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1 七带石斑鱼早期形态发育与仔稚幼鱼培育的观察研究周庆杰 1, 于道德 2, 高天翔 1 2, 官曙光 (1., ; 2., ) 摘要 : 为掌握一套可行的七带石斑鱼 (Epinephelus septemfasciatus) 人工繁育操作规程积累七带石斑鱼生物学资料和苗种培育技术经验, 作者对七带石斑鱼早期发育各个发育阶段的形态特征及苗种培育进行了观察与研究在水温 23.5 ±0.5 盐度 31 ph7.9~8.2 溶氧量 6~8 mg/l 的条件下, 七带石斑鱼的受精卵历时 31 h5 min 完成胚胎发育过胚胎发育可划分 5 个时期 : 卵裂期囊胚期原肠胚期神经胚期和器官形成期根据卵黄囊第二背鳍棘与腹鳍棘鳞片和体色的变化, 七带石斑鱼胚后发育可划分为仔鱼期稚鱼期幼鱼期 ; 仔鱼期又可分为前期仔鱼和后期仔鱼在水温 24~26, 盐度 28 的海水中培育, 初孵至 2 日龄 (day post hatching, dph) 仔鱼是前期仔鱼, 3~30dph 为后期仔鱼, 31~57dph 为稚鱼期, 58dph 进入幼鱼期第二背鳍棘和腹鳍棘的生长与收缩是七带石斑鱼胚后发育过程中最明显的变化特征第二背鳍棘的长度在仔鱼 28~30 dph 达到最大值, 占仔鱼全长的 81% 这些特点增加了苗种培育的难度和障碍关键词 : 七带石斑鱼 (Epinephelus septemfasciatus); 人工繁育 ; 形态发育 ; 苗种培育中图分类号 : S917.4 文献标识码 : A 文章编号 : (2014) doi: /hykx (Epinephelus septemfasciatus) (Perciformes), 鮨 (Serranidae), (Epinephelus),,, [1], 7~8, [2],,, [1, 3-6] [7-, 8] [9-11] [12] [13],,,, 1 材料与方法 1.1 受精卵孵化 ~8 58 / 2014 / 38 / 12,, ~7, (HCG) 700IU/kg, (LRH-A 3 )6µg /kg HCG200 /kg 72 h,,,,, 1.5 ml,,,, 23.5 ± ph7.9~8.2 6~8mg/L ~2 000 lx, 1.2 胚胎发育观察,, Olympus sz61,, 50% : ; : : 948 (2014-Z49); (SDAIT ) : (1988-),,,,, : , shuichanzhou@ 163.com;,, : , shuguang63@126.com

2 1.3 苗种培育及观察 4 m 4 m 1.5 m, 24~26, 28 (Crassostrea gigas) ;, S (Brachionus) L (Artemia salina) (Chlorella),, 3~5 d, 20, Olympus sz61 Nikon TS100,, 2 结果与分析 2.1 胚胎发育,,, (0.854±0.021)mm(n 30), (0.181±0.006)mm(n 30),,, 31,, (23.5±0.5) 31, 31 h5 min, 50%,, 5, : min,, ; 2 h20 min, 6 ( 1-1~ 1-6) 64 ( 1-7),,,, ( 1-7); 2 h50 min,, ( 1-8); 4 h 15 min,,,,, ( 1-9) h 30 min,,,,, ( 1-10);,, 6 h 30 min 表 1 七带石斑鱼胚胎发育过程 Tab.1 Embryonic development of E. septemfasciatus 0, 38 min 2 1, 2 58 min 4 2, 4 1 h 3 min 8 3, 8 1 h 10 min 16 4, 16 1 h 40 min 32 5, 32 2 h 20 min 64 64, 2 h 50 min 4 h 15 min, 5 h 30 min, 6 h 30 min, 7 h 45 min 1 /3, 10 h 45 min 1 /2 11 h 40 min 2 /3, 12 h 45 min, 14 h 35 min 1 15 h 15 min 15 h 45 min 1 17 h 35 min 21 h 25 min 23 h 15 min 26 h 15 min 30 h 30 min 30 h 45 min 31 h 5 min,,,,, ( 1-11) 2.1.3,, ( 1-12) ( 1-13) ( 1-14) h 45 min,,,, ( 1-15) h 35 min, Marine Sciences / Vol. 38, No. 12 /

Fig.1 图 1 七带石斑鱼胚胎发育 Embryonic development of E. septemfasciatus 1-1.受精卵, 40; 1-2.2 细胞期, 40; 1-3.4 细胞期, 40; 1-4.8 细胞期, 40; 1-5.16 细胞期, 40; 1-6.32 细胞期, 40; 1-7.64 细胞期, 40; 1-8. 多细胞期, 40; 1-9.

