滇金丝猴粪便类固醇激素抽提及检测方法评估

Evaluation on Extraction and Assay Methods for Fecal Steroid Hormones in Yunnan Snub-Nosed Monkeys

采用非损伤性取样法监测野生动物的生理状态已被广泛接受,但分析结果却因对粪便样品的抽提和检测方法不同而异。本文以圈养滇金丝猴为研究对象,分别采用乙醇加热法、乙醇-蒸馏水法和乙醇-丙酮法对其粪便样品进行激素抽提,然后采用放射免疫分析法(RIA)和酶联免疫吸附测定法(ELISA)检测其雌二醇(E_2)和孕酮(P_4)水平。结果表明,不论采用RIA还是ELISA检测,选用乙醇加热法和乙醇蒸馏水法所抽提样品的孕酮和雌二醇测定结果均不存在显著差异,而采用乙醇丙酮法所抽提样品的孕酮测定结果显著低于前面两种方法所测定的结果。综合考虑平均含量、相对误差、变异系数及抽提步骤的可行性,建议选用乙醇加热法抽提和RIA检测滇金丝猴粪样中孕酮与雌二醇水平。

Non-invasive sampling has been commonly used in monitoring the endocrine status of wild animals in recent years,yet the results vary greatly with different usage of hormone extracting and assay methods.We collected feces of three captive adult female Yunnan snub-nosed monkeys(Rhinopithecus bieti)at Beijing Zoo,and examined and compared the fecal concentrations of estradiol(E 2)and progesterone(P 4)of 21 fecal samples by using three different steroid hormone extracting methods,i.e.,ethanol-heating(EH),ethanol-distilled-water(EW),and ethanol-acetone methods(EA),and two assay methods of radioimmunoassay(RIA)and enzyme-linked immunosorbent assay(ELISA),respectively.We found no significant difference in the fecal concentrations in E 2 and P 4 levels by either EH or EW regardless of using RIA or ELISA,yet P 4 values were lower when estimated by EA than when measured by either RIA or ELISA.In considering the average concentration,relative error,coefficient variation as well as practicability,we recommend to use the ethanol-heating method(EH)to extract and RIA for examining the fecal concentrations of E 2 and P 4 in Yunnan snub-nosed monkeys.