Serosurvey of Schmallenberg Virus Infection in the Highest Goat‐Specialized Region of France

The monitoring of both the spread and clinical impact of Schmallenberg virus (SBV) infection within its full host range is important for the control of the epidemic and potential new outbreaks. In France, a national surveillance plan based on voluntary notifications of congenital malformations in newborn ruminants revealed that goats were the less affected host species. However, seroprevalence studies only targeted sheep and cattle, preventing accurate estimations of the real impact of SBV infection in goats. Here, a serological survey was conducted in the highest goat‐specialized region of France between June 2012 and January 2013. A total of 1490 goat sera from 50 herds were analysed by ELISA. The between‐herd and within‐herd prevalences were estimated at 62% and 13.1%, respectively. Seroprevalence was not uniformly distributed throughout the territory and markedly differed between intensive and extensive herds. The low within‐herd seroprevalence demonstrates that a large fraction of the French goat population remains susceptible to SBV infection.     

Distribution of Schmallenberg Virus and Seroprevalence in Belgian Sheep and Goats

A serological survey to detect Schmallenberg virus (SBV)‐specific antibodies by ELISA was organized in the Belgian sheep population to study the seroprevalence at the end of the epidemic. One thousand eighty‐two sheep samples which were collected from 83 herds all over Belgium between November 2011 and April 2012 were tested. The overall within‐herd seroprevalence and the intraclass correlation coefficient were estimated at 84.31% (95% CI: 84.19–84.43) and 0.34, respectively. The overall between‐herd seroprevalence was 98.03% (95% CI: 97.86–98.18). A spatial cluster analysis identified a cluster of six farms with significantly lower within‐herd seroprevalence in the south of Belgium compared with the rest of the population (P = 0.04). It was shown that seroprevalence was associated to flock density and that the latter explained the presence of the spatial cluster. Additionally, 142 goat samples from eight different herds were tested for SBV‐specific antibodies. The within‐herd seroprevalence in goats was estimated at 40.68% (95% CI: 23.57–60.4%). The results of the current study provided evidence that almost every Belgian sheep herd has been in contact with SBV during 2011 and should be taken into consideration as part of comprehensive SBV surveillance and control strategies.     

Coxiella burnetii Infections in Small Ruminants and Humans in Switzerland

The recent Q fever epidemic in the Netherlands raised concerns about the potential risk of outbreaks in other European countries. In Switzerland, the prevalence of Q fever in animals and humans has not been studied in recent years. In this study, we describe the current situation with respect to Coxiella (C.) burnetii infections in small ruminants and humans in Switzerland, as a basis for future epidemiological investigations and public health risk assessments. Specific objectives of this cross‐sectional study were to (i) estimate the seroprevalence of C. burnetii in sheep and goats, (ii) quantify the amount of bacteria shed during abortion and (iii) analyse temporal trends in human C. burnetii infections. The seroprevalence of C. burnetii in small ruminants was determined by commercial ELISA from a representative sample of 100 sheep flocks and 72 goat herds. Herd‐level seroprevalence was 5.0% (95% CI: 1.6–11.3) for sheep and 11.1% (95% CI: 4.9–20.7) for goats. Animal‐level seroprevalence was 1.8% (95% CI: 0.8–3.4) for sheep and 3.4% (95% CI: 1.7–6) for goats. The quantification of C. burnetii in 97 ovine and caprine abortion samples by real‐time PCR indicated shedding of >10⁴ bacteria/g in 13.4% of all samples tested. To our knowledge, this is the first study reporting C. burnetii quantities in a large number of small ruminant abortion samples. Annual human Q fever serology data were provided by five major Swiss laboratories. Overall, seroprevalence in humans ranged between 1.7% and 3.5% from 2007 to 2011, and no temporal trends were observed. Interestingly, the two laboratories with significantly higher seroprevalences are located in the regions with the largest goat populations as well as, for one laboratory, with the highest livestock density in Switzerland. However, a direct link between animal and human infection data could not be established in this study.     

