Introduction Of West Nile Virus Research Articles

Impacts of ground ultra-low volume adulticide applications on Culex pipiens and Culex restuans (Diptera: Culicidae) abundance, age structure, and West Nile virus infection in Cook County, Illinois.

Since the introduction of West Nile virus (WNV) to the United States over 20 years ago, thousands of cases of human disease and death have been reported. Yearly seasonal outbreaks continue to persist, and the city and suburbs of Chicago, Illinois, is considered a "hot spot" for WNV activity. To interrupt WNV transmission, ground ultra-low volume (ULV) adulticide applications are regularly used to reduce Culex pipiens L. and Culex restuans Theobold (Diptera: Culicidae) abundance and infection. The real-world effectiveness of adulticide applications has not been comprehensively assessed, and prior studies, including our own investigation, have yielded inconclusive or conflicting results. Therefore, we expanded our prior work and evaluated the effects of 5 sequential weekly truck-mounted ULV adulticide applications in large residential areas in the northern suburbs of Chicago, Illinois, in 2019 and 2020. Each day, Cx. pipiens and Cx. restuans host-seeking and gravid mosquitoes were collected to assess abundance, age structure, and WNV infection rates. Adulticide applications resulted in significant reductions of both host-seeking and gravid abundance on the night of treatment. The reduction in host-seeking mosquitoes was followed by a reduction in gravid mosquitoes trapped 3 and 4 days after adulticide application and an increase in the proportion of nulliparous mosquitoes. WNV infection rates were significantly reduced in treatment sites as compared to untreated sites when infection rates were higher in 2020. This large-scale study provides comprehensive evidence that ground ULV adulticide applications are an effective tool in an integrated mosquito management program for combating WNV vectors and infection risk.

Many-core algorithms for high-dimensional gradients on phylogenetic trees.

Advancements in high-throughput genomic sequencing are delivering genomic pathogen data at an unprecedented rate, positioning statistical phylogenetics as a critical tool to monitor infectious diseases globally. This rapid growth spurs the need for efficient inference techniques, such as Hamiltonian Monte Carlo (HMC) in a Bayesian framework, to estimate parameters of these phylogenetic models where the dimensions of the parameters increase with the number of sequences N. HMC requires repeated calculation of the gradient of the data log-likelihood with respect to (wrt) all branch-length-specific (BLS) parameters that traditionally takes O(N2) operations using the standard pruning algorithm. A recent study proposes an approach to calculate this gradient in O(N), enabling researchers to take advantage of gradient-based samplers such as HMC. The CPU implementation of this approach makes the calculation of the gradient computationally tractable for nucleotide-based models but falls short in performance for larger state-space size models, such as Markov-modulated and codon models. Here, we describe novel massively parallel algorithms to calculate the gradient of the log-likelihood wrt all BLS parameters that take advantage of graphics processing units (GPUs) and result in many fold higher speedups over previous CPU implementations. We benchmark these GPU algorithms on three computing systems using three evolutionary inference examples exploring complete genomes from 997 dengue viruses, 62 carnivore mitochondria and 49 yeasts, and observe a >128-fold speedup over the CPU implementation for codon-based models and >8-fold speedup for nucleotide-based models. As a practical demonstration, we also estimate the timing of the first introduction of West Nile virus into the continental Unites States under a codon model with a relaxed molecular clock from 104 full viral genomes, an inference task previously intractable. We provide an implementation of our GPU algorithms in BEAGLE v4.0.0 (https://github.com/beagle-dev/beagle-lib), an open-source library for statistical phylogenetics that enables parallel calculations on multi-core CPUs and GPUs. We employ a BEAGLE-implementation using the Bayesian phylogenetics framework BEAST (https://github.com/beast-dev/beast-mcmc).

Gone (and spread) with the birds: Can chorotype analysis highlight the spread of West Nile virus within the Afro-Palaearctic flyway?

