West Nile Virus Dissemination Research Articles

Levels of Circulating NS1 Impact West Nile Virus Spread to the Brain.

Dengue virus (DENV) and West Nile virus (WNV) are arthropod-transmitted flaviviruses that cause systemic vascular leakage and encephalitis syndromes, respectively, in humans. However, the viral factors contributing to these specific clinical disorders are not completely understood. Flavivirus nonstructural protein 1 (NS1) is required for replication, expressed on the cell surface, and secreted as a soluble glycoprotein, reaching high levels in the blood of infected individuals. Extracellular DENV NS1 and WNV NS1 interact with host proteins and cells, have immune evasion functions, and promote endothelial dysfunction in a tissue-specific manner. To characterize how differences in DENV NS1 and WNV NS1 might function in pathogenesis, we generated WNV NS1 variants with substitutions corresponding to residues found in DENV NS1. We discovered that the substitution NS1-P101K led to reduced WNV infectivity in the brain and attenuated lethality in infected mice, although the virus replicated efficiently in cell culture and peripheral organs and bound at wild-type levels to brain endothelial cells and complement components. The P101K substitution resulted in reduced NS1 antigenemia in mice, and this was associated with reduced WNV spread to the brain. Because exogenous administration of NS1 protein rescued WNV brain infectivity in mice, we conclude that circulating WNV NS1 facilitates viral dissemination into the central nervous system and impacts disease outcomes. IMPORTANCE Flavivirus NS1 serves as an essential scaffolding molecule during virus replication but also is expressed on the cell surface and is secreted as a soluble glycoprotein that circulates in the blood of infected individuals. Although extracellular forms of NS1 are implicated in immune modulation and in promoting endothelial dysfunction at blood-tissue barriers, it has been challenging to study specific effects of NS1 on pathogenesis without disrupting its key role in virus replication. Here, we assessed WNV NS1 variants that do not affect virus replication and evaluated their effects on pathogenesis in mice. Our characterization of WNV NS1-P101K suggests that the levels of NS1 in the circulation facilitate WNV dissemination to the brain and affect disease outcomes. Our findings facilitate understanding of the role of NS1 during flavivirus infection and support antiviral strategies for targeting circulating forms of NS1.

High Transmission Potential of West Nile Virus Lineage 1 for Cx. pipiens s.l. of Iran.

Vector competence is an important parameter in evaluating whether a species plays a role in transmission of an arbovirus. Although the protocols are similar, interpretation of results is unique given the specific interactions that exist between a mosquito population and a viral genotype. Here, we assessed the infection (IR), dissemination (DR), and transmission (TR) rates of Cx. pipiens s.l., collected from Iran, for West Nile virus (WNV) lineage 1a. We showed that Cx. pipiens s.l. mosquitoes in Iran were susceptible to WNV with IR up to 89.7%, 93.6%, and 83.9% at 7, 14, and 21 days post-infection (dpi) respectively. In addition, DR and TR reached respectively 92.3% and 75.0% at 21 dpi, and the number of viral particles delivered with saliva reached up to 1.33 × 105 particles. Therefore, an unexpected high risk of WNV dissemination in the region where Cx. pipiens s.l. mosquitoes are well established should be considered carefully and surveillance measures implemented accordingly.

Locally temperature - driven mathematical model of West Nile virus spread in Germany

West Nile virus (WNV) is an arthropod-borne virus (arbovirus) transmitted by the bites of infected mosquitoes. WNV can also infect horses and humans, where it may cause serious illness and can be fatal. Birds are the natural reservoir, and humans, equines and probably other mammals are dead-end hosts. In 2018, WNV occurred for the first time in Germany, affecting birds and horses. Seroconversion of an exposed veterinarian has also been reported. It is therefore of importance to evaluate the circumstances, under which WNV may establish in Germany as a whole or in particular favourable regions.In our current work, we formulate a dynamic model to describe the spreading process of West Nile virus in the presence of migratory birds. To investigate the possible role of migratory birds in the dissemination of WNV in Germany, we include the recurring presence of migratory birds through a mechanistic ordinary differential equations (ODE) model system. We also perform a sensitivity analysis of the infection curves. Seasonal impacts are also taken into consideration. As result, we present an analytical expression for the basic reproduction number R0. We find that after introducing WNV into Germany, R0 will be above the critical value in many regions of the country. Furthermore, we observe that in the south of Germany, the disease reoccurs in the following season after the introduction. We include a potential distribution map associated with WNV cases in Germany to illustrate our findings in a spatial scale.

