Characterization of intrauterine cultivable aerobic microbiota at the time of insemination in dairy cows with and without mild endometritis.

It is generally accepted that postpartum uterine infections decrease conception rates in dairy cows. Whereas clinical endometritis (CE) has been studied intensively, only little information about CE at the time of artificial insemination (AI) is available. The aim of this study was to characterize the intrauterine cultivable aerobic microbiota in healthy cows and in cows with mild CE at AI and to investigate its effect on the subsequent insemination success. The vaginal discharge score (VDS) of 120 Holstein Frisian dairy cows was assessed shortly after AI using the Metricheck device and pregnancy diagnosis was performed 39 days after AI. On average, cows received their second insemination (2.0 ± 1.2 standard deviation (SD)) and were 120.5 ± 40.2 days in milk. Intrauterine cytobrush samples were taken from cows with clear mucus (VDS 0, n = 58) and from cows with flecks of pus (VDS 1, n = 62), which was regarded as mild CE. Bacteria collected with the cytobrush were cultivated aerobically and identified by matrix assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS). A total of 358 bacterial isolates were recovered, comprising 49 bacterial genera and 116 species. Bacillus, Staphylococcus, Corynebacterium and Streptococcus were the most prevalent genera with relative abundances of 19.6%, 14.2%, 10.1% and 8.1%, whereas common uterine pathogens, such as Trueperella pyogenes and Escherichia coli, were rarely detected. Bacillus spp. were more frequently detected in group VDS 0 (51.7%) than in VDS 1 (22.6%; P = 0.001) and there was a higher prevalence of Corynebacterium spp. in VDS 0 than in VDS 1 (37.9% vs 19.3%; P = 0.03). Cows with VDS 1 at the time of AI harbored neither Bacillus licheniformis nor Bacillus subtilis, while these species were detected in 27.6% and 8.6% of VDS 0 (P < 0.001 and P = 0.024 respectively). The insemination success in VDS 0 and 1 was 43.1 and 25.8% (P = 0.046). The diagnosis of mild CE at AI and the total bacterial load decreased the chance of pregnancy (P = 0.03) but no significant correlation between a distinctive bacterial species and insemination success was observed. In summary, cultivable uterine bacteria at AI presumably represent a part of the physiological microbiota, which is not directly linked to impaired fertility. These findings question intrauterine antimicrobial treatment shortly after AI and could represent the basis for the development of future treatment strategies.

Field trial on the post-insemination intrauterine treatment of dairy cows with mild endometritis with cephapirin.

Cows in estrus but with signs of clinical endometritis (CE) are often not inseminated or undergo an intrauterine treatment after artificial insemination (AI). Decades ago, the so-called Aström method was described as intrauterine infusion of iodine-potassium solution 2-4 days after AI. Nowadays, it is common to use antibiotics instead of iodine solution and the treatment is performed only a few hours after AI. Although widespread in practice, there is only little information about the efficacy of this treatment. Thus, this study evaluated the effect of a post-breeding intrauterine treatment with cephapirin on insemination success in cows with signs of mild CE. In total, 281 cows subjected to an Ovsynch program with fixed-time AI and examined for vaginal discharge straight after AI by use of the Metricheck device were included. Cows with cloudy discharge or flecks of pus in the mucus were assigned to a treatment or a control group. The treatment group (MET; n = 87) received 6 ± 1 h after AI an intrauterine treatment with 500 mg of cephapirin (Metricure, Intervet Deutschland GmbH). Control cows (CON; n = 91) remained untreated. Animals with clear discharge were assigned to a healthy comparison group (HE; n = 103). Pregnancy diagnosis was performed 39 days after AI. The proportion of pregnant cows after the AI directly preceding the enrollment did not differ significantly (P > 0.05) between HE (35.0%), CON (27.5%) and MET (32.2%). Cephapirin treatment had also no positive effect on other reproductive performance measures, i.e, the percentage of pregnant cows 200 days after enrollment (HE: 64.1%, CON: 73.6%, and MET: 73.6%) or the mean interval from enrollment to conception (HE: 25.4 days, CON: 30.0 days, and MET: 29.7 days). The binary logistic regression showed that the only risk factors with a detrimental effect on fertility were a history of CE 28-34 days postpartum and season. Although cows in MET and HE were 1.74 and 1.37 times more likely to conceive after AI than CON, this effect was not significant. Uterine sampling of a subset of cows with CE (n = 50) revealed 127 bacterial isolates. The most frequently found genera were Staphylococcus (19.7%), Bacillus (12.6%), Streptococcus (10.2%), Corynebacterium (8.7%), and Lysinibacillus (7.9%). The finding that common uterine pathogenic bacteria were rarely detected additionally questions an intrauterine antibiotic treatment of cows with mild CE at AI.