文獻報告已證實人工合成雌激素‐17α乙炔雌二醇(17α-ethynylestradiol, EE2)在環境中廣泛存在並具有持久性以及內分泌干擾效應。EE2 常被應用於口服避孕藥、醫藥、化妝品、個人護理產品中。容易隨著人類活動或廢水排放進入環境水體，常可在旅遊活動興盛的沿岸地區被偵測到，目前已被視為海洋生態系統的新興污染物。許多珊瑚礁魚類獨特的社會結構是由敵對行為表現所調控，敵對行為會受到性荷爾蒙調控，EE2 可能透過擾亂性激素來影響魚類敵對行為，本研究透過環境暴露的方式來探討 EE2 對眼斑海葵魚(Amphiprion ocellaris)的內分泌干擾效應，並著重觀察在敵對行為和社會階級的影響。經過馴養並檢疫後，魚隻暴露在各 EE2 濃度下 28 天，分別為控制組(0 ng/ L )、低濃度組(3 ng/ L)和高濃度組(30 ng/ L)，各組別眼斑海葵魚三隻個體為一組分配在獨立的缸子中以形成小群，每 7 天使用錄影機把其威嚇行為、屈服行為與遮蔽所利用率記錄並量化分析。在暴露實驗結束時，犧牲魚隻並採集海葵魚血漿，測量眼斑海葵魚血漿中的睪丸硬甾酮(11-KT)濃度。本實驗結果顯示，暴露 EE2 會影響眼斑海葵魚的社會表現，不論暴露濃度，皆觀察到行為一致性下降的現象。並在高濃度組 (30 ng/ L EE2)的結果發現，中間地位的個體對於威嚇對象的選擇性降低，位居主導地位的個體在暴露實驗後期展示屈服行為的頻率增加，在暴露實驗後期群體內威嚇行為增加等現象。表明主導階級的海葵魚侵略性弱化，以及中間階級個體對於評估雙方條件的能力下降，因此主導階級的個體可能必須花費更多時間及精力維持其地位與階級穩定。在 EE2 30 ng/L 組別也觀察到與主導地位相關的 11-KT 濃度減少。本實驗顯示EE2在環境濃度暴露下可能會影響群居珊瑚礁魚類的社會行為。

Previous studies have shown that the synthetic estrogen 17α-ethinylestradiol (EE2) can cause endocrine disrupting effects and is environmentally stable and lipophilic. EE2 is extensively used in oral contraceptive pills, medication, cosmetics, and personal care products. EE2 has been frequently detected in coastal areas and is now considered an emerging pollutant in marine ecosystems. Agonistic behavior is crucial to individual fitness and population social structure of coral reef fish. Endocrine disrupting chemicals may disturb fish agonistic behavior and social interactions by interfering sex hormone levels. In this study, we used an experimental approach to characterize the effects of EE2 on social behavior of the false clown anemonefish (Amphiprion ocellaris) via environmental exposure. The fish were exposed to EE2 at doses of 0, 3 and 30 ng/L for 28 days. The fish from each group were randomly distributed to separate tanks to form small colonies consisting of three individuals. During the exposure, agonistic behavior, submissive response and shelter utilization were recorded with video camera weekly and quantitatively analyzed. At the end of the experiment, the 11-ketotestone (11-KT) in the plasma of the fish were measured. Our results show that EE2 exposure affected social behaviors and behavioral consistency of the fish. In the 30 ng/L EE2 group, the second-ranked fish performed more agonistic behavior toward the dominant fish, while the dominant fish responded more tremble behavior. Intra-colonial agonistic interactions also increased in the later period of the exposure. The results suggest that the ability of the second-ranked individuals to assess opponents and the intensity of aggression performed by the dominant individuals were both reduced in the EE2 group. It may cost more time and energy for the dominant fish to maintain their dominancy. Concentrations of plasma 11-KT, which is associated with social hierarchy, were also reduced in the 30 ng/L EE2 group. This study demonstrates that environmental exposure of EE2 can affect social behavior and endocrine levels in coral fish.

