Surveillance and Early Warning of West Nile Virus Lineage 2 Using Backyard Chickens and Correlation to Human Neuroinvasive Cases

In 2010, a West Nile virus (WNV) epidemic was reported in Central Macedonia, Northern Greece, with 197 neuroinvasive disease (WNND) cases in humans. The following 3 years, WNV spreads to new areas of Greece and human cases reoccurred during the transmission periods. After the initial outbreak, a WNV surveillance system using juvenile backyard chickens was established in Central Macedonia (after the 2011 outbreak) and Eastern Macedonia‐Thrace (after the 2012 outbreak). Sera were screened for the presence of antibodies against WNV using cELISA and serum neutralization test, to monitor the spread of WNV and to assess the correlation between the WNV point seroprevalence in chickens and the incidence rates of human WNND cases in the aforementioned areas. WNV seroprevalence in chickens was 10.4% (95% CI: 7–15) in Central Macedonia (2011) and 18.1% (95% CI: 14–23) in Eastern Macedonia‐Thrace (2012). Seroprevalence in chickens and incidence rates of human WNND cases in Eastern Macedonia‐Thrace were strongly positively correlated (ρ = 0.98, P = 0.005) at the regional unit level, with the incidence of WNND in humans increasing with increasing WNV point seroprevalence in chickens. In Central Macedonia, the correlation was weaker (ρ = 0.68, P = 0.20), apparently due to small number of reported human WNND cases. Another study was also conducted using juvenile backyard chickens in Central Macedonia, aiming to detect early WNV enzootic circulation, before the onset of human cases during 2011 and 2013. The first seroconverted chickens were detected about 1.5 months before the laboratory diagnosis of any human WNND cases in Central Macedonia, for both years. WNV surveillance, using juvenile backyard chickens, was reliable for the identification of areas with WNV enzootic and silent transmission, and for early warning. Timely diffusion of information to public health authorities facilitated the successful implementation of preparedness plans to protect public health.     

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.     

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.     

Serologic evidence of West Nile virus and Usutu virus infections in Eurasian coots in the Netherlands

West Nile virus (WNV) and Usutu virus (USUV) are arboviruses that are maintained in enzootic transmission cycles between mosquitoes and birds and are occasionally transmitted to mammals. As arboviruses are currently expanding their geographic range and emerging in often unpredictable locations, surveillance is considered an important element of preparedness. To determine whether sera collected from resident and migratory birds in the Netherlands as part of avian influenza surveillance would also represent an effective source for proactive arbovirus surveillance, a random selection of such sera was screened for WNV antibodies using a commercial ELISA. In addition, sera of jackdaws and carrion crows captured for previous experimental infection studies were added to the selection. Of the 265 screened serum samples, 27 were found to be WNV–antibody‐positive, and subsequent cross‐neutralization experiments using WNV and USUV confirmed that five serum samples were positive for only WNV‐neutralizing antibodies and seven for only USUV. The positive birds consisted of four Eurasian coots (Fulica atra) and one carrion crow (Corvus corone) for WNV, of which the latter may suggest local presence of the virus, and only Eurasian coots for USUV. As a result, the screening of a small selection of serum samples originally collected for avian influenza surveillance demonstrated a seroprevalence of 1.6% for WNV and 2.8% for USUV, suggesting that this sustained infrastructure could serve as a useful source for future surveillance of arboviruses such as WNV and USUV in the Netherlands.     

Spreading of West Nile virus infection in Croatia

West Nile virus (WNV) is an emerging zoonotic pathogen with rapid global expansion. The virus circulation is confirmed in many countries of Mediterranean Basin and Southern and Central Europe. In our study detection of specific WNV antibodies was performed in horses and cattle sera samples collected from October 2010 to April 2011. Serum samples were randomly taken from different parts of Croatia and tested by IgG and IgM ELISA. Positive serological results were confirmed by virus neutralization assay (VN-assay) and plaque reduction neutralization test (PRNT). Results showed that WNV antibodies were present in 72 out of 2098 horse sera (3.43%) and 3 of 2695 cattle sera (0.11%). The highest seroprevalence was found in Eastern Croatia in counties next to Hungarian, Serbian and Bosnia and Herzegovinian state borders. In Adriatic part of Croatia positive animals were found only in the westernmost county, near Slovenian and Italian borders. Geographic distribution and number of positive horses indicated that WNV is highly present in Croatia and spreading from East to West. However, positive horses in westernmost part of country indicate possible second origin of spreading. Location of serological positive cattle supports the hypothesis that seropositive cattle could be indicators of high WNV activity in the respective geographic regions.     

