Simulation of the seasonal cycles of bird, equine and human West Nile virus cases

The West Nile virus (WNV) is an arthropod-borne virus (arbovirus) circulating in a natural transmission cycle between mosquitoes (enzootic vectors) and birds (amplifying hosts). Additionally, mainly horses and humans (dead-end hosts) may be infected by blood-feeding mosquitoes (bridge vectors). We developed an epidemic model for the simulation of the WNV dynamics of birds, horses and humans in the U.S., which we apply to the Minneapolis metropolitan area (Minnesota). The SEIR-type model comprises a total of 19 compartments, that are 4 compartments for mosquitoes and 5 compartments or health states for each of the 3 host species. It is the first WNV model that simulates the seasonal cycle by explicitly considering the environmental temperature. The latter determines model parameters responsible for the population dynamics of the mosquitoes and the extrinsic incubation period. Once initialized, our WNV model runs for the entire period 2002-2009, exclusively forced by environmental temperature. Simulated incidences are mainly determined by host and vector population dynamics, virus transmission and herd immunity, respectively. We adjusted our WNV model to fit monthly totals of reported bird, equine and human cases in the Minneapolis metropolitan area. From this process we estimated that the proportion of actually WNV-induced dead birds reported by the Centers for Disease Control and Prevention is about 0.8%, whereas 7.3% of equine and 10.7% of human cases were reported. This is consistent with referenced expert opinions whereby about 10% of equine and human cases are symptomatic (the other 90% of asymptomatic cases are usually not reported). Despite the restricted completeness of surveillance data and field observations, all major peaks in the observed time series were caught by the simulations. Correlation coefficients between observed and simulated time series were R=0.75 for dead birds, R=0.96 for symptomatic equine cases and R=0.86 for human neuroinvasive cases, respectively.     

West Nile virus and its emergence in the United States of America

Zoonotic West Nile virus (WNV) circulates in natural transmission cycles involving certain mosquitoes and birds, horses, humans, and a range of other vertebrates are incidental hosts. Clinical infections in humans can range in severity from uncomplicated WNV fever to fatal meningoencephalitis. Since its introduction to the Western Hemisphere in 1999, WNV had spread across North America, Central and South America and the Caribbean, although the vast majority of severe human cases have occurred in the United States of America (USA) and Canada. By 2002-2003, the WNV outbreaks have involved thousands of patients causing severe neurologic disease (meningoencephalitis and poliomyelitis-like syndrome) and hundreds of associated fatalities in USA. The purpose of this review is to present recent information on the epidemiology and pathogenicity of WNV since its emergence in North America.     

West Nile virus infection of horses

West Nile virus (WNV) is a flavivirus closely related to Japanese encephalitis and St. Louis encephalitis viruses that is primarily maintained in nature by transmission cycles between mosquitoes and birds. Occasionally, WNV infects and causes disease in other vertebrates, including humans and horses. West Nile virus has re-emerged as an important pathogen as several recent outbreaks of encephalomyelitis have been reported from different parts of Europe in addition to the large epidemic that has swept across North America. This review summarises the main features of WNV infection in the horse, with reference to complementary information from other species, highlighting the most recent scientific findings and identifying areas that require further research.     

Ecological Surveillance for West Nile in Catalonia (Spain), Learning from a Five‐Year Period of Follow‐up

To enhance early detection of West Nile virus (WNV) transmission, an integrated ecological surveillance system was implemented in Catalonia (north‐eastern Spain) from 2007 to 2011. This system incorporated passive and active equine surveillance, periodical testing of chicken sentinels in wetland areas, serosurveillance wild birds and testing of adult mosquitoes. Samples from 298 equines, 100 sentinel chickens, 1086 wild birds and 39 599 mosquitoes were analysed. During these 5 years, no acute WNV infection was detected in humans or domestic animal populations in Catalonia. WNV was not detected in mosquitoes either. Nevertheless, several seroconversions in resident and migrant wild birds indicate that local WNV or other closely related flaviviruses transmission was occurring among bird populations. These data indicate that bird and mosquito surveillance can detect otherwise silent transmission of flaviviruses and give some insights regarding possible avian hosts and vectors in a European setting.     

禽流感感染人的分子机制研究进展

禽流感病毒可以感染多种动物,包括人、猪、鸟、马、海豹、鲸和雪貂等。流感病毒在不同的宿主存在一定的屏障,但禽流感毒株能突破宿主屏障直接感染人,造成死亡。因此流感病毒的变异和病毒如何选择物种跨越物种流行的机制,对预防和控制流感的爆发是非常重要的。本文综述病毒毒力的分子生物学基础、禽流感染人的分子机制研究进展及在控制禽流感方面的研究。     

Avian titer development against West nile virus after extralabel use of an equine vaccine.

