Optimization of artificial membrane feeding system for lone star ticks, Amblyomma americanum (Acari: Ixodidae), and experimental infection with Rickettsia amblyommatis (Rickettsiales: Rickettsiaceae).

With the introduction of siliconized artificial membranes, various artificial feeding systems (AFS) for hard ticks (Ixodidae) have been developed over the last decades. Most AFS utilize similar core components but employ diverse approaches, materials, and experimental conditions. Published work describes different combinations of the core components without experimental optimizations for the artificial feeding of different tick species. Amblyomma americanum L., (Acari: Ixodidae) (lone star tick) is a known vector and reservoir for diverse tick-borne pathogens, such as Rickettsia amblyommatis and Ehrlichia chaffeensis. Ongoing environmental changes have supported the expansion of A. americanum into new habitats, contributing to increased tick-borne diseases in endemic areas. However, a significant knowledge gap exists in understanding the underlying mechanisms involved in A. americanum interactions with tick-borne pathogens. Here, we performed a systematic analysis and developed an optimized AFS for nymphal lone star ticks. Our results demonstrate that Goldbeater's membranes, rabbit hair, hair extract, and adult lone star ticks significantly improved the attachment rate of nymphal ticks, whereas tick frass and frass extract did not. With the optimized conditions, we achieved an attachment rate of 46 ±â€…3% and a success rate of 100% (i.e., one or more attached ticks) in each feeding experiment for nymphal lone star ticks. When fed on sheep blood spiked with R. amblyommatis, both nymphal and adult lone star ticks acquired and maintained R. amblyommatis, demonstrating the feasibility of studying A. americanum-pathogen interactions using AFS. Our study can serve as a roadmap to optimize and improve AFS for other medically relevant tick species.

Evaluation of An Aerial Application of Duet Hd® Against Aedes Dorsalis and Culex Tarsalis in Rural Habitats of the Great Salt Lake, Utah.

The Salt Lake City Mosquito Abatement District (SLCMAD) has been conducting aerial applications using an organophosphate insecticide against adult mosquitoes for several decades. In order to evaluate a potential rotation product, aerial applications of Duet HD™, a pyrethroid, were conducted under operational conditions against wild populations of Aedes dorsalis and Culex tarsalis and against colony strains of Cx. pipiens and Cx. quinquefasciatus. The erratic wind patterns of the greater Salt Lake area did not prevent sufficient droplet deposition flux at 9 monitoring locations spread across a 5,120-acre (2,072 ha) spray block within rural habitats. Three separate aerial application trials showed great efficacy against Ae. dorsalis. In contrast, Cx. tarsalis exhibited inconsistent treatment-associated mortalities, suggesting the presence of less susceptible or resistant field populations as a result of spillover from agricultural or residential pyrethroid usage. Bottle bioassays to diagnose pyrethroid resistance using field-collected Cx. tarsalis indicated that some populations of this species, especially those closest to urban edges, failed to show adequate mortality in resistance assays. Despite challenging weather conditions, Duet HD worked reasonably well against susceptible mosquito species, and it may provide a crucial role as an alternative for organophosphate applications within specific and sensitive areas. However, its area-wide adoption into control applications by the SLCMAD could be problematic due to reduced impacts on the most important arboviral vector species, Cx. tarsalis, in this area. This study demonstrates the importance of testing mosquito control products under different operational environments and against potentially resistant mosquito populations by municipal mosquito control districts.

Evaluation of an open forecasting challenge to assess skill of West Nile virus neuroinvasive disease prediction.

