Using environmental factors to predict Rhipicephalus sanguineus s.l. (Acari: Ixodidae) mortality.

BACKGROUND: Rhipicephalus sanguineus s.l. (Latreille, 1806) can establish indoor populations, which increases the risk of pathogen transmission to humans and companion dogs. Rhipicephalus sanguineus s.l. ticks spend most of their life cycle off the host, which subjects developmental timescale to abiotic factors. Previous studies showed that both temperature and relative humidity (RH) influenced Rhipicephalus sanguineus s.l. survival time across all life stages. However, quantified relationships between environmental factors and Rhipicephalus sanguineus s.l. mortality is not currently available. Here, three Rhipicephalus sanguineus s.l. strains were evaluated for mortality under 20 combinations of five temperatures and four RHs. The data obtained were analyzed to quantify the relationship between environmental factors and Rhipicephalus sanguineus s.l. RESULTS: Mortality probabilities did not show a consistent pattern between the three tick strains. Temperature, RH, and their interaction influenced Rhipicephalus sanguineus s.l. mortality probabilities across all life stages, with mortality probability generally increasing with temperature but decreasing with RH. With 50% and lower RH, larvae cannot survive for more than 1 week. However, mortality probabilities in all strains and stages were more sensitive to temperature than to RH. CONCLUSION: This study identified the predictive relationship between environmental factors and Rhipicephalus sanguineus s.l. survival, which enables estimations of tick survival time under varied residential situations, allows parameterization of population models, and provides guidance for pest control professionals to develop efficient management strategies. © 2023 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Evidence of Permethrin Resistance and Fipronil Tolerance in Rhipicephalus sanguineus s.l. (Acari: Ixodidae) Populations From Florida and California.

Rhipicephalus sanguineus s.l. (Latreille), is a vector of multiple disease-causing pathogens to humans and dogs. Permethrin and fipronil are two acaricides used to manage R. sanguineus s.l. infestations. Homeowners have reported treatment failures in managing brown dog ticks using permethrin and fipronil based products. Previous studies demonstrated that high permethrin resistance in some R. sanguineus s.l. populations was due to metabolic detoxification and target site insensitivity. In this study, three R. sanguineus s.l. strains, one from a laboratory colony (NC) and two colonies originally collected from Florida (FL) and California (CA), were evaluated for resistance expression against permethrin and fipronil. Metabolic detoxification mechanisms were evaluated in the FL strain using three synergists, while a polymerase chain reaction assay was used to detect a resistance mutation in all strains. The NC strain was susceptible to both permethrin and fipronil, while both the FL and CA strains exhibited high resistance to permethrin and tolerance to fipronil. The synergist tests and PCR results indicated that the FL strain utilized both metabolic resistance and target site insensitivity against permethrin, while the CA strain was documented to have the target-site insensitivity resistant allele. This study confirmed permethrin resistance in both California and Florida populations and its persistence in Florida populations, although its susceptibility can potentially be increased by adding a synergist. Fipronil resistance was not detected suggesting this acaricide may provide suitable tick control.

Variable Effects of Temperature and Relative Humidity on Rhipicephalus sanguineus s.l. (Acari: Ixodidae) Development.

Rhipicephalus sanguineus s.l. (Latreille, 1806) can establish populations in residences and may lead to severe domestic and peridomestic infestations. Detection in the early infestation stage is challenging because of their small body size and the lack of visibility when ticks stay in sheltered refugia. The residents may believe that the infestation has been eliminated when no ticks are observed until ticks reappear when seeking hosts. Thus, it is necessary to improve our understanding of tick phenology to achieve more effective infestation management. In this study, the relationships between environmental conditions and tick development were explored in laboratory and using linear and nonlinear models. Three R. sanguineus s.l. strains, from one colony of the temperate lineage and two of the tropical lineage, were evaluated for the development of all life stages and conversion efficiency index (CEI) under five temperatures and four relative humidities (RHs). The development times differed between the three tick strains across stages and were primarily dependent on temperature. The CEIs had little variance explained by temperature, RH, or strains. Compared with the linear and exponential models with temperature as the only variable, the Brière-1 model was the best approximating model for most of the developmental rates. The developmental temperature thresholds for R. sanguineus s.l. development estimated by the Brière-1 model varied inconsistently across strains and life stages. We developed a more predictive relationship between environmental factors and R. sanguineus s.l. development, which can be utilized to predict tick development using temperature and develop appropriate control strategies.

