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研究生:吳祥恩
研究生(外文):Hsiang-En Wu
論文名稱:性類固醇激素對海鱺 (Rachycentron canadum)性別分化調控作用及在育種與養殖上的應用
論文名稱(外文):Effect and application of sex steroids on gonadal differentiation, selective breeding and culture of cobia, Rachycentron canadum
指導教授:李彥宏 博士曾美珍 博士
指導教授(外文):Yan-Horn LeeMei-Chen Tseng
學位類別:碩士
校院名稱:國立屏東科技大學
系所名稱:水產養殖系所
學門:農業科學學門
學類:漁業學類
論文種類:學術論文
論文出版年:2013
畢業學年度:101
語文別:中文
論文頁數:65
中文關鍵詞:海鱺近親交配遠親交配性類固醇激素性別分化
外文關鍵詞:Cobiainbreedingoutbreedingsex steroidsex differentiation
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海鱺是台灣主要箱網養殖魚種之一。但目前面臨近親化的問題,導致魚苗生物特性下降、畸形率過高以及成長緩慢等問題,使產業嚴重受限,故有必要透過育種繁殖方式來改善魚苗品質。本研究是以遠親海鱺配種繁殖,研究藉由增加遺傳歧異度,是否能提升海鱺魚苗品質,並進行後續各項生物特性試驗,另外,本研究也利用不同劑量 (2 mg kg–1, 10 mg kg–1)的雌二醇 (E2)與甲基睪固酮 (MT)添加在飼料中,來進行海鱺的性別控制試驗,並以組織切片來確認海鱺生殖腺分化時間。試驗結果顯示,遠親組子代在受精率、孵化率、卵徑及成長速率表現上皆明顯優於近親組子代;在魚苗飢餓耐受性方面,遠親組魚苗比近親魚苗多活存2天;遠親組剛孵化魚苗之畸形率為1.3±0.2 %,孵化後20天為0.061±0.002 %,均顯著低於近親仔魚。經組織切片觀察,海鱺幼魚性別分化時期介於孵化後84至98天之間。餵食2 mg kg–1雌二醇之雌性化比例為79.3±6.1 %;10 mg kg–1為87.7±10 %,而餵食甲基睪固酮2 mg kg–1與10 mg kg–1之海鱺雄性化比例皆為75 %,顯示性類固醇激素確實能控制海鱺性別。總之,在海鱺繁殖過程中,適當的加入遠親種魚,可以提升子代生物特性的表現,且能藉由餵食類固醇激素來控制海鱺性別,將可應用在養殖產業上。
Cobia (Rachycentron canadum) is one of potential candidates for sea-cage culture species in Taiwan. Recently, due to the inbreeding depression of cobia, the quality of seeds is decreasing, including biological characteristics, deformity, growth rate, etc. We want to solve these problems by using selected breeding method to improve the quality of cobia seeds. In this study, the cobia outbred breeders are ready to propagate for increasing the genetic diversity and quality of seeds. In addition, we use the feeds which contain Estradiol (E2) and Methyltestosterone (MT) in different dose (2 mg kg-1, 10 mg kg-1) to control the sex of cobia. Results showed that the performances of outbred were better than inbred, including egg fertilization rate, hatching rate, egg diameter, yolk-sac larval deformity, 25 days post-hatched larval deformity, larval starvation and larval growth rate. From histological analysis, the gonadal sex differentiation time of cobia larvae is between 84 to 98 days post hatching (dph). Oral administration of sex steroids can promote cobia gonadal differentiation. The feminization rates of 2 mg and 10 mg estradiol are 79.3±6.1 % and 87.7±10 %, respectively. The masculinization of 2 mg and 10 mg methyltestosterone both are 75 %. In conclusion, the study show that outbred cobia progeny is better than inbred’s and sex steroids can control cobia gender effectively. These results will be applied to cobia aquaculture.
