This is the first report of Toxoplasma gondii infection in black vultures (Coragyps atratus), which are obligate scavengers found throughout the Americas. Serum samples from 121 wild black vultures caught in urban areas of the city of Sao Paulo, Brazil, were tested for the presence of T. gondii antibodies using the modified agglutination test (MAT; cutoff point 1:5). T. gondii antibodies were found in 16 birds (13.2%), with titers of 1:5 (6 birds), 1:10 (8 birds), and 1:20 (2 birds).

An evaluation was made of the presence of anti-Leishmania infantum chagasi antibodies in domestic dogs from the urban and rural areas of Brazil’s Pantanal wetland region using serological techniques. A total of 429 dogs were sampled in three areas of the Pantanal biome, including the municipalities of Poconé, Santo Antônio de Leverger, and Barão de Melgaço, in the state of Mato Grosso, and in the municipality of Corumbá, in the state of Mato Grosso do Sul. The immunofluorescence assay (IFA) was used to detect antibodies (cut-off point 40) using Leishmania infantum chagasi antigen. Because of the possibility of cross-reactivity between species of the genus Leishmania, samples that were positive in the IFA against L. infantum chagasi were also tested by IFA in the same conditions, using L. amazonensis and L. braziliensis antigens. IFA-positive samples to L. infantum chagasi were also evaluated using Enzyme-Linked Immunosorbent Assay (ELISA). The results showed the presence of antibodies against L. infantum chagasi in 23 (5.36%; 95% CI: 3.50%-8.05%) dogs and at least one seroreactive dog was found in each of the municipalities evaluated in this study. Antibody titers ranged from 40 to 5,120, and all IFA positive samples were positive in the ELISA. Among the 23 positive dogs, nine were also were seroreactive for L. amazonensis and L. braziliensis antigens. The occurrence of anti- L. infantum chagasi antibodies in dogs was higher in rural areas (7.06%) than in urban areas (2.50%) (P < 0.05). Based on this study, we concluded that dogs from rural areas of the Pantanal wetlands were in contact with Leishmania species, which is relevant information given their importance to public health.

Introduction: Aujeszky’s disease virus (ADV) infects a wide range of animals, including members of the Suidae family, i.e. domestic and wild pigs, as well as wild boar. Since wild boar are a potential ADV reservoir and a source of infection for domestic pigs, the aim of the study was to evaluate ADV antibody prevalence in the Polish wild boar population, during the years 2011 to 2014. Material and Methods: Wild boar blood samples were collected during three consecutive hunting seasons; i.e. 2011/2012, 2012/2013, and 2013/2014, and tested for ADV antibodies by ELISA. Results: ADV antibodies were detected in samples from all tested voivodships. The average seroprevalence reached 32.2%. Seroprevalence, over the examined hunting seasons, was 27.4% in 2011/2012, 32.4% in 2012/2013, and 35.5% in 2013/2014. The highest percentage of seroreagents was detected in four voivodships, situated along the western border of Poland, i.e. Zachodnio-Pomorskie (ZP), Lubuskie (LB), Dolnośląskie (DS), and Opolskie (OP). This area is positively correlated with the highest density of the wild boar population and the highest wild boar hunting bag. Conclusion: The results of this study confirm that the wild boar population may still pose a threat to domestic pigs, which is of special importance at the final stage of Aujeszky’s disease eradication programme in Poland.

Introduction: A novel to Europe Schmallenberg virus (SBV) causes clinical disease manifested by reproduction disorders in farm ruminants. In free-living ruminants, SBV antibodies as well as the virus were detected. Recent studies also revealed SBV antibodies in wild boars. The study investigates SBV antibodies occurring in wild boars in Poland at the peak of recent virus epidemics in the country.Material and Methods: Samples collected from 203 wild boars culled during the 2012/2013 and 2013/2014 hunting season were serologically tested using multi-species cELISA. Attempted neutralisation tests failed due to poor serum quality. RT-PCR was implemented in seropositive and doubtful animals.Results: Two samples collected from wild boar in the winter of 2013 gave a positive result in ELISA, while another two from the 2012/2013 hunting season were doubtful. No SBV RNA was detected in spleen and liver tissues.Conclusion: Low SBV seroprevalence in wild boars, despite high incidence of SBV infections occurring simultaneously in wild ruminants, suggests that boars are unlikely to be a significant reservoir of the virus in the sylvatic environment in Poland.