BACKGROUND: West Nile virus (WNV) is the leading cause of mosquito-borne illness in the continental USA. WNV occurrence has high spatiotemporal variation, and current approaches to targeted control of the virus are limited, making forecasting a public health priority. However, little research has been done to compare strengths and weaknesses of WNV disease forecasting approaches on the national scale. We used forecasts submitted to the 2020 WNV Forecasting Challenge, an open challenge organized by the Centers for Disease Control and Prevention, to assess the status of WNV neuroinvasive disease (WNND) prediction and identify avenues for improvement. METHODS: We performed a multi-model comparative assessment of probabilistic forecasts submitted by 15 teams for annual WNND cases in US counties for 2020 and assessed forecast accuracy, calibration, and discriminatory power. In the evaluation, we included forecasts produced by comparison models of varying complexity as benchmarks of forecast performance. We also used regression analysis to identify modeling approaches and contextual factors that were associated with forecast skill. RESULTS: Simple models based on historical WNND cases generally scored better than more complex models and combined higher discriminatory power with better calibration of uncertainty. Forecast skill improved across updated forecast submissions submitted during the 2020 season. Among models using additional data, inclusion of climate or human demographic data was associated with higher skill, while inclusion of mosquito or land use data was associated with lower skill. We also identified population size, extreme minimum winter temperature, and interannual variation in WNND cases as county-level characteristics associated with variation in forecast skill. CONCLUSIONS: Historical WNND cases were strong predictors of future cases with minimal increase in skill achieved by models that included other factors. Although opportunities might exist to specifically improve predictions for areas with large populations and low or high winter temperatures, areas with high case-count variability are intrinsically more difficult to predict. Also, the prediction of outbreaks, which are outliers relative to typical case numbers, remains difficult. Further improvements to prediction could be obtained with improved calibration of forecast uncertainty and access to real-time data streams (e.g. current weather and preliminary human cases).

Microhabitat modeling of the invasive Asian longhorned tick (Haemaphysalis longicornis) in New Jersey, USA.

The Asian longhorned tick (Haemaphysalis longicornis) is a vector of multiple arboviral and bacterial pathogens in its native East Asia and expanded distribution in Australasia. This species has both bisexual and parthenogenetic populations that can reach high population densities under favorable conditions. Established populations of parthenogenetic H. longicornis were detected in the eastern United States in 2017 and the possible range of this species at the continental level (North America) based on climatic conditions has been modeled. However, little is known about factors influencing the distribution of H. longicornis at geographic scales relevant to local surveillance and control. To examine the importance of local physiogeographic conditions such as geology, soil characteristics, and land cover on the distribution of H. longicornis we employed ecological niche modeling using three machine learning algorithms - Maxent, Random Forest (RF), and Generalized Boosting Method (GBM) to estimate probability of finding H. longicornis in a particular location in New Jersey (USA), based on environmental predictors. The presence of H. longicornis in New Jersey was positively associated with Piedmont physiogeographic province and two soil types - Alfisols and Inceptisols. Soil hydraulic conductivity was the most important predictor explaining H. longicornis habitat suitability, with more permeable sandy soils with higher hydraulic conductivity being less suitable than clay or loam soils. The models were projected over the state of New Jersey creating a probabilistic map of H. longicornis habitat suitability at a high spatial resolution of 90×90 meters. The model's sensitivity was 87% for locations sampled in 2017-2019 adding to the growing evidence of the importance of soil characteristics to the survival of ticks. For the 2020-2022 dataset the model fit was 57%, suggestive of spillover to less optimal habitats or, alternatively, heterogeneity in soil characteristics at the edges of broad physiographic zones. Further modeling should incorporate abundance and life-stage information as well as detailed characterization of the soil at collection sites. Once critical parameters that drive the survival and abundance of H. longicornis are identified they can be used to guide surveillance and control strategies for this invasive species.

Rapid invasion and expansion of the Asian longhorned tick (Haemaphysalis longicornis) into a new area on Long Island, New York, USA.

