Department of Veterinary Experimental Science, University of Padova, Legnaro (PD), Italy.
Theriogenology. 2009 Dec;72(9):1282-92. doi: 10.1016/j.theriogenology.2009.07.025. Epub 2009 Sep 23.
The objective was to develop and test radioimmunoassays (RIAs) to measure fecal progestogens (P) and estrogens (E) to monitor ovarian activity in the bottlenose dolphin (Tursiops truncatus). Fecal samples were collected at least once a week for 20 mo from three peripubertal female bottlenose dolphins. Blood samples were collected at least once a month to compare serum and fecal steroid concentrations. Moreover, random fecal samples from three pregnant females, one lactating female, and one sexually mature female receiving oral altrenogest treatment were also collected. Fecal samples were collected behaviorally with a probe to avoid water contamination and extracted with petroleum ether (for P analysis) or diethyl ether (for E analysis). When possible, vaginal cytology and ovarian ultrasonography were used to monitor the estrous cycle. The RIA for fecal P had good reproducibility and negligible matrix effect. In addition, when fecal samples (N=25) were extracted with ethanol, the results with the two methods of extraction were highly correlated (r=0.923). Therefore, extraction of fecal samples with petroleum ether represented a valid alternative to other, more time-consuming methods of determining fecal P concentrations. In the absence of luteal activity, fecal P concentrations were consistently < 10 pmol/g feces, although they never decreased below 10 pmol/g during pregnancy. Thus, the threshold to confirm the presence of an active corpus luteum was provisionally set at 10 pmol/g. Around the onset of puberty, luteal phases appeared shorter and irregular in the bottlenose dolphin, as in other mammalian species. Additional HPLC-MS studies should be performed to identify predominant P metabolites to be used as fecal indicators of luteal activity in this species.
本研究旨在开发和测试放射免疫分析（RIA），以测量粪便孕激素（P）和雌激素（E），从而监测宽吻海豚（Tursiops truncatus）的卵巢活动。在 20 个月期间，每周至少从 3 只处于性成熟前的雌性宽吻海豚中采集一次粪便样本。每月至少采集一次血液样本，以比较血清和粪便类固醇浓度。此外，还采集了 3 只怀孕雌性、1 只哺乳期雌性和 1 只接受口服阿尔特龙治疗的性成熟雌性的随机粪便样本。通过探针采集粪便样本以避免水污染，并使用石油醚（用于 P 分析）或二乙醚（用于 E 分析）进行提取。当可能时，通过阴道细胞学和卵巢超声检查来监测发情周期。用于粪便 P 的 RIA 具有良好的重现性和可忽略的基质效应。此外，当使用乙醇提取粪便样本（N=25）时，两种提取方法的结果高度相关（r=0.923）。因此，使用石油醚提取粪便样本是一种替代其他更耗时的粪便 P 浓度测定方法的有效方法。在黄体活动不存在的情况下，粪便 P 浓度始终<10 pmol/g 粪便，尽管在怀孕期间从未降至 10 pmol/g 以下。因此，确定黄体活动存在的阈值暂定为 10 pmol/g。在青春期开始时，宽吻海豚的黄体期似乎比其他哺乳动物更短且不规则。应该进行更多的 HPLC-MS 研究，以鉴定主要的 P 代谢物，作为该物种黄体活动的粪便指示剂。