Light pollution increases West Nile virus competence of a ubiquitous passerine reservoir species.

Among the many anthropogenic changes that impact humans and wildlife, one of the most pervasive but least understood is light pollution. Although detrimental physiological and behavioural effects resulting from exposure to light at night are widely appreciated, the impacts of light pollution on infectious disease risk have not been studied. Here, we demonstrate that artificial light at night (ALAN) extends the infectious-to-vector period of the house sparrow (Passer domesticus), an urban-dwelling avian reservoir host of West Nile virus (WNV). Sparrows exposed to ALAN maintained transmissible viral titres for 2 days longer than controls but did not experience greater WNV-induced mortality during this window. Transcriptionally, ALAN altered the expression of gene regulatory networks including key hubs (OASL, PLBD1 and TRAP1) and effector genes known to affect WNV dissemination (SOCS). Despite mounting anti-viral immune responses earlier, transcriptomic signatures indicated that ALAN-exposed individuals probably experienced pathogen-induced damage and immunopathology, potentially due to evasion of immune effectors. A simple mathematical modelling exercise indicated that ALAN-induced increases of host infectious-to-vector period could increase WNV outbreak potential by approximately 41%. ALAN probably affects other host and vector traits relevant to transmission, and additional research is needed to advise the management of zoonotic diseases in light-polluted areas.

Insecticide resistance genes affect Culex quinquefasciatus vector competence for West Nile virus.

Insecticide resistance has been reported to impact the interactions between mosquitoes and the pathogens they transmit. However, the effect on vector competence for arboviruses still remained to be investigated. We examined the influence of two insecticide resistance mechanisms on vector competence of the mosquito Culex quinquefasciatus for two arboviruses, Rift Valley Fever virus (RVFV) and West Nile virus (WNV). Three Cx. quinquefasciatus lines sharing a common genetic background were used: two insecticide-resistant lines, one homozygous for amplification of the Ester2 locus (SA2), the other homozygous for the acetylcholinesterase ace-1 G119S mutation (SR) and the insecticide-susceptible reference line Slab. Statistical analyses revealed no significant effect of insecticide-resistant mechanisms on vector competence for RVFV. However, both insecticide resistance mechanisms significantly influenced the outcome of WNV infections by increasing the dissemination of WNV in the mosquito body, therefore leading to an increase in transmission efficiency by resistant mosquitoes. These results showed that insecticide resistance mechanisms enhanced vector competence for WNV and may have a significant impact on transmission dynamics of arboviruses. Our findings highlight the importance of understanding the impacts of insecticide resistance on the vectorial capacity parameters to assess the overall consequence on transmission.

Infection and dissemination of West Nile virus in China by the potential vector, Culex pipiens pallens.

The distribution of the West Nile virus (WNV) in the organs and tissues of the mosquito Culex pipiens pallens, a potential vector of WNV in China, was investigated up to 14 days after oral infection. The WNV antigen was detected in paraffin-embedded mosquitoes using immunocytochemistry and viral titers of post-infected mosquitoes determined by plaque assay. Viral titers sharply decreased 24 h post-infection, were undetectable for the first few days, then rose over the course of infection. The first midgut infection appeared after one day, and the overall infection rate (based on midgut infection) was 43.9%. Other tissues, including hindgut, foregut, ovarian follicles, Malpighian tubules, and ommatidia, showed weak WNV antigens as early as three days post-infection. Staining in the salivary glands first appeared after seven days, and the salivary gland infection rate on the 14th day was 37.5%. Specimens with no detectable WNV antigens in any tissues, and with positive results confined to the midgut, anterior midgut, and hindgut, were observed on the 14th day. The route of viral dissemination from the midgut, and the relative importance of amplifying tissues in mosquitoes' susceptibility to infection, were evaluated. The results indicate that Cx. p. pallens has the ability to harbor WNV throughout its alimentary system and that midgut epithelial cells may be the initial site of the replication of this virus in this species.

Differential virulence and pathogenesis of West Nile viruses.