West Nile virus (WNV) is a globally significant vector-borne disease that is primarily transmitted between birds and mosquitoes. Recently, there has been an increase in WNV in southern Europe, with new cases reported in more northern regions. Bird migration plays a crucial role in the introduction of WNV in distant areas. To better understand and address this complex issue, we adopted a One Health approach, integrating clinical, zoological, and ecological data. We analyzed the role of migratory birds in the Palaearctic-African region in the spread of WNV across Africa and Europe. We categorized bird species into breeding and wintering chorotypes based on their distribution during the breeding season in the Western Palaearctic and the wintering season in the Afrotropical region, respectively. By linking these chorotypes to the occurrence of WNV outbreaks in both continents throughout the annual bird migration cycle, we investigated the relationship between migratory patterns and virus spread. We demonstrate that WNV-risk areas are interconnected through the migration of birds. We identified a total of 61 species that potentially contribute to the intercontinental spread of the virus or its variants, as well as pinpointed high-risk areas for future outbreaks. This interdisciplinary approach, which considers the interconnectedness of animals, humans, and ecosystems, represents a pioneering effort to establish connections between zoonotic diseases across continents. The findings of our study can aid in anticipating the arrival of new WNV strains and predicting the occurrence of other re-emerging diseases. By incorporating various disciplines, we can enhance our understanding of these complex dynamics and provide valuable insights for proactive and comprehensive disease management strategies.

Rise and fall of St. Louis Encephalitis Virus in Florida

St. Louis encephalitis virus has had a fascinating history in Florida. The virus was introduced into the Miami area in the early 1950s. This introduction resulted in two human outbreaks, one in 1952 and a second in 1958. Three urban SLE epidemics were reported in the Tampa Bay area in 1959, 1961, and 1962. The virus virtually disappeared from the state until 1977 when a widespread rural SLE epidemic was reported in 20 Florida counties with 110 confirmed human cases. An almost identical rural SLE epidemic was reported in 1990 when 226 human SLE cases were reported in 28 Florida counties. Following the introduction of West Nile virus into Florida in 2001, reports of SLEV transmission to sentinel chickens and humans decreased dramatically. Except for a 2014 focal outbreak of two SLE cases in Duval County, only sporadic transmission of SLEV to humans and sentinel chickens was reported in the state between 1998 and 2022.

Comparison of Pathology and Immunohistochemistry in Natural and Experimental West Nile Virus Infections in Ruffed Grouse (Bonasa umbellus).

Ruffed Grouse (Bonasa umbellus) populations have declined in much of the eastern US in recent decades. Research suggests that West Nile virus (WNV) contributed to these declines, based on decreasing population indices temporally correlated to WNV introduction into North America, high morbidity and mortality in experimentally infected Ruffed Grouse, and relatively low statewide seroprevalence concurrent with high WNV vector indices. We describe lesions and relevant diagnostic findings in six, free-ranging Ruffed Grouse that directly or indirectly died of natural WNV infection and compare results to experimentally infected Ruffed Grouse. All naturally infected grouse were found moribund or dead from August to December 2015-18 in the northeastern US; 4/6 grouse were subadults. Necropsy, histopathology, immunohistochemistry, and/or ancillary tests contributed to the diagnosis of WNV-associated disease in all cases. The most common lesions in naturally infected grouse were myocardial inflammation, degeneration, and/or necrosis, consistent with lesions in experimentally infected grouse. Evidence of trauma in 6/6 naturally infected grouse suggests the possibility of WNV-associated morbidity as a predisposing factor. Diagnostic findings in 3/6 naturally infected grouse were consistent with experimentally infected grouse with severe disease 7-8 d postinoculation; the remaining naturally infected birds had similar findings to experimentally infected subclinical grouse. Our results further support the notion that WNV contributes to mortality of free-ranging Ruffed Grouse and may be used to improve surveillance strategies and population-level management approaches.

Susceptibility of wild turkeys (Meleagris gallopavo) to experimental West Nile virus infection

