Evaluation of the application of sequence data to the identification of outbreaks of disease using anomaly detection methods.

Anomaly detection methods have a great potential to assist the detection of diseases in animal production systems. We used sequence data of Porcine Reproductive and Respiratory Syndrome (PRRS) to define the emergence of new strains at the farm level. We evaluated the performance of 24 anomaly detection methods based on machine learning, regression, time series techniques and control charts to identify outbreaks in time series of new strains and compared the best methods using different time series: PCR positives, PCR requests and laboratory requests. We introduced synthetic outbreaks of different size and calculated the probability of detection of outbreaks (POD), sensitivity (Se), probability of detection of outbreaks in the first week of appearance (POD1w) and background alarm rate (BAR). The use of time series of new strains from sequence data outperformed the other types of data but POD, Se, POD1w were only high when outbreaks were large. The methods based on Long Short-Term Memory (LSTM) and Bayesian approaches presented the best performance. Using anomaly detection methods with sequence data may help to identify the emergency of cases in multiple farms, but more work is required to improve the detection with time series of high variability. Our results suggest a promising application of sequence data for early detection of diseases at a production system level. This may provide a simple way to extract additional value from routine laboratory analysis. Next steps should include validation of this approach in different settings and with different diseases.

Evaluation of the efficacy of two postweaning colibacillosis vaccines in a field herd with PRRS circulation during postweaning stage.

Postweaning diarrhea (PWD) and PRRS are two major concerns in swine production, which association has not been consistently explored. In the current scenario of restrictions in the use of antibiotics and ZnO, vaccination is more relevant to control PWD, but PRRS virus circulation may compromise the immune protection conferred by postweaning colibacillosis vaccines. We evaluated the efficacy of two postweaning colibacillosis vaccines (parenteral and oral) in a commercial herd affected by an outbreak of PWD and with PRRS circulation in postweaning. Five groups were studied during the postweaning period: one control (Group 1) and four vaccinated: two with each postweaning colibacillosis vaccine administered alone (Groups 2 and 3) or with sow vaccination against PRRS (Groups 4 and 5). We evaluated the effects on piglet weight, average daily weight gain and in the percentage of piglets with diarrhea, its duration, lethality and mortality. PRRS viremia and anti-PRRS antibodies were evaluated by qPCR and ELISA. Regarding control group, colibacillosis vaccination generally improved most of the measured parameters; but significant improvements were only observed in Groups 4 and 5 (p < 0.05). Moreover, cases of diarrhea occurred at different ages: in Groups 2 and 3 the peak of cases occurred just after ZnO was removed from the feed compared to Group 1, while in Groups 4 and 5 no peak was observed. This suggests that postweaning colibacillosis vaccination may be compromised by the PRRS circulation. In PRRS endemic herds an effective protection against PWD through vaccination may require PRRS vaccination to obtain a better performance.

Presence of Porcine Circovirus Type 2 in the Environment of Farm Facilities without Pigs in Long Term-Vaccinated Farrow-to-Wean Farms.

Vaccination against Porcine Circovirus Type 2 (PCV2) even over several years has proven as an insufficient measure to eradicate the infection from farms, possibly due to not producing sterilizing immunity. Viral persistence in the farm environment has been proposed as a possible cause of reinfection, and for that reason, the main objective of this study was to identify potential critical points where PCV2 could persist in farrow-to-wean farms which had been vaccinating piglets for years. Surface samples were collected from different farm facilities with and without animals and analyzed by qPCR to detect and quantify the viral load. Most of the samples taken in animal housing facilities tested negative (96.6%); however, PCV2 was more frequently detected in samples from the offices (37.5%), the farm staff (25%) and the perimeter (21%). These results indicate that PCV2 contamination is frequent in facilities despite the long-term use of vaccination programs. Therefore, PCV2 control programs should include more exhaustive cleaning and disinfection protocols in non-animal facilities, as well as the implementation of specific biosecurity measures in these areas to minimize the risk of PCV2 introduction from external sources.