West Nile virus (WNV) is a neurotropic flavivirus that cycles between mosquitoes and birds but that can also infect humans, horses, and other vertebrate animals. In most humans, WNV infection remains subclinical. However, 20%–40% of those infected may develop WNV disease, with symptoms ranging from fever to meningoencephalitis. A large variety of WNV strains have been described worldwide. Based on their genetic differences, they have been classified into eight lineages; the pathogenic strains belong to lineages 1 and 2. Ten years ago, Beasley et al. (2002) found that dramatic differences exist in the virulence and neuroinvasion properties of lineage 1 and lineage 2 WNV strains. Further insights on how WNV interacts with its hosts have recently been gained; the virus acts either at the periphery or on the central nervous system (CNS), and these observed differences could help explain the differential virulence and neurovirulence of WNV strains. This review aims to summarize the current state of knowledge on factors that trigger WNV dissemination and CNS invasion as well as on the inflammatory response and CNS damage induced by WNV. Moreover, we will discuss how WNV strains differentially interact with the innate immune system and CNS cells, thus influencing WNV pathogenesis.

West Nile Virus Infection in American Robins: New Insights on Dose Response

West Nile virus (WNV) is a vector-borne pathogen that was first detected in the United States in 1999. The natural transmission cycle of WNV involves mosquito vectors and avian hosts, which vary in their competency to transmit the virus. American robins are an abundant backyard species in the United States and appear to have an important role in the amplification and dissemination of WNV. In this study we examine the response of American robins to infection with various WNV doses within the range of those administered by some natural mosquito vectors. Thirty American robins were assigned a WNV dosage treatment and needle inoculated with 100.95 PFU, 101.26 PFU, 102.15 PFU, or 103.15 PFU. Serum samples were tested for the presence of infectious WNV and/or antibodies, while oral swabs were tested for the presence of WNV RNA. Five of the 30 (17%) robins had neutralizing antibodies to WNV prior to the experiment and none developed viremia or shed WNV RNA. The proportion of WNV-seronegative birds that became viremic after WNV inoculation increased in a dose dependent manner. At the lowest dose, only 40% (2/5) of the inoculated birds developed productive infections while at the highest dose, 100% (7/7) of the birds became viremic. Oral shedding of WNV RNA followed a similar trend where robins inoculated with the lower two doses were less likely to shed viral RNA (25%) than robins inoculated with one of the higher doses (92%). Viremia titers and morbidity did not increase in a dose dependent manner; only two birds succumbed to infection and, interestingly, both were inoculated with the lowest dose of WNV. It is clear that the disease ecology of WNV is a complex interplay of hosts, vectors, and viral dose delivered.

Flaviviruses in Game Birds, Southern Spain, 2011–2012

Flaviviruses in Game Birds, Southern Spain, 2011–2012

The impact of type I interferon on the development of the adaptive immune response (67.14)

Abstract Determining the impact of type I interferon on immune responses using mice missing the common IFN-α/β signaling receptor (IFNAR- /-) is limited to studying the functions of type I IFN at all stages of viral infection. To define the temporal functions of type I IFN signaling in the context of infection by West Nile virus (WNV), we treated mice with MAR1-5A3, a neutralizing, non cell-depleting anti-IFNAR antibody at later stages of infection. Inhibition of type I IFN signaling at or before day 2 after infection was associated with markedly enhanced viral burden, whereas treatment at day 4 altered WNV dissemination less dramatically. Blockade of type I IFN signaling starting at day 4 induced dysfunctional CD8+ T cells with depressed cytokine responses and expression of phenotypic markers suggesting exhaustion. The later maturation phase (days 4-8) of anti-WNV CD8+ T cell development requires type I IFN signaling. Collectively, our results suggest that cell non-autonomous type I IFN signaling shapes maturation of antiviral CD8+ T cell response at a stage distinct from the initial priming event.

West Nile Virus: an Update on Recent Developments

West Nile Virus arrived in North America just over 12 years ago, and since that time, much has been learned about its pathology and transmission of infection and the factors affecting prevalence. Key developments uncovered critical risk factors for severe neurologic disease and revealed successful host defense strategies. At present, only equine vaccines are available; human vaccines remain in various stages of clinical trials. This review discusses the advances in and limitations of the current research related to the emergence and continued dissemination of West Nile virus throughout the Americas.

