Exploring the immunogenicity of an insect-specific virus vectored Zika vaccine candidate.

Zika virus (ZIKV) is an important re-emerging flavivirus that presents a significant threat to human health worldwide. Despite its importance, no vaccines are approved for use in humans. Insect-specific flaviviruses (ISFVs) have recently garnered attention as an antigen presentation platform for vaccine development and diagnostic applications. Here, we further explore the safety, immunogenicity, and efficacy of a chimeric ISFV-Zika vaccine candidate, designated Aripo-Zika (ARPV/ZIKV). Our results show a near-linear relationship between increased dose and immunogenicity, with 1011 genome copies (i.e., 108 focus forming units) being the minimum dose required for protection from ZIKV-induced morbidity and mortality in mice. Including boosters did not significantly increase the short-term efficacy of ARPV/ZIKV-vaccinated mice. We also show that weanling mice derived from ARPV/ZIKV-vaccinated dams were completely protected from ZIKV-induced morbidity and mortality upon challenge, suggesting efficient transfer of maternally-derived protective antibodies. Finally, in vitro coinfection studies of ZIKV with Aripo virus (ARPV) and ARPV/ZIKV in African green monkey kidney cells (i.e., Vero-76) showed that ARPV and ARPV/ZIKV remain incapable of replication in vertebrate cells, despite the presence of active ZIKV replication. Altogether, our data continue to support ISFV-based vaccines, and specifically the ARPV backbone is a safe, immunogenic and effective vaccine strategy for flaviviruses.

Expression of anti-chikungunya single-domain antibodies in transgenic Aedes aegypti reduces vector competence for chikungunya virus and Mayaro virus.

Chikungunya virus (CHIKV) and Mayaro virus (MAYV) are closely related alphaviruses that cause acute febrile illness accompanied by an incapacitating polyarthralgia that can persist for years following initial infection. In conjunction with sporadic outbreaks throughout the sub-tropical regions of the Americas, increased global travel to CHIKV- and MAYV-endemic areas has resulted in imported cases of MAYV, as well as imported cases and autochthonous transmission of CHIKV, within the United States and Europe. With increasing prevalence of CHIKV worldwide and MAYV throughout the Americas within the last decade, a heavy focus has been placed on control and prevention programs. To date, the most effective means of controlling the spread of these viruses is through mosquito control programs. However, current programs have limitations in their effectiveness; therefore, novel approaches are necessary to control the spread of these crippling pathogens and lessen their disease burden. We have previously identified and characterized an anti-CHIKV single-domain antibody (sdAb) that potently neutralizes several alphaviruses including Ross River virus and Mayaro virus. Given the close antigenic relationship between MAYV and CHIKV, we formulated a single defense strategy to combat both emerging arboviruses: we generated transgenic Aedes aegypti mosquitoes that express two camelid-derived anti-CHIKV sdAbs. Following an infectious bloodmeal, we observed significant reduction in CHIKV and MAYV replication and transmission potential in sdAb-expressing transgenic compared to wild-type mosquitoes; thus, this strategy provides a novel approach to controlling and preventing outbreaks of these pathogens that reduce quality of life throughout the tropical regions of the world.

Novel murine models for studying Cache Valley virus pathogenesis and in utero transmission.

Cache Valley virus (CVV) is a prevalent emerging pathogen of significant importance to agricultural and human health in North America. Emergence in livestock can result in substantial agroeconomic losses resulting from the severe embryonic lethality associated with infection during pregnancy. Although CVV pathogenesis has been well described in ruminants, small animal models are still unavailable, which limits our ability to study its pathogenesis and perform preclinical testing of therapeutics. Herein, we explored CVV pathogenesis, tissue tropism, and disease outcomes in a variety of murine models, including immune -competent and -compromised animals. Our results show that development of CVV disease in mice is dependent on innate immune responses, and type I interferon signalling is essential for preventing infection in mice. IFN-αßR-/- mice infected with CVV present with significant disease and lethal infections, with minimal differences in age-dependent pathogenesis, suggesting this model is appropriate for pathogenesis-related, and short- and long-term therapeutic studies. We also developed a novel CVV in utero transmission model that showed high rates of transmission, spontaneous abortions, and congenital malformations during infection. CVV infection presents a wide tissue tropism, with significant amplification in liver, spleen, and placenta tissues. Immune-competent mice are generally resistant to infection, and only show disease in an age dependent manner. Given the high seropositivity rates in regions of North America, and the continuing geographic expansion of competent mosquito vectors, the risk of epidemic and epizootic emergence of CVV is high, and interventions are needed for this important pathogen.

