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李静宇,张启信,黄明竟,夏东坡,李进华.2021.短尾猴粪便雌二醇和孕酮处理与 提取方法比较.动物学杂志,56(2):247-254.
短尾猴粪便雌二醇和孕酮处理与 提取方法比较
Fecal Treatment and Extraction of Estradiol and Progesterone in Tibetan Macaques (Macaca thibetana): Comparison among Different Approaches
投稿时间:2020-06-05  修订日期:2021-02-27
DOI:10.13859/j.cjz.202102010
中文关键词:  短尾猴  粪便  雌二醇  孕酮  乙醇加热法
英文关键词:Tibetan Macaques (Macaca thibetana)  Feces  Estradiol  Progesterone  Ethanol-heating method
基金项目:国家自然科学基金项目(No. 31672307,31971404),安徽省中央引导地方科技发展专项(No. 2019b11030018)
作者单位E-mail
李静宇 安徽大学资源与环境工程学院 合肥 230601黄山生物多样性与短尾猴行为生态学国际联合研究中心 合肥 230601 jyl7991@163.com 
张启信 安徽大学资源与环境工程学院 合肥 230601黄山生物多样性与短尾猴行为生态学国际联合研究中心 合肥 230601 674267835@qq.com 
黄明竟 安徽大学资源与环境工程学院 合肥 230601黄山生物多样性与短尾猴行为生态学国际联合研究中心 合肥 230601 371648725@qq.com 
夏东坡 黄山生物多样性与短尾猴行为生态学国际联合研究中心 合肥 230601安徽大学生命科学学院 合肥 230601 dpxia@163.com 
李进华 安徽大学资源与环境工程学院 合肥 230601黄山生物多样性与短尾猴行为生态学国际联合研究中心 合肥 230601合肥师范学院生命科学学院 合肥 230601 jhli@ahu.edu.cn 
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中文摘要:
      非损伤性的粪便样本已在监测动物性激素水平中得到广泛应用,但粪便处理和激素的提取方法在不同物种中因其食性与代谢不同具有明显的物种特异性。本研究以野生和笼养两种环境下的短尾猴(Macaca thibetana)为研究对象,采集粪便样品,利用放射免疫法(RIA)检测激素含量,比较不同粪便处理方式(湿便法、冷冻干燥粉碎法、冷冻干燥研筛法)和激素提取方法(乙醇加热法、甲醇乙醚法、乙醇丙酮法、甲醇乙醇法)的效果,探讨不同取食条件下短尾猴雌二醇(E2)和孕酮(P4)水平的最佳测定方法。粪便不同处理方法的比较中,冷冻干燥研筛法测得雌二醇和孕酮含量最高,冷冻干燥研筛法与冷冻干燥粉碎法在野生短尾猴中具有显著差异(P < 0.01),但在笼养短尾猴中无显著差异(P > 0.05)。湿便法测得含量最低,与另两种处理方法均具有显著差异性(P < 0.01)。激素提取方法的比较中,乙醇加热法提取雌二醇和孕酮含量最高,与甲醇乙醚法(P < 0.01)、乙醇丙酮法(P < 0.01)、甲醇乙醇法(P < 0.01)之间均具有显著性差异。综合考虑激素平均含量、处理过程的简便性、抽提步骤的可行性,野生短尾猴粪便处理建议选用冷冻干燥研筛法,笼养短尾猴粪便由于冷冻干燥粉碎法操作简便且与冷冻干燥研筛法无显著差异,因此笼养短尾猴建议选用冷冻干燥粉碎法。野生与笼养短尾猴粪便激素的提取均建议采用乙醇加热法。
英文摘要:
      Non-invasive fecal samples have been widely used in monitoring animal sex hormone levels, but fecal treatment and hormone extraction need obvious species-specific manipulations in different species due to their different feeding habits and metabolism. In this study, we used wild and captive macaque (Macaca thibetana) to collect feces samples, and the contents of hormones were detected by using radiation immunoassay (RIA) after feces had been treated differently (wet feces,freeze-dried crushed feces,freeze-dried sifted feces) and the hormones had been extracted by various approaches (ethanol-heating method,methanol-ether method,ethanol-acetone method,methanol-ethanol method), in order to find optimal methods for detecting estradiol (E2) and progesterone (P4) levels in macaques under different feeding conditions. The data were analyzed using Two-Simple-Wilcoxon test. The results showed that the contents of estradiol and progesterone were the highest in feces treated by freeze-drying sifting method (Fig.1). There was significant difference in results between freeze-drying sifting method and freeze-drying crushing method in wild macaques (P < 0.01, Fig. 2), but there was no significant difference in captive macaques (P > 0.05,Fig. 2). The content of hormone in wet feces treatment was the lowest, which was significantly different from the other two treatment methods (P < 0.01, Fig. 1);the content of E2 and P4 extracted by ethanol-heating method was the highest(Table 1), which was significantly different from that of methanol-ether method (P < 0.01, Table 3), ethanol-acetone method (P < 0.01, Table 1) or methanol-ethanol method (P < 0.01, Table 1); Considering the average content of hormones, the simplicity of treatment process and the feasibility of extraction steps, it is recommended to use freeze-drying sifting method for the treatment of wild macaque feces. As the freeze-drying crushing method is easy to operate and there is no significant difference between the two methods, the freeze-drying crushing method is recommended for captive macaque feces. Ethanol-heating method is recommended for the extraction of fecal hormones from wild and captive macaque.
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