Application of a bacteriological on-farm test to reduce antimicrobial usage in dairy cows with purulent vaginal discharge

The objective of this study was to assess the effect of a selective antibiotic treatment strategy based on a quick bacteriological on-farm test (Petrifilm, 3M Corp., St. Paul, MN) compared with the conventional antibiotic treatment of all cows having clinical endometritis (CE) defined by the presence of purulent vaginal discharge on both clinical cure rate and reproductive performance. The study was simultaneously conducted with dairy cows reared under a highly supplemented rotational grazing system in Argentina and in a freestall system in Slovakia. Cows having an abnormal vaginal discharge (VD, indicative of clinical endometritis) on 21 to 35 d in milk (DIM) were randomly allocated to 1 of 2 study groups: selective treatment (ST) or conventional treatment (CT). All cows in the CT group (n = 174) received a single intrauterine administration of 500 mg of cephapirin. In the ST group (n = 178), treatment decision was made according to the results of the bacteriological on-farm test. For this test, we collected intrauterine samples with the cytobrush technique and stroke the brushes onto 2 different Petrifilm plates, one for aerobic count and another for Enterobacteriaceae count, incubated the plates, and counted the number of colonies after 24 h. Positive cows (≥5 colonies in one or both plates) received a single intrauterine treatment with 500 mg of cephapirin, whereas negative cows (<5 colonies) remained untreated. Clinical cure rate was assessed by direct vaginal inspection at 14 d after treatment (VD-0). The odds for conception at first artificial insemination, artificial insemination by 80 DIM, pregnancy by 100 DIM, and for nonpregnancy by 200 DIM were estimated with mixed logistic regression models. The hazard of conception was also assessed with proportional hazard regression model. The selective antibiotic treatment strategy based on the outcome of Petrifilm test reduced the number of required treatments (57%) and maintained similar efficacy in terms of clinical cure and reproductive performance as the conventional antibiotic treatment of all endometritic cows.     

The Role of mTOR and eIF Signaling in Benign Endometrial Diseases

Adenomyosis, endometriosis, endometritis, and typical endometrial hyperplasia are common non-cancerous diseases of the endometrium that afflict many women with life-impacting consequences. The mammalian target of the rapamycin (mTOR) pathway interacts with estrogen signaling and is known to be dysregulated in endometrial cancer. Based on this knowledge, we attempt to investigate the role of mTOR signaling in benign endometrial diseases while focusing on how the interplay between mTOR and eukaryotic translation initiation factors (eIFs) affects their development. In fact, mTOR overactivity is apparent in adenomyosis, endometriosis, and typical endometrial hyperplasia, where it promotes endometrial cell proliferation and invasiveness. Recent data show aberrant expression of various components of the mTOR pathway in both eutopic and ectopic endometrium of patients with adenomyosis or endometriosis and in hyperplastic endometrium as well. Moreover, studies on endometritis show that derangement of mTOR signaling is linked to the establishment of endometrial dysfunction caused by chronic inflammation. This review shows that inhibition of the mTOR pathway has a promising therapeutic effect in benign endometrial conditions, concluding that mTOR signaling dysregulation plays a critical part in their pathogenesis.     

Short communication: Repeat breeder cows with fluid in the uterine lumen had poorer fertility

The objective of the present study was to test the association between the presence of fluid in uterine lumen and the fertility of repeat breeder cows (RBC). Grazing Holstein cows that were clinically normal and were not pregnant after at least 3 consecutive inseminations were defined as RBC and included in the study (n = 358). A cut-point ≥2 mm of uterine lumen visible by ultrasonography was used as indicative of the presence of fluid in the uterus. The effect of this fluid on the fertility of RBC was assessed by mixed logistic regression. Twenty percent of the RBC had ≥2 mm of fluid in their uterus, and this finding was associated with a marked reduction in the odds of conception (odds ratio = 0.46). Additional intrauterine sampling revealed that 24.0% of RBC showed signs of subclinical endometritis [≥5% neutrophils in endometrial cytology (SCE)]. However, we found no agreement between the finding of uterine fluid and the diagnosis of SCE (kappa coefficient = 0.03, 95% confidence interval = ?0.07 to 0.14). In conclusion, the presence of fluid (≥2 mm) in uterine lumen, detected by ultrasonography, was associated with lower fertility in RBC, but was not related to the diagnosis of SCE. Thus, ultrasonographic evaluation is an important tool to assist in assessment of RBC.     

