Cátedra y Servicio de Reproducción Animal, Facultad de Ciencias Veterinarias, Universidad Nacional de La Plata (FCV-UNLP), La Plata, B1900AVW, Argentina.
J Dairy Sci. 2013 Jul;96(7):4333-9. doi: 10.3168/jds.2012-6269. Epub 2013 May 16.
The objectives of this study were to assess the effect of the stage of estrous cycle on the percentage of endometrial polymorphonuclear cells (PMN) obtained by cytobrush to determine cutoff values for the diagnosis of subclinical endometritis under pastoral conditions, to measure the prevalence of subclinical endometritis 21 to 62d in milk (DIM), and to evaluate the effect of subclinical endometritis on reproductive performance in grazing dairy cows. The first experiment was conducted on a commercial dairy farm in Buenos Aires province (Argentina), where 17 postpartum cyclic dairy cows without clinical endometritis were selected and synchronized by Ovsynch protocol. Endometrial cytology (cytobrush technique) and blood (tail vessels) samples were obtained on d0, 4, 11, and 18 of the estrous cycle (corresponding to estrus, metestrus, diestrus, and proestrus, respectively) and used for measuring percentage of PMN and P4 concentration, respectively. The percentage of PMN was determined 3times by blinded count by 2 operators. Data were analyzed with PROC MIXED, PROC GENMOD, and PROC FREQ from SAS 9.1. The percentage of PMN did not vary with the stage of the estrous cycle. In addition, PMN counts were below any of the reported thresholds in this study (4%) for most of the cows. Therefore, the risk for false positive test results as a consequence of physiological changes in the counts of PMN during estrous cycle is low. The second experiment was conducted on 4 commercial dairy farms in Buenos Aires province (Argentina), where lactating Holstein dairy cows (n=418) 21 to 62 DIM without clinical endometritis were studied. Samples of endometrial cytology were collected with the cytobrush technique. Data were analyzed with receiver operator characteristic curves with Sigmaplot 10.0, and with PROC GLIMMIX, PROC PHREG, and PROC LIFETEST from SAS 9.1. Cutoff values for the diagnosis of subclinical endometritis in grazing dairy cows are 8% PMN for 21 to 33 DIM, 6% PMN for 34 to 47 DIM, 4% PMN for 48 to 62 DIM, and overall 5% PMN for 21 to 62 DIM; the prevalence of subclinical endometritis 21 to 62 DIM was 17%. Finally, subclinical endometritis diagnosed at 21 to 62 DIM decreases the hazard for pregnancy (hazard ratio=0.668; 95% confidence interval=0.492-0.909) and increases the calving to conception interval by d30 compared with normal cows (median 95% confidence interval=133 vs. 93, respectively).
本研究的目的是评估发情周期阶段对通过细胞刷获得的子宫内膜多形核细胞(PMN)百分比的影响,以确定在放牧条件下诊断亚临床子宫内膜炎的截断值,测量 21 至 62 天泌乳期(DIM)的亚临床子宫内膜炎患病率,并评估亚临床子宫内膜炎对放牧奶牛繁殖性能的影响。第一项实验在布宜诺斯艾利斯省(阿根廷)的一家商业奶牛场进行,从产后循环奶牛中选择了 17 头无临床子宫内膜炎的奶牛,并通过 Ovsynch 方案进行同步化。在发情周期的 d0、4、11 和 18 时(分别对应发情期、间情期、发情后期和发情前期),采集子宫内膜细胞学(细胞刷技术)和血液(尾静脉)样本,分别用于测量PMN 百分比和 P4 浓度。PMN 百分比由 2 名操作人员进行 3 次盲法计数。数据采用 SAS 9.1 中的 PROC MIXED、PROC GENMOD 和 PROC FREQ 进行分析。PMN 百分比不受发情周期阶段的影响。此外,在大多数奶牛中,PMN 计数低于本研究报道的任何阈值(4%)。因此,由于发情周期中PMN 计数的生理变化导致假阳性测试结果的风险较低。第二项实验在布宜诺斯艾利斯省(阿根廷)的 4 家商业奶牛场进行,研究了 21 至 62 DIM 无临床子宫内膜炎的泌乳荷斯坦奶牛(n=418)。使用细胞刷技术采集子宫内膜细胞学样本。采用 Sigmaplot 10.0 中的接收者操作特征曲线对数据进行分析,并采用 SAS 9.1 中的 PROC GLIMMIX、PROC PHREG 和 PROC LIFETEST 进行分析。放牧奶牛亚临床子宫内膜炎诊断的截断值为 21 至 33 DIM 时为 8%PMN,34 至 47 DIM 时为 6%PMN,48 至 62 DIM 时为 4%PMN,21 至 62 DIM 时总体为 5%PMN;21 至 62 DIM 的亚临床子宫内膜炎患病率为 17%。最后,与正常奶牛相比,21 至 62 DIM 时诊断出的亚临床子宫内膜炎降低了怀孕的风险(风险比=0.668;95%置信区间=0.492-0.909),并使产犊至配种间隔延长了 30 天(中位数 95%置信区间=133 对 93)。