Genomic characterization of Arcanobacterium pyogenes isolates recovered from the uterus of dairy cows with normal puerperium or clinical metritis.

Arcanobacterium pyogenes is considered to be the most relevant bacterium involved in the establishment of puerperal uterine infection in cattle due to its persistence in utero, resistance to treatment and synergic action with Gram negative anaerobes. Once the infection is established, A. pyogenes is responsible for the persistence of the infection. The objective of this study was to characterize A. pyogenes field isolates recovered from the uterus of cows with either normal puerperium or clinical metritis, in an attempt to identify factors that might be associated with the establishment and persistence of the disease. This characterization was based on BOX-PCR typing and on screening of eight virulence factor genes (plo, nanP, nanH, cbpA, fimA, fimC, fimE, fimG) by conventional PCR. Finally, a relationship between clonal types, virulence factors and presence of disease was investigated. A. pyogenes clonal types identified from isolates recovered from the uterus of postpartum dairy cows differed among herds. Although some clonal types were strictly associated with the development of clinical metritis, others were identified from isolates recovered from normal puerperium and clinical metritis cows. Moreover, the presence of the eight virulence factor genes was not related with the ability to induce clinical metritis, suggesting that the type of A. pyogenes may not be a determinant factor in the development of the disease. We suggest that host intrinsic factors, the synergism between A. pyogenes and other bacteria and the differential gene expression of virulence factor genes may play a more relevant role in the establishment of puerperal uterine infections.

Blood COX-2 and PGES gene transcription during the peripartum period of dairy cows with normal puerperium or with uterine infection.

In the dairy cow, puerperal uterine intra-luminal concentrations of PGE(2) are related to the establishment and severity of uterine infections. Here we evaluated whether the blood concentrations of PGE(2) and the gene transcription profiles of enzymes involved in its synthesis (cyclooxygenase-2 and prostaglandin E synthase) could be used as markers of predisposition and/or presence of puerperal uterine infections. We also studied the relationship between the endocrine status and the leukocyte profiles around parturition and the transcription patterns of the genes. Finally, we have characterized the in vitro gene transcription and expression response to a challenge of LPS. Gene transcription profiles, quantified by real-time PCR, were similar in normal puerperium and metritis/endometritis cows, indicating that they are not suitable markers of predisposition to/presence of puerperal uterine infections. Transcription decreased from 2 weeks before parturition until parturition, when a minimum was attained, and then increased during the first week postpartum. The lowest gene transcription, at parturition, was coincidental with the highest total leukocytes, polymorphonuclear neutrophils and CD14 positive cell numbers. It is suggested that by this mechanism, a large number of PMN can be recruited into the uterus after parturition, avoiding an excessive acute inflammatory response. The lowest gene transcription was also coincidental with the surge in cortisol concentrations, indicating that this hormone plays a main immunomodulatory role around parturition. Gene transcription was significantly greater after stimulation with LPS than in non-stimulated blood. We suggest that this PGE(2) producing cells might arrive to the uterine lumen, contributing to the local PGE(2) concentrations and mediating the inflammatory response.