotica virgifera virgifera andAcalymma vittatum to very weak forD. barberi, is the primary synergistic component. Indole combined with 4-methoxycinnamaldehyde was significantly synergistic toD. v. virgifera at a ratio of 1:300 and produced 4-fold synergism at a ratio of 1:1. Indole combined with 4-methoxyphenethanol was less synergistic toD. barberi with 1.5- to 2-fold synergism at a 1:1 ratio. These consistent variations in diabroticite beetle olfactory responses presumably indicate evolutionary divergences in the numbers of relict indole antennal receptors.

Mosquito larvae face numerous biotic and abiotic challenges that affect their development and survivorship, as well as adult fitness. We conducted two experiments under semi-natural conditions to evaluate the effects of intraspecific competition, nutrient limitation and sub-lethal doses of malathion on individual life history traits in adult Culex pipiens (Diptera: Culicidae). In the first experiment, larvae of Cx. pipiens were reared at different intraspecific densities and exposed to sub-lethal doses of malathion. In the second experiment, different intraspecific densities of Cx. pipiens larvae were reared under conditions of low or high larval nutrients, and subsequent adults were fed on either water or 10% sucrose solution. Malathion treatment had relatively minor effects compared with density, which had significant negative effects on development rate, survivorship to adulthood, body size (wing length) and longevity. As larval density increased, a sex ratio distortion in survivorship to adulthood emerged, in which a bias towards males was apparent. Nutrient-rich larval environments alleviated, in part, the effects of increasing density and extended the lifespan of mosquitoes fed on water and 10% sucrose. Density-dependent alterations in adult longevity attributable to the larval environment are complex and show contrasting results depending on interactions with other environmental factors. This study suggests that larval resource availability and competition influence Cx. pipiens population growth correlates and have lasting effects on traits that relate to a mosquito's ability to vector pathogens.

The introduction of Aedes albopictus (Skuse) in the United States has been associated with declines in abundance of resident mosquito species, presumably because of resource competition, as larvae of Ae. albopictus have been illustrated as superior competitors under certain resource conditions. We evaluated the hypothesis that varying the type and ratio of two food resources (Foxtail grass: American elm) alters the competitive outcome of Ae. albopictus and Culex pipiens (L.). We measured survivorship, development time, size, and adult longevity, and estimated the population growth index (A') of populations raised both alone and in equal number with the interspecific competitor, across five ratios of the two food resources. Competition was asymmetric with Ae. albopictus, the superior competitor across all resource treatments; however, the competitive advantage Ae. albopictus had over Cx. pipiens was reduced as grass became the predominant resource. With elm as the predominant resource, the population growth index (A') for both Ae. albopictus and Cx. pipiens was lower in intraspecific and interspecific competition treatments, respectively. The treatments also impacted adult life history, as life spans of both Ae. albopictus and Cx. pipiens varied when they emerged from larval conditions with different resource and competition treatments. We discuss the possible differences in the two species' efficiencies in exploiting the two resource types. Despite some resource conditions alleviating the competitive effects of Ae. albopictus on Cx. pipiens, competition remained asymmetric; thus, additional mechanisms are likely operating under field conditions when the two species coexist.

Mosquito larval development occurs in aquatic habitats that are directly or indirectly exposed to chemical contaminants. Little is known about how interaction of these chemicals with other biotic and abiotic stressors impact mosquito populations. We used two levels of nutrient (low and high) and four larval densities (10, 20, 30, 40) to examine the effects of low concentrations of insecticide malathion, on Culex pipiens L. mosquitoes experiencing stress from larval competition. Addition of malathion at the high nutrient condition enhanced survival with increasing larval densities, but this effect was not observed at low nutrient condition. Males exposed to malathion were significantly larger than those from control treatments while the effect of malathion on size of females varied with larval density and the level of nutrients. Larval exposure to malathion and low nutrient resulted in significantly larger females with increases in larval densities compared with other treatments. The effect of malathion on male longevity varied with larval density and amount of nutrients. At higher densities, male longevity was consistently higher in low nutrient than in high nutrient conditions and addition of malathion in high nutrient treatment increased male longevity at the highest density. These effects are most likely attributable to release from competition among survivors after mortality from malathion and density-dependent effects. We conclude that biotic conditions of the larval habitat can alter the impact of low concentrations of chemical contaminants on mosquito populations in ways that may influence the pattern of disease transmission and the outcome of vector control efforts.