宫腔镜检查在不明原因复发性自然流产中的应用

目的：部分复发性自然流产病因不明确,本研究通过宫腔镜检查,探讨宫腔镜在不明原因复发性自然流产中的病因诊断作用。方法：对2008年1月至2010年1月在我院就诊的原因不明的复发性自然流产患者38例行宫腔镜检查,并取内膜活检。结果：38例患者中,宫腔镜检查4例有轻度宫腔粘连,4例子宫内膜有炎症表现,病理诊断确诊5例有慢性子...     

Presence of Escherichia coli, Trueperella pyogenes, α-hemolytic streptococci,and coagulase-negative staphylococci and prevalence of subclinical endometritis

Subclinical endometritis (SCE) has a severe negative effect on reproductive performance of dairy cows. Different studies have shown a wide variety of cut points for the polymorphonuclear neutrophilic leukocyte (PMNL) percentage to define SCE and differing sampling times. Possible risk factors for SCE, such as low body condition score, hyperketonemia, and increased haptoglobin concentrations have been demonstrated. The aim of this study was to determine the relationship between bacterial findings in the uterus and PMNL in the early postpartum period and to compare 2 classification schemes for the diagnosis of SCE. Cytological and bacteriological samples from the uterus were taken from 149 cows at 2 consecutive times postpartum [10 ± 1 and 24 ± 1 d in milk (DIM)] and additionally, 131 cows were sampled at 21 to 27 DIM. For sample collection, the cytobrush technique was used and vaginal discharge was examined by vaginoscopy. Polymorphonuclear neutrophilic leukocyte cut points were set at 5, 10, and 18%. Cows positive with α-hemolytic streptococci at 10 ± 1 DIM had significant higher PMNL percentages at 21 to 27 DIM and greater median days to pregnancy (193.5 d) than cows not infected (123.0 d). Cows with Trueperella pyogenes at 10 ± 1 DIM had higher odds (5.2) of having more than 18% PMNL at 21 to 27 DIM compared with cows without infection. Escherichia coli had no effect on the prevalence of SCE regardless of the cut point. Primiparous cows with more than 18% PMNL had greater median days to pregnancy (144.5 d) than cows with less than 5% (80.0 d) or cows between 5 and 18% PMNL (68.0 d). Cows between 5 and 18% PMNL or 10 and 18% PMNL showed the best reproductive performance. The results demonstrate that an early infection with α-hemolytic streptococci increased the PMNL percentage 2 wk later and that 18% PMNL at 21 to 27 DIM was a predictive cut point for primiparous cows to diagnose animals with or without SCE.     

Evaluation of the Metricheck device and the gloved hand for the diagnosis of clinical endometritis in dairy cows

The objective of this study was to compare 2 surrogate vaginal examination methods (i.e., gloved hand and a vaginal device) with vaginoscopy as a reference method for diagnosing clinical endometritis in dairy cows. Holstein-Friesian cows (n = 1,002) in 2 commercial dairy herds in Germany were examined for endometritis at 21 to 27 d in milk (DIM) by using 1 of 3 vaginal diagnostic methods. Vaginal examinations were performed either with a speculum (reference method), a vaginal device (Metricheck, Simcro, New Zealand), or a gloved hand. Vaginal discharge adhering to the diagnostic tool was classified according to a vaginal discharge score ranging from 0 to 3 (where 0 = translucent mucus, 1 = mucus containing flecks of white or off-white pus, 2 = less than 50% white or off-white mucopurulent material, and 3 = greater than 50% white or yellow pus that may be sanguineous). Cows with vaginal discharge scores of 1 to 3 received 500 μg of cloprostenol after examination and again 14 d later (35 to 41 DIM). The prevalence of endometritis in both herds was 40.6 and 40.3%, respectively. With the Metricheck device, significantly more cows were diagnosed as affected with endometritis than by examination with a speculum or a gloved hand (47.5 vs. 36.9 and 36.8%). Binary logistic regression for the risk of conception after first AI as an outcome variable, with vaginal discharge score, diagnostic method, and farm as covariates, revealed a significant effect of degree of endometritis, but not of the diagnostic methods. Survival analyses for the hazard of insemination and pregnancy within 200 DIM, respectively, revealed a significant effect of degree of endometritis, herd, and parity, but not of the diagnostic tool. It can be concluded that any one of the 3 vaginal examination methods can be used interchangeably, without a negative effect on reproductive performance.     