ABSTRACT Since the introduction of West Nile virus (WNV) to North America in 1999, WNV is estimated to have contributed to population-level declines in numerous avian species. However, the potential impacts of this virus on many free-ranging upland game bird species, including the wild turkey (Meleagris gallopavo), which is undergoing regional population declines, remain unknown. Herein, two age groups (∼5 to 6 weeks and ∼15 to 16 weeks post-hatch) of juvenile wild turkeys were subcutaneously inoculated with WNV, sampled daily from 1 to 7 days post-inoculation (dpi), and euthanized on 14 dpi. No clinical signs and minimal gross lesions were attributable to WNV infection. Peak viraemia titres were similar between age groups (<101.7 to 104.6 plaque-forming units [PFU]/ml), but the duration of viraemia was longer in the old group (3–4 days) than in the young group (0–3 days). Intermittent oral and/or cloacal viral shedding from 2 to 7 dpi was detected in both age groups. No infectious virus was detected in the heart, brain, kidney, skeletal muscle, spleen, and feathers from WNV-inoculated turkeys euthanized on 14 dpi. All WNV-inoculated birds seroconverted by 14 dpi, as well as two co-housed sham-inoculated birds. The most consistent microscopic lesions among all WNV-inoculated birds were mild lymphoplasmacytic myocarditis and encephalitis. Minimal immunohistochemical labelling was detected in tissues in addition to scant macrophages within the blood, spleen, and bone marrow. These data suggest WNV is unlikely to pose a significant risk to wild turkey populations, although the possibility remains that WNV may indirectly decrease fitness or predispose wild turkeys to other health stressors. RESEARCH HIGHLIGHTS Clinical disease was not observed in wild turkeys experimentally infected with WNV. Pathology attributed to WNV was mild and included brain and heart inflammation. Viraemias suggest WNV-infected wild turkeys do not play a role in WNV transmission. No age-associated differences in WNV clinical disease or pathology were observed.

Fact Sheet: West Nile virus

Introduced to the United States in 1999, West Nile virus (WNV) is transmitted from mosquitoes to humans and equines. With over 52,000 reported human infections, it is the most frequent cause of mosquito-borne disease in the continental United States. Many mosquito species can transmit WNV, and many wildlife species can be infected with the virus and transmit it to biting mosquitoes, which makes identifying WNV's high risk areas or time periods complex. The US equine industry, domestic pets, and livestock are all at risk from harm from WNV. Many equine WNV epizootics have occurred since 1999, with more than 30,000 equine cases reported in the United States. Since WNV's introduction to Florida in 2001, infections have been reported in a total of 460 people and 757 equines. This publication provides information about WNV to the general public, researchers and stakeholders in mosquito control, and public health professionals.

Optimized In Vitro CRISPR/Cas9 Gene Editing Tool in the West Nile Virus Mosquito Vector, Culex quinquefasciatus.

Simple SummaryCulex mosquitoes are responsible for many established and emerging mosquito-borne diseases worldwide. These mosquitoes serve as the principal global transmission vector of West Nile virus, which is the leading cause of mosquito-borne disease in humans in the United States. Unfortunately, effective therapeutic drugs and vaccines are still lacking for most mosquito-borne diseases, and current vector controls (e.g., insecticide use) have fallen short of eradicating the disease burden. These concerns highlight the importance of developing novel strategies to prevent arbovirus transmission by Culex mosquitoes, especially as the prevalence of Culex-borne diseases increases globally. Underlying such approaches is often an improved understanding of virus–mosquito interactions. To investigate the putative antiviral role of Culex immune genes, we developed and characterized a Culex-optimized CRISPR/Cas9 plasmid for use in Culex quinquefasciatus-derived (Hsu) cell cultures. We showed that this newly constructed plasmid allows for efficient and reliable codelivery of all CRISPR reagents in vitro in a single plasmid system. These findings suggest that this tool may serve as a valuable resource for the establishment of mutant Hsu-derived cell populations, enabling the identification of mosquito host genes involved in antiviral response. Elucidating gene functions supports the development of alternative gene-based vector control strategies for Culex mosquitoes.Culex quinquefasciatus mosquitoes are a globally widespread vector of multiple human and animal pathogens, including West Nile virus, Saint Louis encephalitis virus, and lymphatic filariasis. Since the introduction of West Nile virus to the United States in 1999, a cumulative 52,532 cases have been reported to the CDC, including 25,849 (49.2%) neuroinvasive cases and 2456 (5%) deaths. Viral infections elicit immune responses in their mosquito vectors, including the RNA interference (RNAi) pathway considered to be the cornerstone antiviral response in insects. To investigate mosquito host genes involved in pathogen interactions, CRISPR/Cas9-mediated gene-editing can be used for functional studies of mosquito-derived cell lines. Yet, the tools available for the study of Cx. quinquefasciatus-derived (Hsu) cell lines remain largely underdeveloped compared to other mosquito species. In this study, we constructed and characterized a Culex-optimized CRISPR/Cas9 plasmid for use in Hsu cell cultures. By comparing it to the original Drosophila melanogaster CRISPR/Cas9 plasmid, we showed that the Culex-optimized plasmid demonstrated highly efficient editing of the genomic loci of the RNAi proteins Dicer-2 and PIWI4 in Hsu cells. These new tools support our ability to investigate gene targets involved in mosquito antiviral response, and thus the future development of gene-based vector control strategies.