The relationship between wing length and body weight of mosquitoes was examined by comparing the fit of regressions using logarithmic and cubic transformations. A two-parameter line using a double logarithmic transformation provided the best fit, and we found no evidence to justify the use of wing length cubed as a transformation. Wing length was not directly proportional to body weight, but rather increased at a lesser rate than did body weight. Equations that predict body weight from wing length may not be an appropriate substitute for weighing mosquitoes, when studying the extremes of size in populations.

In the absence of contact and visual stimuli, significant numbers of adult striped cucumber beetles, Acalymma vittatum (F.), were caught on veiled sticky cups containing either squash seedlings or blossoms. The role of volatile kairomones in host finding was further demonstrated by the attraction of A. vittatum adults to a synthetic odor mimic of cucurbit blossoms (a mixture of 1,2,4-trimethoxybenzene, indole, and trans -cinnamaldehyde [TIC]. From July to late August 1988, TIC-baited traps caught 4–100 times more adult beetles than controls. Although indole was the most active single component of TIC, the slope of the log-dose response curve for the TIC-baited traps was 7 times greater than the slope for the indole-baited traps. Traps baited with the three-component mixture caught twice as many beetles as the theoretical additive response based on the trap catches of the individual components. Several phenylpropanoid attractants for Diabrotica species and other squash blossom odor components also were bioassayed in field tests, but none was as active as TIC or indole.

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.

Marked spatiotemporal variabilities in mosquito infection of arboviruses, exemplified by the transmission of West Nile virus (WNV) in America, require adaptive strategies for mosquito sampling, pool screening and data analyses. Currently there is a lack of reliable and consistent measures of risk exposure, which may compromise comparison of surveillance data. Based on quantitative reasoning, we critically examined fundamental issues regarding mosquito sampling design and estimation of transmission intensity. Two surveillance strategies were proposed, each with a distinct focus, i.e. targeted surveillance for detection of low rates of mosquito infection and extensive surveillance for evaluation of risk exposure with high levels of mosquito infection. We strongly recommend the use of indicators embodying both mosquito abundance and infection rates as measures of risk exposure. Aggregation of surveillance data over long periods of time and across broad areas obscures patterns of focal arboviral transmission. We believe that these quantitative issues, once addressed by mosquito surveillance programs, can improve the epidemiological intelligence of arbovirus transmission.

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.

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.

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.

Pool testing of vector samples for arboviruses is widely used in surveillance programmes. The proportion of infected mosquitoes (Diptera: Culicidae) is often estimated from the minimum infection rate (MIR), based on the assumption of only one infected mosquito per positive pool. This assumption becomes problematic when pool size is large and/or infection rate is high. By relaxing this constraint, maximum likelihood estimation (MLE) is more useful for a wide range of infection levels that may be encountered in the field. We demonstrate the difference between these two estimation approaches using West Nile virus (WNV) surveillance data from vectors collected by gravid traps in Chicago during 2002. MLE of infection rates of Culex mosquitoes was as high as 60 per 1000 at the peak of transmission in August, whereas MIR was less than 30 per 1000. More importantly, we demonstrate roles of various pooling strategies for better estimation of infection rates based on simulation studies with hypothetical mosquito samples of 18 pools. Variable size pooling (with a serial pool sizes of 5, 10, 20, 30, 40 and 50 individuals) performed consistently better than a constant size pooling of 50 individuals. We conclude that variable pool size coupled with MLE is critical for accurate estimates of mosquito infection rates in WNV epidemic seasons.