Since its discovery in the United States in 2017, the Asian longhorned tick (Haemaphysalis longicornis) has been detected in most eastern states between Rhode Island and Georgia. Long Island, east of New York City, a recognized high-risk area for tick-borne diseases, is geographically close to New Jersey and New York sites where H. longicornis was originally found. However, extensive tick surveys conducted in 2018 did not identify H. longicornis on Long Island. In stark contrast, our 2022 tick survey suggests that H. longicornis has rapidly invaded and expanded in multiple surveying sites on Long Island (12 out of 17 sites). Overall, the relative abundance of H. longicornis was similar to that of lone star ticks, Amblyomma americanum, a previously recognized tick species abundantly present on Long Island. Interestingly, our survey suggests that H. longicornis has expanded within the Appalachian forest ecological zone of Long Island's north shore compared to the Pine Barrens located on the south shore of Long Island. The rapid invasion and expansion of H. longicornis into an insular environment are different from the historical invasion and expansion of two native tick species, Ixodes scapularis (blacklegged tick or deer tick) and A. americanum, in Long Island. The implications of H. longicornis transmitting or introducing tick-borne pathogens of public health importance remain unknown.

Mosquito Surveillance for West Nile Virus.

Identifying the mosquitoes responsible for transmitting human disease-causing pathogens is of critical importance for effective control of mosquito-borne outbreaks. West Nile virus is often transferred by adult female mosquitoes in the genus Culex, which deposit eggs in a variety of aquatic habitats throughout the world. Herein we describe several methodological approaches to monitor these species in nature, as well as offering details for data collection and analysis.

Modeling of historical and current distributions of lone star tick, Amblyomma americanum (Acari: Ixodidae), is consistent with ancestral range recovery.

The lone star tick, Amblyomma americanum L., is a three-host hard tick notorious for aggressive feeding behavior. In the early to mid-20th century, this species' range was mostly limited to the southern USA. Since the 1950s, A. americanum has been detected in many new localities in the western, northcentral, and northeastern regions of the country. To examine the influence of climate on this apparent expansion, we used historical (1748-1950) lone star locations from the literature and museum records to model areas suitable for this species based on past environmental conditions in the late 1800s - early 1900s. We then projected this model forward using present (2011-2020) climatic conditions and compared the two for evidence of climate-associated distributional shifts. A maximum entropy distribution or Maxent model was generated by using a priori selected climatic variables including temperature, precipitation, and vapor pressure deficit. Temperature and vapor pressure deficit were selected as the most important factors in creating a sensitive and specific model (success rate = 82.6 ± 6.1%) that had a good fit to the existing data and was significantly better than a random model [partial ROC (receiver operating characteristic) to AUC (area under the ROC curve) ratio = 1.97 ± 0.07, P < 0.001]. The present projected model was tested with an independent dataset of curated museum records (1952-2020) and found to be 95.6% accurate. Comparison of past and present models revealed > 98% A. americanum niche overlap. The model suggests that some areas along the western fringe are becoming less suitable for A. americanum, whereas areas in some Great Lakes and coastal northeastern regions are becoming more suitable, results that are compatible with possible effects of climate change. However, these changes are minor, and overall climate in North America does not appear to have changed in ways significant to A. americanum's distribution. These findings are consistent with an alternative hypothesis that recent changes in A. americanum's distribution are a result of this species re-occupying its historical range, driven predominantly by factors other than climate, such as shifts in land use and population densities of major hosts.

Field applications of granular and liquid pyrethroids, carbaryl, and IGRs to control the asian longhorned tick (Haemaphysalis longicornis) and impacts on nontarget invertebrates.