West Nile Virus and Eastern Equine Encephalitis Virus High Probability Habitat Identification for the Selection of Sentinel Chicken Surveillance Sites in Florida.

To mitigate the effects of West Nile virus (WNV) and eastern equine encephalitis virus (EEEV), the state of Florida conducts a serosurveillance program that uses sentinel chickens operated by mosquito control programs at numerous locations throughout the state. Coop locations were initially established to detect St. Louis encephalitis virus (SLEV), and coop placement was determined based on the location of human SLEV infections that occurred between 1959 and 1977. Since the introduction of WNV into Florida in 2001, WNV has surpassed SLEV as the primary arbovirus in Florida. Identifying high probability locations for WNV and EEEV transmission and relocating coops to areas of higher arbovirus activity would improve the sensitivity of the sentinel chicken surveillance program. Using 2 existing models, this study conducted an overlay analysis to identify areas with high probability habitats for both WNV and EEEV activity. This analysis identified approximately 7,800 km2 (about 4.5% of the state) as high probability habitat for supporting both WNV and EEEV transmission. Mosquito control programs can use the map resulting from this analysis to improve their sentinel chicken surveillance programs, increase the probability of virus detection, reduce operational costs, and allow for a faster, targeted response to virus detection.

A new cluster of West Nile virus lineage 1 identified in Spain

<h3>Purpose</h3> West Nile Virus (WNV) is a zoonotic arbovirus worldwide spread. In Spain, its active circulation provokes outbreaks in birds, horses and lately in humans. In September 2017, an outbreak affecting raptors, in the province of Málaga (Andalusia), was investigated. One northern goshawk (<i>Accipiter gentilis</i>) died, while two peregrine falcons (<i>Falco peregrinus</i>) recovered. Here we report the isolation and characterization of a new WNV lineage 1 variant from this outbreak. <h3>Methods & Materials</h3> Heart, feathers, brain and kidney samples from the goshawk were collected and firstly analysed by real-time RT-PCR for WNV (Jiménez-Clavero <i>et al</i>., 2006, https://doi.org/doi:10.1177/104063870601800505). Virus isolation was performed in BSR and Vero cells. The whole genome of the isolate was sequenced by Sanger method. Phylogenetic analyses were performed including a wide range of representatives of: 1) all described WNV lineages, and 2) the different clusters of WNV lineage 1. <h3>Results</h3> All samples from the goshawk were positive for WNV lineage 1 by RT-PCR. Virus isolation was successful in heart and brain samples. A full genome sequence of 10,919 nt was obtained for the WNV isolated from the brain (named Spain/2017/NG-b). Phylogenetic analyses confirmed that Spain/2017/NG-b belonged to WNV clade 1a. Surprisingly, this isolate was placed neither in the cluster 2 of this clade, which comprises most of the recent European and Mediterranean strains (including the preceding Spanish ones), nor in other closest clusters. When analysing the polyprotein, amino acid substitutions that define WNV clusters 1-4 were not found in Spain/2017/NG-b isolate (May <i>et al.</i>, 2011, https://doi.org/10.1128/jvi.01963-10). Moreover, it presented 12 unique residues compared to cluster 2 viruses. <h3>Conclusion</h3> These results strongly support that Spain/2017/NG-b represents a new, independent introduction of WNV into the Western Mediterranean since it showed no close relationship to other strains from this region. Therefore, we propose to associate this variant to a new cluster 7 within WNV clade 1a. Finally, a WNV outbreak occurred in equids in December 2017, nearby the raptors´ outbreak above mentioned, suggests the ability of this new variant to infect and cause disease also in horses.

Integrating Spatiotemporal Epidemiology, Eco-Phylogenetics, and Distributional Ecology to Assess West Nile Disease Risk in Horses.

