Alberto Allepuz

Porcine circovirus type 2 (PCV2) vaccination of conventional pigs prevents viremia against PCV2 isolates of different genotypes and geographic origins.

The efficacy of recently developed porcine circovirus type 2 (PCV2) vaccines has not been tested yet against PCV2 isolates of the two proposed genotypes. In the present work, the efficacy of a subunit vaccine containing PCV2 capsid protein was evaluated by using a challenge model with four different PCV2 isolates of different genotype and geographic origin. The vaccine prevented the development of viremia in all cases as well as significantly decreased nasal and faecal shedding of the virus. Also, the vaccine elicited PCV2-specific neutralizing antibodies to PCV2 even in the presence of maternally derived immunity.

Longitudinal study of hepatitis E virus infection in Spanish farrow-to-finish swine herds

Hepatitis E is a zoonotic disease and is highly prevalent in European swine livestock. There is a need to compare the infection dynamics of hepatitis E virus (HEV) between herds with the same production system and determine the percentage of animals that could arrive infected at slaughter age. Therefore, a longitudinal study was performed in six Spanish farrow-to-finish affected farms. Twenty piglets per farm were monitored from nursery to slaughter. RT-PCR and serology techniques were applied to analyze longitudinally collected sera and/or faecal samples. Liver and bile samples were also taken at the abattoir. Anti-HEV IgM were firstly detected at 7 weeks of age in 5 farms whereas at 13 weeks of age in 1 farm (farm 2). At slaughter age 50-100% of pigs had seroconverted to anti-HEV IgG in the former 5 farms whereas in the other herd only 5% of pigs were IgG seropositive (farm 2). Six out of 96 livers and 5 out of 80 biles analyzed were HEV positive at the abattoir (total percentage of infected animals: 11.5%). All these positive animals had already seroconverted except 2 pigs of farm 2. Hence, pigs can be seronegative at slaughter age being infected during the latest fattening period. Manipulation of HEV-infected livers or other organs from pigs could be considered a possible route of transmission in Spanish abattoirs. This study represents the first longitudinal survey on swine HEV infection dynamics conducted in different herds.

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.

Pig-major acute phase protein and haptoglobin serum concentrations correlate with PCV2 viremia and the clinical course of postweaning multisystemic wasting syndrome.

The aim of the present longitudinal study was to assess the evolution of two acute phase proteins (APPs), pig-major acute phase protein (pig-MAP) and haptoglobin (HPT), in serum from pigs that developed postweaning multisystemic wasting syndrome (PMWS) in comparison to healthy and wasted non-PMWS affected pigs. In addition, evidence of infection with other pathogens and its relation with variations in APPs concentrations was also assessed. Fourteen independent batches of 100-154 pigs were monitored from birth to PMWS outbreak occurrence in 11 PMWS affected farms. Pigs displaying PMWS-like signs and age-matched healthy controls were euthanized during the clinical outbreak. PMWS was diagnosed according to internationally accepted criteria and pigs were classified as: (i) PMWS cases, (ii) wasted non-PMWS cases and (iii) healthy pigs. At the moment of PMWS occurrence, pig-MAP and HPT concentration in PMWS affected pigs were higher than in healthy ones (p<0.0001). No differences in APPs serum concentrations between subclinically PCV2-infected pigs and healthy non-PCV2-infected pigs (based on quantitative PCR on serum results) were detected. Results showed a significant correlation between PCV2 loads and both pig-MAP (R=0.487-0.602, p<0.0001) and HPT (R=0.326-0.550, p<0.05-0.0001) concentrations in serum of PMWS affected pigs, indicating that the acute phase response in PMWS affected pigs occurred concomitantly to PCV2 viremia. No other pathogen, apart from PCV2, was consistently related with variations in APPs concentrations. A ROC analysis, made to determine the capacity of discrimination of both APPs between PMWS affected and non-affected pigs, showed higher sensitivity and specificity values using pig-MAP compared to HPT. These results suggest that pig-MAP might be a better indicator of PMWS status than HPT. Moreover, the fact that APR occurred some weeks before the start of clinical signs suggests that APPs could provide valuable prognostic information for PMWS development.

Evidence of goats acting as domestic reservoirs of bovine tuberculosis

Measures, such as the increase in the frequency of controls, the use of the gamma-interferon assay as ancillary test, the improvement of slaughterhouse surveillance, or the implementation of premovement testing have helped to progressively reduce the prevalence of bovine tuberculosis (bTB) in Spain in the last decade (Anonymous 2012a). However, the objective of eradication is far from being achieved, and this failure has been mainly attributed to the role of wild ungulate species (mainly wild boar and red deer) as reservoirs of bTB (Vicente and others 2006, Naranjo and others 2008). In Spain, with the second largest goat population in the European Union – about 2.9 million heads in 2010 (Anonymous 2012b) – the presence of tuberculosis in goats may also compromise the bTB eradication campaign (Liebana and others 1998, Humblet and others 2009). Goats seem to be very susceptible to infection by Mycobacterium bovis and Mycobacterium caprae , which result in the development of disseminated lesions and a fast transmission within the herd (Crawshaw and others 2008, Rodriguez and others 2011). The reporting of caprine tuberculosis outbreaks has increased in recent years, not only in southern European countries where the disease is considered to be endemic, but also in countries such as the UK or Ireland (Daniel and others 2009, Quintas and others 2010, Shanahan and others 2011). Besides the important economic losses it may cause to the livestock sector (Daniel and others 2009), caprine tuberculosis represents a zoonotic risk (Gutierrez and others 1997, Rodriguez and others 2009). Even though TB in goats in Spain is considered widespread, there is no official data on the prevalence of caprine TB (Liebana and others 1998, Domingo and others 2009), and the …

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.

Seroprevalence and risk factors associated with Toxoplasma gondii infection in pig farms from Catalonia, north-eastern Spain

Seroprevalence and associated risk factors for Toxoplasma gondii infection in pigs were analyzed in 1202 sera samples, including sows and pigs of three, seven, 11, 15 and 20 weeks of age, from 23 farms in Catalonia, north-eastern Spain. Antibodies were tested by the modified agglutination test (MAT) at titers > or =1:25. Antibodies to T. gondii were found in 228 samples (19.0%; 95% CI: 16.8-21.2). The individual prevalence in animals higher than 7 weeks of age was 22.8% (174/762; 95% CI: 16.6-29.0) and the within-farm prevalence ranged from 7.1% to 36.4%. Statistically significant differences were found among age classes. The risk factors significantly associated with T. gondii seroprevalence were the presence of cats, percentage of mortality at weaning and the presence of outdoor facilities in the farms. The seroprevalence observed in the present study indicates widespread exposure to T. gondii among domestic pigs in Catalonia, which may have important implications for public health.

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.

Comparison of bovine cysticercosis prevalence detected by antigen ELISA and visual inspection in the North East of Spain.

A cross-sectional serological study on cattle less than 2 years old, using an antigen ELISA for the detection of bovine cysticercosis was carried out between November 2009 and February 2010 in 10 slaughterhouses from the Catalonia region (North-Eastern Spain). Circulating antigen was detected in 23 of 2073 animals, i.e. a sero-prevalence of 1.11% (CI95%: 0.76-1.75%). The determined sero-prevalence was about 50 times higher than the prevalence obtained by visual inspection within the same period: 19 positive animals of 90,891 slaughtered animals (0.02%) in the same slaughterhouses. None of the animals with positive result in the Ag-ELISA was detected by meat inspection.