Effects of Mosquito Biology on Modeled Chikungunya Virus Invasion Potential in Florida.

Arboviruses transmitted by Aedes aegypti and Aedes albopictus have been introduced to Florida on many occasions. Infrequently, these introductions lead to sporadic local transmission and, more rarely, sustained local transmission. Both mosquito species are present in Florida, with spatio-temporal variation in population composition. We developed a two-vector compartmental, deterministic model to investigate factors influencing Chikungunya virus (CHIKV) establishment. The model includes a nonlinear, temperature-dependent mosquito mortality function based on minimum mortality in a central temperature region. Latin Hypercube sampling was used to generate parameter sets used to simulate transmission dynamics, following the introduction of one infected human. The analysis was repeated for three values of the mortality function central temperature. Mean annual temperature was consistently important in the likelihood of epidemics, and epidemics increased as the central temperature increased. Ae. albopictus recruitment was influential at the lowest central temperature while Ae. aegypti recruitment was influential at higher central temperatures. Our results indicate that the likelihood of CHIKV establishment may vary, but overall Florida is permissive for introductions. Model outcomes were sensitive to the specifics of mosquito mortality. Mosquito biology parameters are variable, and improved understanding of this variation will improve our ability to predict the outcome of introductions.

Wild pigs as sentinels for hard ticks: A case study from south-central Florida.

As a result of shifts in the habitable range of ticks due to climate change and the ongoing threat of exotic tick species introductions, efficient surveillance tools for these pests and disease vectors are needed. Wild pigs are habitat generalists, distributed throughout most of the United States, and often hunted recreationally or removed as part of management programs, making them potentially useful sentinel hosts for ticks. We compared ticks collected from captured wild pigs and standard tick dragging methods on a south-central Florida cattle ranch from May 2015-August 2017. Three hundred and sixteen wild pigs were surveyed, and 84 km spanning three habitat types (seminative pasture, improved pasture, and hammock) were dragged. In total, 1023 adults of four species (Amblyomma auricularium, Amblyomma maculatum, Dermacentor variabilis, and Ixodes scapularis) were collected from wild pigs, while 39 adults of three species (A. auricularium, A. maculatum, and I. scapularis) were collected from drags. Only one immature specimen, a nymph, was collected from a pig, while dragging collected 2808 larvae and 150 nymphs. Amblyomma maculatum comprised 96% of adults collected from pigs, while A. maculatum, I. scapularis, and A. auricularium comprised 38%, 33%, and 28% of adults collected from drags, respectively. Adults of all tick species found on drags were found on pigs, and wild pig surveillance detected adults of an additional species not found on drags. Dragging was far superior for collection of immatures but not for adults of most species found in this study. These findings suggest wild pigs could be used as a sentinel for the detection of tick species. When combined with ongoing wild pig research, hunting, or management, wild pig surveillance can provide an effective method to survey for adult tick presence of some species of interest and may assist in tracking the range expansion of some tick species.

Distribution and host associations of ixodid ticks collected from wildlife in Florida, USA.

A tick survey was conducted to document tick-host associations with Florida (USA) wildlife, and to determine the relative abundance and distribution of ixodid ticks throughout the state. The survey was conducted using collection kits distributed to licensed Florida hunters as well as the examination of archived specimens from ongoing state wildlife research programs. Collected tick samples were obtained from 66% of Florida counties and were collected from nine wildlife hosts, including black bear, bobcat, coyote, deer, gray fox, Florida panther, raccoon, swine, and wild turkey. In total, 4176 ticks were identified, of which 75% were Amblyomma americanum, 14% Ixodes scapularis, 8% A. maculatum, 3% Dermacentor variabilis, and < 1% were I. affinis and I. texanus. americanum, D. variabilis, and I. scapularis had the broadest host range, while A. maculatum, D. variabilis, and I. scapularis had the widest geographic distribution. While the survey data contribute to an understanding of tick-host associations in Florida, they also provide insight into the seasonal and geographic distribution of several important vector species in the southeastern USA.

Environmental Drivers of Seasonal Patterns of Host Utilization by Culiseta melanura (Diptera: Culicidae) in Florida.

