Anna Alba

Bluetongue epidemiology in wild ruminants from Southern Spain

Serum samples from 210 wild ruminants collected between 2006 and 2007 in southern Spain were tested for antibodies against bluetongue virus (BTV) by means of a competitive ELISA assay. Eighty-seven of the 210 wild ruminants analysed (41%) showed antibodies against BTV. Statistically significant differences were found in the seroprevalence among species: 66% (65 of 98) for red deer (Cervus elaphus), 50% (ten of 20) for fallow deer (Dama dama), 33% (three of nine) for mouflon (Ovis aries musimon) and 11% (nine of 83) for Spanish ibex (Capra pyrenaica). Overall, the sites where seropositive wild ruminants were found coincide with the areas where BTV had been detected in livestock, but in eastern Sierra Morena, the virus circulated in wild ruminants, although it had not been detected in domestic ruminants in the same areas. Wild ruminants over 1-year of age (sub-adults and adults) had significantly higher seroprevalences than juvenile animals. Statistically significant differences were also observed between BTV seroprevalence and management (free-ranging vs. captivity) with higher prevalence in free-ranging animals. The high seroprevalences obtained suggest that BTV is widespread in wild ruminants in southern Spain. This factor could have an important influence on the evolution of the infection in domestic livestock and indicates the need to include wild ruminant species in BTV surveillance or control programs.

Analysis of the spatial variation of Bovine tuberculosis disease risk in Spain (2006-2009).

In this study we explored the spatial variation of Bovine tuberculosis (BTB) risk of being positive, new positive or persistently positive, as well as the risk of eliminating BTB in positive herds throughout Spain from 2006 to 2009 by means of hierarchical Bayesian models. The results of the models showed that the risk of infection (positive or new positive herds), persistence and elimination was lower in counties located in north and north-eastern of Spain, and in the Balearic and Canary islands than in the rest of the country. In some counties the risk of positivity was high during the four years of study, whereas there were others where the risk of positivity was high only in some of the years. With regard to the risk of persistence of BTB positive herds, counties located in the central, western and south-western part of the country had a higher risk in the three studied periods. This study has identified some specific areas of increased BTB risk in Spain, information that is useful for disease management.

Descriptive and spatial epidemiology of bovine cysticercosis in North-Eastern Spain (Catalonia).

From March 2005 to December 2007, 284 animals from 67 cattle farms (24 dairy and 43 beef) affected by bovine cysticercosis were detected in the region of Catalonia (North-Eastern Spain). Dairy farms were almost twice more likely to be affected than beef farms (OR=1.79, 95% CI=1.08-2.96, p<0.05), and infected premises have a statistically significant (p<0.05) larger number of animals when compared to uninfected farms in Catalonia. The geographical distribution of the infected farms was evaluated and two statistically significant clusters were identified. The most likely cluster was located in the western part of the study region, with 8 out of 10 farms infected. Epidemiological investigations revealed that the 8 farms belonged to the same company. The secondary cluster was located in Eastern Catalonia with 12 infected farms out of 167 cattle farms. No epidemiological links were found among the 12 infected premises. A questionnaire, based on the EFSA risk assessment, was used to assess the most likely route of introduction into each affected farm. Water supply for animals was the route with the highest score in 41.8% of the cases.

Persistence of highly pathogenic avian influenza virus (H7N1) in infected chickens: feather as a suitable sample for diagnosis.

Selection of an ideal sample is a vital element in early detection of influenza infection. Rapid identification of infectious individuals or animals is crucial not only for avian influenza virus (AIV) surveillance programmes, but also for treatment and containment strategies. This study used a combination of quantitative real-time RT-PCR with an internal positive control and a cell-titration system to examine the presence of virus in different samples during active experimental AIV infection and its persistence in the infected carcasses. Oropharyngeal/cloacal swabs as well as feather pulp and blood samples were collected from 15-day-old chicks infected with H7N1 highly pathogenic AIV (HPAIV) and the kinetics of virus shedding during active infection were evaluated. Additionally, several samples (muscle, skin, brain, feather pulp and oropharyngeal and cloacal swabs) were examined to assess the persistence of virus in the HPAIV-infected carcasses. Based on the results, feather pulp was found to be the best sample to detect and isolate HPAIV from infected chicks from 24 h after inoculation onwards. Kinetic studies on the persistence of virus in infected carcasses revealed that tissues such as muscle could potentially transmit infectious virus for 3 days post-mortem (p.m.), whilst other tissues such as skin, feather pulp and brain retained their infectivity for as long as 5-6 days p.m. at environmental temperature (22-23 degrees C). These results strongly favour feather as a useful sample for HPAIV diagnosis in infected chickens as well as in carcasses.

