密點少棘胡椒鯛(Diagramma pictum)為廣泛分布於印度-西太平洋海域的魚類，有關其完整的產卵和初期生活史資訊尚付之闕如。本研究主要針對密點少棘胡椒鯛之仔稚魚的初期生活史進行探討，記錄產卵週期，並利用光學顯微鏡、解剖顯微鏡及掃描式電子顯微鏡對魚卵和仔稚魚進行形質記錄並探索微細構造，同時製作透明骨骼標本觀察仔稚魚的骨骼發育情形。最後也藉由溫度和鹽度實驗來探討兩者對受精卵孵化率、畸形率及SAI值的影響。本研究的親魚比例不詳，於2019年6月1日至2021年7月31日期間，在水溫18.1 - 33.0 °C、鹽度28.03 - 35.34 psu下自然產卵，實際產卵日有127天，總產卵量為140111粒，平均產卵量為1103 ± 1171顆卵；孵化率為13.3 % - 96.6 % (59.3 ± 22 %)。受精卵為透明球形的浮性卵，卵徑為0.78 ± 0.02 mm，油球徑為0.178 ± 0.013 mm，在水溫28 ± 0.2 °C、鹽度33.4-33.5 psu的條件下，孵化時間為14小時30分鐘。剛孵化仔魚之體全長為1.81 ± 0.19 mm，黃色素胞廣佈於肌節上；孵化後第三天的仔魚體全長2.27 ± 0.30 mm，口部、眼睛及肛門發育完成，開始攝食；孵化後第九天脊索上屈完畢；孵化後第十八天進入變態期；孵化後第二十五天鰭條達成魚定數(背鰭 IX, 23-25；臀鰭 III, 7；尾鰭 17；胸鰭 16；腹鰭 I, 5)，進入稚魚期。在電子顯微鏡的觀察下，可以發現密點少棘胡椒鯛的受精卵外觀呈圓球狀，壁孔大小一致，密度約為38個 /100 μm2。卵門通道呈圓柱型，直徑約為5.24 μm，卵門周圍有副孔呈放射狀散佈，大小不一，數量約為125個。神經丘在仔魚剛孵化時出現，隨著成長數量會增加；牙齒和頭部棘刺在孵化後第四天開始發育。透過透明骨骼標本可以發現密點少棘胡椒鯛仔魚在孵化後第二天開始發育頭部和脊索骨骼；在孵化後第七天開始發育尾鰭骨骼；在孵化後第十天開始發育背鰭和臀鰭骨骼；在孵化後第十一天開始發育腹鰭骨骼；脊索骨骼在孵化後第十三天骨化完成，背鰭、臀鰭及尾鰭骨骼皆在孵化後第十六天骨化完畢；腹鰭骨骼在孵化後第二十天骨化完成。在溫鹽實驗中，最佳的孵化溫度和鹽度分別為27 °C和37 psu，孵化率顯著高於其它的組別(P < 0.05)；畸形率最低的鹽度組別則是29 psu，溫度處理組間的畸形率皆為0 %；生存活力指數(Survival activity index, SAI)最佳的溫度和鹽度組別為27 °C和33 psu。本研究不僅能提供密點少棘胡椒鯛初期生活史的完整資訊，也能做為日後開發人工繁養殖的參考依據。

Painted sweetlips, Diagramma pictum is distributed in Indo- West Pacific, and knowledge of the integral spawning and early development of this species is particularly limited. This study focused on early life stage, spawning cycle, early ontogeny, amd microstructure of D. pictum using optical microscope, dissecting microscope and scanning electron microscope. At the same time, transparent skeletal specimens were made to observe the skeletal development of larvae and juveniles. The effects of temperature and salinity on hatching rate, deformity rate, and SAI were also demonstrated. Natural spawning of D. pictum (female: male ratio unknown) in capacity was ovserved from June 1, 2019 to July 31, 2021. Throughout the entire spawning period, the water temperature fluctuated between 18.1 and 33 °C and the salinity fluctuated between 28.03 and 35.34 psu. Daily fecundity for the 127 days in which a total of 140111 eggs were collected, averaged 1103 ± 1171 (mean ± SD) eggs per female. Hatching rates ranged from 13.3 % to 96.6 % (59.3 ± 22 %). Fertilized eggs were spherical, transparent and buoyant and had a diameter of 0.780 ± 0.02 mm, with oil globule in of 0.178 ± 0.013 mm diameter. Embryonic development lasted 14h 30 min at 28 ± 0.2 °C, 33.4-33.5 psu. The newly hatched larvae were 1.81 ± 0.19 mm in total length (TL) with xanthophore evenly distributed on meomeres. Larvae at three days post hatching (dph) were 2.27 ± 0.30 mm TL, and eyes, mouth, anus were fully developed, and start feeding. At 9 dph, larvae into flexion stage. At 18 dph, larvae into transformation stage. At 25 dph, juvenile stage was completed, and all fins had the adult complement of ray and spines (D. IX, 23-25; A. III, 7; C. 17; P 16; V I, 5). Under scanning electron microscope, the fertilized eggs were spherical, the pores on the egg surface were uniform in size and evenly distributed with a density about 38 /100 μm2. The micropyle was cylindrical, the size was about 5.24 μm in diameter, and there are accessory opening evenly distributed near the micropyle with a number about 125. The neuromasts appeared at newly hatched larvae, and the number increase with the larvae growth; at 4 dph, the formation of teeth and spines of head were observed. For the skeletal ontogeny of D. pictum, the skull and vertebral column start developing at 2 dph; the caudal fin start developing at 7 dph; the dorsal and anal fin start developing at 10 dph; the pelvic fin start developing at 11 dph. Expect vertebral column ossification completed at 13 dph, skull, dorsal fin, anal fin and caudal fin ossification completed at 16 dph; pelvic fin ossification completed at 20 . In the temperature and salinity experiment, the best hatching treatment was temperature 27 °C and salinity 37 psu; the hatching rate was significantly higher than other treatments (P < 0.05). The salinity 29 psu show the lowest deformity rate, and all the temperature group the deformity rate were 0 %. The best SAI treatment was temperature 27 °C and salinity 33 psu. In this study, we not only provide the whole information of early life stage, but also be a reference for aquaculture in the future.