Field studies were carried out in four Florida counties to investigate winter and spring ecology of host use by Culiseta melanura (Coquillet), the primary vector of eastern equine encephalomyelitis virus (EEEV) in North America. Bloodmeal analysis by PCR was used to identify 233 host bloodmeals, which mainly originated from birds (78.5%) and reptiles (17.2%), primarily Anolis spp. lizards. Across counties, the percentage of bloodmeals from reptiles (7-37% depending upon county) increased with increasing day length and temperature in the spring. Multiple logistic regression revealed that differences in reptile host use across collection sites were largely explained by differences in average day length and temperature on the day of collection, and is probably owing to environment-driven behavioral patterns of ectothermic animals. Although past studies have demonstrated reptile biting by epizootic vectors of EEEV, including Culex (Melanoconion) spp., this is the first study to demonstrate widespread and common feeding upon ectothermic hosts by Cs. melanura. This work suggests that reptiles, particularly anole lizards, play a role in the ecology of EEEV in Florida either as amplifying hosts or as noncompetent hosts which dilute vector feedings thereby suppressing transmission. Detailed laboratory studies investigating impacts of environmental variables (temperature and photoperiod) on EEEV competence of anoles are needed to assess whether these animals support virus amplification.

Transmission risk of two chikungunya lineages by invasive mosquito vectors from Florida and the Dominican Republic.

Between 2014 and 2016 more than 3,800 imported human cases of chikungunya fever in Florida highlight the high risk for local transmission. To examine the potential for sustained local transmission of chikungunya virus (CHIKV) in Florida we tested whether local populations of Aedes aegypti and Aedes albopictus show differences in susceptibility to infection and transmission to two emergent lineages of CHIKV, Indian Ocean (IOC) and Asian genotypes (AC) in laboratory experiments. All examined populations of Ae. aegypti and Ae. albopictus mosquitoes displayed susceptibility to infection, rapid viral dissemination into the hemocoel, and transmission for both emergent lineages of CHIKV. Aedes albopictus had higher disseminated infection and transmission of IOC sooner after ingesting CHIKV infected blood than Ae. aegypti. Aedes aegypti had higher disseminated infection and transmission later during infection with AC than Ae. albopictus. Viral dissemination and transmission of AC declined during the extrinsic incubation period, suggesting that transmission risk declines with length of infection. Interestingly, the reduction in transmission of AC was less in Ae. aegypti than Ae. albopictus, suggesting that older Ae. aegypti females are relatively more competent vectors than similar aged Ae. albopictus females. Aedes aegypti originating from the Dominican Republic had viral dissemination and transmission rates for IOC and AC strains that were lower than for Florida vectors. We identified small-scale geographic variation in vector competence among Ae. aegypti and Ae. albopictus that may contribute to regional differences in risk of CHIKV transmission in Florida.

Transstadial Effects of Bti on Traits of Aedes aegypti and Infection with Dengue Virus.

Most mosquito control efforts are primarily focused on reducing the adult population size mediated by reductions in the larval population, which should lower risk of disease transmission. Although the aim of larviciding is to reduce larval abundance and thus recruitment of adults, nonlethal effects on adults are possible, including transstadial effects on phenotypes of adults such as survival and pathogen infection and transmission. In addition, the mortality induced by control efforts may act in conjunction with other sources of mosquito mortality in nature. The consequences of these effects and interactions may alter the potential of the population to transmit pathogens. We tested experimentally the combined effects of a larvicide (Bacillus thuringiensis ssp. israelensis, Bti) and competition during the larval stages on subsequent Aedes aegypti (Linnaeus) traits, population performance, and susceptibility to dengue-1 virus infection. Ae. aegypti that survived exposure to Bti experienced accelerated development, were larger, and produced more eggs with increasing amounts of Bti, consistent with competitive release among surviving mosquitoes. Changing larval density had no significant interactive effect with Bti treatment on development and growth to adulthood. Larval density, but not Bti or treatment interaction, had a strong effect on survival of adult Ae. aegypti females. There were sharper declines in cumulative daily survival of adults from crowded than uncrowded larval conditions, suggesting that high competition conditions of larvae may be an impediment to transmission of dengue viruses. Rates of infection and dengue-1 virus disseminated infections were found to be 87±13% and 88±12%, respectively. There were no significant treatment effects on infection measurements. Our findings suggest that larvicide campaigns using Bti may reduce the number of emerged adults, but survivors will have a fitness advantage (growth, development, enhanced production of eggs) relative to conspecifics that are not under larvicide pressure. However, under most circumstances, these transstadial effects are unlikely to outweigh reductions in the adult population by Bti and altered risk of disease transmission.

Vector competence of Culex pipiens quinquefasciatus (Diptera: Culicidae) for West Nile virus isolates from Florida.

