Enzyme immunoassay of progesterone in the feces from beef cattle to monitor the ovarian cycle
Enzyme immunoassay of progesterone in the feces from beef cattle to monitor the ovarian cycle
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DOI：
10.1016/j.anireprosci.2004.08.006
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发表时间：
2005-06-01
影响因子：
2.2
通讯作者：
Shimada, M
中科院分区：
文献类型：
--
作者：
Isobe, N;Nakao, T;Shimada, M
本研究利用抗孕酮抗体测定肉牛粪便孕酮浓度，探讨该方法能否用于肉牛卵巢周期的监测。将从14头肉牛中采集的直肠粪便样品与6 ml 100%甲醇混合并振荡15 min。离心后，用石油醚提取上清液，然后进行孕酮的酶免疫测定（EIA）。通过HPLC分离粪便溶液，然后在每个馏分中进行EIA，检查测定的专属性。本试验仅鉴别了用石油醚提取的粪便样品中的孕酮，而未鉴别其他代谢物。该测定法的灵敏度估计为0.0055 ng/ml（0.11 ng/g）。批内和批间变异系数分别为9.6-10.9%和10.8- 16.6%。回收率在73%至84%之间。在卵巢周期的粪便孕酮浓度的模式几乎平行于血浆浓度。粪便孕酮浓度与血浆孕酮浓度呈显著正相关（r = 0.59-0.84，P < 0.001，n = 10），合并数据也呈显著正相关（r = 0.70，P < 0.001，n = 65）。卵巢周期第0天粪孕酮浓度低于50 ng/g，第9天粪孕酮浓度显著升高（P < 0.01）。第14 ~ 18天粪孕酮浓度显著降低（P < 0.01）。卵巢周期的最小和最大粪便孕酮浓度之间的差异至少为48 ng/g（平均值= 74 ng/g）。与第0天相比，所有牛在第9、11和14天的粪便孕酮浓度均高于20 ng/g。结果表明，本方法可用于牛粪便中孕酮的测定，并可用于卵巢周期的监测。(c)2004 Elsevier B. V.保留所有权利。
The present study was undertaken to measure fecal progesterone concentration of beef cattle using antibody against authentic progesterone and to examine whether this method can monitor the ovarian cycle in beef cattle. Rectal fecal samples collected from 14 beef cattle were mixed with 6 ml of 100% methanol and shaken for 15 min. After centrifugation, supernatant was extracted with petroleum ether followed by an enzyme immunoassay (EIA) for progesterone. Specificity of the assay was examined by HPLC separation of fecal solution followed by the EIA in each fraction. The present assay identified only progesterone but not other metabolites in the feces sample that was extracted with petroleum ether. Sensitivity of the assay was estimated to be 0.0055 ng/ml (0.11 ng/g). Coefficient variations of intra- and inter-assay were 9.6-10.9% and 10.8-16.6%, respectively. Recovery rates ranged between 73 and 84%. Patterns in the fecal progesterone concentrations during the ovarian cycle were almost parallel to the plasma concentrations. A significant positive correlation was established between the fecal and plasma progesterone concentrations in individual animal (r = 0.59-0.84, P < 0.001, n = 10) as well as pooled data (r = 0.70, P < 0.001, n = 65). Fecal progesterone concentrations of day 0 (showing the nadir of concentration) of the ovarian cycle were less than 50 ng/g, which increased significantly toward day 9 (P < 0.01). From days 14 to 18, there was significant reduction of fecal progesterone concentration (P < 0.01). Ovarian cycles had at least 48 ng/g (mean = 74 ng/g) of difference between minimum and maximum fecal progesterone concentrations. All cattle at days 9, 11 and 14 had higher fecal progesterone concentrations by more than 20 ng/g compared with day 0. These results suggest that the present EIA is suitable to measure the progesterone in cattle feces and can monitor ovarian cycle. (c) 2004 Elsevier B.V. All rights reserved.