Application of Bayesian Regression for the Identification of a Catchment Area for Cancer Cases in Dogs and Cats.

Research on cancer in dogs and cats, among other diseases, finds an important source of information in registry data collected from hospitals. These sources have proved to be decisive in establishing incidences and identifying temporal patterns and risk factors. However, the attendance of patients is not random, so the correct delimitation of the hospital catchment area (CA) as well as the identification of the factors influencing its shape is relevant to prevent possible biases in posterior inferences. Despite this, there is a lack of data-driven approaches in veterinary epidemiology to establish CA. Therefore, our aim here was to apply a Bayesian method to estimate the CA of a hospital. We obtained cancer (n = 27,390) and visit (n = 232,014) registries of dogs and cats attending the Veterinary Medical Teaching Hospital of the University of California, Davis from 2000 to 2019 with 2,707 census tracts (CTs) of 40 neighboring counties. We ran hierarchical Bayesian models with different likelihood distributions to define CA for cancer cases and visits based on the exceedance probabilities for CT random effects, adjusting for species and period (2000-2004, 2005-2009, 2010-2014, and 2015-2019). The identified CAs of cancer cases and visits represented 75.4 and 83.1% of the records, respectively, including only 34.6 and 39.3% of the CT in the study area. The models detected variation by species (higher number of records in dogs) and period. We also found that distance to hospital and average household income were important predictors of the inclusion of a CT in the CA. Our results show that the application of this methodology is useful for obtaining data-driven CA and evaluating the factors that influence and predict data collection. Therefore, this could be useful to improve the accuracy of analysis and inferences based on registry data.

Cardiorespiratory nematodes and co-infections with gastrointestinal parasites in new arrivals at dog and cat shelters in north-western Spain.

Metastrongyloid and trichuroid cardiorespiratory nematodes of dogs and cats are considered emergent in several European countries, and an increase in the number and extent of endemic foci has been described. Since data on their prevalence are limited in this continent, faecal samples from new arrivals (365 dogs and 65 cats) at two animal shelters in North-western Spain were analysed using both floatation and Baermann techniques. In order to confirm the microscopic identification of Metastrongylidae first stage larvae, molecular characterization based on the sequence of the ITS-2 was performed. The possible influence of some variables such as the species, sex and age of the animals and the co-infection with other gastrointestinal parasites on the prevalence of cardiorespiratory nematodes was analysed. The most prevalent metastrongylid was Aelurostrongylus abstrusus (15.4%) followed by Angiostrongylus vasorum (4.1%) and Crenosoma vulpis (1.1%). Regarding trichuroids, Eucoleus aerophilus and/or Eucoleus boehmi eggs were detected in 28 dogs (7.7%) and four cats (6.2%). Almost all animals positive to cardiorespiratory nematodes (86.8%) were co-infected with gastrointestinal parasites. The prevalence of Metastrongylidae and respiratory trichuroids was significantly higher in dogs co-infected with Taenia spp. and Toxocara canis or Giardia duodenalis and Sarcocystis spp., respectively. In cats, a significant higher prevalence of Metastrongylidae nematodes was found in animals co-infected with Toxocara cati. Our results reveal that cardiorespiratory nematodes are common in companion animals from north-western Spain, showing higher prevalences than those previously reported from this country. This investigation represents the first report of C. vulpis, E. aerophilus and E. boehmi in dogs from Spain. The identification of a number of zoonotic parasites is of public health concern. Our results indicate that these nematodes should be included in the differential diagnosis of dogs and cats from north-western Spain showing respiratory or cardiac clinical signs.

Antimicrobial Susceptibility of Enterotoxigenic Escherichia coli from Diarrhoeic Neonatal Calves in Spain.