West Nile virus affects many animals, but the highest prevalence of morbidity and mortality is observed in birds, horses, and humans. The purpose of this study was to determine a protocol in birds of prey and corvids, using a vaccine developed for horses. The birds were assigned to five groups. Groups 1-4 received 0.25 ml, 0.5 ml, 0.75 ml, and 1.0 ml, respectively, and group 5 served as a control group. The greatest percentage of seroconversion (58.3%) was observed in the vaccine group that received a dose of 1.0 ml administered thrice, 3 wk apart. This report demonstrates that a vaccine developed for equines against West Nile virus can be administered to birds.     

Explaining Usutu virus dynamics in Austria: model development and calibration

Usutu virus (USUV), a flavivirus of the Japanese encephalitis virus complex, was for the first time detected outside Africa in the region around Vienna (Austria) in 2001 by Weissenb?ck et al. [Weissenb?ck, H., Kolodziejek, J., Url, A., Lussy, H., Rebel-Bauder, B., Nowotny, N., 2002. Emergence of Usutu virus, an African mosquito-borne flavivirus of the Japanese encephalitis virus group, central Europe. Emerg. Infect. Dis. 8, 652-656]. USUV is an arthropod-borne virus (arbovirus) circulating between arthropod vectors (mainly mosquitoes of the Culex pipiens complex) and avian amplification hosts. Infections of mammalian hosts or humans, as observed for the related West Nile virus (WNV), are rare. However, USUV infection leads to a high mortality in birds, especially blackbirds (Turdus merula), and has similar dynamics with the WNV in North America, which, amongst others, caused mortality in American robins (Turdus migratorius). We hypothesized that the transmission of USUV is determined by an interaction of developing proportion of the avian hosts immune and climatic factors affecting the mosquito population. This mechanism is implemented into the present model that simulates the seasonal cycles of mosquito and bird populations as well as USUV cross-infections. Observed monthly climate data are specified for the temperature-dependent development rates of the mosquitoes as well as the temperature-dependent extrinsic-incubation period. Our model reproduced the observed number of dead birds in Austria between 2001 and 2005, including the peaks in the relevant years. The high number of USUV cases in 2003 seems to be a response to the early beginning of the extraordinary hot summer in that year. The predictions indicate that >70% of the bird population acquired immunity, but also that the percentage would drop rapidly within only a couple of years. We estimated annually averaged basic reproduction numbers between R (0)=0.54 (2004) and 1.35 (2003). Finally, extrapolation from our model suggests that only 0.2% of the blackbirds killed by USUV were detected by the Austrian USUV monitoring program [Chvala, S., Bakonyi, T., Bukovsky, C., Meister, T., Brugger, K., Rubel, F., Nowotny, N., Weissenb?ck, H., 2007. Monitoring of Usutu virus activity and spread by using dead bird surveillance in Austria, 2003-2005. Vet. Microbiol. 122, 237-245]. These results suggest that the model presented is able to quantitatively describe the process of USUV dynamics.     

Hendra and Nipah virus infections.

The most important clinical and pathological manifestation of Hendra virus infection in horses and humans is that of severe interstitial pneumonia caused by viral infection of small blood vessels. The virus is also capable of causing nervous disease. Hendra virus is not contagious in horses and is spread by close contact with body fluids, such as froth from infected lungs. Diagnosis should be based on the laboratory examination of blood, lung, kidney, spleen, and, if nervous signs are present, also of the brain. Evidence of infection with the more recently identified and related Nipah virus was found in the brain of one horse in which there was inflammation of the meningeal blood vessels. Fruit bats, especially Pteropus s., have been incriminated as the natural and reservoir hosts of both Hendra and Nipah viruses.     

