微塑膠汙染在全球廣泛分佈，微塑膠普遍地被魚類所攝入，但目前關於探討魚類攝入微塑膠的可能因子相關文獻仍相當稀少。本研究以珊瑚礁魚類眼斑海葵魚 (Amphiprion ocellaris)作為實驗物種，及500-850 μm的聚對苯二甲酸乙二酯(PET)微塑膠作為實驗材料，以三個實驗分別探討(一)眼斑海葵魚在食物的有無下對微塑膠之行為反應及攝入情形、(二)與食物共同存在下不同微塑膠濃度對海葵魚行為與攝入風險、(三)氣味(生物膜的有無)及光照條件(明亮與昏暗)下海葵魚對微塑膠行為反應及攝入風險。所有實驗皆進行微塑膠攝食暴露1小時，接著在48小時的淨化時間後收集魚隻排出的微塑膠，以計數微塑膠的攝入量。實驗一組別包括食物組、微塑膠組、混合組，實驗二組別包括低、中、高濃度組(食物與微塑膠配比為1:0.1、1:1、1:10)。實驗三以氣味及光照條件兩個因子進行雙因子實驗(共四個實驗組)。實驗一結果顯示，魚隻主要攝食活動發生在暴露前期(前15分鐘)，後隨著食慾的減少而行為下降。在過了主要攝食時間後(15-30分鐘)，微塑膠組的攝食行為與活動能力下降，混合組則較慢延緩攝食行為並保持一定游動能力，微塑膠組及混合組在排遺部分皆有發現微塑膠，但混合組顯著較高。實驗二結果顯示，食物與微塑膠比例相當的中濃度組在攝食區的探索行為仍持續發生，而在低濃度與高濃度組攝食探索行為隨暴露時間增加而下降，而眼斑海葵魚對高濃度組表現明顯的迴避反應，排遺顯示隨著暴露濃度增加微塑膠攝入量也會增加，但中濃度組與高濃度組間無顯著差異。實驗三結果顯示，明亮條件組隨暴露時間而活動行為下降，昏暗條件組仍持續活動並探索接觸攝食區，但在排遺顯示明亮條件組別微塑膠攝入量顯著高於昏暗條件組，微塑膠表面有無生物膜則不影響魚的行為與微塑膠攝入。總結以上結果，眼斑海葵魚在只有微塑膠存在時仍會攝入微塑膠，若有食物同時存在更會增加其攝入微塑膠的機率，且隨微塑膠濃度升高增加其攝入量，攝入量達一定程度具有極限而不持續增加。周遭光照條件顯著影響魚隻的行為表現及其微塑膠攝入，但微塑膠因老化產生的氣味則無顯著影響。

Microplastic pollution is widely distributed globally, and microplastics are commonly ingested by marine fish. However, there is still a scarcity of literature on the potential factors related to the ingestion of microplastics by fish. In this study, the anemonefish Amphiprion ocellaris, was used as the experimental species, and polyethylene terephthalate (PET) microplastics ranging from 500-850 μm in size were used as the experimental material. This study examines (1) the behavioral responses and ingestion patterns of A.ocellaris towards microplastics in the presence or absence of food, (2) the effects of different concentrations of microplastics on the behavior and ingestion risk of anemonefish when food is present, and (3) In the presence of feed, the effects of odor (presence or absence of biofilm) and light conditions (bright and dark) on the behavioral responses and ingestion risks of anemonefish. All experiments involved a 1-hour exposure to microplastic ingestion, followed by a 48-hour purification period to collect the excreted microplastics from the fish for quantifying the ingestion levels. Experiment 1 consisted of three groups: food group, microplastic group, and mixed group. Experiment 2 included low, medium, and high-concentration groups (food to microplastics ratios were 1:0.1, 1:1, and 1:10). In Experiment 3, the odor of the microplastics and ambient light condition were chosen as the two factors for a two-by-two experiment, resulting 4 experimental groups in total. The results of Experiment 1 showed that the main feeding activity of the fish occurred during the early exposure period (first 15 minutes), followed by a decline in behavior as appetite decreased. After the primary feeding time (15-30 minutes), the feeding behavior and locomotor activity decreased in the microplastic group, while the mixed group showed a slower decline in feeding behavior and maintained a certain level of swimming ability. Microplastics were found in the fecal samples of both the microplastic group and the mixed group, with a significantly higher presence in the mixed group.The results of Experiment 2 showed that the moderate concentration group, with a comparable ratio of food to microplastics, continued to exhibit exploratory behavior in the feeding zone. However, in the low and high concentration groups, the feeding exploratory behavior decreased with increasing exposure time. The A.ocellaris exhibited a noticeable avoidance response to the high concentration group. Fecal analysis indicated that as the exposure concentration increased, the intake of microplastics also increased, but there was no significant difference between the moderate and high concentration groups. The results of Experiment 3 showed that the activity behavior of the Bright condition group decreased over time, while the Dim condition group continued to be active and explored the feeding area. However, the excretion results indicated that the Bright condition group had significantly higher microplastic ingestion compared to the Dim condition group, and the odor produced by the biofilm on the surface of microplastics did not affect the fish's behavior and microplastic ingestion. To conclude, anemonefish would still ingest microplastics when they are present alone, and the presence of food increases their likelihood of ingesting microplastics. The ingestion quantity increases with increasing microplastic concentration, but there is a certain limit to the increase in ingestion, and the fish's behavior and microplastic ingestion are mainly influenced by light conditions rather than the odor of the microplastics produced by biofilm.