Effects of Plant Substrate, Insecticide, and Blood Meal Status on the Efficacy of Barrier Treatments Against Aedes albopictus.

The effects of 3 plant species (Cornus florida [dogwood], rhododendron X 'Chionoides' [rhododendron], and Ilex opaca [American holly]), 4 insecticide treatments (Demand® CS [lambda-cyhalothrin] at 6.25 ml[AI]/liter; Talstar® Professional [bifenthrin] at 7.81 ml[AI]/liter, and Suspend® Polyzone® [deltamethrin] at 11.72 ml[AI]/liter, and water), and 2 physiological states (blood-fed and unfed) were evaluated for knockdown (1 h) and mortality (24 h) against female Aedes albopictus over an 8-wk sampling period. Analyses determined that there was a significant interaction between the tested plant species and the insecticides evaluated. Significant differences were likewise observed between the insecticide treatments for unfed Ae. albopictus females, with Demand CS demonstrating the highest knockdown and mortality rates (from >90% to >10% at wk 8 and >95% to ∼50% at wk 8, respectively), followed by Talstar Professional (from >75% to <10% at wk 2 and >90% to <10% at wk 2, respectively) and Suspend Polyzone (from >20% to <10% at wk 8 and >25% to >50% at wk 8, respectively). All treatments were no longer significant for knockdown or mortality at the end of the 8-wk timeframe. Significant differences were also observed between insecticide treatments for blood-fed Ae. albopictus females; Demand CS showed high knockdown and mortality rates (from 100% to ∼50% at wk 8 and 100% to >60% at wk 8, respectively), Suspend Polyzone rates were similar to Demand CS (from >80% to ∼50% at wk 8 and ∼90% to >65% at wk 8, respectively), and both were followed by Talstar Professional (from 100% to <10% at wk 4 and 100% to <20% at wk 4, respectively). All tested pyrethroid sprays showed a significant increase in effectiveness against recently blood-fed Ae. albopictus females, as compared to the unfed females. These results suggest that Demand CS can be used as an effective barrier spray against Ae. albopictus adults due to the limited impact of target foliage, its long-term efficacy under environmental conditions, and its continued effectiveness regardless of the blood meal status of the target mosquito.

Vector competence of Virginia mosquitoes for Zika and Cache Valley viruses.

BACKGROUND: Vector-borne diseases are a major public health concern and cause significant morbidity and mortality. Zika virus (ZIKV) is the etiologic agent of a massive outbreak in the Americas that originated in Brazil in 2015 and shows a strong association with congenital ZIKV syndrome in newborns. Cache Valley virus (CVV) is a bunyavirus that causes mild to severe illness in humans and ruminants. In this study, we investigated the vector competence of Virginia mosquitoes for ZIKV and CVV to explore their abilities to contribute to potential outbreaks. METHODS: To determine vector competence, mosquitoes were fed a blood meal comprised of defibrinated sheep blood and virus. The presence of midgut or salivary gland barriers to ZIKV infection were determined by intrathoracic inoculation vs oral infection. After 14-days post-exposure, individual mosquitoes were separated into bodies, legs and wings, and saliva expectorant. Virus presence was detected by plaque assay to determine midgut infection, dissemination, and transmission rates. RESULTS: Transmission rates for Ae. albopictus orally infected (24%) and intrathoracically inoculated (63%) with ZIKV was similar to Ae. aegypti (48% and 71%, respectively). Transmission rates of ZIKV in Ae. japonicus were low, and showed evidence of a midgut infection barrier demonstrated by low midgut infection and dissemination rates from oral infection (3%), but increased transmission rates after intrathoracic inoculation (19%). Aedes triseriatus was unable to transmit ZIKV following oral infection or intrathoracic inoculation. CVV transmission was dose-dependent where mosquitoes fed high titer (ht) virus blood meals developed higher rates of midgut infection, dissemination, and transmission compared to low titer (lt) virus blood meals. CVV was detected in the saliva of Ae. albopictus (ht: 68%, lt: 24%), Ae. triseriatus (ht: 52%, lt: 7%), Ae. japonicus (ht: 22%, lt: 0%) and Ae. aegypti (ht: 10%; lt: 7%). Culex pipiens and Cx. restuans were not competent for ZIKV or CVV. CONCLUSIONS: This laboratory transmission study provided further understanding of potential ZIKV and CVV transmission cycles with Aedes mosquitoes from Virginia. The ability for these mosquitoes to transmit ZIKV and CVV make them a public health concern and suggest targeted control programs by mosquito and vector abatement districts.