OBJECTIVES: To assess vector competence (infection, dissemination and transmission) of Culex pipiens quinquefasciatus for Florida (FL) West Nile virus (WNV) isolates. METHODS: West Nile virus isolates (WN-FL-03: NY99 genotype; WN-FL-05-558, WN-FL-05-2186, WN-FL-05-510: WN02 genotype) collected from different regions of FL were used for vector competence experiments in Cx. p. quinquefasciatus from Alachua County and Indian River County in FL. Mosquitoes from both colonies were fed blood containing 7.9 ± 0.2 log10 plaque-forming units WNV/ml ± SE and incubated at 28 °C for 14 days. Vector competence, including rates of infection, dissemination, and transmission, was compared between colonies for WN-FL-03 using chi-squared. Virus titres in bodies, legs and saliva were compared using anova. Daily measurements of in vitro replication of WNV isolates were evaluated in Vero cells so that a standardised virus dose for each isolate could be delivered to mosquitoes. RESULTS: Infection and dissemination rates were high (≥ 95%) and not affected by isolate or colony (infection, P = 0.679; dissemination, P = 0.799). Transmission rates were low (≤ 20%), detected in one colony and affected by isolate (P = 0.008). Body and leg titres differed between isolates (body titre, P = 0.031; leg titre, P = 0.044) and colonies (body titre, P = 0.001; leg titre, P = 0.013) while saliva titre did not differ between isolates (P = 0.462). CONCLUSIONS: Variation in vector competence of mosquito populations may be attributed, in part, to exposures to WNV with genetic differences leading to different rates of replication in mosquitoes. Evaluation of vector competence for different WNV isolates may help us understand vector-virus interactions and, hence, the role of vectors in complex virus transmission cycles in nature.

Survival of West Nile virus-challenged Southern house mosquitoes, Culex pipiens quinquefasciatus, in relation to environmental temperatures.

We investigated the effect of West Nile virus (WNV) infection on survival in two colonies of Culex pipiens quinquefasciatus Say (Diptera: Culicidae) originating from Vero Beach and Gainesville, FL. Mosquitoes were fed West Nile virus-infected blood and checked daily for survival. Exposure to WNV decreased survival among Cx. p. quinquefasciatus from Gainesville relative to unexposed individuals at 31° C. In contrast, exposure to WNV enhanced survival among Cx. p. quinquefasciatus from Vero Beach relative to unexposed individuals at 27° C. These results may suggest that exposure to WNV and associated infection could increase or decrease components of fitness, dependent on environmental temperature and intraspecific variation in Cx. p. quinquefasciatus. The relationship between lifespan (time of death in days) and WNV titer differed in the colonies at 31° C and 27° C, suggesting that intraspecific species variation in response to temperature impacts interactions with WNV. While further work is needed to determine if similar effects occur under field conditions, this suggests intraspecific variation in vector competence for WNV and adult survival of Cx. p. quinquefasciatus, both aspects of vectorial capacity.

Effects of virus dose and extrinsic incubation temperature on vector competence of Culex nigripalpus (Diptera: Culicidae) for St. Louis encephalitis virus.

Culex nigripalpus Theobald is a primary vector of St. Louis encephalitis virus in the southeastern United States. Cx. nigripalpus females were fed blood containing a low (4.0 +/- 0.01 log10 plaque-forming unit equivalents (PFUeq) /ml) or high (4.7 +/- 0.1 log10 PFUeq/ml) St. Louis encephalitis virus dose and maintained at extrinsic incubation temperatures (EIT) of 25 or 28 degrees C for 12 d. Vector competence was measured via quantitative real-time reverse transcriptase polymerase chain reaction to estimate PFUeq using rates of infection, dissemination, and transmission. There were no differences in infection rates between the two EITs at either dose. The low dose had higher infection rates at both EITs. Dissemination rates were significantly higher at 28 degrees C compared with 25 degrees C at both doses. Virus transmission was observed (<7%) only at 28 degrees C for both doses. The virus titer in body tissues was greater at 28 degrees C compared with 25 degrees C at both doses. The difference between the EITs was greater at the low dose, resulting in a higher titer for the low dose than the high dose at 28 degrees C. Virus titers in leg tissues were greater in mosquitoes fed the high versus low dose, but were not influenced by EIT. Further investigations using a variety of environmental and biological factors would be useful in exploring the complexity of vector competence.

Relationships between infection, dissemination, and transmission of West Nile virus RNA in Culex pipiens quinquefasciatus (Diptera: Culicidae).