Flavivirus infection induces indoleamine 2,3-dioxygenase in human monocyte-derived macrophages via tumor necrosis factor and NF-κB

Infection with West Nile virus (WNV) via a mosquito bite results in local viral replication in the skin, followed by viremia. Thus, tissue macrophages are ideally located to prevent the dissemination of WNV throughout the host. The current study shows that WNV infection of human monocyte-derived macrophages (MDM) results in increased WNV mRNA, protein, and infectious virions at 24 h p.i. with a decline in titer after 48 h. Concomitant with viral control was the robust induction of indoleamine 2,3-dioxygenase (IDO) and resultant metabolism of L-tryptophan (L-Trp) to kynurenine. In WNV-exposed cultures, IDO protein was induced primarily in noninfected versus viral-infected MDM. Whereas WNV infection increased the production of IFN-α, IFN-β, and TNF, only antibody neutralization of TNF attenuated IDO expression and activity. WNV infection also activated NF-κB, and inhibition of this pathway with BMS-345541 abrogated IDO induction. Similar results were also obtained with MDM infected with the related flavivirus, Japanese encephalitis virus. Whereas IDO-mediated L-Trp metabolism can exhibit antiviral properties, inhibition of IDO activity in MDM with L-1-MT or the addition of excess L-Trp did not affect viral control. However, culturing MDM in L-Trp-deficient medium or overexpression of IDO in cells prior to infection significantly attenuated WNV replication, which was reversed by adding excess L-Trp. Together, these data support that although IDO is not required by MDM for the clearance of established viral infection, the spread of flavivirus infection is limited by IDO expressed in uninfected, neighboring cells.

Environmental Factors and Risk Areas of West Nile Virus in Southern California, 2007–2009

The West Nile virus (WNV) may post a significant health risk for mammals, including humans and insects. This study examines the spatial–temporal effects of environmental factors on WNV dissemination with a case study of ten counties in the southern California, where the epidemic was recently most prevalent within the USA. WNV surveillance data were obtained from the California Vectorborne Disease Surveillance System and Centers for Disease Control and Prevention. Remote sensing and Geographic Information Systems (GIS) techniques were combined to derive environmental variables. Principal component analysis was performed to select the most relevant environmental variables. Two ecological zones were identified based on the selected variables. Identification of risk areas for WNV was limited to a zone with 95% mosquitoes surveillance records. Three time windows, the epidemiological weeks 18–26, 27–35, and 36–44 in each year of 2007–2009, were examined in details with risk area mapping. It is found that the southern part of San Joaquin Valley in Kern County and Los Angeles County (especially its southern part) were the most vulnerable locations for WNV outbreak. Main factors contributing to the WNV propagation included summer mean temperature, annual mean deviation from the mean temperature, land surface temperature, elevation, landscape complexity, landscape diversity, and vegetation water content. The result of this study improves understanding of WNV ecology and provides tools for detecting, tracking, and predicting the epidemic. The holistic approach developed for this study, which integrated remotely sensed, GIS-based, and in situ-measured environmental data with landscape metrics, may be applied to studies of other vector-borne diseases.

A temporal role of type I interferon signaling in CD8+ T cell maturation during acute West Nile virus infection.

A genetic absence of the common IFN- α/β signaling receptor (IFNAR) in mice is associated with enhanced viral replication and altered adaptive immune responses. However, analysis of IFNAR-/- mice is limited for studying the functions of type I IFN at discrete stages of viral infection. To define the temporal functions of type I IFN signaling in the context of infection by West Nile virus (WNV), we treated mice with MAR1-5A3, a neutralizing, non cell-depleting anti-IFNAR antibody. Inhibition of type I IFN signaling at or before day 2 after infection was associated with markedly enhanced viral burden, whereas treatment at day 4 had substantially less effect on WNV dissemination. While antibody treatment prior to infection resulted in massive expansion of virus-specific CD8+ T cells, blockade of type I IFN signaling starting at day 4 induced dysfunctional CD8+ T cells with depressed cytokine responses and expression of phenotypic markers suggesting exhaustion. Thus, only the later maturation phase of anti-WNV CD8+ T cell development requires type I IFN signaling. WNV infection experiments in BATF3 -/- mice, which lack CD8-α dendritic cells and have impaired priming due to inefficient antigen cross-presentation, revealed a similar effect of blocking IFN signaling on CD8+ T cell maturation. Collectively, our results suggest that cell non-autonomous type I IFN signaling shapes maturation of antiviral CD8+ T cell response at a stage distinct from the initial priming event.