摘要 I
目錄 III
圖表目錄 VII
第1章 前言 1
第2章 文獻回顧 3
2.1 海鱺之生態習性 3
2.2. 種魚養殖與魚苗培育 3
2.3 海鱺養殖沿革及現況 4
2.4 近親交配對魚類之影響 5
2.5 魚類的性別決定機制 7
2.6 卵巢發育及成熟過程 8
2.6.1 第一次成長期 8
2.6.2 第二次成長期 8
2.6.3 成熟期 8
2.7 精巢發育及成熟過程 9
2.7.1 第一期精巢 9
2.7.2 第二期精巢 9
2.7.3 第三期精巢 9
2.7.4 成熟期 9
2.7 性類固醇激素對魚類的作用 10
2.7.1 雌二醇(17β–Estradiol, E2) 10
2.7.2 甲基睪固酮 (Methyltestosterone, MT) 10
2.7.3 11–酮基睪酮(11–Keto Testosterone, 11–KT) 11
第3章 材料與方法 1
3.1 遠親與近親海鱺生殖與子代之特性比較 1
3.1.1海鱺種魚來源 1
3.1.2 種魚分組方式 1
3.1.3 種魚管理 1
3.1.4 產卵及受精卵的收集 2
3.1.5 孵化率的計算 2
3.1.6 畸形率的計算 2
3.1.7 飢餓耐受性的測定 2
3.1.8 魚苗培育 3
3.1.9 育苗期間水質監控 3
3.1.10 魚苗成長速率比較 3
3.1.11 海上箱網養殖試驗 3
3.2 以性類固醇激素控制海鱺性別分化 5
3.2.1 實驗分組 5
3.2.2 類固醇溶液製備 5
3.2.3 飼料製作 6
3.2.4 海鱺之雌雄辨別 7
3.2.5 海鱺生殖腺固定 7
3.2.6 海鱺生殖腺的組織切片 7
3.2.7 海鱺血液中性類固醇激素之測定 8
3.3 統計分析 8
第4章 結果 9
4.1 遠親與近親海鱺生殖與子代之特性比較 9
4.1.1 海鱺繁殖及受精卵特性 9
4.1.2 海鱺魚苗之畸形率比較 9
4.1.3 魚苗飢餓耐受性 12
4.1.4育苗期間水質監控 13
4.1.5 遠親與近親海鱺魚苗之成長速率比較 13
4.1.6 海鱺養殖於海上箱網之體長變化 14
4.2以性類固醇激素控制海鱺性別分化 15
4.2.1 海鱺之性別分化時期 15
4.2.2 性別比例 16
4.2.3 性別控制期間之體重變化 19
4.2.4 海鱺生殖腺發育 21
4.2.5投餵實驗飼料後血清性類固醇含量變化 27
第5章 討論 32
5.1遠親與近親海鱺生殖與子代之特性比較 32
5.1.1海鱺繁殖及受精卵特性 32
5.1.3 魚苗之飢餓耐受性 33
5.1.4 遠親與近親海鱺魚苗之成長速率比較 34
5.1.5海鱺養殖於海上箱網之體長變化 34
5.2 以性類固醇激素控制海鱺性別分化 34
5.2.1 海鱺之性別分化時期 34
5.2.2 性別比例 36
5.2.3 性別控制期間之體重變化 36
5.2.4 投餵實驗飼料後血清性類固醇含量變化 37
第6章 文獻回顧 41
附錄 50
作者簡介 51


圖表目錄
圖 1 近親組與遠親組海鱺魚苗畸形率;(A)剛孵化時畸形率;(B)孵化後25天畸形率;Outbred, 遠親組;Inbred, 近親組;組間差異以*代表 (P<0.05) 11
圖 2 海鱺近親組及遠親組之受精卵孵化後,魚苗飢餓耐受性存活率之比較。(Outbred, 遠親組;Inbred, 近親組;組間差異以*代表(P<0.05) 12
圖 3 海鱺遠親組及近親組繁殖之魚苗,在孵化後20天成長體長之比較。Outbred, 遠親組;Inbred, 近親組;組間差異以*代表(P<0.05) 13
圖 4 海鱺遠親及近親繁殖之幼魚,於海上箱網養殖時體重變化之比較。Outbred, 遠親組;Inbred, 近親組 14
圖 5 對照組與投餵雌二醇海鱺的性別分化時間比較。(Control, 對照組; LE2, E2, 2 mg/feed;HE2, E2, 10 mg/feed;gonadal sex differentiation, GSD) 16