Serosurvey of Schmallenberg Virus Infections in Sheep and Goat Flocks in Lower Saxony,Germany

Schmallenberg virus (SBV) infections can cause congenital musculoskeletal and vertebral malformations as well as neurological failures in foetuses of several ruminant species if susceptible mother animals were infected during early gestation. Blood samples gained from 17 goat and 64 sheep flocks in Lower Saxony (LS), Germany (January–May 2012), which is located in the core region of the 2011/2012 epidemic were tested for antibodies against SBV by ELISA to detect past exposure to SBV. A SBV‐specific questionnaire was raised in all flocks. The calculated median within‐herd prevalence was 43.8% (min–max: 5.6–93.3%) for goats and 58.7% (min–max: 6.5–100%) for sheep, showing that small ruminants in LS, especially goats, are still at risk of novel SBV infections in the following lambing seasons as not all animals have seroconverted yet. Statistical analysis revealed that goats have a significantly lower risk of SBV infections than sheep which might be explained by different host preferences of Culicoides ssp. as main vectors for SBV and different housing conditions.     

Evidence for Circulation of Bovine Viral Diarrhoea Virus Type 2c in Ruminants in Southern Italy

Recently, bovine viral diarrhoea virus type 2c (BVDV‐2c) was responsible for a severe outbreak in cattle in northern Europe. Here, we present the results of an epidemiological survey for pestiviruses in ruminants in southern Italy. Pooled serum samples were obtained from 997 bovine, 800 ovine, 431 caprine and eight bubaline farms, and pestiviral RNA was detected by molecular methods in 44 farms consisting of 16 cattle and one buffalo herds and of 21 sheep and six goat flocks. Twenty‐nine and 15 farms were infected by BVDV‐1 and BVDV‐2 strains, respectively. BVDV‐1 strains were recovered mainly from cattle and were heterogeneous, belonging to the subtypes 1b, 1u, 1e, 1g and 1h. In contrast, all BVDV‐2 viruses but two were detected in sheep or goats and were characterized as BVDV‐2c by sequence analysis of 5′UTR. These strains displayed high genetic identity to BVDV‐2c circulating in cattle in northern Europe and were more distantly related to a BVDV‐2c isolate recovered from a cattle herd in southern Italy more than 10 years before. The circulation of a BVDV‐2c in small ruminants suggests the need for a continuous surveillance for the emergence of pestivirus‐induced clinical signs in southern Italian farms.     

Control of Bovine Brucellosis from Persistently Infected Holdings Using RB51 Vaccination with Test‐and‐Slaughter: A Comparative Case Report from a High Incidence Area in Portugal

Bovine brucellosis due to Brucella abortus infection causes significant reproductive and production losses in cattle and is a major zoonosis. Eradication of this disease has proved difficult to achieve in Portugal where it still occurs in some regions despite an ongoing national eradication programme. In 2004, the Alentejo region, a major cattle producing area, reported one of the highest levels of bovine brucellosis in the country, especially in one divisional area. In that area, bovine brucellosis was particularly problematic in a holding of ten herds, the largest extensive cattle unit in the country, which remained infected despite an extensive test‐and‐slaughter programme and depopulation of five herds. A 5‐year programme of RB51 vaccination with biannual test‐and‐slaughter was thus implemented in 2004. The apparent animal seroprevalence decreased from 19% (646/3,400) to 3% (88/2930) on the third herd‐level test and remained below 0.8% (27/3324) after the fourth test. After the tenth test, the holding had a prevalence of 0.1% (2/2332) and only one herd remained positive with a within‐herd prevalence of 1.1% (2/177). The results were compared to all other herds (n = 10) in the divisional area that were also persistently infected but were subject only to test‐and‐slaughter before being depopulated. In these herds, the strategy of test‐and‐slaughter did not reduce the prevalence, which remained significantly higher than the vaccinated group (median = 0.48% and 8.5% in vaccinated versus non‐vaccinated herds; Wilcoxon rank sum test; P < 0.01). The success of this pilot programme in continental Portugal provided a valuable case study to the official veterinary services by illustrating the value of RB51 vaccination with parallel testing and improved biosecurity as a comprehensive and sustainable strategy for bovine brucellosis control in persistently infected herds.     