Mosquito-borne West Nile virus (WNV) is the causative agent of West Nile disease in humans, horses, and some bird species. Since the initial introduction of WNV to the United States (US), approximately 30,000 horses have been impacted by West Nile neurologic disease and hundreds of additional horses are infected each year. Research describing the drivers of West Nile disease in horses is greatly needed to better anticipate the spatial and temporal extent of disease risk, improve disease surveillance, and alleviate future economic impacts to the equine industry and private horse owners. To help meet this need, we integrated techniques from spatiotemporal epidemiology, eco-phylogenetics, and distributional ecology to assess West Nile disease risk in horses throughout the contiguous US. Our integrated approach considered horse abundance and virus exposure, vector and host distributions, and a variety of extrinsic climatic, socio-economic, and environmental risk factors. Birds are WNV reservoir hosts, and therefore we quantified avian host community dynamics across the continental US to show intra-annual variability in host phylogenetic structure and demonstrate host phylodiversity as a mechanism for virus amplification in time and virus dilution in space. We identified drought as a potential amplifier of virus transmission and demonstrated the importance of accounting for spatial non-stationarity when quantifying interaction between disease risk and meteorological influences such as temperature and precipitation. Our results delineated the timing and location of several areas at high risk of West Nile disease and can be used to prioritize vaccination programs and optimize virus surveillance and monitoring.

West Nile Virus Epidemic in Germany Triggered by Epizootic Emergence, 2019.

One year after the first autochthonous transmission of West Nile virus (WNV) to birds and horses in Germany, an epizootic emergence of WNV was again observed in 2019. The number of infected birds and horses was considerably higher compared to 2018 (12 birds, two horses), resulting in the observation of the first WNV epidemy in Germany: 76 cases in birds, 36 in horses and five confirmed mosquito-borne, autochthonous human cases. We demonstrated that Germany experienced several WNV introduction events and that strains of a distinct group (Eastern German WNV clade), which was introduced to Germany as a single introduction event, dominated mosquito, birds, horse and human-related virus variants in 2018 and 2019. Virus strains in this clade are characterized by a specific-Lys2114Arg mutation, which might lead to an increase in viral fitness. Extraordinary high temperatures in 2018/2019 allowed a low extrinsic incubation period (EIP), which drove the epizootic emergence and, in the end, most likely triggered the 2019 epidemic. Spatiotemporal EIP values correlated with the geographical WNV incidence. This study highlights the risk of a further spread in Germany in the next years with additional human WNV infections. Thus, surveillance of birds is essential to provide an early epidemic warning and thus, initiate targeted control measures.

Population structure and Admixture of Culex pipiens complex in New York City using Genotyping By Sequencing

The first cases of encephalitis associated with West Nile Virus (WNV) in New York City presented in 1999. Birds serve as the primary host for WNV and transmit the pathogen through mosquitoes of the Culex pipiens complex. The Culex pipiens form pipiens (Cx. pipiens f. pipiens) are exclusively ornithophilic (avian feeding) and require blood meal in order to reproduce. Cx. pipiens f. molestus are mammophilic and feed almost exclusively on mammals. WNV transmission to mammals occurs through a succession of feeding between an infected bird and subsequent mammal. While the two forms of Cx. pipiens have preferential food sources, they are capable of hybridizing and become indiscriminate in blood meal. Biosurveillance and clinical presentation in humans has been consistent since the introduction of WNV 20 years ago. While these two forms are physically identical, they differ fundamentally in behavior. Traditionally, a single locus STR (CQ11) has been the defining disambiguator of the two forms. However, this does not illustrate any level of hybridization that would provide insight to bloodmeal preference. Understanding the amount of admixture within NYC populations can aid in the identification of these forms and in vector control processes. This study uses Genotyping by Sequencing (GBS) through Restriction site associated DNA Sequencing (RAD‐Seq) in order to identify the level of admixture within the NYC Culex populations as well as the levels of ancestral states. Admixture analysis has identified unique SNPs that illuminates some underlying behavioral differences between the two forms, as well as evolutionary divergence.

Introduction, Spread, and Establishment of West Nile Virus in the Americas.