Enhancing temporal resolution of satellite imagery for public health studies: A case study of West Nile Virus outbreak in Los Angeles in 2007

Although remotely sensed data have been used in public health studies, these studies are often limited by the critical choice that has to be made in data selection: either using data with high spatial but low temporal resolution, or data with high temporal but low spatial resolution. This choice creates significant limitations for time-dependent epidemiological studies, since it is often essential to have datasets with high spatial and temporal resolution. Effectively synthesizing high temporal resolution imagery with high spatial resolution imagery can potentially ease this limitation. To this end, we conducted an experiment by creating a series of simulated ASTER datasets by fusing ASTER and MODIS data with the STARFM image fusion model. These simulated datasets are then used to derive the following urban environmental variables: normalized difference vegetation index, normalized difference water index, and land surface temperature. These variables are used to quantitatively examine the effects of urban environmental characteristics on West Nile Virus dissemination in Los Angeles County, California, where the epidemic was most prevalent in the United States in 2007. Mosquito surveillance data were collected from the weekly summary reports published in the California West Nile Virus website. A spatial–temporal analysis of WNV dissemination was conducted by synthesizing remote sensing variables and mosquito surveillance records. We focused on assessment of WNV risk areas in July through September due to data sufficiency of tested-positive mosquito pools. Moderate- and high-risk areas of WNV infections in mosquitoes were identified for five selected time windows, i.e., epidemiological weeks 30–31, 32–33, 34–35, 36–37, and 38–39. The results show that elevation and urban built-up conditions were negatively associated with the WNV propagation, while LST positively correlated with viral transmission. NDVI was not significantly associated with WNV transmission during the studied time intervals. San Fernando Valley was found to be the most vulnerable to mosquito infections of WNV within the City of Los Angeles. With the complementation of high-spatial resolution ASTER and high-temporal resolution MODIS images, the fused image datasets allow for estimating environmental parameters at desired epidemiological weeks. This paper provides important insights into how high temporal resolution remote sensing imagery might be used to study time-dependant events in public health, especially in the operational surveillance and control of vector-borne or other epidemic diseases.

Prevalence of West Nile Virus in Migratory Birds during Spring and Fall Migration

To investigate the role of migratory birds in the dissemination of West Nile virus (WNV), we measured the prevalence of infectious WNV and specific WNV neutralizing antibodies in birds, principally Passeriformes, during spring and fall migrations in the Atlantic and Mississippi flyways from 2001-2003. Blood samples were obtained from 13,403 birds, representing 133 species. Specific WNV neutralizing antibody was detected in 254 resident and migratory birds, representing 39 species, and was most commonly detected in northern cardinals (Cardinalis cardinalis) (9.8%, N = 762) and gray catbirds (Dumetella carolinensis) (3.2%, N = 3188). West Nile virus viremias were detected in 19 birds, including 8 gray catbirds, and only during the fall migratory period. These results provide additional evidence that migratory birds may have been a principal agent for the spread of WNV in North America and provide data on the occurrence of WNV in a variety of bird species.

An examination of the effect of landscape pattern, land surface temperature, and socioeconomic conditions on WNV dissemination in Chicago

This paper developed an approach by the synthesis of remote sensing, landscape metrics, and statistical methods to examine the effects of landscape pattern, land surface temperature, and socioeconomic conditions on the spread of West Nile virus (WNV) caused by mosquitoes and animal hosts in Chicago, USA. Land use/land cover and land surface temperature images were derived from Terra's Advanced Spaceborne Thermal Emission and Reflection Radiometer imagery. An analytical procedure using landscape metrics was developed, applying configuration analysis of landscape patterns in the study area. The positive reports of mosquitoes and animal hosts for WNV in fall, 2001-2006, were collected from the Cook County Public Health Department. Forty-nine municipalities were found to have WNV-positive records in mosquitoes and animal hosts in fall 2004. Socioeconomic data were obtained from the 2000 US Census. Statistical analysis was applied to WNV data in fall 2004 to identify the relationship between potential predictors and WNV spread. As a result, landscape factors, such as landscape aggregation index and the urban areas and areas of grass and water, showed strong correlations with the WNV-positive records. Socioeconomic conditions, such as the population over 65 years old, also showed a strong correlation with WNV-positive records. Thermal conditions of water showed a less but still considerable correlation to WNV-positive records. This research offers an opportunity to explore the effects of landscape pattern, land surface temperature, and socioeconomic conditions on the spread of WNV caused by mosquitoes and animal hosts. Results can contribute to public health and environmental management in the study area.