Assessment of the risk of a bluetongue outbreak in Europe caused by Culicoides midges introduced through intracontinental transport and trade networks.

The importation of infected hosts and the arrival of windborne infected Culicoides (Diptera: Ceratopogonidae) were considered unlikely mechanisms for bluetongue virus (BTV) incursion into a BTV‐free area during the recent BTV serotype 8 (BTV‐8) epidemic in northern Europe. Therefore, alternative mechanisms need to be considered. Air, sea and land transport networks continue to expand, and an important consequence of this is vector‐borne pathogen importation. One important aspect of bluetongue (BT) epidemiology not yet addressed is the potential movement of infected Culicoides via transport and trade networks. Therefore, a risk assessment model was constructed to assess the probability of a BTV outbreak as a consequence of the introduction of Culicoides via these networks. The model was applied to calculate the risk for a BTV‐8 epidemic in Spain in 2007 caused by the introduction of Culicoides from affected northern European countries. The mean weighted annual risk for an outbreak caused by transportation of a single vector from an affected northern European country varied from 1.8 × 10−7 to 3.0 × 10−13, with the highest risks associated with Culicoides imported from Belgium, the Netherlands, Germany and France. For this mechanism to pose a significant risk to BTV‐free countries, a large number of vectors would have to be transported.

Influenza A virus subtypes in wild birds in North-Eastern Spain (Catalonia).

Since the spread of H5N1 highly pathogenic avian influenza virus in 2005, many surveillance programmes have been initiated in poultry and wild birds worldwide. This study describes for the first time the detection of different subtypes of avian influenza viruses (AIV) in wild birds in the West Mediterranean area (Catalonia, North-Eastern Spain). During a 3-year period (from mid-2006 to mid-2009), 1374 birds from 16 different families were examined, and a total of 62 AIV were detected by means of a real-time reverse transcriptase PCR assay. AIV were more frequently detected in Anatidae, Phoenicopteridae, Rallidae and Laridae families. Of the 62 positive samples, 28 AIV could be isolated in embryonated eggs. All isolates were subtyped by haemagglutinin and neuraminidase inhibition techniques and 10 different haemagglutinins (HA) and 7 neuraminidases (NA) were found in 13 different subtype combinations. The most common combinations were H4N6 (22.2%) and H1N1 (18.5%). The HA and NA gene sequences of different AIV subtypes were compared and aligned with those available AIV strains from genome databases. Our studies on AIV phylogenetic analysis revealed that all AIV genes sequenced from wild birds in North-Eastern Spain clustered within Eurasian avian clades, including the sequences of H8, N4 and N5 genes analyzed for the first time in Europe. The results contribute to the understanding of AIV in the Mediterranean area and in Europe.

Quantitative assessment of the probability of bluetongue virus overwintering by horizontal transmission: application to Germany

Even though bluetongue virus (BTV) transmission is apparently interrupted during winter, bluetongue outbreaks often reappear in the next season (overwintering). Several mechanisms for BTV overwintering have been proposed, but to date, their relative importance remain unclear. In order to assess the probability of BTV overwintering by persistence in adult vectors, ruminants (through prolonged viraemia) or a combination of both, a quantitative risk assessment model was developed. Furthermore, the model allowed the role played by the residual number of vectors present during winter to be examined, and the effect of a proportion of Culicoides living inside buildings (endophilic behaviour) to be explored. The model was then applied to a real scenario: overwintering in Germany between 2006 and 2007. The results showed that the limited number of vectors active during winter seemed to allow the transmission of BTV during this period, and that while transmission was favoured by the endophilic behaviour of some Culicoides, its effect was limited. Even though transmission was possible, the likelihood of BTV overwintering by the mechanisms studied seemed too low to explain the observed re-emergence of the disease. Therefore, other overwintering mechanisms not considered in the model are likely to have played a significant role in BTV overwintering in Germany between 2006 and 2007.

