雌性台灣梅花鹿排遺滯放於環境溫度中對糞孕酮濃度之影響與對策

Effect of ambient temperature exposure on the fecal progesterone concentration in female Formosan sika deer and its countermeasures

鄒易晨(Yi-Chen Tzou)

指導教授 ： 裴家騏

國立屏東科技大學/農學院/野生動物保育研究所/碩士(2016年)

摘要

糞孕酮（fecal progesterone）的檢測是研究鹿科動物繁殖生理的利器，然而排遺中的性固醇類荷爾蒙濃度，往往會隨著排出後的滯放時間而改變，而在野外研究時，常會收集到新鮮度不同的排遺，潛在的誤差不應忽視。本研究以台灣梅花鹿（Cervus nippon taiouanus）為對象，以酵素連結免疫吸附分析檢視雌性排遺中的糞孕酮隨時間變化的情形，並找出適當的樣本處理方式以避免產生檢測誤差。結果顯示，母梅花鹿糞孕酮濃度於最初的248±113 ng/g顯著提升至滯放半天的588±245 ng/g（t-test; p=0.006），接著濃度將再度顯著提升至滯放一天的1011±230 ng/g（t-test; p=0.006），之後在滯放第七天內維持在高值不再有顯著變化。滯放前後的濃度呈現高度正相關（rs =0.8, n=12），顯示樣本經滯放後的糞孕酮濃度，依舊能高度地反映出剛排出時的濃度高低。本研究也顯示，孕二醇葡萄醣醛酸的酵素免疫分析比起糞孕酮雖能減少排遺排出後一天內滯放時間所造成的濃度檢測誤差，然而各樣本間仍會隨滯放時間有相當的變異。總結以上，建議以酵素免疫分析測試梅花鹿的糞孕酮時，除了可以在排遺被排出後即迅速保存，也可將收集的排遺滯放於室溫中至少2天再行檢測，以減少排遺新鮮度不同所造成的誤差。

關鍵字

台灣梅花鹿 ； 繁殖生理學 ； 糞孕酮 ； 酵素連結免疫吸附分析

並列摘要

Measurement of fecal progesterone (fP4) is widely used in studying reproductive biology of deer. However, the concentrations of fecal sexual steroid hormones often vary with time after defecation, which should not be ignored because collection of fresh fecal samples would be difficult in the wild. This study examined how fecal steroid hormone concentrations in female Formosan sika deer （Cervus nippon taiouanus）vary with time by using enzyme-linked immunosorbent assay (ELISA). The results showed that the mean fP4 concentration increased significantly (t-test; p<0.05) from 248±113 ng/g in D0 (day 0, exposed at ambient temperature for 0 day) to 588±245 ng/g in D0.5 and 1011±230 ng/g in D1, and it remained at the same level for 6 days at least. The mean fP4 concentration at D0 was highly correlated (rs =0.8) to those after exposure, suggested that the after exposure measurement can represent the original value. The results also showed that using the ELISA which immunoreacts with both conjugated- and freed-pregnanediol can reduce the variation of progestogen measurement between D0 and D1, but the sample variation was still notable. This study suggested that, for fP4 measurement, it is better to collect and analyze fecal samples immediately after defecation, otherwise to expose unknown-aged fecal samples at ambient temperature for at least 2 days before analysis will also help to reduce the bias.

並列關鍵字

Formosan sika deer ； reproductive biology ； fecal progesterone ； enzyme-linked immunosorbent assay

參考文獻

蘇南伃 （2011） 雌性台灣水鹿之動情週期及懷孕期之糞孕酮濃度變化暨22例難產病例之回溯性調查。國立中興大學獸醫學系。86頁。

McCullough, E. D. R. (2009) Sika deer in Taiwan. In: McCullough, D.R., S. Takatsuki, and K. Kaji (eds.) Sika deer. Springer, Japan.

Bamberg, E., R. Palme, and J. G. Meingassner (2001). Excretion of corticosteroid metabolites in urine and faeces of rats. Lab. Anim., 35(4), 307-314.

Beehner, J. C., and P. L. Whitten (2004). Modifications of a field method for fecal steroid analysis in baboons. Physiol. Behav., 82(2), 269-277.

Cross, N., M. K. Pines, and L. J. Rogers (2004) Saliva sampling to assess cortisol levels in unrestrained common marmosets and the effect of behavioral stress. Am. J. Primatol. 62(2): 107-114.