Effects of La Crosse virus infection on the host-seeking behavior and levels of two neurotransmitters in Aedes triseriatus.

BACKGROUND: La Crosse virus (LACV) infection has been shown to manipulate the blood-feeding behaviors of its main vector, Aedes triseriatus. Here, we investigated the effects of virus infection on serotonin and dopamine and their potential roles in host-seeking. In mosquitoes, serotonin depletion has been shown to interfere with blood-feeding but not host-seeking. Dopamine depletion does not affect either blood-feeding or host-seeking; elevations of dopamine, however, has been shown to inhibit host-seeking. The purpose of this study was to determine the effects of LACV infection on the host-seeking behavior of and neurotransmitter levels in Ae. triseriatus. METHODS: Host-seeking behavior was evaluated using a uni-port olfactometer and a membrane feeder assay. Levels of serotonin and dopamine in infected and control mosquito heads were measured using HPLC-ED. RESULTS: Infection with LACV significantly inhibited the activation and attraction of Ae. triseriatus females to a host. A higher proportion of uninfected Ae. triseriatus females were activated by the presence of a host compared to infected mosquitoes and more uninfected mosquitoes were full responders (95.7%) compared to infected ones (91.1%). However, infection with LACV did not significantly affect the landing, probing, or blood-feeding rates of female mosquitoes. LACV-infected mosquitoes had lower serotonin levels than controls (104.5 vs 138.3 pg/head) while the dopamine levels were not affected by infection status (282.3 vs 237 pg/head). CONCLUSIONS: Our work suggests that virus-induced reduction of serotonin is related to previously reported blood-feeding alterations in LACV-infected mosquitoes and could lead to enhanced transmission and increased vectorial capacity. In addition, some aspects of host-seeking were inhibited by virus infection.

Influence of Forest Disturbance on La Crosse Virus Risk in Southwestern Virginia.

Forest disturbance effects on La Crosse virus (LACV) are currently unknown. We determined the abundance of three LACV accessory vectors (Aedes albopictus, Ae. canadensis, and Ae. vexans) and the primary amplifying host (Eastern chipmunk; Tamias striatus), and tested for LACV prevalence in both vectors and chipmunks, across a gradient of experimental forest disturbance treatments in southwest Virginia. Forest disturbance significantly affected the abundance of LACV accessory vectors, with a higher abundance on disturbed sites for Ae. canadensis and Ae. vexans. However, there was no significant disturbance effect on chipmunk abundance. Forest disturbance significantly affected LACV prevalence in mosquito vectors, with most (80%) detections on unlogged control sites, which past work showed harbor the highest abundance of the two most common LACV vectors (the primary vector Aedes triseriatus, and Ae. japonicus). Interestingly, LACV nucleic acid was only detected in Ae. japonicus and Culex pipiens/restuans, with no detections in the primary vector, Ae. triseriatus. In contrast to the vector results, antibodies were only found in chipmunks on logged sites, but this result was not statistically significant. Overall, our results suggest that human LACV risk should generally decline with logging, and reveal the potential importance of accessory vectors in LACV maintenance in Appalachian forests.

Cache Valley Virus in Aedes japonicus japonicus Mosquitoes, Appalachian Region, United States.

We detected Cache Valley virus in Aedes japonicus, a widely distributed invasive mosquito species, in an Appalachian forest in the United States. The forest contained abundant white-tailed deer, a major host of the mosquito and virus. Vector competence trials indicated that Ae. j. japonicus mosquitoes can transmit this virus in this region.

Effects of Plant Species, Insecticide, and Exposure Time On the Efficacy Of Barrier Treatments Against Aedes albopictus.