Introduction: The aim of this study was to assess the seroprevalence of swine influenza A virus (SIV) in Polish farrow-to-finish pig herds.Material and Methods: Serum samples collected from 5,952 pigs, from 145 farrow-to-finish herds were tested for the presence of antibodies against H1N1, H1N1pdm09, H1N2, and H3N2 SIV subtypes using haemagglutination inhibition (HI) test. Samples with HI titres equal or higher than 20 were considered positive.Results: HI antibodies to at least one of the analysed SIV subtypes were detected in 129 (89%) herds and in 2,263 (38%) serum samples. Antibodies to multiple SIV subtypes were detected in 104 (71.7%) herds and in 996 (16.7%) serum samples. Concerning the seroprevalence rate, according to age category, the highest prevalence of the antibodies was detected in weaners, with regard to the H1N1, H1N2, and H3N2, and in sows, with regard to the H1N1pdm09. The lowest seroprevalence for all evaluated SIV subtypes was detected in finishers.Conclusion: The study indicates that antibodies against single and multiple SIV subtypes are circulating in Polish farrow-to-finish herds and highlights the importance of conducting a molecular surveillance programme in future studies.

Introduction: Infections with bovine foamy virus (BFV) were found in many countries but there is a lack of large-scale surveys on the prevalence of BFV among dairy cattle. The aim of this study was to develop and validate the recombinant Gag protein-based ELISA and to estimate the prevalence of antibodies against BFV. Material and Methods: Gag coding region from BFV was cloned into expression vector pT7Arg-STOP, which expressed a high level of recombinant Gag protein from E.coli. The ELISA was standardised, and the cut-off value and sensitivity and specificity of the test were calculated using a receiver operating characteristic and Bayesian estimation. Results: A total of 3,051 serum samples were tested by ELISA and 939 (30.8%) sera were recognised as positive. When Bayesian approach was used, the overall true BFV prevalence was 29.7% (95% CI: 25.9-33.4%). Conclusion: Expressed Gag protein of BFV has been used successfully as an antigen for ELISA. Eventually, this study provides basic information about the epidemiological status of infection with BFV in dairy cattle in Poland, which can be used for further studies on dissemination and transmission of BFV infection.

Introduction: The prevention and control of Actinobacillus pleuropneumoniae in commercial production settings is based on serological monitoring. Enzyme-linked immunosorbent assays (ELISAs) have been developed to detect specific antibodies against a variety of A. pleuropneumoniae antigens, including long-chain lipopolysaccharides (LPS) and the ApxIV toxin, a repeats-in-toxin (RTX) exotoxin unique to A. pleuropneumoniae and produced by all serovars. The objective of this study was to describe ApxIV antibody responses in serum and oral fluid of pigs.Material and Methods: Four groups of pigs (six pigs per group) were inoculated with A. pleuropneumoniae serovars 1, 5, 7, or 12. Weekly serum samples and daily oral fluid samples were collected from individual pigs for 56 days post inoculation (DPI) and tested by LPS and ApxIV ELISAs. The ApxIV ELISA was run in three formats to detect immunlgobulins M, G, and A (IgM, IgG and IgA) while the LPS ELISA detected only IgG.Results: All pigs inoculated with A. pleuropneumoniae serovars 1 and 7 were LPS ELISA serum antibody positive from DPI 14 to 56. A transient and weak LPS ELISA antibody response was observed in pigs inoculated with serovar 5 and a single antibody positive pig was observed in serovar 12 at ≥35 DPI. Notably, ApxIV serum and oral fluid antibody responses in pig inoculated with serovars 1 and 7 reflected the patterns observed for LPS antibody, albeit with a 14 to 21 day delay.Conclusion: This work suggests that ELISAs based on ApxIV antibody detection in oral fluid samples could be effective in population monitoring for A. pleuropneumoniae.