圖 6 對照組與投餵甲基睪固酮海鱺的性別分化時間比較。(Control, 對照組;LMT, MT, 2 mg/feed;HMT, MT, 10 mg/feed;gonadal sex differentiation, GSD) 16
圖 7 海鱺幼魚在投餵雌二醇飼料與一般商業飼料時的體重變化。Control, 對照組;LE2, E2, 2 mg/feed;HE2, E2, 10 mg/feed;數值以平均數±SEM表示 (n=8),組間差異以不同小寫英文字母代表(P<0.05)。 20
圖 8 海鱺幼魚在投餵甲基睪固酮飼料與一般商業飼料時的體重變化。Control, 對照組;LMT, MT, 2 mg/feed;HMT, MT, 10 mg/feed 20
圖 9 對照組海鱺孵化後(A)84天、(B)第98天、(C)第112天之生殖腺切片圖。 23
圖 10 投餵含有性類固醇飼料的第70天(DPH=154)時卵巢切片圖。 24
圖 11. 投餵含有性類固醇飼料的第70天(DPH=154)時精巢切片圖。 25
圖 12 投餵高劑量甲基睪固酮組之海鱺精巢成熟各階段的組織切片圖。 26
圖 13 餵食第42天時(A)對照組;(B)HE2組;(C)LE2組海鱺之卵巢切片。 27
圖 14 海鱺幼魚經投餵性類固醇激素飼料後,血清雌二醇濃度之變化。Control, 對照組;LE2, E2, 2 mg/feed;HE2, E2, 10 mg/feed;LMTMT,2 mg/feed;HMT, MT, 10 mg/feed;數值以平均數±SEM表示 (n=8),組間差異以不同小寫英文字母代表(P<0.05)。 29
圖 15 海鱺幼魚經投餵性類固醇激素飼料後,血清睪固酮濃度之變化。Control, 對照組;LE2, E2, 2 mg/feed;HE2, E2, 10 mg/feed; LMT, MT, 2 mg/feed;HMT, MT, 10 mg/feed;數值以平均數±SEM表示(n=8),組間差異以不同小寫英文字母代表(P<0.05)。 30
圖 16 海鱺幼魚經投餵性類固醇激素飼料後,血清11–酮基睪酮濃度之變化。Control, 對照組;LE2, E2, 2 mg/feed;HE2, E2, 10 mg/feed;LMT, MT, 2 mg/feed;HMT, MT, 10 mg/feed;數值以平均數±SEM表示(n=8),組間差異以不同小寫英文字母代表 (P<0.05)。 32
表 1 本實驗之海上箱網規格及養殖環境介紹 4
表 2. 海鱺之分組及各組別之類固醇濃度及樣本尾數 (E2, Estradiol;MT, Methyltestosteron)。 5
表 3 本實驗所使用飼料之配方 (每公斤飼料所需之原料量)。 6
表 4 投餵含有性類固醇激素飼料期間之 飼料粒徑 6
表 5 自100年7月至101年11月,遠親組海鱺種魚產卵日期、總卵量、受精率、孵化率及卵徑(網底部分為本次育苗實驗所採用的受精卵,孵化率標有” – ”代表此批魚卵未計算孵化率)。 10
表 6 自100年7月至101年11月,近親組海鱺種魚產卵日期、總卵量、受精率、孵化率及卵徑(網底部分為本次育苗實驗所採用的受精卵,因近親組之受精卵來自業界養殖戶,因此無法計算總產卵量)。 11
表 7 海鱺經投餵性類固醇激素飼料後,在不同孵化天數(餵食天數)之性別分化及雌雄比例 (control, 對照組;le2, e2, 2 mg/feed;he2, e2, 10 mg/feed;lmt, mt, 2 mg/feed;hmt, mt, 10 mg/feed;網底部份表示不同孵化後天數時,各組之性別比例;dph, days post hatching;gsd, gonadal sex differentiation)。 18
附圖 1 蓄養遠親種魚之80噸水泥池。 51
附圖 2 剛孵化之畸形海鱺。 51

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