The effect of 5 plant species (arborvitae [Thuja occidentalis], boxwood [Buxus sp., Japanese honeysuckle [Lonicera japonica], rhododendron [Rhododendron sp.], and zebra grass [Miscanthus sinensis]) and 2 rates of lambda-cyhalothrin (3.13 ml and 6.25 ml active ingredient [AI]/liter) on knockdown (1 h) and mortality (24 h) of adult female Aedes albopictus was evaluated over an 8-wk period. A significant difference in knockdown was observed between the 2 rates of lambda-cyhalothrin on the 5 plant species, with the highest proportion of knockdown observed on zebra grass and rhododendron treated at the higher rate. Although mortality was ≥60% and 85% on the 5 plant species at the low and high rates of lambda-cyhalothrin, respectively, a significant difference between the 2 rates was only observed on boxwood and Japanese honeysuckle (P < 0.0001). We also tested the residual toxicity of 3 barrier sprays (lambda-cyhalothrin, bifenthrin, and deltamethrin) and evaluated the efficacy of a short (5-min) exposure to the insecticides on knockdown and mortality of adults over time. Significantly higher knockdown was observed with lambda-cyhalothrin compared with bifenthrin and deltamethrin (P < 0.0001). Mean knockdown was ∼98%, 92%, and 20% for lambda-cyhalothrin, bifenthrin, and deltamethrin, respectively, at week 2, and ∼98%, 0%, and 44%, respectively, 8 wk after treatments were applied. Adult mortality from the 3 chemical treatments, however, remained above 90% throughout the study. Lastly, the trends in mean proportion of knockdown were similar for mosquitoes exposed for either 5 min or 24 h to the 3 chemicals. An overall decline in mean mortality over time, however, was observed for mosquitoes exposed for 5 min to the chemicals compared with mortality from the 24-h exposure. The results suggest that lambda-cyhalothrin can be an effective barrier spray treatment against Ae. albopictus adults because its efficacy is limited little by plant species, it has long residual toxicity, and it is effective following only 5 min of exposure.

La Crosse Virus Field Detection and Vector Competence of Culex Mosquitoes.

La Crosse virus (LACV), a leading cause of arboviral pediatric encephalitis in the United States, is emerging in Appalachia. Here, we report field and laboratory evidence that suggest LACV may be using Culex mosquitoes as additional vectors in this region. This bunyavirus was detected by reverse-transcriptase polymerase chain reaction in two pools of Culex mosquitoes in southwestern Virginia and in six pools in West Virginia. To assess vector competence, we offered LACV blood meals to field-collected Culex restuans Theobald, Cx. pipiens L., and Aedes triseriatus (Say). Both Culex species were susceptible to infection. LACV-positive salivary expectorate, indicative of the ability to transmit, was detected in a small proportion of Cx. restuans (9%) and Cx. pipiens (4%) compared with Ae. triseriatus (40%). In a companion study of Cx. restuans only, we found that adults derived from nutritionally stressed larvae were significantly more likely to disseminate and transmit LACV. Our results indicate a potential role of Culex spp. in LACV dynamics that should be explored further in endemic areas.

La Crosse Virus in Aedes japonicus japonicus mosquitoes in the Appalachian Region, United States.

La Crosse virus (LACV), a leading cause of arboviral encephalitis in children in the United States, is emerging in Appalachia. For local arboviral surveillance, mosquitoes were tested. LACV RNA was detected and isolated from Aedes japonicus mosquitoes. These invasive mosquitoes may significantly affect LACV range expansion and dynamics.

La Crosse virus infection alters blood feeding behavior in Aedes triseriatus and Aedes albopictus (Diptera: Culicidae).

The effects of La Crosse virus (LACV) infection on blood feeding behavior in Aedes triseriatus (Say) and Aedes albopictus (Skuse) were investigated in the laboratory by measuring the size of the bloodmeal imbibed and the extent of refeeding by virus-infected and uninfected mosquitoes. LACV-infected Ae. triseriatus and Ae. albopictus took significantly less blood compared with uninfected mosquitoes. Twice as many virus-infected Ae. triseriatus mosquitoes refed compared with uninfected individuals (18 vs. 9%; P < 0.05); however, virus infection had no significant effect on the refeeding rate of Ae. albopictus. Reduction in bloodmeal size followed by an increased avidity for refeeding may lead to enhanced horizontal transmission of the LACV by its principal vector, Ae. triseriatus.

Select small core structure carbamates exhibit high contact toxicity to "carbamate-resistant" strain malaria mosquitoes, Anopheles gambiae (Akron).