Few documented control strategies exist for the invasive tick, Haemaphysalis longicornis, despite its potential to reach extremely high numbers and vector human and animal pathogens. In 2020, we evaluated the effects of single applications of five granular and liquid acaricides on H. longicornis in a public park in northern New Jersey. Acaricides tested included pyrethroids (lambda-cyhalothrin, bifenthrin), a carbamate (carbaryl), and the insect growth regulators (IGRs) pyriproxyfen and novaluron. We also monitored the impact of each treatment on non-target soil and above-ground invertebrate species using pitfall and sticky traps, respectively. We recorded over 70,000 H. longicornis ticks in the study area from July to October 2020. An average of 99% control was achieved with lambda-cyhalothrin spray and 95% with granular bifenthrin. In contrast, granular carbaryl did not significantly reduce any life stages of H. longicornis. The IGR (pyriproxyfen/novaluron) resulted in a significant 45% reduction of the larval stage following treatments in July. No other stages were significantly impacted by pyriproxyfen alone or in combination with novaluron. Analysis of non-target species revealed that the community composition of soil-dwelling arthropods was strongly impacted by pyrethroid treatments and, to a lesser extent, by the carbamate treatment. The granular pyrethroid bifenthrin had more pronounced effects and impacted a broader range of non-target groups in the pitfall traps than the liquid pyrethroid lambda-cyhalothrin. Arthropod groups that were negatively impacted included Isopoda, Formicidae, Coleoptera, Araneae, Acari, and Grylloidea. Collembola numbers, however, were elevated in both pyrethroid treatments. The community composition of arthropods collected on the above-ground sticky traps was strongly impacted only in the liquid lambda-cyhalothrin treatment. The primary groups impacted in the sticky trap analysis were Collembola and Hemiptera. Community composition in traps remained distinct in the pyrethroid treatments through the entire survey period up to 62 days post-treatment. The results of this study indicate that pyrethroid acaricides were highly effective at controlling H. longicornis, while other compounds, including carbaryl and IGRs, did not achieve consistent levels of control. Further research is needed to find effective and environmentally sustainable alternatives. Integrated management programs can include the judicious use of pyrethroids to control H. longicornis.

Laboratory evaluation of sugar alcohols for control of mosquitoes and other medically important flies.

Insecticide application for vector control is the most controversial component of a public health program due to concerns about environmental and human health safety. One approach to overcome this challenge is the use of environmentally benign active ingredients. Among the most promising emerging strategies are attractive toxic sugar baits. Sugar alcohols-naturally occurring molecules safe for human consumption but potentially toxic to insects when ingested, have received increased attention for use with this approach. For this study, we screened the toxicity of four different sugar alcohols on several mosquito species, a biting midge, and a filth fly. Sugar alcohol mortalities exceeded those in the sucrose (positive control) only group. However, only erythritol and highly concentrated xylitol induced mortalities exceeding those in the water only (negative control) treatment ranging from approximately 40-75%. Formulations containing erythritol and xylitol should be further investigated under field conditions for efficacy in reducing populations of biting flies and for assessing potential non-target impacts.

The Original Scientific Description of the Lone Star Tick (Amblyomma americanum, Acari: Ixodidae) and Implications for the Species' Past and Future Geographic Distributions.

Amblyomma americanum L. is an important vector in North America originally described by Linnaeus based on Pehr Kalm's 1754 report. While Kalm's 'Travels into North America' is well known, his 1754 report remains obscure. Some authors were skeptical that Kalm referred to A. americanum because he encountered them at sites farther north outside of the species' range. However, the details in 1754 report leave no doubt that Kalm described lone star ticks. In this historical review, we provide support for Kalm's identification using a modern translation of his 1754 report and other sources. We also delineate distributional changes of lone star ticks from the pre-colonization era to the present and interpret them in the context of large-scale anthropogenic changes in the landscape. In this framework, the lone star tick's current northward expansion is a recolonization of their former range. Extensive deforestation and extirpation of their principal host species, white-tailed deer, led to A. americanum's disappearance from the northern parts of its range by the 20th century. Subsequent recolonization by second-growth forest and increases in white-tailed deer populations by the mid-20th century is now allowing A. americanum to reclaim its former range. These changes in the land appear to be the driving force behind A. americanum's present expansion. Understanding this species' history and the factors contributing to its current expansion will enable better predictions about its future distribution and potential to transmit human pathogens.

A proposed framework for the development and qualitative evaluation of West Nile virus models and their application to local public health decision-making.