The introduction of West Nile virus (WNV) to North America in 1999 and its subsequent rapid spread across the Americas demonstrated the potential impact of arboviral introductions to new regions, and this was reinforced by the subsequent introductions of chikungunya and Zika viruses. Extensive studies of host-pathogen-vector-environment interactions over the past two decades have illuminated many aspects of the ecology and evolution of WNV and other arboviruses, including the potential for pathogen adaptation to hosts and vectors, the influence of climate, land use and host immunity on transmission ecology, and the difficulty in preventing the establishment of a zoonotic pathogen with abundant wildlife reservoirs. Here, we focus on outstanding questions concerning the introduction, spread, and establishment of WNV in the Americas, and what it can teach us about the future of arboviral introductions. Key gaps in our knowledge include the following: viral adaptation and coevolution of hosts, vectors and the virus; the mechanisms and species involved in the large-scale spatial spread of WNV; how weather modulates WNV transmission; the drivers of large-scale variation in enzootic transmission; the ecology of WNV transmission in Latin America; and the relative roles of each component of host-virus-vector interactions in spatial and temporal variation in WNV transmission. Integrative studies that examine multiple factors and mechanisms simultaneously are needed to advance our knowledge of mechanisms driving transmission.

West Nile Virus: Veterinary Health and Vaccine Development.

West Nile virus (WNV) (Flaviviridae: Flavivirus) was discovered in Africa more than 80 yr ago and became recognized as an avian pathogen and a cause of neurologic disease in horses largely during periodic incursions into Europe. Introduction of WNV into North America stimulated great anxiety, particularly in the equine industry, but also for pet owners and livestock producers concerned about the effect of WNV on other domestic animals. Numerous subsequent studies of naturally occurring and experimentally induced disease greatly expanded our understanding of the host range and clinical consequences of WNV infection in diverse species and led to rapid development and deployment of efficacious vaccines for horses. In addition to humans, horses are clearly the animals most frequently affected by serious, sometimes lethal disease following infection with WNV, but are dead-end hosts due to the low-magnitude viremia they develop. Dogs, cats, and livestock species including chickens are readily infected with WNV, but only occasionally develop clinical disease and are considered dead-end hosts for the virus.

Impact of West Nile Virus on Bird Populations: Limited Lasting Effects, Evidence for Recovery, and Gaps in Our Understanding of Impacts on Ecosystems.

The introduction of West Nile virus to North America in 1999 had profound impacts on human and wildlife health. Here, we review studies of WNV impacts on bird populations and find that overall impacts have been less than initially anticipated, with few species showing sustained changes in population size or demographic rates across multiple regions. This raises four questions: 1) What is the evidence for WNV impact on bird populations and how can we strengthen future analyses? We argue that future studies of WNV impacts should explicitly incorporate temporal variation in WNV transmission intensity, integrate field data with laboratory experimental infection studies, and correct for multiple comparisons. 2) What mechanisms might explain the relatively modest impact of WNV on most bird populations? We suggest that spatial and temporal variation in WNV transmission moderates WNV impacts on species that occur in multiple habitats, some of which provide refugia from infection. 3) Have species recovered from the initial invasion of WNV? We find evidence that many species and populations have recovered from initial WNV impact, but a few have not. 4) Did WNV cause cascading effects on other species and ecosystems? Unfortunately, few studies have examined the cascading effects of WNV population declines, but evidence suggests that some species may have been released from predation or competition. We close by discussing potentially overlooked groups of birds that may have been affected by WNV, and one highlight species, the yellow-billed magpie (Pica nutalli Audubon, 1837 [Passeriformes: Corvidae]), that appears to have suffered the largest range-wide impact from WNV.

Evidence of West Nile Virus (WNV) Circulation in Wild Birds and WNV RNA Negativity in Mosquitoes of the Danube Delta Biosphere Reserve, Romania, 2016.

West Nile virus (WNV) is a zoonotic flavivirus whose transmission cycle in nature includes wild birds as amplifying hosts and ornithophilic mosquito vectors. Bridge vectors can transmit WNV to mammal species potentially causing West Nile Fever. Wild bird migration is a mode of WNV introduction into new areas. The Danube Delta Biosphere Reserve (DDBR) is a major stopover of wild birds migrating between Europe and Africa. The aim of this study was to investigate the presence of WNV in the DDBR during the 2016 transmission season in wild birds and mosquitoes. Blood from 68 wild birds (nine different species) trapped at four different locations was analyzed by competitive ELISA and Virus Neutralization Test (VNT), revealing positive results in 8/68 (11.8%) of the wild birds by ELISA of which six samples (three from juvenile birds) were confirmed seropositive by VNT. Mosquitoes (n = 6523, 5 genera) were trapped with CDC Mini Light traps at two locations and in one location resting mosquitoes were caught. The presence of WNV RNA was tested in 134 pools by reverse transcription quantitative PCR (RT-qPCR). None of the pools was positive for WNV-specific RNA. Based on the obtained results, WNV was circulating in the DDBR during 2016.