Spatio-temporal analysis of the relationship between WNV dissemination and environmental variables in Indianapolis, USA

BackgroundThis study developed a multi-temporal analysis on the relationship between West Nile Virus (WNV) dissemination and environmental variables by using an integrated approach of remote sensing, GIS, and statistical techniques. WNV mosquito cases in seven months (April-October) of the six years (2002–2007) were collected in Indianapolis, USA. Epidemic curves were plotted to identify the temporal outbreaks of WNV. Spatial-temporal analysis and k-mean cluster analysis were further applied to determine the high-risk areas. Finally, the relationship between environmental variables and WNV outbreaks were examined by using Discriminant Analysis.ResultsThe results show that the WNV epidemic curve reached its peak in August for all years in the study area except in 2007, where the peak was reached in July. WNV dissemination started from the central longitudinal corridor of the city and spread out to the east and west. Different years and seasons had different high-risk areas, but the southwest and southeast corners show the highest risk for WNV infection due to their high percentages of agriculture and water sources.ConclusionMajor environmental factors contributing to the outbreak of WNV in Indianapolis were the percentages of agriculture and water, total length of streams, and total size of wetlands. This study provides important information for urban public health prevention and management. It also contributes to the optimization of mosquito control and arrangement of future sampling efforts.

West Nile virus dissemination and tissue tropisms in orally infected Culex pipiens quinquefasciatus.

We investigated the spatial and temporal distribution of West Nile virus (WNV) in organs and tissues of Culex pipiens quinquefasciatus mosquitoes for up to 27 days following oral infection. WNV antigen was detected in paraffin-embedded mosquitoes by immunohistochemistry. Immunofluorescence assays were performed on dissected salivary glands and midguts and analyzed by confocal microscopy. We evaluated the route of virus dissemination following midgut escape and the relative importance of amplifying tissues in mosquito susceptibility to infection. WNV infection was persistent in all tissues analyzed including the midgut, salivary glands, nervous system, and fat body and only declined in the cytoplasm of posterior midgut epithelial cells over time. The phenomenon of cell-to-cell spread was observed in the midgut epithelium and WNV intensely infected both circular and longitudinal muscles of the same organ. It is possible that muscle tissue serves as a conduit for virus dissemination and contributes to WNV amplification, particularly late in infection. These findings provide insight into WNV infection dynamics in a highly susceptible, epidemiologically important, North American vector. Our results pave the way for future studies to analyze physical and biological barriers to WNV dissemination in less competent mosquitoes.

Rôle des oiseaux migrateurs dans l'épidémiologie du virus West Nile

West Nile (WN) virus, first isolated from a human in Uganda, is a common arboviral disease in sub-Saharan Africa and India. The viral cycle of transmission, which involves mosquitoes and birds, usually inducing a non-symptomatic disease, was extensively studied in Egypt 50 years ago. Humans and horses are sensitive to WN virus infection. Most cases are non-symptomatic, but encephalitis cases are also reported as well as some fatalities, mostly in old patients. For a few years, West Nile has appeared as a reemerging virus with several outbreaks recorded in the Mediterranean region and Southern and Eastern Europe (Algeria 1994, Romania 1996, Israel and France 2000). West Nile anti-bodies were detected in many bird species, and the virus was isolated from birds like a turtle dove in Slovakia, a barred warbler in Cyprus, a rock pigeon in Egypt or a dead white stork in Israel. Birds are suspected for multiple introductions of WN virus in Europe from endemic countries in tropical Africa. Usually outbreaks occurred in temperate countries during the summer period, and mostly in humid regions (deltas, etc.) where large colonies of migrating and resident birds and populations of potential vectors like Culex modestus are present. Several factors are necessary for an outbreak to occur: the arrival of migrating infected birds presenting a high viremia susceptible of infecting potential vectors, replication in mosquitoes and amplification of the virus within susceptible birds and mosquitoes, and transmission to humans and/or horses. West Nile virus emergence for the first time in the Americas in 1999 in New York was revealed by the death of several thousand native birds, mostly crows and blue jays. However, bird migrations do not seem to be involved in the recent WN introduction in the United States. A better knowledge of the different factors involved in the WN virus transmission cycle are needed, especially the possible persistence of viral infection in birds and the role of ticks in WN virus dissemination. Combinations of surveys of potential regions of reemergence of WN virus in Europe in connexion with other places in the Middle East, North Africa and tropical Africa would be important for a better understanding of the epidemiology of this arbovirosis.