3 Fig.1 图 1 七带石斑鱼胚胎发育 Embryonic development of E. septemfasciatus 1-1.受精卵, 40; 细胞期, 40; 细胞期, 40; 细胞期, 40; 细胞期, 40; 细胞期, 40; 细胞期, 40; 1-8. 多细胞期, 40; 1-9.桑葚期, 40; 1-10.高囊胚期, 40; 1-11.低囊胚期, 40; 1-12.原肠早期, 40; 1-13.原肠中期, 40; 1-14.原肠晚期, 40; 1-15.胚体形成期, 40; 1-16.视囊形成期, 40; 1-17.肌节形成期, 40; 1-18.听囊形成期, 40; 1-19.脑泡形成期, 40; 1-20.尾芽期, 40; 1-21.晶体形成期, 40; 1-22.心跳期, 40; 1-23.出膜前期, 40; 1-24.初孵仔鱼, Fertilized egg, 40; cell stage, 40; cell stage, 40; cell stage, 40 ; cell stage, 40 ; cell stage, 40 ; cell stage, 40; 1-8.Multi-cell stage, 40; 1-9. Murula, 40; 1-10.High blastula, 40;1-11. Low blastula, 40; Early gastrula, 40; Middle gastrula, 40; 1-14.Late gastrula, 40; Embryo body stage, 40; Optic capsule stage, 40; 1-17.Muscle burl stage, 40; Otocyst stage, 40; 1-19.Brain vesicle stage, 40; 1-20.Tail-bud stage, 40; 1-21.Crystal stage, 40; 1-22.Heart-beating stage, 40; 1-23.Pre-hatching, 40; 1-24.Newly hatched larvae, 40 进入视囊形成期(图1-16); 受精后15 h15 min 脊索两体背面的视囊之间出现椭圆形板状脑泡, 脑泡尚未侧出现长方形的体节, 胚体进入肌节出现期, 视囊分室, 胚体进入脑泡形成期 (图 1-19); 受精后 21 h 轮廓变得更加清晰(图1-17); 受精后15 h45 min 在胚 25 min 胚体背腹两处逐渐形成鳍褶, 尾部有少部分体头部视囊靠后的位置出现１对听泡, 肌节数增多, 与卵黄囊分离, 尾鳍褶可见, 胚体进入尾芽期 (图胚体进入听囊形成期(图1-18); 受精后17 h 35 min 胚 1-20); 受精后23 h15 min 胚体开始间歇抽动, 视囊 60 海洋科学 / 2014 年 / 第 38 卷 / 第 12 期

4 内可见折光性强的晶体(图1-21); 受精后26 h15min 0.014mm 油球 1 个, 圆球形, 油球直径 0.18 mm± 心脏开始微弱跳动, 胚体进入心跳器(图1-22); 受精 0.012mm, 大部分仔鱼油球位于卵黄囊后端, 一部分后30 h30 min 胚体的扭动更频繁有力, 进入出膜前仔鱼的油球位于卵黄囊下端, 极少数仔鱼的油球位期(图1-23); 受精后30 h45 min 进入出膜期, 胚体头于卵黄囊前端刚孵出的仔鱼, 鱼体透明, 脊索略有部先顶破卵膜, 然后尾部用力摆动, 脱去卵膜弯曲, 肌节 22~24 对, 肛前肌节 8~9 对, 肛后肌节 ~15 对头部上方, 背腹部, 尾部有无色透明的鳍仔稚幼鱼形态发育特征七带石斑鱼仔稚鱼各期的发育和形态特征如图 2 所示仔鱼期初孵仔鱼全长 1.99 mm±0.041 mm, 卵黄囊呈椭球形, 长径 0.92 mm±0.012mm, 短径 0.55 mm± Fig.2 膜连成一体肛门稍离开卵黄囊, 位于体中央偏后, 尚未开口于体外孵化后 5h 的仔鱼, 消化管后方背面出现黑色素细胞, 鱼体靠近尾部出现两个上下对称的黑色素细胞初孵仔鱼腹部朝上, 悬浮于水中, 游泳能力较弱, 尾部偶尔快速扭动图 2 七带石斑鱼仔稚幼鱼 Larva, juvenile, and young fish of E. septemfasciatus 日龄仔鱼, 35; 日龄仔鱼, 35; 日龄仔鱼, 30; 日龄仔鱼, 30; 日龄仔鱼, 30; 日龄仔鱼, 30; 日龄仔鱼, 25; 日龄仔鱼, 25; 日龄仔鱼, 25; 日龄仔鱼, 25; 日龄仔鱼, 25; 日龄仔鱼, 20; 日龄仔鱼, 20; 日龄仔鱼, 10; 日龄仔鱼, 10; 日龄稚鱼, 10; 日龄稚鱼, d larva, 35; d larva, 35; d larva, 30; d larva, 30; d larva, 30; d larva, 30; d larva, 25; d larva, 25; d larva, 25; d larva, 25; d larva, 25; d larva, 20; d larva, 20; d larva, 10; d larva, 10; d juvenile, 10; d juvenile, 日龄仔鱼全长 2.48 mm±0.030mm, 卵黄囊长形, 23~25 对油球边缘出现一圈黑色素, 卵黄囊前径 0.70 mm±0.010 mm, 短径 0.24 mm±0.012 mm 油端和眼后方出现星状黑色素, 仔鱼背部也新出现黑球 1 个, 圆球形, 油球直径 0.14 mm±0.012 mm, 卵黄色素细胞仔鱼游泳能力增强囊与油球逐渐缩小脊索稍伸直, 头部增大, 大部分 2 日龄仔鱼全长 2.41 mm±0.024 mm, 卵黄囊长仔鱼头部已脱离卵黄囊并抬起消化道开始膨大, 肛径 0.29 mm±0.021 mm, 短径 0.16 mm±0.014 mm 油门位置略前移, 位于鱼体中央, 输尿管细小透明, 后球直径 0.09 mm±0.024 mm, 卵黄囊与油球进一步被端膨大为膀胱仔鱼眼部晶体透明, 可清楚看到耳囊, 吸收, 部分仔鱼卵黄囊明显萎缩口裂已经形成, 但耳囊内有 2 个耳石, 很小, 前后排列肌节呈 V 字口不能活动, 未开口消化管变粗, 前端膨大形成胃, Marine Sciences / Vol. 38, No. 12 /