Enterotoxigenic Escherichia coli (ETEC) is one of the major pathogens involved in neonatal calf diarrhoea (NCD) causing high economic losses in dairy farms. Antibiotic treatment is common in cases of systemic illness caused by NCD, but antimicrobial susceptibility tests (AST) are usually not performed. Thus, the aim of this study was to characterize the antimicrobial susceptibility of ETEC strains obtained from calves with diarrhoea between 2018-2020. Faecal samples (n = 420) were analyzed to detect the typical ETEC virulence factors F5 and STa. Positive samples were cultured to identify and isolate ETEC strains (n = 41) and ASTs were performed. Our results are alarming since ETEC strains resistant to three or more families of antimicrobials were detected in all isolates. Only four antibiotics (ceftiofur, cefoperazone, cefquinome and gentamicin) presented efficacy against more than 90% of the ETEC strains, while the other ten antibiotics were effective against less than 40% of the strains. In addition, a high number of strains were resistant to most first-line antimicrobials used in veterinary practice. For this reason, when ETEC infection is suspected, an AST must always be performed to select the most appropriate antimicrobial in each case and to avoid the emergence of new resistance mechanisms.

Prevalence of selected tick-borne pathogens in wild ungulates and ticks in southern Spain.

A survey study was carried out to assess the occurrence of selected tick-borne pathogens (TBP) in wild ungulates in Mediterranean ecosystems in southern Spain. Spleen samples were collected from 1,132 wild ungulates, including 578 red deer, 269 wild boar, 135 mouflon, 121 fallow deer and 29 roe deer, between 2009 and 2015. Eighty-nine ticks collected from TBP-positive animals were also analysed. Samples were tested by PCR and sequenced whenever possible. TBP DNA was detected in 127 of 863 wild ruminants (14.7%; 95% CI: 12.4-17.3) including the following: Anaplasma phagocytophilum (9.2%), Babesia divergens (2.9%), Theileria sp. OT3 (1.7%), Borrelia afzelii (0.7%) and Theileria capreoli (0.2%), but no positive samples were detected in wild boar (0/269). All the strains from mouflon were identified as Theileria sp. OT3, while B. divergens and T. capreoli were mainly found in red deer. Co-infection with A. phagocytophilum and B. divergens, and A. phagocytophilum and Theileria spp. was detected in red deer and mouflon, respectively. The risk factor analysis showed that the prevalences of A. phagocytophilum and piroplasms were species-related. Eighty-nine tick specimens collected from ungulates found to be infected with the selected TBP were identified as Hyalomma lusitanicum (95.5%) and Ixodes ricinus (4.5%). Thirty ticks were positive for Anaplasma/Ehrlichia spp. (33.7%), 25 for Babesia/Theileria (28.1%) and two for B. burgdorferi s.l. (2.3%). Eleven specimens showed co-infections with Anaplasma/Ehrlichia and Babesia/Theileria (10.1%) or Anaplasma/Ehrlichia and B. burgdorferi s.l. (2.3%). The estimated prevalences obtained in the present study suggest the possible contribution of wild ruminants to the maintenance of some selected TBP in Mediterranean ecosystems in southern Spain, while the role of wild boar in the epidemiology of these pathogens seems to be limited in this region.

Monitoring of porcine circovirus type 2 infection through air and surface samples in vaccinated and unvaccinated fattening farms.

Air and surfaces of swine farms are the two alternative samples to obtain information about the health status of the herd. The aim of this study was to assess air and surface sampling for the detection of porcine circovirus type 2 (PCV2) in vaccinated and unvaccinated fattening farms, studying the relationship between the viral load in these samples with the viremia at herd level. Three swine fattening batches (one unvaccinated; two vaccinated) were monitored at 10, 12, 14, 16 and 18 weeks old; at each stage, blood, air and different surfaces were sampled and analysed by qPCR. In all herds, PCV2 was detected in all types of samples. Whenever viremia was detected, PCV2 was also detected in air and surface samples, even in those cases with a low estimated prevalence (1.6%); moreover, in two out of the three herds, PCV2 was detected in air and surface samples earlier than in the blood of the sampled population. In addition, a good correlation between the viremia of pig population and the PCV2 load in air and surface samples was found in both cases (τ = 0.672 and 0.746, respectively; p <0.05). These results show that air and surface samples could be useful tools to monitor PCV2 infection, being suitable for detecting the virus in cases of low prevalence and even before pigs develop viremia; therefore, these sampling techniques would speed up the implementation of the required measures to prevent productive and economic losses due to PCV2 infection.

