BackgroundYersiniosis is a zoonosis widely distributed in Europe and swine carry different serotypes of Yersinia enterocolitica and Y. pseudotuberculosis. The aim of this study was to determine the prevalence of Y. enterocolitica and Y. pseudotuberculosis in wild boars in northern Spain. The blood of wild boars (n = 505) was sampled between 2001 and 2012. Seroprevalence was determined in 490 serum samples with an indirect enzyme-linked immunosorbent assay. Seventy-two of the animals were also examined for the presence of Y. enterocolitica or Y. pseudotuberculosis in the tonsils with real-time polymerase chain reaction. All the tonsils were analysed twice, directly and after cold enrichment in phosphate-buffered saline supplemented with 1 % mannitol and 0.15 % bile salts.ResultsAntibodies directed against Y. enterocolitica and Y. pseudotuberculosis were detected in 52.5 % of the animals. Yersinia enterocolitica was detected with real-time polymerase chain reaction in 33.3 % of the wild boars and Y. pseudotuberculosis in 25 %. Significant differences were observed according to the sampling year, and the highest prevalence was during winter and spring. The highest antibody levels and Y. enterocolitica prevalence were observed in mountainous areas at altitudes higher than 600 m, with very cold winters, and with the highest annual rainfall for each dominant climate. Areas with low and medium livestock populations were associated with the highest seroprevalence of Yersinia spp. in wild boars, whereas areas with high ovine populations had the highest prevalence of Y. enterocolitica.ConclusionsThis study shows that Y. enterocolitica and Y. pseudotuberculosis are highly prevalent among wild boars in the Basque country, with Y. enterocolitica most prevalent. The risk of infection among wild boars is influenced by the season and the area in which they live.Electronic supplementary materialThe online version of this article (doi:10.1186/s13028-016-0184-9) contains supplementary material, which is available to authorized users.

BackgroundYersiniosis is a zoonotic disease reported worldwide. Culture and PCR based protocols are the most common used methods for detection of pathogenic Yersinia species in animal samples. PCR sensitivity could be increased by an initial enrichment step. This step is particularly useful in surveillance programs, where PCR is applied to samples from asymptomatic animals. The aim of this study was to evaluate the improvement in pathogenic Yersinia species detection using a suitable enrichment method prior to the real time PCR (rtPCR). Nine different enrichment protocols were evaluated including six different broth mediums (CASO, ITC, PSB, PBS, PBSMSB and PBSSSB).ResultsThe analysis of variance showed significant differences in Yersinia detection by rtPCR according to the enrichment protocol used. These differences were higher for Y. pseudotuberculosis than for Y. enterocolitica. In general, samples incubated at lower temperatures yielded the highest detection rates. The best results were obtained with PBSMSB and PBS2. Application of PBSMSB protocol to free-ranging wild board samples improved the detection of Y. enterocolitica by 21.2% when compared with direct rtPCR. Y. pseudotuberculosis detection was improved by 10.6% when results obtained by direct rtPCR and by PBSMSB enrichment before rtPCR were analyzed in combination.ConclusionsThe data obtained in the present study indicate a difference in Yersinia detection by rtPCR related to the enrichment protocol used, being PBSMSB enrichment during 15 days at 4°C and PBS during 7 days at 4°C the most efficient. The use of direct rtPCR in combination with PBSMSB enrichment prior to rtPCR resulted in an improvement in the detection rates of pathogenic Yersinia in wild boar and could be useful for application in other animal samples.

A Pasteurella multocida strain affecting nulliparous heifers and calves in different ways.Veterinary Microbiology http://dx.doi.org/10. 1016/j.vetmic.2016.08.022 This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers we are providing this early version of the manuscript. The manuscript will undergo copyediting, typesetting, and review of the resulting proof before it is published in its final form. Please note that during the production process errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain. ABSTRACTPasteurella multocida isolates from dairy cattle on a farm in Spain were associated with pneumonia of calves (six isolates) and mastitis of heifers (five isolates). The objective was to determine if the P. multocida isolates retrieved from both disease scenarios were the same strain or whether more than one strain was present. The isolates were identified by a species-specific polymerase chain (PCR) assay, serotyped by the Heddleston scheme and then typed by a number of molecular genotyping assays including multi-locus sequence typing (MLST). The 11 isolates were confirmed as P. multocida but failed to react with any of the 16 Heddleston antisera. The PCR targeting the genes associated with the lipopolysaccharide outer core biosynthesis locus assigned all the isolates to L3 -the type that containsHeddleston serovars 3 and 4. The MLST analysis showed all isolates belonging to ST 79 within the clonal complex of ST13. Only one of the isolates showed a slight different profile by the repetitive extragenic palindromic PCR. The conclusion was that the same strain was associated with pneumonia in calves and mastitis in heifers.

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