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BackgroundCorynebacterium pseudotuberculosis can be classified into two biovars or biovars based on their nitrate-reducing ability. Strains isolated from sheep and goats show negative nitrate reduction and are termed biovar Ovis, while strains from horse and cattle exhibit positive nitrate reduction and are called biovar Equi. However, molecular evidence has not been established so far to understand this difference, specifically if these C. pseudotuberculosis strains are under an evolutionary process.ResultsThe ERIC 1 + 2 Minimum-spanning tree from 367 strains of C. pseudotuberculosis showed that the great majority of biovar Ovis strains clustered together, but separately from biovar Equi strains that also clustered amongst themselves. Using evolutionarily conserved genes (rpoB, gapA, fusA, and rsmE) and their corresponding amino acid sequences, we analyzed the phylogenetic relationship among eighteen strains of C. pseudotuberculosis belonging to both biovars Ovis and Equi. Additionally, conserved point mutation based on structural variation analysis was also carried out to elucidate the genotype-phenotype correlations and speciation. We observed that the biovars are different at the molecular phylogenetic level and a probable anagenesis is occurring slowly within the species C. pseudotuberculosis.ConclusionsTaken together the results suggest that biovar Equi is forming the biovar Ovis. However, additional analyses using other genes and other bacterial strains are required to further support our anagenesis hypothesis in C. pseudotuberculosis.Electronic supplementary materialThe online version of this article (doi:10.1186/s12866-016-0717-4) contains supplementary material, which is available to authorized users.

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The aims of the present study were: to estimate the seroprevalence of Bluetongue vírus (BTV), bovine alphaherpesvirus 1 (BoHV-1), bovine viral diarrhea virus (BVDV), Brucella abortus and Leptospira spp. in cattle from family farms in North of Minas Gerais; to determine the intercurrence and association among these pathogens; and to assess the possible associated factors to seropositive herds and animals. For this, 476 cows from 46 farms were sampled and evaluated serologically. The seroprevalence in herds and cattle was, respectively, for BTV 100% and 52.0%, for BoHV-1 95.7% and 48.6%, for BVDV 78.3% and 46.1%, for Leptospira spp. 76.1% and 29.1%, and for B. abortus was 0% in herd and cattle. More than 65% of the herds was simultaneously seropositive for BTV, BoHV-1, BVDV and Leptospira spp. Seropositivity for BoHV-1, Leptospira spp. serovar Autumnalis and serovar Hardjoprajitno were associated with abortion, whereas seropositivity for BVDV was associated with BoHV-1. Moreover, association among seropositivity for BVDV, BoHV-1 and Leptospira spp. was also observed. In conclusion, BTV, BoHV-1, BVDV and Leptospira spp. are highly seroprevalent and occurred simultaneously in cattle from family farms in Minas Gerais, indicating the need for the implementation of control measures to avoid economic losses related to these diseases.

Biochemical, serological, and molecular methods have been developed for the laboratory diagnosis of diseases caused by C. pseudotuberculosis (CP), but the identification of the pathogen and biovars differentiation may be time-consuming, expensive, and confusing compared with other bacteria. This study aimed to evaluate MALDI Biotyper and Overall Genome Relatedness Index (OGRI) analysis to optimize the identification and differentiation of biovars of C. pseudotuberculosis. Out of 230 strains isolated from several hosts and countries, 202 (87.8%) were precisely classified using MALDI Biotyper and the BioNumerics platform. The classification accuracies for the Ovis and Equi biovars were 80 (88.75%) and 82 (92.68%), respectively. When analyzing a sampling of these strains by Average Nucleotide Identity based on BLAST and TETRA analyses using genomic sequence data, it was possible to differentiate 100% of the strains in Equi and Ovis. Our data show that MALDI Biotyper and OGRI analysis help identify C. pseudotuberculosis at the species and biovar levels.

O objetivo deste estudo foi analisar as doenças de notificação obrigatória no Brasil no período de 1999 a 2019 em caprinos (Capra hircus) e ovinos (Ovis aries). Para isso, foi realizado um estudo longitudinal descritivo e retrospectivo nos casos notificados entre os anos de 1999 e 2019 em todo Brasil. Os dados foram obtidos do Sistema Nacional de Informações Zoossanitárias (SIZ), que é gerenciado pela Coordenação de Informação e Epidemiologia (CIEP) do Ministério da Agricultura, Pecuária e Abastecimento (MAPA). Foram analisadas as frequências absolutas das doenças listadas nas categorias 1, 2 e 3 da Instrução Normativa nº 50, de 24 de setembro de 2013, e a presença ou ausência das doenças da categoria 4 da mesma lista. Língua Azul, Raiva, Febre Aftosa e Scrapie foram as doenças mais notificadas em pequenos ruminantes no Brasil no período de 1999 a 2019. Carbúnculo Hemático, Tuberculose e Estomatite Vesicular também foram confirmadas neste período no País, porém em frequências muito baixas. Completam o painel de doenças notificáveis presentes, Agalaxia Contagiosa, Artrite-Encefalite Caprina, Clamidofilose Ovina, Epididimite Ovina e Maedi-Visna. Globalmente, os achados revelam o panorama das doenças de notificação obrigatória no Brasil em pequenos ruminantes.

The aims of the present study were (i) to genotype Corynebacterium pseudotuberculosis, C. silvaticum, and C. auriscanis strains using enterobacterial repetitive intergenic consensus (ERIC-PCR), and (ii) to analyze the epidemiological relationships among isolates according to biovar (Equi and Ovis), species, host, and geographical origin of the C. pseudotuberculosis strains. Sixty-eight C. pseudotuberculosis, nine C. silvaticum, and one C. auriscanis, C. pseudotuberculosis ATCC® 19410™ strain and the attenuated C. pseudotuberculosis 1002 vaccinal strain were fingerprinted by ERIC 1+2-PCR. Field strains were isolated from various hosts (cattle, buffaloes, sheep, goats, horses, dogs, and pigs) in six countries (Mexico, Portugal, Brazil, Equatorial Guinea, Egypt, and Israel). High genetic diversity was found among the studied Corynebacterium spp. isolates, clustering in 24 genotypes with a Hunter & Gaston diversity index (HGDI) of 0.937. The minimal spanning tree of Corynebacterium spp. revealed three clonal complexes, each associated with one bacterial species. Twenty-two genotypes were observed among C. pseudotuberculosis isolates, with an HGDI of 0.934. Three major clonal complexes were formed at the minimal spanning tree, grouped around the geographic origin of C. pseudotuberculosis isolates. These results reinforce the high typeability, epidemiological concordance, and discriminatory power of ERIC-PCR as a consistent genotyping method for C. pseudotuberculosis, which could be useful as an epidemiological tool to control caseous lymphadenitis. Moreover, our results also indicate the potential of ERIC 1+2-PCR for the genotyping of other species of Corynebacterium other than C. pseudotuberculosis.