第一章 前言.…………………………………………………..…………………….....1
1.1人工雌激素17α-ethynylestradiol…..……………………..1
1.2動物的社會行為與社會階級……………………….………………....4
1.3實驗動物………………………..……………...................4
1.4實驗目標………………………..…….....................5
第二章 材料及方法………………………..….................7
2.1實驗動物………………………..……………………….................7
2.2眼斑海葵魚 EE2 暴露實驗………………………..……………………….....7
2.2.1儲備溶液配置………………………..……………….................7
2.2.2暴露實驗設計………………………..…....................7
2.3行為分析………………………..………………......................8
2.4血漿中荷爾蒙測定……………………….......................9
2.5水中EE2濃度化學分析………………………..……………………….........10
2.6統計分析………………………..………………………......……...……...…..10
第三章 結果………………………..………………………......……...……...……..11
3.1暴露EE2存活率……………..………………………......……...……...…….11
3.1.1暴露3 ng/L EE2存活率……………..………………….……......…..……..11
3.1.2暴露30 ng/L EE2存活率……………..………………………......……...…12
3.2各階級體型……………..………………………......……...……...…………..13
3.2.1暴露3 ng/L EE2各階級體型……………..………………………..........13
3.2.2暴露30 ng/L EE2各階級體型…………..………………………............13
3.3 暴露3 ng/L EE2行為實驗…………..……………………….................15
3.3.1暴露3 ng/L EE2 總威嚇行為分析…………..………………………..........15
3.3.2暴露3 ng/L EE2 α階級威嚇行為分析………..……………………….......15
3.3.3暴露3 ng/L EE2 β階級威嚇行為分析………..……………………….......16
3.3.4暴露3 ng/L EE2 γ階級威嚇行為分析………..……………………….......16
3.3.7暴露3 ng/L EE2 總逃離行為分析 ………………………………………..21
3.3.7暴露3 ng/L EE2 α階級逃離行為分析...…………………………………..21
3.3.7暴露3 ng/L EE2 β階級逃離行為分析...…………………………………..22
3.3.8暴露3 ng/L EE2 γ階級逃離行為分析...…………………………………..22
3.3.9暴露3 ng/L EE2 總顫抖行為分析...………………………………………27
3.3.10暴露3 ng/L EE2 α階級顫抖行為分析…………………………………...27
3.3.11暴露3 ng/L EE2 β階級顫抖行為分析……………………………….…...28
3.3.12暴露3 ng/L EE2 γ階級顫抖行為分析…………………………………....28
3.3.13暴露3 ng/L EE2 進入遮蔽所持續時間………………………………......33
3.4 暴露30 ng/L EE2行為實驗.......35
3.4.1暴露30 ng/L EE2 總威嚇行為分析.....35
3.4.2暴露30 ng/L EE2 α階級威嚇行為分析....35
3.4.3暴露30 ng/L EE2 β階級威嚇行為分析.....36
3.4.4暴露30 ng/L EE2 γ階級威嚇行為分析......36
3.4.5暴露30 ng/L EE2 總逃離行為分析.......41
3.4.6暴露30 ng/L EE2 α階級逃離行為分析....41
3.4.7暴露30 ng/L EE2 β階級逃離行為分析....42
3.4.8暴露30 ng/L EE2 γ階級逃離行為分析.....42
3.4.9暴露30 ng/L EE2 總顫抖行為分析........47
3.4.10暴露30 ng/L EE2 α階級顫抖行為分析....47
3.4.11暴露30 ng/L EE2 β階級顫抖行為分析…...48
3.4.12暴露30 ng/L EE2 γ階級顫抖行為分析.....48
3.4.13暴露30 ng/L EE2 進入遮蔽所持續時間....53
3.5群體內社會行為表現趨勢....................55
3.6魚類內分泌生理分析.......................59
3.6.1暴露3 ng/L EE2睪丸硬甾酮(11-KT)濃度...59
3.6.2暴露30 ng/L EE2睪丸硬甾酮(11-KT)濃度..59
3.7水體中EE2濃度分析.......................61
第四章 討論…...............................63
4.1死亡率與體型差異........................63
4.2眼斑海葵魚社會行為......................65
4.2.1階級對行為的影響......................65
4.2.2暴露 EE2對眼斑海葵魚行為的影響.........66
第五章 結論................................71
參考文獻...................................73

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