Development of a Multiplex PCR for the Identification of Major Pathogenic Bacteria of Post‐Partum Endometritis in Dairy Cows

Abstract

Post-partum period has an important role in cows' breeding due to its effects on reproductive efficiency and subsequent pregnancy. Escherichia coli, Trueperella pyogenes (Arcanobacterium pyogenes), Fusobacterium necrophorum and Prevotella melaninogenicus are recognized as major pathogens associated with uterine endometrial lesions. The objective of this study was to identify these pathogens using the polymerase chain reaction (PCR) as a culture-independent sensitive method. A total of 172 cows were examined 25-35 days post-partum, and 128 cows were examined at 2 weeks later (39-49 days post-partum). Uterine discharges were collected by covered plastic infusion pipettes. The prevalence of endometritis was greater in the first examination than the second (35.5% vs. 16%). E. coli was detected in eight of the samples, T. pyogenes was detected in 13 of the samples and F. necrophorum was detected in 11 of the samples. There was no positive sample of P. melaninogenicus. Uterine contamination by T. pyogenes and F. necrophorum in the first examination was higher than the second examination. T. pyogenes affected as a tendency the prevalence of clinical endometritis in first examination. Primiparous cows showed 4.02 times higher odds of clinical endometritis compared with second-parity cows in first examination. A multiplex PCR protocol as a simple, less expensive, fast assay was introduced to identify E. coli, T. pyogenes and F. necrophorum.

A Discussion on Risk Factors, Therapeutic Approach of Endometritis and Metritis in Cattle

“…Puerperal metritis tends to develop in cows having parity less than 3 because of uterine tissue damage during dystocia (Bruun et al, 2002). Primiparous cows having higher risk of clinical endometritis compared to toward second-parity cows (Aghamiri et al, 2014). Reports also support the risk of puerperal metritis was highest following first calving and lowest in second parity cows (Markusfeld, 1987).…”

Lactoferrin suppresses lipopolysaccharide-induced endometritis in mice via down-regulation of the NF-κB pathway

“…However, many researchers have used mice to investigate diseases that affect cows, particularly mastitis [3,4]. Gram-negative bacteria, such as Escherichia coli, Arcanobacterium pyogenes, Fusobacterium necrophorum and Prevotella melaninogenicus, are recognized as major pathogens associated with uterine endometrial lesions [5,6]. LPS, a main component of the outer membrane of gram-negative bacteria, is an important factor in induced inflammatory responses in cultured cells and live animals [7,8].…”

Development of a multiplex PCR for simultaneous detection of Pasteurella multocida, Mannheimia haemolytica and Trueperella pyogenes

“…These genes were reported to be highly conserved among strains of these three bacteria. In addition, some studies have enrolled these genes as a target for establishing the detection method for the three bacteria (Townsend et al, 1998;Angen et al, 2009;Aghamiri et al, 2014). Therefore, choosing these genes as the targets of the multiplex PCR would guarantee the credibility and reproducibility of the method.…”