West Nile virus infection in horses,Indian ocean

The circulation of West Nile virus (WNV) in horses was investigated in the Southwest Indian ocean. In 2010, blood samples were collected from a total of 303 horses originating from Madagascar, Mauritius, Reunion and the Seychelles and tested for WNV-specific antibodies. An overall seroprevalence of 27.39% was detected in the Indian Ocean with the highest WNV antibody prevalence of 46.22% (95% CI: [37.4–55.2%]) in Madagascar. The age and origin of the horses were found to be associated with the WNV infection risk. This paper presents the first seroprevalence study investigating WN fever in horses in the Southwest Indian Ocean area and indicates a potential risk of infection for humans and animals. In order to gain a better understanding of WN transmission cycles, WNV surveillance needs to be implemented in each of the countries.     

Recent rapid changes in the spatio‐temporal distribution of West Nile Neuro‐invasive Disease in Italy

In Italy, the first human case of West Nile Virus (WNV) infection was reported in 2008 and, since then, the number of cases has been steadily increasing. In this study, we describe the temporal and spatial pattern of WNV infection risk among humans in Italy, focusing on the human cases of West Nile Neuro‐invasive Disease (WNND) observed between 2008 and 2017. Incidence rates are estimated for each year and province under study. The incidence temporal trend is estimated using Poisson regression, and a spatio‐temporal cluster detection analysis is performed to detect high‐risk areas. In total, 231 WNND cases were notified in Italy between 2008 and 2017. The annual incidence rates increased during the study period (annual percentage change: 11.7%; 95%CI: ?0.9%; 26.1%). A geographical spread of the disease was observed during the study period throughout Northern Italy, with an increasing number of affected provinces. Provinces close to the Po River (the main river in the north of Italy) and the Oristano province (in the Sardinia Island) experienced the highest incidence rates during the study period. Our study shows a gradual, but rapid spread of WNND across Northern Italy from east to west and suggests the hypothesis that provinces close to Po River might present ecological and climatic conditions favourable to the virus circulation.     

The first detection of anti-West Nile virus antibody in domestic ruminants in Egypt

West Nile virus (WNV) is a mosquito-borne disease, usually present as a symptomatic disease but can cause various clinical signs ranged from mild fever to severe encephalitis and death in various animals and humans. In Egypt, the epidemiological data about WNV infection in different animal species particularly in domestic ruminants are scarce. The present study aimed to investigate the seroprevalence of WNV in cattle, buffalo, camel, sheep, and goats at some Governorates northern Egypt. In total, 360 serum samples (100 cattle, 50 buffalo, 50 camels, 85 sheep, and 75 goats) were examined using ELISA. The results revealed that the seroprevalence of WNV among ruminants was highly significant (P = 0.03) at Kafr El Sheikh Governorate (17.6%) in comparison with other the Governorates. Besides, the seroprevalence of WNV antibodies significantly differed between the examined species (P = 0.0001); it was 22%, 0%, 40%, 3.5%, and 5.3% in cattle, buffalo, camel, sheep, and goats, respectively. This is the first study to confirm that domestic ruminants act as a reservoir in the epidemiology of WNV infection and represent a risk for human and equine infections in Egypt.

     

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.     