Diversity of Anaplasma species and importance of mixed infections in roe deer from Spain.

Although wildlife can act as reservoirs of some Anaplasma species, studies on the presence and distribution of Anaplasma spp. in wild cervids are mainly limited and focused on zoonotic species. In order to identify the Anaplasma species in roe deer from Spain and to detect co-infections, 224 spleen samples were tested for Anaplasma spp. using a commercial qPCR; positive samples were further characterized using generic 16S rRNA primers and species-specific primers targeting the msp2 and groEL genes. Anaplasma DNA was detected in the 50.9% of samples, and four Anaplasma species were identified. Anaplasma phagocytophilum (43.8%) was predominant, followed by Anaplasma bovis (13.8%), Anaplasma capra (5.8%) and Anaplasma ovis (2.2%). In addition, strains similar to Anaplasma platys were found in nine animals. Most positive roe deer (71.9%) were infected with a single Anaplasma species, whereas co-infections with two (19.3%) or three (8.8%) Anaplasma species were also found. This study confirms the widespread occurrence of Anaplasma spp. in roe deer from Spain, being the first report of A. platys-like strains and A. capra in this cervid; it is also the first report of A. capra in Spain. The detection of Anaplasma species pathogenic for humans and/or domestic animals in roe deer suggests that this cervid may play a role in the sylvatic cycle of these bacteria contributing to the appearance of clinical anaplasmosis cases. In addition, co-infections are common in roe deer revealing that Anaplasma species specific PCR assays are essential for a reliable identification as well as for determining their real prevalence.

Seroprevalence of Toxoplasma gondii and associated risk factors in domestic pigs raised from Cuba.

A cross-sectional study was carried out to determine the seroprevalence of Toxoplasma gondii and associated risk factors in pigs in the largest pork-producing region in Cuba. Serum samples from 420 pigs, including 210 sows and 210 post-weaning pigs, were tested for antibodies against T. gondii using a commercial indirect enzyme-linked immunosorbent assay. Anti-T. gondii antibodies were detected in 56 animals (13.3%, 95% CI: 10.1-16.6). A generalized estimating equations model revealed that the risk factors associated with higher seropositivity in pigs were altitude (higher in farm's location < 250 m above sea level (masl) versus ≥ 250 masl) and age (higher in sows compared to post-weaning pigs). The results indicated that this protozoan parasite is widely distributed on pig farms in the study area, which is a public health concern since the consumption of raw or undercooked pork meat products containing tissue cysts is considered one of the main routes of T. gondii transmission worldwide. Control measures should be implemented to reduce the risk of exposure to T. gondii in pigs in Cuba.

Efficacy of Two Commercial Ready-To-Use PCV2 and Mycoplasma hyopneumoniae Vaccines under Field Conditions.

Porcine Circovirus Type 2 (PCV2) and Mycoplasma hyopneumoniae are economically important pathogens in swine farms. Vaccination is the main preventive measure for both infections. In order to test two ready-to-use bivalent vaccines, 646 piglets from a herd actively infected with both pathogens were stratified according to the sow parity number and randomly assigned to three groups: A and B were vaccinated with two different vaccines, respectively, while C remained as the unvaccinated control. Vaccine efficacy was assessed based on the weight, average daily weight gain (ADWG), degree of lung lesions, presence of PCV2 viremia by qPCR and presence of PCV2 and M. hyopneumoniae antibody levels by ELISA. Our data revealed that the sow parity did not influence the vaccine outcomes. Good results for most of the analyzed parameters were observed in both vaccinated groups. ADGW and final weight were higher and lung lesions were less evident in both vaccinated groups than in the control one, but only Group A showed a significant improvement. PCV2 viremia was not detected in Group A, but it did appear in Group B coinciding with its peak in Group C. Finally, both the PCV2 and M. hyopneumoniae serological patterns differed depending on the employed vaccine.

