| Estrous and mating is an important way to enlarge the population and improve its fitness.In primates,the study of female sex hormone secretion level and physiological cycle information can not only reveal the reproductive law,but also provide an important way to further understand female sexual behavior,sexual strategy,male mating investment and species evolution.Macaca thibetana is a second-class protected animal in China,which is listed as a vulnerable species by the Nature Conservation Union.Due to the fact that there is no obvious sexual skin swelling during the breeding period,the body size is relatively large,and the abdominal changes after pregnancy are not obvious,and the complex habitat environment and other reasons,researchers have been relatively backward in the research on female physiology of Macaca thibetana for a long time.There are few studies on the secretion of sex hormones and basic physiological information such as menstrual cycle and pregnancy in female macaque under different physiological conditions.In this study,the adult female of Huangshan wild macaque and captive macaque in Tongling zoo were selected as the research objects.633 fresh feces of target female individuals were continuously collected by non-invasive sampling method from October 10,2018 to October 3,2019.Behavior sampling method was used to record mating behavior,and all occurrence recording method was used to record female estrus status and birth date Interest.Fecal samples from wild and captive macaque were used to evaluate the fecal sex hormone levels in different fecal treatment methods(Wet-feces,Freeze-drying crushing,Freeze-drying screening),different hormone extraction(Ethanol-heating method,Methanol-ether method,Ethanol-acetone method,Methanolethanol method)and detection methods(Radioimmunoassay,Enzyme-linked immunosorbent assay).The best method was applied to the study of physiological cycle of wild female macaque.The sex hormone levels of female macaque under different physiological conditions were analyzed.Combined with the indication effect of different hormone indexes(Estrone,Estradiol,Progesterone,Pregnediol)on ovulation and pregnancy diagnosis,as well as the female mating behavior,estrus state,birth date,etc.,were used to infer the macaque Menstrual cycle and length of pregnancy.The main results are as follows:1.The fluctuation of estrone(E1C)and estradiol(E2)are similar to each other,but the fluctuation period is very small and the ovulation signal cannot be judged,and the physiological cycle of female macaque cannot be clearly indicated.The content of progesterone(P4)was higher than that of progesterone(P4),but the level of progesterone tended to be linear.E2 and P4 are more suitable as diagnostic indicators of ovulation and pregnancy.2.The content of E2 and P4 in the saliva of female macaque collected by using cotton swab to dip Kool Aid beverage powder was low,and the hormone level had no obvious regularity.The results of hormone detection showed that the feces in the field were better than saliva in monitoring the sex hormone level of wild macaque.3.The contents of E2 and P4 in faeces treated by freeze-drying screening method were the highest.There were significant differences between freeze-drying screening method and freeze-drying crushing method in wild macaque(P < 0.01),but there was no significant difference in captive macaque(P > 0.05).The content of hormone extracted directly from wet stool was the lowest,which was significantly different from the other two treatments(P < 0.01).The content of E2 and P4 extracted by ethanol heating method was the highest,which was significantly different from that of methanol ether method(P < 0.01),ethanol acetone method(P < 0.01)and methanol ethanol method(P < 0.01).4.The fluctuation of fecal estradiol(E2)content has obvious ovulation peak value and pregnancy marker,which can be used for ovulation period,early pregnancy diagnosis,inferring fertilization date and birth date of macaque species.However,the effect of progesterone(P4)on the diagnosis of pregnancy is not as good as that of estradiol.RIA and ELISA were used to detect estradiol in feces of pregnant individuals.The results showed that radioimmunoassay(E2: 5000?50000 pg/g)was more sensitive than enzyme-linked immunosorbent assay(E2 < 700 pg/g).5.Female macaque menstruation feces estradiol levels of about 2578.87 ± 168.99 pg/g,basic value range of 0?5000 pg/g,two ovulation peak estradiol levels were 4906.9 pg/g and 4962.9 pg/g,basic value is about 1.90 times and 1.92 times of pregnancy is slowly rising,rising along with the sharp fluctuations in the process of,late pregnancy at a higher level,litter fell significantly after hormone to unfertilized preliminary judgment benchmarks for female macaque menstrual cycle is about 30 d,the length is about 158.6 ± 2.9 days during pregnancy(n = 3).In this study,we evaluated the method of fecal sex hormone determination in Macaca thibetana.The results showed that the best method was freeze-drying method for female macaque feces,ethanol heating method for extracting sex hormone and radioimmunoassay for detection.This method was first applied to the analysis of sex hormone level in the physiological cycle of macaque,and the secretion level of sex hormone in different physiological state of macaque was discussed The results showed that the menstrual cycle of female macaque was about 30 days,and the length of pregnancy was about 158.6 ± 2.9 days(n = 3),which provided a method support for obtaining more basic data of female menstrual cycle and pregnancy period,and provided theoretical basis for further study on the relationship between behavior mechanism and endocrine,protection and management of wild Macaca thibetana in Huangshan. |
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