Association between interleukin-8 receptor-α (CXCR1) polymorphism and disease incidence, production, reproduction, and survival in Holstein cows

The objective was to evaluate the association between the single nucleotide polymorphism at position +735 in the interleukin-8 receptor-α (CXCR1) gene (CXCR1c.735) and disease incidence, milk production, reproductive performance, and survival in Holstein cows. Three-hundred fifty Holstein cows were enrolled. No association was found between CXCR1c.735 genotype and retained fetal membranes, metritis, or endometritis. Incidence rate of clinical mastitis was associated with CXCR1c.735 genotype; cows with genotypes CC and GC had a decreased incidence rate of clinical mastitis compared with GG cows. Milk yield was associated with CXCR1c.735 genotype; cows with genotype GC had greater milk yield than GG cows. Hazard of pregnancy was not associated with CXCR1c.735 genotype. Cows that had clinical mastitis had decreased hazard of pregnancy, and cows that had endometritis tended to have a decreased hazard of pregnancy. Hazard of death or culling was not associated with CXCR1c.735 genotype. Multiparous cows and cows that had mastitis had increased hazard of death or culling. In contrast to what we expected, cows with the genotype GG had an increased incidence rate of clinical mastitis and decreased milk yield.     

Effects of Substance P and Neurokinin A on the Contractile Activity of Inflamed Porcine Uterus

Disturbances in uterine contractile activity contribute to the development of inflammation, and recent evidence indicates that tachykinins, including substance P (SP) and neurokinin A (NKA), are involved in controlling uterine function. Here, we determined the effect of Escherichia coli (E. coli)-induced inflammation on expression of protein receptor subtypes for substance P (NK1R) and neurokinin A (NK2R) in the pig myometrium as well as their role in contractility of inflamed uterus. The severe acute endometritis developed in the E. coli group and the expression of NK1R and NK2R proteins increased in the myometrium. Compared to the pre-administration period, SP (10⁻⁶ M) reduced the amplitude and frequency in the myometrium of the E. coli group and the amplitude was higher and the frequency was lower versus other groups. NKA reduced the amplitude and increased the frequency in endometrium/myometrium of the E. coli group. In this group, the amplitude was lower and the frequency was higher than in the CON and SAL groups. Our research showed that NK2R (10⁻⁶ M) antagonist application abolished the NKA inhibitory effect on uterine amplitude. The application of the NK1R (10⁻⁵ M) antagonist together with SP revealed that the inhibitory effect of SP on uterine contractility is achieved independently of the NKR1. Additionally, taking into account the fact that NKA shows an inhibitory effect with the use of NK2R on uterine amplitude suggests the possibility of therapeutic use of the antagonist as a drug increasing uterine contractility in inflammation.     

Relationships between endometritis and metabolic state during the transition period in pasture-grazed dairy cows

The primary objective of this study was to identify relationships between endometritis and metabolic state during the calving transition and early lactation periods. A subset of mixed age and breed dairy cows (n = 78) from a seasonal, pasture-grazed herd of 389 cows was examined. The selected cows were grouped as having endometritis at d 42 postpartum or being unaffected by endometritis. Endometritis was defined as >6% (upper quartile) of uterine nucleated cells being polymorphonuclear cells (H-PMN; n = 38); unaffected by endometritis was defined as ≤1% of nucleated cells being polymorphonuclear (L-PMN; n = 40). Milk yield was determined at each milking, and milk composition (fat and protein) was determined at 2-wk intervals. Blood samples collected on d −14, 0 (d of calving), 4, 7, 14, 28, and 42 were analyzed for indicators of energy status (nonesterified fatty acids, glucose, and urea), liver function (albumin, globulin, glutamate dehydrogenase, and aspartate aminotransferase), inflammation (haptoglobin), and mineral status (Ca and Mg). Samples collected weekly from d 21 to 63 or 70 were analyzed for progesterone content. The postpartum anovulatory interval was defined to end on the first day postpartum that plasma progesterone concentration was ≥1 ng/mL. A greater percentage of H-PMN cows failed to ovulate before d 63 or 70 (34%) compared with L-PMN cows (10%), although the proportions of cows ovulating within either polymorphonuclear group was similar through d 56 postpartum. Plasma concentrations of albumin and the albumin:globulin ratio were consistently lower in H-PMN cows. Plasma Mg was lower, whereas glutamate dehydrogenase and aspartate aminotransferase were higher, in H-PMN cows during early lactation compared with L-PMN cows. Circulating metabolites indicative of energy status (nonesterified fatty acids, glucose, and urea) were not different between polymorphonuclear groups. Among 3- to 5-yr-old cows, daily milk yield for the first 42 d after calving was lower for H-PMN cows than for L-PMN cows. Among cows >5 yr old, protein percentage was lower in H-PMN cows compared with L-PMN cows. In summary, endometritis at 42 d postpartum in the herd studied was associated with an increased likelihood of remaining anovulatory. These cows had lower albumin concentrations throughout the calving transition period, perhaps indicating impaired liver function, with lower plasma Mg and evidence of hepatocellular damage in early lactation. Similar profiles of nonesterified fatty acids and glucose indicated that energy status was not a risk factor for endometritis.     

