Barbato O, Menchetti L, Sousa N M, Malfatti A, Brecchia G, Canali C, Beckers J F, Barile V L
Department of Veterinary Medicine, University of Perugia, 06126, Italy.
Department of Veterinary Medicine, University of Perugia, 06126, Italy.
Theriogenology. 2017 Jul 15;97:73-77. doi: 10.1016/j.theriogenology.2017.04.021. Epub 2017 Apr 13.
For the first time in literature this study describes the pregnancy-associated glycoprotein (PAG) profile of buffalo cows during gestation and the post-partum period using antiserum raised against PAG-molecules purified from buffalo placenta (AS#860). Ninety-eight buffalo cows, belonging to a buffalo herd subjected to a synchronization and artificial insemination (AI) program, were enrolled in this study. Blood samples were taken on days 0 (AI), 23, 25, 28, 30 and then biweekly until the end of pregnancy. Pregnancy was confirmed by ultrasonography on days 28 and 45, and by rectal palpation from day 60 onwards. Blood samples were suspended for the non-pregnant cows on day 45, while the blood of 20 buffaloes that had calved was tested every five days from the day of calving until day 50 post-calving. A cut-off value of 1.0 ng/mL was used in order to discriminate between pregnant and non-pregnant buffaloes. We used Linear Mixed models after Log(x+1) transformation to analyse the PAG concentrations. Fifty-two buffalo cows had become pregnant out of 98 synchronized (53%) and 46 remained non-pregnant (47%) as shown by ultrasonography and the PAG analysis. Significant differences (P < 0.001) in PAG concentrations were observed between the pregnant and non-pregnant buffaloes from day 23 as the PAG of the non-pregnant cows was always close to zero. Conversely, the PAG of the pregnant cows increased progressively from day AI until day 105 post-insemination and then stabilized until the end of pregnancy. Regarding pregnancy diagnosis, the sensitivity of PAG-RIA 860 system (ability of the test to correctly identify pregnant buffalo) ranged from 23% on day 23-98% on day 28 post AI; the specificity (ability to correctly identify non-pregnant buffaloes) was 100% throughout the sampling period. PAG progressively decreased from parturition to day 25 post-partum; from day 30 post-partum, the concentrations fell below 1 ng/mL and were close to 0 on the last day of observation (50 d post-partum). In conclusion, our results showed that RIA-860 is highly accurate for diagnosing pregnancy in buffaloes starting from day 28 of gestation. Furthermore, the rapid disappearance of PAG concentration after calving means that a cut-off limit in post-partum for detecting a new pregnancy is not required.
本研究首次在文献中描述了水牛妊娠期间及产后阶段的妊娠相关糖蛋白(PAG)谱,研究使用了针对从水牛胎盘纯化的PAG分子制备的抗血清(AS#860)。本研究纳入了98头属于同一水牛群的母牛,该水牛群参与了同期发情和人工授精(AI)计划。在第0天(人工授精日)、第23天、第25天、第28天、第30天采集血样,之后每两周采集一次直至妊娠结束。在第28天和第45天通过超声检查确认妊娠,从第60天起通过直肠触诊确认妊娠。在第45天采集未怀孕母牛的血样,而对20头已产犊的水牛,从产犊日起至产后第50天,每五天检测一次血液。使用1.0 ng/mL的临界值来区分怀孕和未怀孕的水牛。我们在对PAG浓度进行Log(x+1)变换后,使用线性混合模型进行分析。超声检查和PAG分析显示,98头参与同期发情的母牛中有52头怀孕(53%),46头未怀孕(47%)。从第23天起,怀孕和未怀孕的水牛之间PAG浓度存在显著差异(P < 0.001),因为未怀孕母牛的PAG始终接近零。相反,怀孕母牛的PAG从人工授精日开始逐渐升高,直至授精后第105天,然后稳定直至妊娠结束。关于妊娠诊断,PAG-RIA 860系统的敏感性(正确识别怀孕水牛的能力)在人工授精后第23天为23%,在第28天为98%;特异性(正确识别未怀孕水牛的能力)在整个采样期间均为100%。PAG从分娩至产后第25天逐渐下降;从产后第30天起,浓度降至1 ng/mL以下,在观察的最后一天(产后50天)接近0。总之,我们的结果表明,从妊娠第28天起,RIA-860在诊断水牛妊娠方面具有高度准确性。此外,产后PAG浓度迅速消失意味着无需设定产后检测新妊娠的临界值。