Data-Driven Network Modeling as a Framework to Evaluate the Transmission of Piscine Myocarditis Virus (PMCV) in the Irish Farmed Atlantic Salmon Population and the Impact of Different Mitigation Measures.

Cardiomyopathy syndrome (CMS) is a severe cardiac disease of Atlantic salmon caused by the piscine myocarditis virus (PMCV), which was first reported in Ireland in 2012. In this paper, we describe the use of data-driven network modeling as a framework to evaluate the transmission of PMCV in the Irish farmed Atlantic salmon population and the impact of different mitigation measures. Input data included live fish movement data from 2009 to 2017, population dynamics events and the spatial location of the farms. With these inputs, we fitted a network-based stochastic infection spread model. After assumed initial introduction of the agent in 2009, our results indicate that it took 5 years to reach a between-farm prevalence of 100% in late 2014, with older fish being most affected. Local spread accounted for only a small proportion of new infections, being more important for sustained infection in a given area. Spread via movement of subclinically infected fish was most important for explaining the observed countrywide spread of the agent. Of the targeted intervention strategies evaluated, the most effective were those that target those fish farms in Ireland that can be considered the most connected, based on the number of farm-to-farm linkages in a specific time period through outward fish movements. The application of these interventions in a proactive way (before the first reported outbreak of the disease in 2012), assuming an active testing of fish consignments to and from the top 8 ranked farms in terms of outward fish movement, would have yielded the most protection for the Irish salmon farming industry. Using this approach, the between-farm PMCV prevalence never exceeded 20% throughout the simulation time (as opposed to the simulated 100% when no interventions are applied). We argue that the Irish salmon farming industry would benefit from this approach in the future, as it would help in early detection and prevention of the spread of viral agents currently exotic to the country.

Molecular epidemiology of Aleutian mink disease virus causing outbreaks in mink farms from Southwestern Europe: a retrospective study from 2012 to 2019.

BACKGROUND: Aleutian mink disease virus (AMDV) causes major economic losses in fur-bearing animal production. The control of most AMDV outbreaks is complex due to the difficulties of establishing the source of infection based only on the available on-farm epidemiological data. In this sense, phylogenetic analysis of the strains present in a farm may help elucidate the origin of the infection and improve the control and biosecurity measures. OBJECTIVES: This study had the following aims: characterize the AMDV strains from most outbreaks produced at Spanish farms between 2012-2019 at the molecular level, and assess the utility of the combined use of molecular and epidemiological data to track the possible routes of infection. METHODS: Thirty-seven strains from 17 farms were partially sequenced for the NS1 and VP2 genes and analyzed phylogenetically with other strains described worldwide. RESULTS: Spanish AMDV strains are clustered in four major clades that generally show a good geographical correlation, confirming that most had been established in Spain a long time ago. The combined study of phylogenetic results and epidemiological information of each farm suggests that most of the AMDV outbreaks since 2012 had been produced by within-farm reservoirs, while a few of them may have been due to the introduction of the virus through international trade. CONCLUSIONS: The combination of phylogenetic inference, together with epidemiological data, helps assess the possible origin of AMDV infections in mink farms and improving the control and prevention of this disease.

Evaluation of two enzyme-linked immunosorbent assays for diagnosis of bluetongue virus in wild ruminants.

Bluetongue (BT) is a reportable re-emerging vector-borne disease of animal health concern. Enzyme-linked immunosorbent assays (ELISA) are frequently used in BT surveillance programs in domestic ruminants, but their diagnostic accuracy has not been evaluated for wild ruminants, which can play an important role as natural reservoirs of bluetongue virus (BTV). The aim of this study was to assess two commercial ELISAs for BT diagnosis in wild ruminants using control sera of known BTV infection status and field samples. When control sera were tested, the double recognition ELISA (DR-ELISA) showed 100 % sensitivity (Se) and specificity (Sp), while the competitive ELISA (C-ELISA) had 86.4 % Se and 97.1 % Sp. Using field samples, the selected latent-class analysis model showed 95.7 % Se and 85.9 % Sp for DR-ELISA, 58.2 % Se and 95.8 % Sp for C-ELISA and 84.2 % Se for the serum neutralization test (SNT). Our results indicate that the DR-ELISA may be a useful diagnostic method to assess BTV circulation in endemic areas, while the C-ELISA should be selected when free-areas are surveyed. The discrepancy between control and field samples point out that the inclusion of field samples is required to assess the accuracy of commercial ELISAs for the serological diagnosis of BTV in wild ruminants.