謝辭 I
摘要 III
Abstract V
目錄 VII
圖目錄 IX
表目錄 XI
附圖目錄 XIII
附表目錄 XV
第一章 前言 1
1.1. 海洋中的塑膠汙染 1
1.2. 微塑膠對魚類的影響 2
1.3. 影響魚類攝入微塑膠的可能因素 3
1.4. 行為生態毒理學 5
1.5. 實驗物種：眼斑海葵魚 6
1.6. 研究目的 7
第二章 材料與方法 9
2.1. 實驗設計 9
2.2. 實驗動物照護 10
2.3. 暴露實驗環境設置 10
2.4. 實驗飼料與微塑膠製備 11
2.5. 老化微塑膠生物膜(Biofilm)質量量化 12
2.6. 微塑膠與飼料表徵觀察與分析 13
2.7. 實驗一：海葵魚對微塑膠行為表現與攝食反應 13
2.8. 實驗二：微塑膠濃度對海葵魚攝食行為與游泳行為之影響 15
2.9. 實驗三：氣味與光照雙因子實驗對海葵魚之行為影響與攝入風險 16
2.10. 眼斑海葵魚的排遺收集與分析量化 18
2.11. 行為影片分析 19
2.12. 汙染控制 20
2.13. 統計分析 21
第三章 結果 23
3.1. 實驗一：海葵魚對於微塑膠行為反應及微塑膠攝入影響 23
3.1.1 不同攝食選擇因素下對海葵魚行為活動反應 23
3.1.2 不同攝食選擇因素下對海葵魚微塑膠攝入情形 26
3.2. 實驗二：不同微塑膠濃度對海葵魚的行為反應及微塑膠攝入影響 28
3.2.1 三種微塑膠攝食濃度對海葵魚行為活動反應 28
3.2.2 不同微塑膠濃度下海葵魚微塑膠攝入情形 31
3.3. 實驗三：氣味與光照雙因子對海葵魚的行為反應及微塑膠攝入影響 33
3.3.1 老化微塑膠表徵與生長質量 33
3.3.2 氣味與光照雙因子對海葵魚行為活動反應 35
3.3.3 氣味與光照雙因子對海葵魚行為參數之GLM分析 39
3.3.4 氣味與光照雙因子對海葵魚微塑膠攝入情形 42
第四章 討論 45
4.1. 海葵魚對不同微塑膠攝食品質選擇之行為反應與攝入情形 45
4.2. 不同微塑膠濃度對眼斑海葵魚之行為與攝入風險 46
4.3. 微塑膠氣味與環境光照條件對眼斑海葵魚之行為反應與攝入風險 48
第五章 結論與建議 51
參考文獻 53
附錄 71

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