Ongoing Activity of Toscana Virus Genotype A and West Nile Virus Lineage 1 Strains in Turkey: A Clinical and Field Survey

Toscana virus (TOSV), West Nile virus (WNV) and tickborne encephalitis virus (TBEV) are among major viral pathogens causing febrile disease and meningitis/encephalitis. The impact of these viruses was investigated at a referral centre in Ankara Province, Central Anatolia in 2012, where previous reports suggested virus circulation but with scarce information on clinical cases and vector activity. Serum and/or cerebrospinal fluid samples from 94 individuals were evaluated, in addition to field‐collected arthropod specimens that included 767 sandflies and 239 mosquitoes. Viral nucleic acids in clinical samples and arthropods were sought via specific and generic nested/real‐time PCRs, and antibody responses in clinical samples were investigated via commercial indirect immunofluorescence tests (IIFTs) and virus neutralization. A WNV antigen assay was also employed for mosquitoes. WNV neuroinvasive disease has been identified in a 63‐year‐old male via RNA detection, and the WNV strain was characterized as lineage 1. TOSV infections were diagnosed in six individuals (6.3%) via RNA or IgM detection. Partial sequences in a 23‐year‐old female, presented with fever and transient pancytopenia, were characterized as TOSV genotype A. Febrile disease with arthralgia and/or peripheral cranial nerve involvement was noted in cases with TOSV infections. Previous WNV and TOSV exposures have been observed in 5.3% and 2.1% of the subjects, respectively. No confirmed TBEV exposure could be identified. Morphological identification of the field‐collected mosquitoes revealed Culex pipiens sensu lato (74.4%), Anopheles maculipennis (20.9%), An. claviger (2.1%) and others. Sandfly species were determined as Phlebotomus papatasi (36.2%), P. halepensis (27.3%), P. major s. l. (19.3%), P. sergenti (8.9%), P. perfiliewi (4.4%), P. simici (2.6%) and others. Viral infections in arthropods could not be demonstrated. TOSV genotype A and WNV lineage 1 activity have been demonstrated as well as serologically proven exposure in patients. Presence of sandfly and mosquito species capable of virus transmission has also been revealed.     

First serological evidence of West Nile virus infection in wild birds in Northern Algeria

While the epidemiology of Flaviviruses has been extensively studied in most of the Mediterranean basin, little is known about the current situation in Algeria. In order to detect the circulation of West Nile (WNV) and Usutu viruses (USUV) in Kabylia, 165 sera were collected from two wild birds species, namely the long distance migrant Turdus philomelos (song thrush) (n = 92) and the resident Passer domesticus (house sparrow) (n = 73). A total of 154 sera were first analyzed by commercial competition ELISA. WNV and USUV micro-neutralization tests were performed on all c-ELISA positive sera and all samples with poor volume. Overall, 7.8 % (CI95 %: 3.5–11.9) were positive by c-ELISA. Positive results were detected in 12.5 % (CI95 %:5.6–19.4) of song thrushes and 1.5 % (CI95 %: 0.0–4.5) for sparrow.Micro-neutralization tests revealed an overall seroprevalence of 6.7 % for WNV (CI95 %: 2.9–10.3), Neutralizing antibodies were found in 8.7 % (CI95 %: 3.0–14.4) for song thrushes and in 4.1 % (CI95 %: 0.0–8.7) of sparrows. The current study demonstrates significant seroprevalence of WNV antibodies in wild birds in Algeria.     

Serosurveillance of Schmallenberg virus in Switzerland using bulk tank milk samples