Prevalence of caprine brucellosis on herds of toba communities in Villa Río Bermejito,Chaco, Argentina (October 2010)

The purpose of this study was to evaluate the prevalence of caprine brucellosis in goat herds in a region known as ‘The Impenetrable’ in Chaco Province, Argentina, in order to prevent the occurrence of the disease in people consuming goats’ milk. There are high levels of child malnutrition and difficulties in accessing high‐quality protein in this region; therefore, goats’ milk is a valuable nutrition source. We travelled with Patricia Sosa's foundation ‘Little gestures, big results’ to Villa Río Bermejito, Chaco, in October 2010 with the laboratory equipment needed to carry out field tests. The foundation is staffed by voluntary workers, including doctors, dentists, experts in social issues, psychologists, who encourage the community to consume goats’ milk in their meals after cooking or pasteurizing it to avoid contamination. Blood samples were taken from 273 goats distributed in 13 herds established in the depth of Chaco dry forest that had not been vaccinated against brucellosis. Samples were analysed using the buffered plate antigen test, and positive results were confirmed using the slow agglutination test. Animals were identified using small plastic ear tags. We found only one positive case, with a titre of 1/50. This was considered suspicious and the owner was advised to take the animal out of the herd. Our results indicate that the isolation of these herds in the depths of the forest has protected them from brucellosis, and therefore, they are a good source of high‐quality protein, which the people of this region need.     

Association of brucellosis to abortions in humans and domestic ruminants in Kagera ecosystem,Tanzania

Brucellosis is a worldwide zoonotic disease of socio‐economic importance. Understanding the association of this disease with pregnancy outcome has the potential of contributing to the reduction of its reproductive burden in humans and animals among pastoral communities in Tanzania. A prospective cohort study was conducted in Kagera Region on pregnant women (n = 76) and gravid ruminants (121 cattle, 125 goats and 111 sheep). Exposed and non‐exposed groups to brucellosis were followed for 6 months (from 15 November 2017 to 15 April 2018). Sera were collected and analysed using Rose Bengal Test (RBT) and Fluorescence polarization assay (FPA) test. Measures of effect, univariable and multivariable logistic regression analyses were computed. Positivity to both RBT and FPA tests was 21% (95% CI: 12.5–32) in pregnant women and 5% (95% CI: 3.1–8) in gravid ruminants. Among aborted cases, four women (out of nine), two cows (out of seven), two goats (out of 26) and zero sheep (out of 11) were positive to brucellosis. The abortion rate in humans and ruminants was 11.8% and 12.3%, respectively. Seropositivity to brucellosis was similar in aborted and non‐aborted cases in humans (p = .08) and in ruminants (p = .2). At the population level, brucellosis was associated with abortions (population attributable risk: PAR) at 3.5% in pregnant women and at 0.5% in gravid ruminants in the study area. Infections to brucellosis were increased in exposed pregnant women (OR = 19; 95% CI: 1.8–203, p = .01) and in cattle (OR = 11; 95% CI: 1.3–88, p = .02). There is an indication that brucellosis could be contributing to abortions in pregnant women and domestic ruminants Kagera Region. Molecular tools could support more the results from serological tests to avoid cross‐reaction with other pathogen agents. Control of brucellosis in animals is likely to reduce the threat of abortions in humans.     

Serological Detection of Antibodies to Peste des Petits Ruminants Virus in Large Ruminants

Peste des petits ruminants (PPR) is an economically important disease of small ruminants with a rapidly expanding geographical distribution. Peste des petits ruminants virus may manifest in a variety of ways with disease ranging from acute to subclinical. We investigated the exposure of large ruminants to PPRV in areas where the virus is endemic in the small ruminant population by assessing the serological status of groups of animals. This study focused on the Punjab province of Pakistan as an area where the virus is endemic and where mixed farming practices occur enabling close interactions between small and large ruminant populations. An overall PPR seropositivity was detected in 10.0% of cattle and 14.16% of buffaloes. Following an assessment of serological profiles in large ruminants within different age groups, a maximum seroprevalence was observed in cattle (17.5%) and buffaloes (22.5%) over 2 years of age indicating the potential utility of sampling large ruminant populations for PPR serosurveillance. The large ruminants sampled between one and two years of age had similar levels of seropositivity within populations with 11.2% and 16.2% of animals being seropositive, respectively. Current PPR vaccination strategies do not enable the differentiation between infected and vaccinated small ruminants, and as such, the serological surveillance of sheep and goats is of little value. When considering eradication programmes for PPRV, this factor is of great significance. However, where large and small ruminants are farmed together, serological surveillance of large ruminants may provide a snapshot of virus infection within populations where mild disease is present or where small ruminants are regularly vaccinated.     