Assessment of different surveillance systems for avian influenza in commercial poultry in Catalonia (North-Eastern Spain)

Compulsory surveillance programmes for avian influenza (AI) have been implemented in domestic poultry and wild birds in all the European Member States since 2005. The implementation of these programmes is complex and requires a close evaluation. A good indicator to assess their efficacy is the sensitivity (Se) of the surveillance system. In this study, the sensitivities for different sampling designs proposed by the Spanish authorities for the commercial poultry population of Catalonia were assessed, using the scenario tree model methodology. These samplings were stratified throughout the territory of Spain and took into account the species, the types of production and their specific risks. The probabilities of detecting infection at different prevalences at both individual and holding level were estimated. Furthermore, those subpopulations that contributed more to the Se of the system were identified. The model estimated that all the designs met the requirements of the European Commission. The probability of detecting AI circulating in Catalonian poultry did not change significantly when the within-holding design prevalence varied from 30% to 10%. In contrast, when the among-holding design prevalence decreased from 5% to 1%, the probability of detecting AI was drastically reduced. The sampling of duck and goose holdings, and to a lesser extent the sampling of turkey and game bird holdings, increased the Se substantially. The Se of passive surveillance in chickens for highly pathogenic avian influenza (HPAI) and low pathogenicity avian influenza (LPAI) were also assessed. The probability of the infected birds manifesting apparent clinical signs and the awareness of veterinarians and farmers had great influence on the probability of detecting AI. In order to increase the probability of an early detection of HPAI in chicken, the probability of performing AI specific tests when AI is suspected would need to be increased.

Animal diseases caused by orbiviruses, Algeria.

Antibodies against bluetongue virus were detected in cattle, sheep, goats, and camels in Algeria in 2008. Antibodies against epizootic hemorrhagic disease virus were detected in cattle, but antibodies against African horse sickness virus were not detected in horses and mules. Epizootic hemorrhagic disease in northern Africa poses a major risk for the European Union.

Exploring the Surveillance Potential of Mortality Data: Nine Years of Bovine Fallen Stock Data Collected in Catalonia (Spain)

The potential of fallen stock data to monitor the health status of animal populations has been noted in previous studies. However, further research is required to implement these systems for surveillance. This work presents a novel approach to determining the baselines associated with bovine fallen stock, comparing patterns between subpopulations and identifying subpopulations in which an abnormal event may occur. This study was based on data from 193,873 disposal visits carried out between 2004 and 2012 across a total of 2,991 bovine farms. Proxy measurements such as the number of collections carried out and the weight of carcasses collected were used. Both outcomes were aggregated weekly at different geographical scales for three production types (beef cattle, dairy cattle and heifer fattening). The analysis of these data combined autoregressive integrated moving average modelling and hierarchical time series methods.The three production types exhibited historical baselines that differed notably from one another. Based on the 757 beef cattle farms monitored, the mean number of collections registered per week at the regional level was 37 (range: 10–83). This series was relatively constant over time and showed a marked yearly seasonality. In contrast, for the 426 dairy cattle farms the mean number of disposal visits registered weekly was 121 (range: 71–180), showing half-yearly and yearly seasonality and a marked increase over the period monitored. From the 1,808 heifer fattening farms the mean number of disposal visits was 248 (range: 166–357) and the pattern presented a marked alternating trend over time. These patterns were assessed and compared at regional, provincial, county and municipal levels. The use of hierarchical time series approaches appeared to be a useful tool for comparing the patterns within different subpopulations over time as well as for assessing the spatial extent to which various abnormal events could be detected.