Infections with Schmallenberg virus (SBV), a novel Orthobunyavirus transmitted by biting midges, can cause abortions and malformations of newborns and severe symptoms in adults of domestic and wild ruminants. Understanding the temporal and spatial distribution of the virus in a certain territory is important for the control and prevention of the disease. In this study, seroprevalence of antibodies against SBV and the spatial spread of the virus was investigated in Swiss dairy cattle applying a milk serology technique on bulk milk samples. The seroprevalence in cattle herds was significantly higher in December 2012 (99.5%) compared to July 2012 (19.7%). This high between-herd seroprevalence in cattle herds was observed shortly after the first detection of viral infections. Milk samples originating from farms with seropositive animals taken in December 2012 (n = 209; mean 160%) revealed significantly higher S/P% ratios than samples collected in July 2012 (n = 48; mean 103.6%). This finding suggests a high within-herd seroprevalence in infected herds which makes testing of bulk tank milk samples for the identification farms with past exposures to SBV a sensitive method. It suggests also that within-herd transmission followed by seroconversion still occurred between July and December. In July 2012, positive bulk tank milk samples were mainly restricted to the western part of Switzerland whereas in December 2012, all samples except one were positive. A spatial analysis revealed a separation of regions with and without positive farms in July 2012 and no spatial clustering within the regions with positive farms. In contrast to the spatial dispersion of bluetongue virus, a virus that is also transmitted by Culicoides midges, in 2008 in Switzerland, the spread of SBV occurred from the western to the eastern part of the country. The dispersed incursion of SBV took place in the western part of Switzerland and the virus spread rapidly to the remaining territory. This spatial pattern is consistent with the hypothesis that transmission by Culicoides midges was the main way of spreading.     

Flaviviruses in migratory passerines during spring stopover in a desert oasis

Bird migration has long been hypothesized as the main mechanism for long‐distance dispersal of flaviviruses, but the role of migratory birds in flaviviruses spillover is not well documented. In this study, we investigated the eco‐epidemiology of West Nile virus (WNV) and Usutu virus (USUV) in trans‐Saharan passerines during their spring stopover in southern Tunisian oases. To do, we combined oral swab analysis and serological tools to assess whether migratory birds could be reaching these stopover sites while infectious or have been previously exposed to viruses. All sampled birds tested negative for oral swab analysis. However, anti‐WNV and anti‐USUV antibodies were detected in 32% and 1% of tested birds, respectively. Among WNV‐seropositive species, the Golden oriole (Oriolus oriolus) showed the highest anti‐WNV occurrence probability. In this species, anti‐WNV occurrence was twice larger in males than females. Inter‐specific and intraspecific morphological, physiological and behavioural differences could explain these results. Although our findings did not show evidence for passerines migrating while infectious, they did not exclude an existing enzootic WNV transmission cycle in Tunisian oases. Further investigations including larger samples of migratory birds are needed for a better understanding of this issue.     

Fecal carriage of multi-drug resistant and extended spectrum β-lactamases producing E. coli in household pigeons,Bangladesh

Antibiotic resistance and ESBL constitute a risk to human and animal health. Birds residing close to humans could mirror the spectrum of human associated antibiotic resistance. Household pigeons were screened in Bangladesh to shed light on human associated, as well as, environmental antibiotic resistance. Escherichia coli from pigeons (n = 150) were tested against 11 antibiotics. 89% E. coli isolates were resistant to one or more critically important human antibiotics like ampicillin, cefadroxil, mecillinam, ciprofloxacin, gentamicin and tigecycline. No carbapenamase-producers were detected and the lower ESBL prevalence (5%) in pigeons. ESBL-producing E. coli isolates had bla_CTX-M-15 genes. Pigeons shared some bacterial clones and had bird associated sequence types like E. coli ST1408. Fecal carriage of bacteria resistance of critically important human antibiotics, together with examples of shared genotypes among pigeons, indicate the human-birds and bird to bird transmissions are important in the epidemiology of antibiotic resistance.     

Evidence of West Nile virus seropositivity in wild birds on the island of Cyprus

West Nile Virus (WNV)¹ is an emerging pathogen in Cyprus, with the first human case of infection reported in 2016, and another documented in 2018. A cluster of cases in humans was then reported in 2019. However, little is known regarding which avian species might bring WNV to Cyprus. Here, we investigated seroprevalence of WNV antibodies in migratory and resident birds, captured across Cyprus to assess to what extent human populations might be exposed to WNV. We used Enzyme-Linked Immunosorbent Assay (ELISA)² to test for the presence of WNV antibodies in 836 avian blood samples of 44 species captured between 2015 and 2020. A seropositivity rate of 1.3 % was found. The majority of seropositive wild birds belonged to the migratory species Sylvia atricapilla, a common and widespread migrant, implying a high risk of WNV being introduced throughout Cyprus.     