Acetylcholinesterase (AChE) is a proven target for control of the malaria mosquito (Anopheles gambiae). Unfortunately, a single amino acid mutation (G119S) in An. gambiae AChE-1 (AgAChE) confers resistance to the AChE inhibitors currently approved by the World Health Organization for indoor residual spraying. In this report, we describe several carbamate inhibitors that potently inhibit G119S AgAChE and that are contact-toxic to carbamate-resistant An. gambiae. PCR-RFLP analysis was used to confirm that carbamate-susceptible G3 and carbamate-resistant Akron strains of An. gambiae carry wild-type (WT) and G119S AChE, respectively. G119S AgAChE was expressed and purified for the first time, and was shown to have only 3% of the turnover number (k(cat)) of the WT enzyme. Twelve carbamates were then assayed for inhibition of these enzymes. High resistance ratios (>2,500-fold) were observed for carbamates bearing a benzene ring core, consistent with the carbamate-resistant phenotype of the G119S enzyme. Interestingly, resistance ratios for two oxime methylcarbamates, and for five pyrazol-4-yl methylcarbamates were found to be much lower (4- to 65-fold). The toxicities of these carbamates to live G3 and Akron strain An. gambiae were determined. As expected from the enzyme resistance ratios, carbamates bearing a benzene ring core showed low toxicity to Akron strain An. gambiae (LC(50)>5,000 µg/mL). However, one oxime methylcarbamate (aldicarb) and five pyrazol-4-yl methylcarbamates (4a-e) showed good to excellent toxicity to the Akron strain (LC(50) = 32-650 µg/mL). These results suggest that appropriately functionalized "small-core" carbamates could function as a resistance-breaking anticholinesterase insecticides against the malaria mosquito.

Induction and inhibition of an apparent neuronal phenotype in Spodoptera frugiperda insect cells (Sf21) by chemical agents.

The goal of this research was to induce neuron-like properties in Sf21 cells, an insect ovarian cell line, which could lead to a new high-throughput insecticide screening method and a way to mass produce insect neuronal material for basic research. This study applied differentiation agents to produce viable neuron-like cells. In the presence of the molting hormone 20-hydroxyecdysone (20-HE), or insulin, in the growth medium, a maximum of ca. 30 % of Sf21 cells expressed an apparent neuronal morphology of unipolar, bipolar, or multipolar axon-like processes within 2-3 days. Maximal differentiation occurred after 2 days in the presence of 50 µM 20-HE or 3 days in 10 µM insulin. Both 20-HE and insulin displayed time- and concentration-dependent differentiation with biphasic curves, suggesting that two binding sites or processes were contributing to the observed effects. In addition, combinations of 20-HE and insulin produced apparent synergistic effects on differentiation. Caffeine, a central nervous system stimulant, inhibited induction of elongated processes by 20-HE and/or insulin, with an IC(50) of 9 nM for 20-HE, and the inhibition was incomplete, resulting in about one-quarter of the differentiated cells remaining, even at high concentrations (up to 1 mM). The ability to induce a neural phenotype simplifies the studies of insect cells, compared to either the use of primary nervous tissue or genetic engineering techniques. The presence of ion channels or receptors in the differentiated cells remains to be determined.

Re-engineering aryl methylcarbamates to confer high selectivity for inhibition of Anopheles gambiae versus human acetylcholinesterase.

To identify potential human-safe insecticides against the malaria mosquito we undertook an investigation of the structure-activity relationship of aryl methylcarbamates inhibitors of acetylcholinesterase (AChE). Compounds bearing a ß-branched 2-alkoxy or 2-thioalkyl group were found to possess good selectivity for inhibition of Anopheles gambiae AChE over human AChE; up to 530-fold selectivity was achieved with carbamate 11d. A 3D QSAR model is presented that is reasonably consistent with log inhibition selectivity of 34 carbamates. Toxicity of these compounds to live Anopheles gambiae was demonstrated using both tarsal contact (filter paper) and topical application protocols.

The potential for emergence of Chagas disease in the United States.

To determine the risk for Chagas disease (American trypanosomiasis) in the United States, the characteristics that make the triatomine vector effective and the areas most at risk for transmission were delineated. In addition, the status of Chagas disease awareness among physicians in areas with a potential risk for the disease was determined. A geographical information system (GIS) was used to analyze three triatomine species within the United States known to harbor Trypanosoma cruzi and that exhibit qualities of domesticity. An analysis of the minimum temperature threshold for increased triatomine activity delineates the current population at increased risk, and by incorporating temperature predictions for 2030, the population at risk under a future climate scenario was also delineated. Considering both environmental and social factors, a vignette-based physician survey, based on the results of the GIS analysis, was used to gauge the level of awareness of Chagas disease within the delineated higher risk range. The current area at increased risk for Chagas disease includes much of the southern United States, and the higher risk range is expected to expand into the central United States based upon the 1 degrees C (1.8 degrees F) increase in temperature predicted by the Intergovernmental Panel on Climate Change (IPCC) by the year 2030. Survey results indicate a limited consideration of Chagas disease during differential diagnosis, illustrating that the low number of Chagas disease cases discovered in the United States may be attributable to a lack of disease awareness as opposed to a lack of disease threat. This study combines GIS and survey analyses to evaluate the role that temperature variability and disease awareness among physicians play in the potential emergence of Chagas disease in the United States. This approach indicates that there is a potential for Chagas disease to emerge in the United States.