Monitoring of West Nile virus infections in Germany

West Nile virus (WNV) is a flavivirus that is maintained in an enzootic cycle between ornithophilic mosquitoes, mainly of the Culex genus, and certain wild bird species. Other bird species like ravens, jays and raptors are highly susceptible to the infection and may develop deadly encephalitis, while further species of birds are only going through subclinical infection. The objective of this study was to continue in years 2009-2011 the serological and molecular surveillance in wild birds in Germany (see Vector Borne Zoonotic Dis. 10, 639) and to expand these investigations for the first time also to sera from domestic poultry and horses collected between 2005 and 2009. All three cohorts function as indicators for the endemic circulation of WNV. The presence of WNV-specific antibodies was detected in all samples by virus neutralization test (VNT), indirect immunofluorescence test (IFT) and/or enzyme-linked immunosorbent assay (ELISA). The presence of WNV genomes was monitored in relevant sera using two qRT-PCRs that amplify lineage 1 and 2 strains. A total of 364 migratory and resident wild bird serum samples (with emphasis on Passeriformes and Falconiformes) as well as 1119 serum samples from domestic poultry and 1282 sera from horses were analysed. With the exception of one hooded crow, antibody carriers were exclusively found in migratory birds, but not in resident birds/domestic poultry or in local horses. Crows are facultative, short-distance winter migrants in Germany. WNV-specific nucleic acids could not be demonstrated in any of the samples. According to these data, there is no convincing evidence for indigenous WNV infections in equines and in wild/domestic birds in Germany. However, since a few years, WNV infections are endemic in other European countries such as Austria, Hungary, Greece and Italy, a state-of-the-art surveillance system for the detection of incursions of WNV into Germany deems mandatory.     

Isolation of Ross River virus from mosquitoes and from horses with signs of musculo-skeletal disease

OBJECTIVE: To report clinical and clinicopathological findings in horses naturally infected with Ross River virus (RRV) and identify likely mosquito arbovirus vector species. PROCEDURES: Veterinarians submitted serum samples from 750 horses because they suspected Ross River virus (RRV) infection. The samples were tested for the presence of IgM and IgG antibody to RRV and for the presence of virus. Mosquitoes were trapped, differentiated to species level and tested for the presence of RRV by virus isolation. RESULTS: RRV was isolated from six species of mosquitoes (Ochlerotatus camptorhyncus, Culex globocoxitus, Cx. australicus, Cx. annulirostris, Cx. quinquefasciatus, Anopheles annulipes) and from 13 horses with clinical signs of musculo-skeletal disease. Antibody to RRV was detected in 420 of the 750 serum samples; 307 contained IgG only; 76 contained both IgM and IgG and 37 contained only IgM antibody to RRV. Virus was isolated from horses with IgM antibody only. CONCLUSIONS: RRV can be isolated from infected horses during the short time period when there is an overlap of clinical signs, positive IgM serology and viraemia. Early spring infections of horses may occur if RRV infected mosquito vectors are present. RRV has not been shown to cause clinical disease in horses. This is the first report of isolation of RRV from Oc. camptorhyncus in the Murray region and indicates a potential for infection of humans and animals in autumn as well as in spring.     

Assessment of the efficacy of a single dose of a recombinant vaccine against West Nile virus in response to natural challenge with West Nile virus-infected mosquitoes in horses

OBJECTIVE: To determine the onset of immunity after IM administration of a single dose of a recombinant canarypox virus vaccine against West Nile virus (WNV) in horses in a blind challenge trial. ANIMALS: 20 mixed-breed horses. PROCEDURE: Horses with no prior exposure to WNV were randomly assigned to 1 of 2 groups (10 horses/group). In 1 group, a recombinant canarypox virus vaccine against WNV was administered to each horse once (day 0). The other 10 control horses were untreated. On day 26, 9 treated and 10 control horses were challenged via the bites of mosquitoes (Aedes albopictus) infected with WNV. Clinical responses and WNV isolation were monitored for 14 days after challenge exposure; antibody responses against WNV after administration of the vaccine and challenge were also assessed in both groups. RESULTS: Following challenge via WNV-infected mosquitoes, 1 of 9 treated horses developed viremia. In contrast, 8 of 10 control horses developed viremia after challenge exposure to WNV-infected mosquitoes. All horses seroconverted after WNV challenge; compared with control horses, antibody responses in the horses that received the vaccine were detected earlier. CONCLUSIONS AND CLINICAL RELEVANCe: In horses, a single dose of the recombinant canarypox virus-WNV vaccine appears to provide early protection against development of viremia after challenge with WNV-infected mosquitoes, even in the absence of measurable antibody titers in some horses. This vaccine may provide veterinarians with an important tool in controlling WNV infection during a natural outbreak or under conditions in which a rapid onset of protection is required.     