Myeloperoxidase (MPO) is a leukocyte-derived enzyme involved in the process of heart failure and is found to have good diagnostic and prognostic values in humans with chronic heart failure. This study evaluated the relationship between serum MPO levels and the severity of heart failure (HF) due to chronic mitral valvular insufficiency (CMVI) in dogs. Eighty-two client-owned dogs consisting of 69 dogs with different stages of HF due to CMVI and 13 age-matched healthy dogs were enrolled in this study. Serum MPO concentrations in the healthy and CMVI groups were determined by enzyme-linked immunosorbent assay (ELISA) using a canine-specific monoclonal anti-MPO antibody. Serum MPO concentrations were 273.3 ± 179.6 ng/L in the controls, 140.8 ± 114.1 ng/L in the International Small Animal Cardiac Health Council (ISACHC) I group, 109.0 ± 85.2 ng/L in the ISACHC II group, and 106.0 ± 42.3 ng/L in the ISACHC III group. Close negative correlation to serum MPO concentration was found in the severity of heart failure (ISACHC stage). Although this study found a modest relationship between serum MPO levels and the severity of HF due to CMVI in dogs, it also suggested that serum MPO levels decreased as the severity of HF increased.

The objective of the present study was to determine the effect of concurrent vaccination of boars with type 1 and type 2 porcine reproductive and respiratory syndrome virus (PRRSV) on seminal shedding of both genotypes. The boars tolerated well concurrent administration of 2 commercial PRRSV vaccines, and no adverse reactions were observed. No interference in the humoral immune response (measured as the level of anti-PRRSV antibodies) or the cell-mediated immune response (measured as the level of PRRSV-specific interferon-γ-secreting cells) was observed after concurrent administration compared with single administration of the same vaccines. Concurrent vaccination significantly reduced the load of type 1 and type 2 PRRSV in blood and semen after singular (type 1 or type 2) and dual (type 1 and type 2) PRRSV challenge, and it did not significantly affect the efficacy of each vaccine. The results demonstrate that concurrent vaccination of boars with type 1 and type 2 PRRSV reduces shedding of both genotypes in semen.

OBJECTIVE To determine whether canine protein C (CnPC) had antichemotactic effects on canine neutrophils, whether endothelial protein C receptor (EPCR) was expressed on canine neutrophils, and the role of EPCR in neutrophil chemotaxis. SAMPLE Neutrophils isolated from blood samples from healthy dogs (n = 6) and sick dogs with (2) or without (3) an inflammatory leukogram. PROCEDURES Neutrophils were analyzed by reverse transcriptase PCR assay and flow cytometry for detection of EPCR mRNA and protein expression, respectively. Neutrophils were incubated with CnPC zymogen or canine activated protein C (CnAPC), with or without RCR-379 (an anti–human EPCR antibody). Neutrophils were then allowed to migrate through a filter membrane toward a chemokine. Untreated neutrophils served as positive control samples. Migration was quantified by fluorescence measurement, and chemotaxis index (Chx) values (fluorescence of test sample/fluorescence of positive control sample) were computed. RESULTS The cDNA for EPCR was amplified, and EPCR expression was detected on neutrophil surfaces. Obtained Chx values were significantly higher in cells treated with RCR-379 than in cells treated with CnPC or CnAPC alone. The Chx values for neutrophils treated with RCR-379 were not significantly different from 1, whereas those for neutrophils treated without RCR-379 were significantly less than 1. The effects of RCR-379 on neutrophil migration were independent of concentration or activation status of protein C. CONCLUSIONS AND CLINICAL RELEVANCE Canine neutrophils expressed EPCR, and inhibition of neutrophil chemotaxis by CnPC and CnAPC depended on EPCR. Interventions with EPCR signaling may have therapeutic application in dogs.