Publisher Summary This chapter discusses synthetic insecticides. It reviews public health pesticides, covering topics such as common terms and concepts in medical entomology to introduce the diverse relationships among humans, arthropods, and disease pathogens; the historical and current impact of arthropods on human health; the basic concepts of vector management; a brief list of noninsecticidal alternatives; the classification schemes and general properties of public health pesticides and repellents; and specific examples of pesticides in vector management, particularly mosquito control. The spectacular success of synthetic insecticides in the decade immediately following World War II generated widespread enthusiasm that the major scourges of humankind, such as yellow fever, malaria, and typhus, could be conquered. However, these short-term successes resulted in major increases in disease transmission primarily due to resistance in both the arthropod to insecticides and to drugs by the pathogen. The result of these failures has forced the public health community to reevaluate strategies and has resulted in the development of evidence-based management or integrated disease management. Employing this management strategy, both insecticides and drugs are used in a targeted manner that results in a significant increase in efficacy, environmental safety, and cost-effectiveness.

Determining the role of various mosquito species in the transmission of arboviruses relies on an accurate estimate of the proportion of infected individuals in an area. Basically, there are two methods for estimating the proportion of infected individuals from pooled samples. One is the widely used minimum infection rate (MIR), which is calculated as the ratio of the number of positive pools to the total number of mosquitoes tested. The underlying assumption of the MIR is that only one infected individual exists in a positive pool. This assumption is valid for arboviruses that are relatively rare, but it becomes problematic when infection rates are high and/or pool sizes are large. The other method of estimating the proportion of infected individuals in pooled samples is the maximum likelihood estimation (MLE), which is defined as the infection rate most likely observed given the testing results and an assumed probabilistic model (i.e., binomial distribution of infected individuals in a positive …

The diabroticite rootworm beetles coevolved with plants of the family Cucurbitaceae as demonstrated by their feeding dependence on the tetracyclic triterpenoid cucurbitacins. These beetles also exhibit strong attraction to phenylpropanoid volatile components of Cucurbita blossoms. A mixture of 1,2,4-trimethoxybenzene, indole, and (E)-cinnamaldehyde, all blossom components, is highly attractive to the several species of diabroticite cucumber beetles and corn rootworms and is considered a simplified Cucurbita blossom kairomone odor. The evolutionary divergence in antennal receptor complementarity is best understood by comparing the species-specific responses of several Diabrotica to structural analogues of (E)-cinnamaldehyde, the major attractant for Diabrotica undecimpunctata howardi. Cinnamyl alcohol is a strong attractant for Diabrotica barberi, and 4-methoxycinnamaldehyde is an exceptional attractant for Diabrotica virgifera. The very closely related species D. barberi and Diabrotica cristata are most strongly attracted to 4-methoxyphenethanol, which is unattractive to the other species studied.

Hydrocarbons of worker honeybees of known pedigree were extracted and analyzed using gas chromatography and mass spectrometry. Variability in hydrocarbon extracts of individual workers is determined at least in part genetically. Correlations in hydrocarbon composition of extracts were highest among more closely related individuals. Individuals maintained in groups exchange hydrocarbons but still maintain enough self-produced compounds to retain genetically determined individual characteristics. These results demonstrate that extractable hydrocarbons of bees provide sufficiently reliable genetic information to function as labels for use during kin recognition.

Field tests in corn with baited sticky traps focused on changes in species specificity and magnitude of response for Diabrotica barberi Smith and Lawrence and D. virgifera virgifera LeConte when candidate lures varied by functional group, chain length, aryl substitents, saturated and unsaturated sidechains, and isosteric replacement. Cinnamyl alcohol, a major component of the blossom volatiles of Cucurbita maxima Duchesne, and its parakairomone (active synthetic analog), 3-phenyl-1-propanol, were significantly more attractive to D. barberi than eugenol in corn and were also attractive to D. barberi than eugenol in corn and were also attractive to D. cristata (Harris) in a reclaimed prairie. Neither compound was appreciably attractive to adult western corn rootwoorms D. v. virgifera and, as previously reported, cinnamyl alcohol is only slightly attractive to adult southern corn rootworms, D. undecimpunctata howardi Barber. The spectrum of rootworm responses to cucurbit blossom components and structurally related compounds amplifies the coevolutionary relationships between Diabroticites and Cucurbitaceae, shows the chemosensory specificity of odor-conditioned behavior in Diabrotica ssp., and illustrates the potential of parakairomones as practical insect lures