West Nile Virus in North‐Eastern Italy, 2011: Entomological and Equine IgM‐Based Surveillance to Detect Active Virus Circulation

Since 2008, West Nile Virus (WNV) has expanded its range in several Italian regions, and its yearly recurrence suggests the virus may have become endemic in some areas. In 2011, a new plan based also on the detection of IgM antibodies was implemented in the north‐eastern Italian regions of Veneto and Friuli Venezia Giulia, aiming to early detect WNV infections in areas where the virus had already circulated during the previous summers, and in adjacent zones. From July to November 2011, 1880 sera from 521 equine premises were screened by a commercial IgM capture ELISA. Mosquitoes were captured by CDC‐CO₂ traps at 61 locations in the two regions. Collected mosquitoes were identified, pooled by species/date/location and examined by real‐time RT‐PCR and sequencing. Passive surveillance was carried out on clinically affected horses and non‐migratory wild birds found dead. IgM sero‐positive equines were detected in 19 holdings, five in the area with WNV circulation (AWC) and 14 in the surveillance area (SA); 10 more horse premises tested positive to further serological controls within 4 km of the positive holdings. A total of 85 398 mosquitoes of 15 species were collected and 2732 pools examined. Five Culex pipiens pools tested positive for the presence of WNV. Passive surveillance on non‐migratory wild birds allowed detection of the virus only in one found dead collared dove (Streptopelia decaocto), of 82 birds sampled. The WNV belonged to the lineage 2, which had been isolated for the first time in Italy earlier in 2011. By the first week of October, nine human cases had been confirmed in the same area. The implementation of a protocol combining IgM screening of horses with surveillance on mosquito vectors proved to be valuable for early detecting WNV circulation.     

Waterbird movement across the Great Dividing Range and implications for arbovirus irruption into southern Victoria

Background Waterbirds are the major hosts of various arboviruses. Murray Valley encephalitis virus (MVEV) is an arbovirus native to northern Australia, the major hosts of which are Phalacrocoraciformes (cormorants), Ciconiiformes (herons) and other waterbirds. MVEV is transmitted to humans by mosquitoes and can cause acute encephalomyelitis. In Victoria, MVEV is restricted to the northern side of the Great Dividing Range (GDR), suggesting that waterbirds cannot cross the high country. Methods and results We tested this hypothesis by analysing data on waterbird banding and recovery and discovered that 12 species can cross the GDR. Conclusion Waterbirds have the potential to carry arboviruses, including MVEV, into southern Victoria.     

Seroconversion of anti-Getah virus antibody among Japanese native Noma horses around 2012

Getah virus (GETV), an arthropod-borne virus transmitted by mosquitoes, has been isolated from several animals. GETV infection in horses shows clinical signs such as fever, rash, and edema in the leg. Noma horses are one of the eight Japanese native horses. The present study aimed to clarify the occurrence of GETV infection in Noma horses. Serum samples collected from Noma horses were analyzed using a virus neutralization test and enzyme-linked immunosorbent assay and showed that the anti-GETV antibody titers in the samples collected in 2017 were significantly higher than those collected in 2012. We concluded that a seroconversion of anti-GETV antibodies was occurred in the Noma horse population around 2012, providing evidence of the GETV epidemic in Japan circa 2012.     

Serological evidence of West Nile virus infection in human populations and domestic birds in the Northwest of Morocco

West Nile virus (WNV) was recently detected in Culex pipiens mosquitoes in Morocco. The aim of this study was to evaluate the seroprevalence of WNV in humans and in domestic birds in two regions of Morocco by the detection of IgG antibodies. Blood samples were obtained from 91 human patients and 92 domestic birds from September to December 2019. All study samples were tested using competitive enzyme-linked immunosorbent assay (cELISA) and WNV neutralization tests (VNT) were performed on positive sera. Of all samples, 4 (4.39 %) humans and 4 (4.34 %) birds were found to be seropositive for flaviviruses by the cELISA test. The VNT revealed that three of the four human samples detected positive by cELISA contained neutralizing antibodies against WNV. Two bird samples were confirmed positive by VNT. These results show a significant seroprevalence of anti-WNV antibodies and therefore suggest the active circulation and exposure of human and bird populations in the northwest of Morocco.     