Diagnosis of Peste des Petits Ruminants in Wild and Domestic Animals in Xinjiang,China, 2013–2016

Peste des petits ruminants viruses (PPRVs) re‐emerged in China at the end of 2013 and then spread rapidly into 22 provinces through movement of live goats and sheep. In this study, 96 samples of domestic animals and 13 samples of wildlife were analysed for the presence of PPRV infection by ELISA or RT‐PCR. Of 96 samples from sheep and goats, 91 were PPRV positive, whereas all of the 13 samples from three wild species, Capra ibex (Capra ibex sibirica), argali (Ovis ammon) and Goitered gazelle (Gazella subgutturosa), were found to be positive. Five wildlife‐origin isolates from the above samples were identified as the lineage IV by a multiple alignment of the partial sequences in N gene.     

Follow‐up of the Schmallenberg Virus Seroprevalence in Belgian Cattle

Schmallenberg virus (SBV), which emerged in Northwestern Europe in 2011, is an arthropod‐borne virus affecting primarily ruminants. Based on the results of two cross‐sectional studies conducted in the Belgian ruminant population during winter 2011–2012, we concluded that at the end of 2011, almost the whole population had already been infected by SBV. A second cross‐sectional serological study was conducted in the Belgian cattle population during winter 2012–2013 to examine the situation after the 2012 transmission period and to analyse the change in immunity after 1 year. A total of 7130 blood samples collected between 1st January and 28 February 2013 in 188 herds were tested for the presence of SBV‐specific antibodies. All sampled herds tested positive and within‐herd seroprevalence was estimated at 65.66% (95% CI: 62.28–69.04). A statistically significant decrease was observed between the beginning and the end of 2012. On the other hand, age‐cohort‐specific seroprevalence stayed stable from 1 year to the other. During winter 2012–2013, calves between 6 and 12 months had a seroprevalence of 20.59% (95% CI: 15.34–25.83), which seems to be an indication that SBV was still circulating at least in some parts of Belgium during summer–early autumn 2012. Results showed that the level of immunity against SBV of the animals infected has not decreased and remained high after 1 year and that the spread of the virus has slowed down considerably during 2012. This study also indicated that in the coming years, there are likely to be age cohorts of unprotected animals.     

Post‐epidemic investigation of Schmallenberg virus in wild ruminants in Slovenia

Schmallenberg virus (SBV) is a vector‐borne virus belonging to the genus Orthobunyavirus within the Bunyaviridae family. SBV emerged in Europe in 2011 and was characterized by epidemics of abortions, stillbirths and congenital malformations in domestic ruminants. The first evidence of SBV infection in Slovenia was from an ELISA‐positive sample from a cow collected in August 2012; clinical manifestations of SBV disease in sheep and cattle were observed in 2013, with SBV RNA detected in samples collected from a total of 28 herds. A potential re‐emergence of SBV in Europe is predicted to occur when population‐level immunity declines. SBV is also capable of infecting several wild ruminant species, although clinical disease has not yet been described in these species. Data on SBV‐positive wild ruminants suggest that these species might be possible sources for the re‐emergence of SBV. The aim of this study was to investigate whether SBV was circulating among wild ruminants in Slovenia and whether these species can act as a virus reservoir. A total of 281 blood and spleen samples from wild ruminants, including roe deer, red deer, chamois and European mouflon, were collected during the 2017–2018 hunting season. Serum samples were tested for antibodies against SBV by ELISA; the overall seroprevalence was 18.1%. Seropositive samples were reported from all over the country in examined animal species from 1 to 15 years of age. Spleen samples from the seropositive animals and serum samples from the seronegative animals were tested for the presence of SBV RNA using real‐time RT‐PCR; all the samples tested negative. Based on the results of the seropositive animals, it was demonstrated that SBV was circulating in wild ruminant populations in Slovenia even after the epidemic, as almost half (23/51) of the seropositive animals were 1 or 2 years old.     

First detection and molecular identification of Sarcocystis spp. in small ruminants in North‐West Tunisia

Sarcocystosis is a parasitic disease caused by varying Sarcocystis species infecting humans and animals. It is commonly found in small ruminants causing pathogenic effects. This contributes to detrimental economic loss for local farmers and the local economy due this disease. Although the distribution of Sarcocystis can be found all over the world, the species infecting small ruminants in Tunisia is still unknown. Through this study, we aim to estimate the molecular prevalence of natural infection with Sarcocystis spp. in sheep and goats using molecular identification. Also, phylogenetic analyses were used to identify the different species of this parasite infecting small ruminants in northern Tunisia for the first time. DNA was extracted from 198 and 121, sheep and goats meat samples, respectively. The molecular prevalence of Sarcocystis spp. in sheep and goats was 58.6% (116/198) and 50.4% (61/121), respectively. Compared to the Noire de Thibar and cross‐breeds, the Barbarine sheep had the highest infection prevalence (63.4%) (p  =  .004). Five of the 116 positive samples were sequenced identifying Sarcocystis tenella from sheep. For goats, the sequencing results showed that five positive PCR products belonged to Sarcocystis capracanis species.     