The attraction of Diabrotica spp. (Coleoptera: Chrysomelidae) to single-component and multicomponent lures was evaluated in corn fields by comparing the relative number of beetles caught on sticky traps over a 24-h period. Initial tests in 1985 showed that a mixture of veratrole (V), indole (I), phenylacetaldehyde (P), trans -anethole (A), and eugenol (E) (=VIPAE mixture) caught 26 times more Diabrotica undecimpunctata howardi Barber, southern corn rootworms (SCR), than untreated control traps and was at least 3 times more active than any of the traps baited with a single component. Significantly fewer D. virgifera virgifera LeConte, western corn rootworms (WCR), were caught on the traps baited with the VIPAE mixture than those baited with 100 mg of trans -anethole. Furthermore, only SCR adults exhibited a concentration-dependent response to the VIPAE mixture. SCR response to the sequential removal of the individual components from the VIPAE mixture suggested the primary attractants to be veratrole, indole, and phenylacetaldehyde. Additional tests showed traps baited with a three-component VIP mixture (veratrole, indole, and phenylacetaldehyde at 20 mg per component) caught ca. 8 times as many beetles as the expected mean additive response with the individual components; therefore, the SCR response was synergistic. The attraction and synergistic response of SCR adults to the VIP mixture was verified by similar tests conducted in 1986. The VIP mixture was also compared with a chemically related mixture consisting of 1,2,4-trimethoxybenzene (substituted for veratrole or ortho-dimethoxybenzene), indole, and trans -cinnamaldehyde (substituted for phenyla-cetaldehyde) (= TIC mixture) at dosages ranging from 1 to 30 mg per trap. Although both mixtures exhibited approximately the same activity for SCR adults, only the new TIC mixture attracted WCR adults. TIC baited traps caught ca. 6 times more WCR adults than did untreated controls at 1 mg per trap and ca. 29 times more WCR adults than controls at 30 mg per trap.

Field tests conducted in a relict prairie using sticky traps baited with a broad range of adult diabroticine attractants showed Diabrotica cristata (Harris) shares some chemosensory responses with both D. barberi Smith and Lawrence and D. virgifera virgifera LeConte. D. cristata was attracted to estragole, β-ionone, para -methoxycinnamaldehyde, and a mixture of trimethoxybenzene, indole, and cinnamaldehyde (=TIC mixture) (attractants for adults of D. virgifera virgifera in corn tests), as well as eugenol, isoeugenol, and cinnamyl alcohol (attractants for adults of D. barheri in corn tests). Several of the active compounds, such as cinnamyl alcohol and β-ionone, are structurally distinct from the previously described eugenol-type attractants for D. barberi and D. cristata and represent new semiochemicals for the genus. In the prairie trials, D. barberi adults were anomalously attracted to two lures (estragole and the TIC mixture) that were not active in corn evaluations.

Adults of two closely related corn rootworms, Diabrotiea barberi Smith and Lawrence and D. virgifera virgifera LeConte, are strongly attracted to two naturally occurring phenylpropanoids, eugenol and estragole respectively. The close chemical relationship of these two kairomones suggested that structure-activity studies of estragole analogues as rootworm attractants would be profitable. During the summers of 1986 and 1987, evaluations of 21 chemical compounds related to estragole revealed several exceptional attractants: 4-methoxycinnamaldehyde and 4-methoxycinnamonitrile for D. v. virgifera , cinnamyl alcohol for D. barberi , and cinnamaldehyde for the southern corn rootworm, D. undecimpunctata howardi Barber.