West Nile virus infection in horses in Saudi Arabia (in 2013–2015)

West Nile virus (WNV) is an important emerging zoonotic arbovirus giving rise to clinical syndromes of varying severity in humans and horses. Culex mosquitoes are the main vector. Although WNV has been reported in many countries in the Middle East and Asia, little is known about its prevalence in equine populations in the Arabian Peninsula. We have carried out a serological study on 200 horses to assess WNV infection in the Eastern and Central regions of Saudi Arabia in 2013–2015. Sera were tested for the presence of WNV antibodies in parallel using a commercial enzyme‐linked immunosorbent assay (ELISA) kit and microneutralization (MN) tests. In comparison with the MN assay used as “gold standard,” we find the ELISA had a sensitivity of 94.7% and specificity of 80.1%. The prevalence of WNV neutralizing antibody ranged from 5 (17.3%) of 29 sera collected in Riyadh up to 15 (55.6%) of 27 sera collected from Al‐Qateef. These findings highlight the need to be aware of the possibility of WNV disease in humans and horses presenting with central nervous system disease in the Kingdom of Saudi Arabia.     

A metapopulation model to simulate West Nile virus circulation in Western Africa, Southern Europe and the Mediterranean basin

In Europe, virological and epidemiological data collected in wild birds and horses suggest that a recurrent circulation of West Nile virus (WNV) could exist in some areas. Whether this circulation is permanent (due to overwintering mechanisms) or not remains unknown. The current conception of WNV epidemiology suggests that it is not: this conception combines an enzootic WNV circulation in tropical Africa with seasonal introductions of the virus in Europe by migratory birds. The objectives of this work were to (i) model this conception of WNV global circulation; and (ii) evaluate whether the model could reproduce data and patterns observed in Europe and Africa in vectors, horses, and birds. The model was calibrated using published seroprevalence data obtained from African (Senegal) and European (Spain) wild birds, and validated using independent, published data: seroprevalence rates in migratory and resident wild birds, minimal infection rates in vectors, as well as seroprevalence and incidence rates in horses. According to this model, overwintering mechanisms are not needed to reproduce the observed data. However, the existence of such mechanisms cannot be ruled out.     

The experimental infection of horses with Murray Valley encephalitis and Ross River viruses

Eleven weanling horses were inoculated with Murray Valley encephalitis and Ross River viruses either by intravenous injection or by the bite of Culex annulirostris or Aedes vigilax mosquitoes infected orally. Five of the 11 horses circulated trace amounts of MVE virus for 1 to 5d and they infected 7/408 Cx annulirostris which subsequently fed on them. Haemagglutination-inhibiting antibody persisted at detectable levels for the 24-week observation period. With Ross River virus, only one of 11 horses inoculated developed a viraemia detectable by inoculation of suckling mice but 5 horses contained virus sufficient to infect 41/383 Cx annulirostris that fed on them 3 to 4 days after inoculation. On primary inoculation with Ross River virus, only 2 horses developed HI antibody but late responses occurred in 3 horses following probable naturally acquired re-infections. With both viruses, most horses remained normal, some developed mild pyrexia and transient clinical signs. This paper, therefore, indicates that horses are unlikely to be efficient amplifiers of either MVE or RR viruses and does little to incriminate them as important pathogens.     

Pathogenesis of West Nile virus lineage 1 and 2 in experimentally infected large falcons

West Nile virus (WNV) is a zoonotic flavivirus that is transmitted by blood-suckling mosquitoes with birds serving as the primary vertebrate reservoir hosts (enzootic cycle). Some bird species like ravens, raptors and jays are highly susceptible and develop deadly encephalitis while others are infected subclinically only. Birds of prey are highly susceptible and show substantial mortality rates following infection. To investigate the WNV pathogenesis in falcons we inoculated twelve large falcons, 6 birds per group, subcutaneously with viruses belonging to two different lineages (lineage 1 strain NY 99 and lineage 2 strain Austria). Three different infection doses were utilized: low (approx. 500 TCID50), intermediate (approx. 4 log10 TCID50) and high (approx. 6 log10 TCID50). Clinical signs were monitored during the course of the experiments lasting 14 and 21 days. All falcons developed viremia for two weeks and shed virus for almost the same period of time. Using quantitative real-time RT-PCR WNV was detected in blood, in cloacal and oropharyngeal swabs and following euthanasia and necropsy of the animals in a variety of neuronal and extraneuronal organs. Antibodies to WNV were first time detected by ELISA and neutralization assay after 6 days post infection (dpi). Pathological findings consistently included splenomegaly, non-suppurative myocarditis, meningoencephalitis and vasculitis. By immunohistochemistry WNV-antigens were demonstrated intralesionally. These results impressively illustrate the devastating and possibly deadly effects of WNV infection in falcons, independent of the genetic lineage and dose of the challenge virus used. Due to the relatively high virus load and long duration of viremia falcons may also be considered competent WNV amplifying hosts, and thus may play a role in the transmission cycle of this zoonotic virus.