West Nile virus (WNV) is a globally distributed mosquito-borne virus of great public health concern. The number of WNV human cases and mosquito infection patterns vary in space and time. Many statistical models have been developed to understand and predict WNV geographic and temporal dynamics. However, these modeling efforts have been disjointed with little model comparison and inconsistent validation. In this paper, we describe a framework to unify and standardize WNV modeling efforts nationwide. WNV risk, detection, or warning models for this review were solicited from active research groups working in different regions of the United States. A total of 13 models were selected and described. The spatial and temporal scales of each model were compared to guide the timing and the locations for mosquito and virus surveillance, to support mosquito vector control decisions, and to assist in conducting public health outreach campaigns at multiple scales of decision-making. Our overarching goal is to bridge the existing gap between model development, which is usually conducted as an academic exercise, and practical model applications, which occur at state, tribal, local, or territorial public health and mosquito control agency levels. The proposed model assessment and comparison framework helps clarify the value of individual models for decision-making and identifies the appropriate temporal and spatial scope of each model. This qualitative evaluation clearly identifies gaps in linking models to applied decisions and sets the stage for a quantitative comparison of models. Specifically, whereas many coarse-grained models (county resolution or greater) have been developed, the greatest need is for fine-grained, short-term planning models (m-km, days-weeks) that remain scarce. We further recommend quantifying the value of information for each decision to identify decisions that would benefit most from model input.

Heterodissemination: precision insecticide delivery to mosquito larval habitats by cohabiting vertebrates.

Conventional larvicide delivery strategies originally developed for permanent and floodwater mosquitoes have proved suboptimal in the small, scattered, and cryptic larval habitats preferred by container-inhabiting Aedes mosquitoes. New methods such as autodissemination, wherein adult mosquitoes spread insecticides to their own larval habitats, have been under study. Another novel delivery method termed heterodissemination, i.e. larvicide delivery by other species sharing the same habitats, has also been proposed. We conducted a proof-of-concept study with four independent experiments using American bullfrogs (Lithobates catesbeianus) and green frogs Lithobates clamitans as carriers of pyriproxyfen, an insect growth regulator, under semi-field conditions in three different locations, two in New Jersey, and one in Utah. Frogs with attached slow-release pyriproxyfen tablets were introduced into outdoor enclosures with water containers. Water samples from the containers were periodically tested using larval Aedes albopictus and Culex pipiens mosquitoes to assess mortality and percent eclosure inhibition. Overall pupal mortality [95% credible intervals] estimated by Bayesian analysis for the treatment group was 73.4% [71.3-75.2] compared to 4.1% [2.9-5.5] for the control group. Mortality within treatment groups in four different experiments ranged from 41 to 100%, whereas control mortalities ranged from 0.5% to 11%. We conclude that heterodissemination is a promising and effective approach deserving of further study.

InsectChange: a global database of temporal changes in insect and arachnid assemblages.

Insects are the most ubiquitous and diverse group of eukaryotic organisms on Earth, forming a crucial link in terrestrial and freshwater food webs. They have recently become the subject of headlines because of observations of dramatic declines in some places. Although there are hundreds of long-term insect monitoring programs, a global database for long-term data on insect assemblages has so far remained unavailable. In order to facilitate synthetic analyses of insect abundance changes, we compiled a database of long-term (≥10 yr) studies of assemblages of insects (many also including arachnids) in the terrestrial and freshwater realms. We searched the scientific literature and public repositories for data on insect and arachnid monitoring using standardized protocols over a time span of 10 yr or longer, with at least two sampling events. We focused on studies that presented or allowed calculation of total community abundance or biomass. We extracted data from tables, figures, and appendices, and, for data sets that provided raw data, we standardized trapping effort over space and time when necessary. For each site, we extracted provenance details (such as country, state, and continent) as well as information on protection status, land use, and climatic details from publicly available GIS sources. In all, the database contains 1,668 plot-level time series sourced from 165 studies with samples collected between 1925 and 2018. Sixteen data sets provided here were previously unpublished. Studies were separated into those collected in the terrestrial realm (103 studies with a total of 1,053 plots) and those collected in the freshwater realm (62 studies with 615 plots). Most studies were from Europe (48%) and North America (29%), with 34% of the plots located in protected areas. The median monitoring time span was 19 yr, with 12 sampling years. The number of individuals was reported in 129 studies, the total biomass was reported in 13 studies, and both abundance and biomass were reported in 23 studies. This data set is published under a CC-BY license, requiring attribution of the data source. Please cite this paper if the data are used in publications, and respect the licenses of the original sources when using (part of) their data as detailed in Metadata S1: Table 1.