Serological Screening Suggests Presence of Schmallenberg Virus in Cattle,Sheep and Goat in the Zambezia Province,Mozambique

Schmallenberg virus (SBV) is a novel Orthobunyavirus within the family Bunyaviridae belonging to the Simbu serogroup. Schmallenberg virus infects ruminants and has since its discovery in the autumn 2011 been detected/spread to large parts of Europe. Most bunyaviruses are arboviruses, and SBV has been detected in biting midges in different European countries, suggesting that they may play a role in the transmission of the virus. It is not known how SBV was introduced to Europe and if SBV is present in countries outside of Europe. Thus, in this study, we conducted a serological screening for SBV antibodies in cattle (no. 79), sheep (no. 145) and goat (no. 141) in the Zambezia Province in Mozambique during September 2013. The results show a high percentage of antibody‐positive animals. All farms tested had seropositive animals; cattle displayed the highest prevalence with 100% positive animals. Sheep and goat also displayed high number of positive animals with a 43–97% and 72–100% within‐herd seroprevalence, respectively. This initial serological screening suggests that SBV is present on the African continent. However, cross‐reactivity with other members of the Simbu serogroup cannot be ruled out, and further studies are needed to identify and characterize the virus responsible for the antibody‐positive results.     

Evidence for Tunisian‐Like Pestiviruses Presence in Small Ruminants in Italy Since 2007

The genus Pestivirus, which belongs to the Flaviviridae family, includes ssRNA+ viruses responsible for infectious diseases in pigs, cattle, sheep, goats and other domestic and wild ruminants. Like most of the RNA viruses, pestivirus has high genome variability with practical consequences on disease epidemiology, diagnosis and control. In addition to the officially recognized species in the genus Pestivirus, such as BVDV‐1, BVDV‐2, BDV and CSFV, other pestiviruses have been detected. Furthermore, most of the ruminant pestiviruses show low or absent species specificity observed in serological tests and are able to infect multiple species. Particularly, small ruminants are receptive hosts of the most heterogeneous group of pestiviruses. The aim of this study was to carry out the molecular characterization of pestiviruses isolated from sheep and goats in Sicily, Italy. Phylogenetic analysis of two viral genomic regions (a fragment of 5′‐UTR and the whole N^pro regions) revealed the presence of different pestivirus genotypes in the analysed goat and sheep herds. Two of five viral isolates were clustered with BVDV‐1d viruses, a strain widespread in Italy, but never reported in Sicily. The other three isolates formed a distinct cluster with high similarity to Tunisian isolates, recently proposed as a new pestivirus species. This represents the first evidence for Tunisian‐like pestivirus presence in small ruminants in Italy. Furthermore, one of the isolates was collected from a goat, representing the first isolation of Tunisian‐like pestivirus from this species.     

The impact of changing farm structure on foot‐and‐mouth disease spread and control: A simulation study

Foot‐and‐mouth disease (FMD) is a highly contagious viral disease that affects ruminants and pigs. Countries with large exports of livestock products are highly vulnerable to economic damage following an FMD incursion. The faster disease spread is controlled, the lower the economic damage. During the past decades, the structure of livestock production has dramatically changed. To maintain the relevance of contingency plans, it is important to understand the effects of changes in herd structure on the spread and control of infectious diseases. In this study, we compare the spread and control of FMD based on 2006/2007 and 2018 livestock data. Spread of FMD in Denmark was simulated using the DTU‐DADS model, applying different control measures. The number of cattle, swine and sheep/goat herds reduced from about 50,000 in total in 2006/2007 to about 33,000 in 2018. During this period, the average number of outgoing animal movements and the exports of swine and swine products increased by about 35% and 22%, respectively. This coincided with an overall increase in herd size of 14%. Using the EU and national control measures (Basic: 3 days standstill, depopulation of detected herds followed by cleaning and disinfection and establishment of control zones, where tracing, surveillance and contact restrictions are implemented), we found that the simulated epidemics in 2018 would be about 50% shorter in duration, affect about 50% fewer herds but cause more economic damage, compared to epidemics using 2006/2007 data. When 2006/2007 data were used, Basic + pre‐emptive depopulation (Depop) overall was the optimal control strategy. When 2018 data were used, this was the case only when epidemics were initiated in cattle herds, whereas when epidemics were initiated in sow or sheep/goats herds, basic performed as well as Depop. The results demonstrate that regular assessment of measures to control the spread of infectious diseases is necessary for contingency planning.     