Dry ground baits impregnated with insecticide and containing cucurbitacins as arrestants and feeding stimulants are effective in controlling adult western corn rootworms, Diabrotica virgifera virgifera LeConte, and southern corn rootworms, Diabrotica undecimpunctata Howardi Barber. Bait effectiveness was proportional to the concentration of cucurbitacins, but differences between different bitter cucurbit sources were negligible. Carbamate insecticides, particularly methomyl, were generally judged superior for bait efficacy compared with organophosphorus and pyrethroid insecticides. Addition of eugenol to the dried cucurbitacin bait increased response of D. u. howardi adults to the bait by ca. 4-fold, but had no effect on response of D. v. virgifera. Initial studies suggest that substantial reductions in adult corn rootworm populations may be obtained with broadcast applications of baits containing ca. 0.1% cucurbitacins and 0.1% insecticide applied at 11–33 kg/ha in corn or cucurbits.

The environmental persistence of pesticides, their terminal residues and their contaminants, has played a major role in environmental regulations. To demonstrate degradation in field studies is difficult, both because of the length of time required and because of the complexity of demonstrating degradation of metabolites as well as of the pesticide itself. These difficulties can be circumvented by evaluating radiolabeled pesticides in a terrestrial aquatic laboratory model ecosystem which provides qualitative and quantitative data on persistence, bioaccumulation, and degradation both of the parent compound and of its metabolites. This microcosm was used to obtain data on the environmental degradation of pesticides with major roles in conservation tillage: paraquat, atrazine, linuron, oryzalin, alachlor, and paraquat paired with each of the other herbicides. The results suggest that alachlor, atrazine, linuron, and oryzalin do not pose serious risks of environmental accumulation when applied alone. The results of paired-herbicide eco-system evaluations compared to evaluations of each herbicide separately demonstrate that: paraquat is only minimally transported to aquatic organisms or to water; differences in herbicide application are more important in determining residue levels for these herbicides than are interactions between the herbicide and paraquat, although the latter do occur; and the addition of paraquat does not significantly alter the capacity of these herbicides for bioaccumulation or ecological magnification.

During the summers of 1984 and 1985, a variety of structurally related benzenoid compounds was evaluated in sweet corn plots as attractants for adult southern corn rootworms (SCR), western com rootworms (WCR), and northern corn rootworms (NCR). Field response to the volatiles was measured by beetle counts on baited cylindrical sticky traps placed inside the corn plots at a height of l m above ground level. SCR adults were attracted late in the season (last week of August through September, 1984 and 1985) to numerous aromatic compounds, including phenylacetaldehyde, benzyl acetone, phenethyl alcohol, phenyl acetate, indole, veratrole, methyl eugenol, methyl isoeugenol, eugenol, and isoeugenol. Although many compounds attracted SCR adults late in the season, only veratrole, phenylacetaldehyde, and chavicol were significantly active in early and middle August 1985. WCR adults were attracted to a different group of compounds, namely estragole,trans-anethole, and indole. Estragole (4-methoxy-1-allylbenzene) was an effective WCR attractant from corn tasseling in early August 1985, until the end of the trapping period in late September and early October 1985. Indole andtrans-anethole (4-methoxy-1-propenylbenzene) were less effective attractants than estragole and were most active at the beginning and/or end of the corn season. Traps baited with 100 mg of estragole caught an average of 20 times more WCR adults than unbaited control traps, and the females outnumbered the males in the baited traps. Estragole dosage tests were conducted in three sweet corn plots on different dates in 1985 and the minimum effective dose ranged between 5 and 30 mg/trap. Field tests with structural analogs revealed the importance of the site of unsaturation in the allylic side chain of estragole and the effect of different ring substituents on WCR response. The phenylpropanoids, eugenol and isoeugenol, significantly attracted NCR adults, even though these beetles were in low abundance in the test corn plots. Field tests indicate there is no cross-species response by WCR and NCR adults to their related phenylpropanoid attractants. However, in late August, SCR adults do respond to some WCR and NCR attractants (indole and several eugenol analogs). Electroantennographic analysis of SCR males revealed they can perceive peripherally a wide range of benzenoid compounds.

A synomone present in hexane extracts of corn silks was found to attractOrius insidiosus (Say). The attraction was a diurnal, innate behavior, independant of sex. A portion of the prey-searching behavior ofO. insidiosus apparently relies on olfactory perception when corn is the prey's host.