Seasonal abundance of Culex restuans and Culex pipiens in southwestern Virginia through ovitrapping.

Ovitrap collections were done to determine the seasonal distribution and ovipositional activity of Culex restuans and Culex pipiens in southwestern Virginia during summers 2002 and 2003. In both years, Cx. restuans was the first species collected and was the dominant species throughout the collecting period. More than 90% of the collections were comprised of Cx. restuans. Two oviposition peaks occurred in both years for Cx. restuans, the first in June and second in the middle of July. Culex pipiens made up less than 7% of the total catch and started to occur in the traps in the middle of the trapping season with a peak occurring in August. Although ovipositional activity began to decline for Cx. restuans, at the end of the trapping period, there was a slight increase in ovipositional activity for Cx. pipiens, but not enough to result in a crossover. A third species, Culex salinarius, was collected in the traps, but the total number of egg rafts accounted for less than 1% of the total collection.

Oviposition preferences of Culex restuans and Culex pipiens (Diptera: Culicidae) for selected infusions in oviposition traps and gravid traps.

Field studies were conducted in southwestern Virginia to determine the ovipositional preferences of Culex restuans and Culex pipiens by using ovitraps and gravid traps baited with selected infusions. For the ovitrap collections, 4 different infusions (manure, hay, grass, and rabbit chow) were used. Significant differences among infusions were detected on most sample dates for both species. For 3 of the first 4 wk of collections, the manure infusion collected significantly more Cx. restuans than all the other infusions. The hay and grass infusions collected the majority of the egg rafts during weeks 5-9. Cx. pipiens egg rafts were absent from the first 3 wk of collections. Of the remaining 6 wk, 4 showed significant differences in attractiveness of infusions, with the hay and grass infusions preferred by Cx. pipiens. Two infusions, manure and hay, were used for the gravid trap experiment and both Cx. restuans and Cx. pipiens data were combined for analysis. Only the first 2 wk showed significance, with manure being preferred over hay in both weeks. In later collections, the relative attractiveness of the hay infusion increased. A seasonal shift in infusion preference may be related to incubation temperature during preparation of the infusions. New infusions were prepared each week and incubation was done outside. Increased attractiveness of the hay infusion coincided with higher average temperatures in July and August. Hay infusion was very effective for trapping both Cx. pipiens and Cx. restuans in southwestern Virginia and is more convenient to use than manure. However, cool outside temperatures in the early season may interfere with the fermentation process and thus incubation should be done for a longer time or brought indoors.

Spatiotemporal oviposition and habitat preferences of Ochlerotatus triseriatus and Aedes albopictus in an emerging focus of La Crosse virus.

The number of cases of encephalitis caused by La Crosse virus recently has increased in southwestern Virginia counties. This article presents results of a study conducted from May to September 2000 in Wise County, VA, that examined the area-wide oviposition activity and habitat preferences of Ochlerotatus triseriatus and Aedes albopictus, potential vectors of La Crosse virus in the region. Data from 490 ovitrap collections made throughout the county showed that mean oviposition activity throughout the study was higher for Oc. triseriatus (20.4 eggs/trap-day) than for Ae. albopictus (3.7 eggs/trap-day). The 2 species also had distinct habitat preferences for oviposition, with Oc. triseriatus favoring forested habitats and Ae. albopictus favoring urban/residential habitats. A landcover map of 6 habitat types derived from Landsat satellite imagery of the county showed that 63% of the county was forested and 18% was urban/residential. A Bayesian decision-rule model that incorporated the ovitrap data and landcover map was moderately successful at predicting the occurrence of high oviposition activity and abundance of the 2 species. The predictions reflected seasonal and spatial fluctuations in oviposition activity, with accuracies between 55 and 79% for Oc. triseriatus and 70 and 94% for Ae. albopictus. Kappa (K), a measure of the predictive power of the model, varied from poor (K < 0.4) to good (0.4 < K < 0.75) for both species, and was highest during periods when actual egg abundance was high. This suggests that the predictions were most accurate during periods when the risk for La Crosse virus transmission is greatest. Limitations and suggestions for improving the model are discussed.