A case of pediatric northern black widow spider (Latrodectus variolus) bite in New York, USA.

Black widow spiders (Latrodectus spp.) are generally perceived as inhabiting southern or western United States. However, some species, such as the northern black widow, Latrodectus variolus, are also found in more temperate regions of USA and adjacent Canada. This species also appears to expand its range northward with the climate change increasing the risk of human encounters. To our knowledge, this is the first case report of a pediatric envenomation from a northern black widow bite in northeastern US. Medical practitioners often fail to recognize the signs and symptoms of latrodectism delaying effective treatment. Even in more northern locales, healthcare workers should take black widow spider envenomation under consideration in the differential diagnosis particularly with suspected exposure.

Serologic Survey of Mosquito-Borne Viruses in Hunter-Harvested White-Tailed Deer (Odocoileus virginianus), New York State.

Sera from white-tailed deer (WTD, Odocoileus virginianus) hunter-harvested throughout New York State (NYS), 2007-2015, were tested by plaque reduction neutralization for antibodies against nine mosquito-borne viruses from the families Peribunyaviridae, Flaviviridae, and Togaviridae. Overall, 76.1% (373/490) of sampled WTD were seropositive against at least one virus, and 38.8% were exposed to multiple viruses. The seropositivity rate in adult WTD (78.0%) was significantly greater (P < 0.0001) than that in fawns (47.7%). Neutralizing antibodies against California serogroup viruses were most common in WTD sampled across all regions (67.1%), followed by the Bunyamwera serogroup (BUN) (37.6%). Jamestown Canyon and Cache Valley orthobunyaviruses were responsible for most California and BUN infections, respectively. Seroprevalence rates to West Nile virus were higher in samples originating from Long Island (LI) (19.0%) than in those originating from the central (7.3%), western (5.0%), and Hudson Valley (4.4%) regions of NYS. Antibodies to Eastern equine encephalitis virus were seen primarily in WTD from central NYS (5.1%), where annual enzootic activity occurs, but low rates were documented in western NYS (1.4%) and LI (1.7%). Low rates of Potosi and LaCrosse orthobunyavirus, and Highlands J virus antibodies were detected over the course of this investigation. St. Louis encephalitis virus (or a closely related virus) antibodies were detected in samples collected from central and western NYS, suggesting local virus transmission despite a lack of evidence from routine mosquito surveillance. Serologic results demonstrate the value of WTD in NYS as an indicator of arbovirus distribution and recent transmission on a relatively fine spatial scale.

Emerging tick-borne pathogens of public health importance: a mini-review.

Ticks are the most important vectors of human pathogens, leading to increased public health burdens worldwide. Tick-borne pathogens include viruses (e.g. tick-borne encephalitis and Powassan); bacteria, such as the causative agents of Lyme disease, spotted fever rickettsiosis and human anaplasmosis; and malaria-like protozoan parasites causing babesiosis. Tick-borne diseases are emerging due to the geographical expansion of their tick vectors, especially in the northern hemisphere. Two examples of this phenomenon are Ixodes scapularis and Amblyomma americanum, which have expanded their ranges in the USA in recent decades and are responsible for the continuous emergence of Lyme disease and human ehrlichiosis, respectively. This phenomenon is also occurring worldwide and is reflected by the increasing number of tick-borne encephalitis and haemorrhagic fever cases in Europe and Asia. In this review, we provide a concise synopsis of the most medically important tick-borne pathogen worldwide, with a particular emphasis on emerging public health threats.

Large-Scale Operational Pyriproxyfen Autodissemination Deployment to Suppress the Immature Asian Tiger Mosquito (Diptera: Culicidae) Populations.