Environmental distribution of Porcine Circovirus Type 2 (PCV2) in swine herds with natural infection.

Porcine circovirus type 2 (PCV2) is the aetiological agent of PCV2-Systemic Disease (PCV2-SD) and PCV2-Subclinical Infection (PCV2-SI). PCV2 is highly resistant to environmental conditions, being able to remain in the farm environment and thus represent a risk for infection maintenance. The aim of this study was to identify, under field conditions, the possible critical points in the environment of non-vaccinated farrow-to-weaning swine farms where PCV2 could accumulate and persist. For that, environmental samples from five swine farms with PCV2-SD or PCV2-SI were taken and analysed by qPCR, including different farm areas, farm personnel and management implements. PCV2 DNA was detected in the environment of all farms (42.9% of positive samples). Overall, the PCV2-SD herd seemed to present more positive samples and higher viral loads than the PCV2-SI herds. At individual farm level, weaning areas appeared to be the most contaminated facilities. In addition, PCV2 was found at high levels in most samples from farm workers, especially work boots, suggesting that they may play a role in within-farm transmission. In addition, PCV2 was detected in areas without animals the like warehouses, offices and farm perimeter. Therefore, this study is helpful to improve measures to reduce within-farm PCV2 dissemination.

Prevalence and distribution of Babesia and Theileria species in roe deer from Spain.

Babesiosis and Theileriosis are important worldwide-distributed tick-borne diseases for human and animals. Their presence in a particular area depends on the presence of suitable tick-vector and host species as well as competent reservoirs such as roe deer, one of the most abundant wild cervids in Spain. Spleen samples from 174 roe deer hunted in Spain were analysed to determine the prevalence of Babesia and Theileria species. DNA of both piroplasms was firstly detected using a commercial qPCR. Then, positive samples were molecularly characterized at the 18S rRNA and ITS1 genes of Babesia spp. and Theileria spp. The possible influence of some factors such as ecological area, age and sex was also assessed. Overall, 89.7% of roe deer were positive to any of the two piroplasms. Theileria spp. was more prevalent (60.9%) than Babesia spp. (19.0%); species identification could not be achieved in 17.3% of positive samples. Babesia prevalence was significantly higher in young animals and in roe deer from Oceanic regions, in contrast to Theileria spp. Five species were identified: Theileria sp. OT3 (60.3%), Babesia capreoli (15.5%), Babesia venatorum (2.9%), Theileria sp. 3185/02 (0.6%) and Babesia bigemina (0.6%). The coinfection B. capreoli/T. sp. OT3 was the most common (4.6%) followed by B. venatorum/T. sp. OT3 (0.6%) and B. bigemina/T. sp. OT3 (0.6%). Our results reveal that Theileria spp. and Babesia spp. are prevalent piroplasms in roe deer from Spain. These cervids can act as reservoirs for several Babesia and Theileria species, including the zoonotic B. venatorum. This study represents the first description of B. venatorum and B. bigemina in roe deer from Spain.

Molecular assessment of visitor personal protective equipment contamination with the Aleutian mink disease virus and porcine circovirus-2 in mink and porcine farms.