Reproductive tract disease in Irish grazing dairy cows: Retrospective observational study examining its association with reproductive performance and accuracy of 2 diagnostic tests

The detection of reproductive tract disease (RTD) 3 wk postpartum is important because of its effect on subsequent reproductive outcomes. Numerous methods for the diagnosis of RTD are described, some of which are more practical and instantaneous in terms of diagnosis. Two of these methods involve identification of purulent vaginal discharge (PVD) and evidence of ultrasonographic uterine changes indicative of endometritis (UE). The objectives of our retrospective observational study were (1) to assess the association of PVD or UE score at the prebreeding examination (PBE) with the hazard of pregnancy within the subsequent breeding season; (2) to determine the test sensitivity (Se) and specificity (Sp) at the point of sampling of both tests using a Bayesian latent class model; and (3) to determine the effect of varying positivity thresholds on test accuracy. To achieve these objectives, we analyzed an initial data set of 5,049 PBE from 2,460 spring-calved cows in 8 herds between 2014 and 2018. Each PBE was conducted once between 25 and 86 d in milk. At each PBE, vaginal discharge was obtained with a Metricheck device (Simcro) whereas uterine contents were assessed using transrectal ultrasonography. Purulent vaginal discharge was scored on a scale of 0 to 3 depending on discharge character, and UE was scored on a scale of 0 to 4 depending on the presence and consistency of intraluminal fluid. Cows with scores of ≥2 in either test had received treatment. Fertility data were available from 4,756 PBE after data exclusion. The association between PVD or UE score at the PBE and subsequent hazard of pregnancy was analyzed using a Cox proportional hazards model. Cows with a PVD score of 2 or 3 were less likely to conceive than cows with a PVD score 0 [score 2 hazard ratio (HR) = 0.74; 95% confidence interval (CI): 0.59–0.94; score 3 HR = 0.65; 95% CI: 0.51–0.84]. Cows with a UE score of 1, 2, 3, or 4 were less likely to conceive than cows with a UE score of 0 (score 1 HR = 0.82; 95% CI: 0.73–0.93; score 2 HR = 0.79; 95% CI: 0.62–1.00; score 3 HR = 0.43; 95% CI: 0.43–0.90; score 4 HR = 0.39; 95% CI: 0.26–0.58). To determine the Se and Sp of PVD or UE score for diagnosis of RTD at the time of PBE, a Bayesian latent class model was fitted on 2,460 individual cow PBE. Flat priors were used for the Se and Sp of UE, whereas informative priors were used for PVD Se (mode = 65%, 5th percentile = 45%) and Sp (mode = 90%, 5th percentile = 80%) and RTD prevalence (mode = 20%, 5th percentile = 10%). Posterior estimates (median and 95% Bayesian probability intervals; BPI) were obtained using ‘rjags' (R Studio). The optimal test thresholds (PVD and UE score ≥1) were selected by assessing the effect of different thresholds on test estimates and using a misclassification cost analysis. Based on these, median (95% BPI) Se for PVD and UE score ≥1 were 44% (29–60%) and 67% (33–100%), respectively. Median Sp for PVD and UE score ≥1 were 90% (86–93%) and 91% (86–93%), respectively. Higher scores in both tests were associated with impaired fertility, and UE scoring with a threshold of ≥1 had the highest test Se and Sp estimates although test Se was conditional on days in milk when the PBE occurred.