A molecular and pathological study of peste des petits ruminants virus (PPRV) from field outbreaks in Palestine, 2017–2019

Peste des petits ruminants (PPR) is a severe, highly contagious and fatal viral disease of small ruminants causing high economic losses. Therefore, the disease was targeted for eradication by 2030. The aim of this study was to investigate for the first time the molecular and pathological characterization of the circulating PPR virus (PPRV) in sheep and goat in Palestine. Samples were collected from suspected necropsy cases of sheep and goats during recent outbreaks in two provinces in Palestine between 2017 and 2019. In this study, severe PPR outbreaks occurred in sheep and goats causing typical lesions which include erosive and ulcerative stomatitis, bronchointerstitial pneumonia and severe enteritis. For the molecular investigation of PPRV, suspected animals were examined for the presence of PPRV by RT‐PCR. PPRV genome was detected in all samples. Subsequently, two samples were used for N gene sequencing and phylogenetic analysis of PPRV isolates. The nucleotide sequence and phylogenetic analysis indicated that the Palestinian PPRV isolates were genetically clustered within the lineage IV isolates of the virus among populations of sheep and goats which most prevalent in Asia, the Middle East and recently Africa. Further analysis showed that the Palestinian isolates were closely related to those described in Turkey and Iraq, suggesting a common origin of PPRV isolates in the region. This information is critical to understand the molecular epidemiology of this disease in the region and helps to develop appropriate control measures for eradication of this disease.     

Evidence of extensive renewed Schmallenberg virus circulation in Belgium during summer of 2016 – increase in arthrogryposis‐hydranencephaly cases expected

A seroprevalence study carried out between June and September 2016 in the Belgian sheep population showed a significant increase in overall (from 25% to 62%) and between‐herd (from 60% to 96%) seroprevalence against Schmallenberg virus (SBV) during this period, indicating the most extensive recirculation of SBV since its original emergence in 2011. SBV recirculation was confirmed by the detection of SBV RNA‐positive Culicoides obsoletus complex midges collected in the region of Antwerp in August 2016, reaching a minimum infection rate of 3%. The recirculation of SBV in the largely unprotected ruminant population during summer 2016 will likely cause an increase in the number of arthrogryposis‐hydranencephaly cases in newborn ruminants during the coming months.     

Monitoring of the West Nile Virus epidemic in Spain between 2010 and 2011

West Nile virus (WNV) is a mosquito‐transmitted flavivirus recognized as an emerging and re‐emerging pathogen in different countries. This study describes the monitoring of the first WNV epidemic in Spain between 2010 and 2011. Between September and December 2010, 36 outbreaks of WNV in horses were reported in three different provinces of Andalusia (southern Spain), with no apparent spread outside this area. The temporal distribution and the clinical signs observed during the WNV epidemic in Spain were, in general, similar to those reported in Europe and in the Mediterranean Basin. Morbidity, mortality and fatality rate in the affected herds were 4.6, 1.4 and 35.3%, respectively. Thirty‐six of 75 (47.4%) suspected herds investigated presented at least one IgM seropositive animal. The individual seroprevalence in unvaccinated animals from the infected holdings was 51.7%. RNA WNV lineage 1 virus was confirmed from blood and cerebrospinal fluid samples in a lethally infected horse. The entomological survey showed that the most abundant mosquito species detected in the affected area was Culex pipiens. A cross‐sectional study was carried out in non‐suspected herds between April 2010 and February 2011 in the affected area. The individual seroprevalence was 11.0%, and six of the 38 herds sampled (15.8%) presented at least one seropositive animal. The results showed active WNV circulation several months before the first outbreak was reported in horses. The seropositivity found in municipalities where clinical cases were not reported indicates a higher geographical dissemination of the virus. Significantly higher seroprevalences were detected in areas close to Morocco. Furthermore, 90 wild ruminants were tested for the presence of antibodies against WNV, but the results were all negative.