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.     

Epidemic West Nile virus encephalomyelitis: a temperature-dependent, spatial model of disease dynamics

Since first being detected in New York in 1999, West Nile virus (WNV) has spread throughout the United States and more than 20,000 cases of equine WNV encephalomyelitis have been reported. A spatial model of disease occurrence was developed, using data from an outbreak of serologically confirmed disease in an unvaccinated population of horses at 108 locations in northern Indiana between 3 August and 17 October 2002. Daily maximum temperature data were recorded at meteorological stations surrounding the study area. The distribution of the total number of degree-days elapsing between July 4 and the date of diagnosis of each case was best described by a normal distribution (mean = 5243 °F, S.D. = 1047). The days on which the average risk was >25, >50 and >75% were predicted (versus observed) to occur on August 23 (August 9), August 31 (September 2) and September 9 (September 9). The epidemic was predicted to occur 3 days earlier, or 4 days later, than observed if temperatures in the study area were uniformly increased, or decreased, by 5 °F, respectively. Maps indicated that WNV encephalomyelitis risk always remained greater in the northwest quadrant of the study area. Since WNV might exist at a hypoendemic level of infection, and occasionally re-emerge as a cause of epidemics in equine populations, by identifying factors that contributed to this epidemic, the potential impact of future epidemics can be reduced. Such studies rely on a GIS framework, availability of meteorological and possibly remotely sensed data and information on host and landscape factors. An early-warning system for WNV transmission in equine populations could be developed.     

Seroprevalence and risk factors of Mycoplasma suis infection in pig farms in central China

Mycoplasma suis, the causative agent of porcine infectious anemia, causes large economic losses to the swine industry worldwide. A questionnaire-based survey was conducted in 69 pig farms in Hubei Province, China, from November 2011 to August 2013 to ascertain the prevalence and associated risk factors of M. suis. Four thousand and four blood samples from pigs of all the age groups were tested for M. suis antibodies using the established rMSG1–ELISA assay. Among these 4004 samples, 1615 blood samples from multiparous sows were examined to identify the association between seroprevalence and different seasons. Information on risk factors collected from farmers or attending veterinarians was recorded on a pre-designed questionnaire. The overall test seroprevalence of M. suis infection at the animal level was 31.9% (1277/4004; 95% CI: 30.5%, 33.4%), whereas at the farm level, this value was 95.65% (66/69; 95% CI: 87.8%, 99.1%). The seroprevalence of M. suis was higher in replacement gilts (40.6%; 95% CI: 35.1%, 46.3%), multiparous sows (48.2%; 95% CI: 45.8%, 50.7%) and boars (44.4%; 95% CI: 34.5%, 54.8%), as compared to piglets (13.0%; 95% CI: 9.4%, 17.3%), weaned-piglets (10.8%; 95% CI: 8.9%, 13.0%), and growing-finishing pigs (25.0%; 95% CI: 22.0%, 28.3%). In terms of seasons, the prevalence of M. suis in pigs was significantly higher in summer (65.3%; 95% CI: 61.0%, 69.5%) and autumn (65.0%; 95% CI: 59.0%, 70.6%) compared to spring (30.1%; 95% CI: 26.0%, 34.4%) and winter (36.4%; 95% CI: 31.4%, 41.5%). Farm-level risk factors were identified by multivariable logistic regression analysis. The associated factors retained in the final multivariable logistic regression model were drug treatment, presence of mosquitoes and flies, and frequency of disinfection. Drug treatment (OR = 0.24; 95% CI: 0.07, 0.88; P = 0.031) and frequency of disinfection (OR = 0.23; 95% CI: 0.06, 0.90; P = 0.035) were protective factors, and the presence of mosquitoes and flies (OR = 5.994; 95% CI: 1.56, 23.00; P = 0.009) was a risk factor for M. suis infection on farms. The results of the present study provide the first insight into the impact of associated determinants on M. suis infection in central China.