Culex pipiens quinquefasciatus Say fed blood containing 6.8 +/- 0.3 logs (mean +/- SE) plaque-forming units of West Nile virus (WNV)/ml were maintained at 28 degrees C for incubation periods (IP) of 7, 14, or 21 d. Several attributes of vector competence were determined at each IP using quantitative real-time reverse transcriptase polymerase chain reaction to estimate plaque forming unit equivalents including: infection rate (WNV-positive abdomens), dissemination rate (WNV-positive legs or thoraces), combined dissemination rate (WNV-positive legs and thoraces), transmission rate (WNV-positive saliva), and WNV titers in abdomens, legs, thoraces, and saliva. Each rate increased or was equivalent with increasing IP. Mosquitoes transmitting WNV in saliva also had significantly higher IP-dependent WNV titers in abdomens, legs, and thoraces. Titers of WNV in abdomens were significantly correlated with titers in legs and thoraces, but the degree of association changed with IP. However, titers of abdomens, legs, and thoraces were not correlated with WNV presence or titer in the saliva. The results show that WNV presence or titer in the saliva of infected Cx. p. quinquefasciatus was not directly influenced by processes involved in WNV replication in other tissues. The processes controlling midgut infection and escape are, in part, independent from the infection processes in other tissues. The relationship between infection, dissemination, and transmission varied over time. The infection and replication of WNV in different tissues is likely influenced by different barriers encountered during the extrinsic incubation period. The significance of these observations for understanding vector competence is discussed.

Impact of West Nile virus dose and incubation period on vector competence of Culex nigripalpus (Diptera: Culicidae).

Female Culex nigripalpus were fed blood containing a low dose (6.3±0.01 logs plaque-forming units (PFU)/mL) or high dose (7.3±0.1 logs PFU/mL) of West Nile virus (WNV) and maintained at 28°C for incubation periods (IPs) of 6 or 12 days. Vector competence was measured using rates of infection (% with WNV-positive bodies), dissemination (% infected with WNV-positive legs), and transmission (% infected with WNV-positive saliva). Infection rates were not influenced by dose or IP. Dissemination rates were significantly higher at the high dose, and this was dependent on IP. Despite 100% infection and 90% dissemination in the most permissive treatment of high dose and 12 days, only 11% transmission was observed. Virus titers in body and leg tissues were significantly lower at the low dose and the titers were not influenced by IP. We show that not all mosquitoes with infections and/or disseminated infections transmit WNV under the conditions of this test. Therefore, characterizing the transmission ability of a vector population using infection or dissemination as indicators of transmission may provide inaccurate information. The complex relationships between infection, dissemination, and transmission must be evaluated under a variety of biological and environmental conditions to begin to assess the epidemiological risk of natural mosquito populations.

Environmental and biological factors influencing Culex pipiens quinquefasciatus (Diptera: Culicidae) vector competence for West Nile Virus.

Interactions between environmental and biological factors affect the vector competence of Culex pipiens quinquefasciatus for West Nile virus. Three age cohorts from two Cx. p. quinquefasciatus colonies were fed blood containing a low- or high-virus dose, and each group was held at two different extrinsic incubation temperatures (EIT) for 13 days. The colonies differed in the way that they responded to the effects of the environment on vector competence. The effects of mosquito age on aspects of vector competence were dependent on the EIT and dose, and they changed depending on the colony. Complex interactions must be considered in laboratory studies of vector competence, because the extent of the genetic and environmental variation controlling vector competence in nature is largely unknown. Differences in the environmental (EIT and dose) and biological (mosquito age and colony) effects from previous studies of Cx. p. quinquefasciatus vector competence for St. Louis encephalitis virus are discussed.

Sources of error in the estimation of mosquito infection rates used to assess risk of arbovirus transmission.

Infection rate is an estimate of the prevalence of arbovirus infection in a mosquito population. It is assumed that when infection rate increases, the risk of arbovirus transmission to humans and animals also increases. We examined some of the factors that can invalidate this assumption. First, we used a model to illustrate how the proportion of mosquitoes capable of virus transmission, or infectious, is not a constant fraction of the number of infected mosquitoes. Thus, infection rate is not always a straightforward indicator of risk. Second, we used a model that simulated the process of mosquito sampling, pooling, and virus testing and found that mosquito infection rates commonly underestimate the prevalence of arbovirus infection in a mosquito population. Infection rate should always be used in conjunction with other surveillance indicators (mosquito population size, age structure, weather) and historical baseline data when assessing the risk of arbovirus transmission.