GENETIC DIVERSITY OF YELLOW-BILLED MAGPIES ( PICA NUTALLI) BEFORE AND AFTER A WEST NILE VIRUS EPIDEMIC.

The appearance of West Nile virus (WNV) coincided with declines in California, US bird populations beginning in 2004, and particularly affected corvid populations, including Yellow-billed Magpies ( Pica nutalli), an endemic species to California. Our objective was to determine if the timing of the WNV epidemic correlated with changes in the genetic diversity or population structure of magpies. We hypothesized the declines in magpie abundance from WNV would lead to genetic bottlenecks and reduced genetic diversity, but not to changes in population genetic structure. To test these hypotheses, we genetically typed magpie samples collected during the Dead Bird Survey before WNV arrived (2002-04), immediately after WNV arrived in late 2004 (2006-08), and several generations after the onset of the epidemic (2009-11). For each of these three time periods, we tested for genetic bottlenecks, estimated genetic heterozygosity, allelic richness, relatedness, effective population sizes, and genetic structure, with the use of 10 nuclear microsatellite loci. Although there was no evidence for spatial or temporal genetic structure, genetic-diversity estimates were similar or below estimates for endangered corvid species. Measures of genetic diversity were consistent across time periods. In contrast to our expectation, we detected a genetic bottleneck prior to the WNV epidemic, which may have coincided with severe drought conditions in California, increasing human population size in magpie range, and an estimated 33% decrease in population size. We found weak evidence to support a bottleneck after the introduction of WNV in California. Our results suggest the WNV epidemic did not have additional catastrophic effects on the neutral genetic diversity of P. nutalli in the sampled areas. However, because we detected lower heterozygosity in Yellow-billed Magpies than has been reported in closely related endangered species, this species is of conservation concern and should be monitored to detect further population declines or loss of genetic diversity.

Reemergence of St. Louis Encephalitis Virus in the Americas.

We summarize and analyze historical and current data regarding the reemergence of St. Louis encephalitis virus (SLEV; genus Flavivirus) in the Americas. Historically, SLEV caused encephalitis outbreaks in the United States; however, it was not considered a public health concern in the rest of the Americas. After the introduction of West Nile virus in 1999, activity of SLEV decreased considerably in the United States. During 2014-2015, SLEV caused a human outbreak in Arizona and caused isolated human cases in California in 2016 and 2017. Phylogenetic analyses indicate that the emerging SLEV in the western United States is related to the epidemic strains isolated during a human encephalitis outbreak in Córdoba, Argentina, in 2005. Ecoepidemiologic studies suggest that the emergence of SLEV in Argentina was caused by the introduction of a more pathogenic strain and increasing populations of the eared dove (amplifying host).

Overwintering of West Nile virus in a bird community with a communal crow roost

In temperate climates, transmission of West Nile virus (WNV) is detectable rarely during the coldest months (late fall through early spring), yet the virus has reappeared consistently during the next warm season. Several mechanisms may contribute to WNV persistence through winter, including bird-to-bird transmission among highly viremic species. Here we consider whether, under realistic scenarios supported by field and laboratory evidence, a winter bird community could sustain WNV through the winter in the absence of mosquitoes. With this purpose we constructed a deterministic model for a community of susceptible birds consisting of communally roosting crows, raptors and other birds. We simulated WNV introduction and subsequent transmission dynamics during the winter under realistic initial conditions and model parameterizations, including plausible contact rates for roosting crows. Model results were used to determine whether the bird community could yield realistic outbreaks that would result in WNV infectious individuals at the end of the winter, which would set up the potential for onward horizontal transmission into summer. Our findings strongly suggest that winter crow roosts could allow for WNV persistence through the winter, and our model results provide synthesis to explain inconclusive results from field studies on WNV overwintering in crow roosts.