Effective suppression of container-inhabiting Asian Tiger [Aedes albopictus (Skuse)] (Diptera: Culicidae) and yellow fever [Aedes aegypti (L.)] (Diptera: Culicidae) mosquitoes presents one of the most intractable problems for modern mosquito control. Traditional tools often fail to control populations of these mosquito species, and are prohibitively expensive or have negative environmental impacts. Novel approaches and tools are urgently needed for integrated container-inhabiting mosquito management programs. One of the most promising techniques is autodissemination. We present the results of a long-term large-scale study conducted in a temperate urbanized environment representing typical Ae. albopictus habitats. Three treatment sites with autodissemination stations and three nearby reference sites were monitored for eggs, immature, and adult mosquitoes over a period of 3 yr from 2014 to 2016. Elevated larval and pupal mortality of 12-19% on average was the most notable outcome in sentinel cups of the treatment sites. The number of eggs in the treatment sites was significantly reduced in 2014, but not in 2015 or 2016. Adult populations remained similar in treatment and reference sites throughout the study. The impact of autodissemination on mosquito populations was lower than reported by previous investigations. Technical and logistical problems associated with wider coverage and working in multiple urban neighborhoods contributed to reduced efficacy. Incorporating autodissemination with routine mosquito control operations and commercializing this methodology for general public use will require further research on combining this tool with other novel or conventional technologies.

Behavior manipulation of mosquitoes by a mermithid nematode.

We examined manipulation of mosquito behavior by the parasitic mermithid nematode, Strelkovimermis spiculatus. This nematode species typically infects early instar host larvae and emerges after parasitic development to kill last-instar larvae. Parasitized adults, however, have occasionally been reported from field collections. We obtained low rates (1.7-11.5%) of parasitized adults in laboratory exposures only when Culex pipiens pipiens fourth-instar larvae nearing pupation were exposed to infective nematodes. This did not allow an adequate interval for parasitic development in immature host stages. Parasitized adult females in a multiple-choice assay were three times more likely to seek water than a blood source (63.1 vs. 20.5%), whereas uninfected females were twice as likely to seek blood than water (64%3.9 vs. 32.6%). This altered host behavior benefits the parasite by providing the only mechanism for dispersal and colonization of new host habitats while concurrently avoiding risks from the defensive behaviors associated with blood-feeding. Behavioral alternation in Cx. p. pipiens larval hosts was also examined using larvae infected as second instars to allow for a normal duration of parasitic development. As larvae neared pupation and parasite emergence, parasitized larvae became more spatially aggregated than unparasitized larvae. This altered host behavior benefits the parasite by providing a corresponding increase in post-parasite aggregation, which facilitates formation of large mating clusters and concomitantly reproductive success. Parasites derive fitness gains by overriding host autonomy, whereas hosts have zero fitness once parasitism is established, suggesting a coevolutionary response is inoperative and that the behavioral modifications may be adaptive.

West Nile Virus Mosquito Vectors in North America.

In North America, the geographic distribution, ecology, and vectorial capacity of a diverse assemblage of mosquito species belonging to the genus Culex determine patterns of West Nile virus transmission and disease risk. East of the Mississippi River, mostly ornithophagic Culex pipiens L. complex mosquitoes drive intense enzootic transmission with relatively small numbers of human cases. Westward, the presence of highly competent Culex tarsalis (Coquillett) under arid climate and hot summers defines the regions with the highest human risk. West Nile virus human risk distribution is not uniform geographically or temporally within all regions. Notable geographic 'hotspots' persist with occasional severe outbreaks. Despite two decades of comprehensive research, several questions remain unresolved, such as the role of non-Culex bridge vectors, which are not involved in the enzootic cycle, but may be involved in virus transmission to humans. The absence of bridge vectors also may help to explain the frequent lack of West Nile virus 'spillover' into human populations despite very intense enzootic amplification in the eastern United States. This article examines vectorial capacity and the eco-epidemiology of West Nile virus mosquito vectors in four geographic regions of North America and presents some of the unresolved questions.