謝辭 I
摘要 III
Abstract IV
第一章 前言 1
1.1 緒言 1
1.2 文獻回顧 1
1.2.1 石鱸科 ( Haemulidae) 1
1.2.2 石鱸科魚類的生殖生物學 2
1.2.3 石鱸科魚類的人工繁殖研究 3
1.2.4 密點少棘胡椒鯛 (Diagramma pictum) 4
1.2.5 魚類的初期生活史研究 5
1.2.6 掃描式電子顯微鏡觀察(Scanning electron microscopy, SEM) 6
1.2.7 骨骼發育觀察 7
1.2.8 溫度與鹽度對海水魚類的初期發育階段之影響 9
1.2.9 生存活力指數 9
1.3 研究目的 10
第二章 材料與方法 11
2.1 實驗設計 11
2.2 親魚培育 11
2.3 自然產卵調查 11
2.3.1 魚卵收集 11
2.3.2 產卵量計算 11
2.3.3 受精率和孵化率計算 12
2.3.4 產卵週期觀察 12
2.4 餌料生物培育 12
2.4.1 S型輪蟲 12
2.4.2 橈足類 13
2.5 仔稚魚培育 13
2.5.1 餌料供給 13
2.5.2 水質檢測 13
2.6 胚胎及仔稚魚形態發育之光學顯微鏡觀察 15
2.6.1 胚胎發育觀察 15
2.6.2仔稚魚發育觀察 15
2.7 胚胎及仔稚魚形態發育之掃描式電子顯微鏡觀察 16
2.7.1固定 16
2.7.2 脫水 16
2.7.3 臨界乾燥 16
2.7.4 離子覆膜 17
2.8 初期骨骼發育觀察 17
2.8.1 透明骨骼製作 17
2.8.2 標本拍攝與觀察 18
2.9 不同溫度對孵化率、畸形率及SAI的影響 18
2.10 不同鹽度對孵化率、畸形率及SAI的影響 19
2.11 統計分析 19
第三張 結果 21
3.1 自然產卵 21
3.2 胚胎與仔稚魚形態發育 21
3.2.1 胚胎發育 21
3.2.2 仔稚魚發育 22
3.2.3卵黃囊和油球的消耗 24
3.2.4 仔魚的口徑發育 25
3.2.5 仔稚魚成長 25
3.2.6 仔稚魚的行為觀察 25
3.2.7 魚苗育成率 26
3.3 微細構造 26
3.3.1 魚卵的微細構造 26
3.3.2 仔魚的微細構造 26
3.4 骨骼發育觀察 27
3.5 不同溫度對孵化率、畸形率及SAI的影響 29
3.6 不同鹽度對孵化率、畸形率及SAI的影響 29
第四章 討論 31
4.1 自然產卵 31
4.2 二性特徵 31
4.3 密點少棘胡椒鯛的初期生活史 32
4.3.1 石鱸科的卵徑大小 32
4.3.2 仔稚魚的形態 32
4.3.3 內因性營養的消耗與仔魚初期攝食的口徑大小 33
4.3.4 仔稚魚行為 34
4.3.5 石鱸科仔稚魚培育 34
4.4 魚卵和仔魚的微細構造 34
4.4.1 魚卵的微細構造 34
4.4.2 仔魚的微細構造 36
4.5 仔稚魚的骨骼發育 37
4.6 溫度與鹽度對魚卵、胚胎及仔魚的影響 39
第五章 結論 43
參考文獻 45
附錄 103

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