The Effect of Multiple Vectors on Arbovirus Transmission.

Many mosquito-borne arboviruses have more than one competent vector. These vectors may or may not overlap in space and time, and may interact differently with vertebrate hosts. The presence of multiple vectors for a particular virus at one location over time will influence the epidemiology of the system, and could be important in the design of intervention strategies to protect particular hosts. A simulation model previously developed for West Nile and St. Louis encephalitis viruses and Culex nigripalpus was expanded to consider two vector species. These vectors differed in their abundance through the year, but were otherwise similar. The model was used to examine the consequences of different combinations of abundance patterns on the transmission dynamics of the virus. The abundance pattern based on Cx. nigripalpus dominated the system and was a key factor in generating epidemics in the wild bird population. The presence of two vectors often resulted in multiple epidemic peaks of transmission. A species which was active in the winter could enable virus persistence until another vector became active in the spring, summer, or fall. The day the virus was introduced into the system was critical in determining how many epidemic peaks were observed and when the first peak occurred. The number of epidemic peaks influenced the overall proportion of birds infected. The implications of these results for assessing the relative importance of different vector species are discussed.

Environmental and biological factors influencing Culex pipiens quinquefasciatus Say (Diptera: Culicidae) vector competence for Saint Louis encephalitis virus.

Complex interactions between environmental and biological factors influence the susceptibility of Culex pipiens quinquefasciatus to St. Louis encephalitis virus and could affect the epidemiology of virus transmission. Similar interactions could have epidemiologic implications for other vector-virus systems. We conducted an experiment to examine four such factors in combination: mosquito age, extrinsic incubation temperature (EIT), virus dose, and colony. The proportion of mosquitoes with body infections or disseminated infections varied between colonies, and was dependant on age, EIT, and dose. We also show that the probability of a body or leg infection interacted in complex ways between colonies, ages, EITs, and doses. The complex interactive effects of environmental and biological factors must be taken into account for studies of vector competence and epidemiology, especially when laboratory studies are used to generalize to natural transmission dynamics where the extent of variation is largely unknown.

Age modifies the effect of body size on fecundity in Culex quinquefasciatus Say (Diptera: Culicidae).

Fecundity of mosquitoes can vary with many factors and can have a strong effect on population growth. This study reports the effects of body size, blood meal size, and age on the reproductive output of nulliparous Culex quinquefasciatus, a vector of arboviruses and other pathogens. Mated adult female mosquitoes from a colony were reared under standard conditions and fed on chickens at different ages post-eclosion. Blood meal size and wing length were recorded, as well as the number of eggs in the first-cycle egg raft. Each of these factors had a significant influence on fecundity considered in a simple regression context. Multiple regression analysis revealed a significant interaction effect between age and body size on fecundity. Up to 13 days of age, fecundity was positively correlated with body size, but in mosquitoes older than 13 days, this relationship was not significantly different from zero. These results are discussed in terms of the known physiology of this and other species.

Arbovirus transmission by Culex nigripalpus in Florida, 2005.

Understanding the transmission patterns of West Nile and St. Louis encephalitis viruses (family Flaviviridae, genus Flavivirus, WNV and SLEV) could result in an increased ability to predict transmission risk to humans. To examine transmission patterns between vector and host, we trapped mosquitoes in three Florida counties from June to November 2005 by using chicken-baited lard can mosquito traps. These traps were used to monitor for presence of WNV and SLEV in mosquitoes and subsequent transmission of these viruses to chickens. In total, 166,615 female mosquitoes were sorted into 4,009 pools based on species and bloodfed status, and they were tested for presence of WNV and SLEV. Sera from 209 chickens were tested for WNV and SLEV antibodies. We detected eight WNV-positive Culex nigripalpus Theobald mosquito pools; SLEV was not detected in any pools. Six positive pools were collected in August and September from Duval County, one pool in September from Manatee County, and one pool in November from Indian River County. Of the eight chickens potentially exposed to WNV, antibodies were detected in only one chicken, indicating a low rate of transmission relative to the observed mosquito infection rates. Low virus transmission rates relative to infection rates would suggest that using sentinel chicken seroconversion data as a means of arbovirus surveillance may underestimate the prevalence of WNV in the mosquito population. However, using mosquito infection rates may overestimate the risk of arboviral transmission. A variety of factors might account for the observed low level of transmission including a lack of viral dissemination in mosquito vectors.