Personal protective equipment (PPE) is an element of biosecurity intended to prevent the access or spread of diseases in farms. Nevertheless, to date no extensive reports exist about the effectiveness of different available PPE on farms. Thus, our aim was to estimate the degree of protection of PPE from viral contamination during farm visits. Two farms, infected with Aleutian mink disease virus and porcine circovirus-2 respectively, were visited by six visitors wearing different combinations of PPE: coveralls with hood and bootcovers, both with a certified barrier to infective agents (certified PPE group) and non-certified bootcover and coverall without hood (non-certified PPE group). Seventy-two swab samples from PPE and both hair and street clothes under PPE were taken after the visit and analysed by qPCR. Our results reveal viral exposure during visits, and the external protections of body and shoes were contaminated in all cases (24/24). In addition, protection from viral contamination varied noticeably according to the biosecurity elements used. A higher number of positives were detected in the non-certified PPE group than in the certified PPE group, both in elements under external protections (14/18 vs 3/18) and also in hair (4/6 vs 0/6). In fact, non-certified bootcovers broke during visits, resulting in viral contamination of the internal elements under them; these are consequently not suitable for using with wrinkled surfaces usually found in farm facilities. Thus, certified coveralls should be used in order to prevent contaminations, and workers and personnel of farms should be trained in their proper use. qPCR is a useful tool in the risk management of biosecurity programmes, and our results may serve as a model to evaluate different biosecurity measures.

Seroprevalence of Toxoplasma gondii and Neospora caninum in goats from north-western Spain.

INTRODUCTION AND OBJECTIVE: Toxoplasma gondii and Neospora caninum are protozoans involved in reproductive failure especially in ruminant livestock. The objective was to estimate the seroprevalence of both parasites in goats from north-western Spain and to study the influence of some factors on seropositivity. MATERIALS AND METHOD: Blood samples from 638 goats were collected in 50 farms. Presence of T. gondii and N. caninum antibodies were detected by direct agglutination and competitiveELISA techniques, respectively. The risk factor analysis was performed using a mixed-effects logistic regression. RESULTS: Individual (48%) and herd-level (74%) T. gondii seroprevalence values were high; the within-herd prevalence was 53%. In contrast, 6% of animals tested positive to N. caninum and 38% of the herds had at least one positive animal, with a true within-herd prevalence of 10%. Mixed infections were limited; 91% of N. caninum seropositive goats were also positive to T. gondii. The risk factor analysis showed that T. gondii seroprevalence is influenced by the presence of sheep in the farm (OR=4.9) and the seropositivity to N. caninum (OR=16.5); goats from the Central-coastal area, more humid and warm, had a 15.7-fold probability of being seropositive to T. gondii than those from the Mountainous area. Cross-breed goats (OR=4.5) and the seropositivity to T. gondii (OR= 9.5) were factors associated with N. caninum seropositivity. CONCLUSIONS: The high T. gondii seroprevalence in goats constitute a noticeable zoonotic risk. The consideration of the risk factors identified in designing T. gondii and N. caninum control programs in goat herds should allow the implementation of more efficient measures, avoiding the appearance of outbreaks of reproductive disorders by both protozoans in goats.

Application of real-time PCR to detect Aleutian Mink Disease Virus on environmental farm sources.

The Aleutian Mink Virus (AMDV) causes the Aleutian Mink Disease (AMD) or Mink Plasmacytosis, a disease responsible of high economic losses for industry worldwide. Despite there is evidence of the environmental persistence of the virus, there is not literature on the detection of this virus in environmental samples in farms and this fact would have great importance in the control programs of the disease. In order to detect contamination caused by AMDV on farms, several environmental samples were taken and examined using qPCR. 93.9% of samples taken from farms confirmed to be infected tested positive. The virus was also detected on a farm which, despite having no previous positive results, was sharing personnel with an infected farm. All samples taken from AMD-free farms tested negative, including a farm where an eradication procedure by stamping out had been performed during the preceding months. Higher contamination levels were observed in samples from those surfaces in direct contact with animals. These results are the first demonstration of environmental contamination in farms, hitherto suggested by epidemiological evidences, caused by AMDV on surfaces, furniture and equipments inside mink farms. qPCR is an useful tool for evaluating the spread of AMDV into the environment, and it may have important applications within the disease control programs.