5 ,, ;,, mm±0.082 mm,, 0.38 mm,,, 4~ mm±0.068 mm,,,,, D,, mm±0.072 mm,, 20 D,, D, D, D,, mm±0.052 mm, D mm±0.096 mm,,, D,, mm±0.088 mm, 0.13 mm,,,,, mm±0.094 mm,,,,,,,, mm±0.098 mm,,,, 0.31mm,,, mm±0.086 mm,,,,, 5~ mm±0.074 mm, 2.90 mm, 6.50 mm, 鰓,,, 2, mm±0.462 mm, 2,,,,, 0.11 mm,,, mm±0.531mm,,,,,,,,,,,, 62 / 2014 / 38 / 12

6 mm±0.681 mm 1/2,, 3, 1, 2 ; 4, 2, mm±0.642 mm,,,,,,, mm±0.724 mm,,, 10,, 2,, V, 21, mm±0.874 mm, 1,, 2/3, 1/2, mm±0.883 mm,,,,,, W, 22 21~ mm±1.120 mm,,,, 10,,,,,, 3, 2,,,,, 3/4, 1/2,,,, 17,, 26~ mm±1.461 mm,,,, 10, 6,, 14 ; 1 4 ;, 3 9 ;, mm, mm, 3,,,,,, ~ mm±1.862 mm,,,,,, 1/3,,,, 1/4,, 9, 15 ; 1, 5 ; 3, 9 ; 17, 8, 9,,,, 41~ mm±1.461mm, mm,,,,,, Marine Sciences / Vol. 38, No. 12 /

7 ,,,, 2, mm±1.383 mm 28.5 mm,, mm,,,, mm±1.962 mm,,, 7,, 2.3 仔稚幼鱼的生长与培育管理 3 Fig.3 3 Growth of larva, juvenile, and young fish of E. septemfasciatus (Chrysophyta) :, (3011) ml 5~6 d 0.4 m 3 7 d (4 m 3 )5~6 d, (15 m 3 ) 3 d, 300 /ml :, 3~4 :, 24, 80 cm, ~2 000 lx, 300 /ml 0.5 m ~6 000 /m 3 3,,, 4, 26 (Ostrea gigas) :, 10: 1, 120, 5,,, (24 22 h D, ) 0.5~1 /ml D, 10 cm, 6, S 9,,,,,, 10, L,, D, 1~3, 1~2 :,, ;,,,,,, 5 /ml 11 80, 10% 15~20,,, :, 30~40 cm; ~1 500 lx,,,,, 17,,,, 64 / 2014 / 38 / 12

8 0.1 /ml, 1 /ml 26, 0.5~1 /ml,,,, 20% cm, 1,, 5~7 d 1 ;,,, 3 讨论与小结 3.1 胚胎发育 mm ±0.021mm, [14] ( mm ±0.024 mm), ( Epinephelus lanceolatus) (0.79 mm± 0.85mm) [15] ( Epinephelus malabaricus) (0.75 mm±077 mm) [16], (Epinephelus moara) (0.871 mm±0.013 mm) [17] 2640 /g, [16] [18] 1500 /g,,,,, 31 h5 min, [18] (22±0.5), 30~35 32 h36 min~32 h42 min,, [15-17, 19] 3.2 七带石斑鱼胚后发育阶段的划分 [20-23],, ;,, [20] ; [23],, 30 dph, [23],,,, 3.3 七带石斑鱼仔稚幼鱼的生长与培育 1.99 mm± 0.041mm, [24] 1.36 mm±0.07 mm, [15], ; [25] ;,,,, (0~3 dph),, (4~30dph),,,, [15],,, [26] S L Marine Sciences / Vol. 38, No. 12 /

9 , (Schizochytrium limacinum),, 2~3 d, ;,,,,,,, (31~57 dph),,,,, [18],, [27],,,,,,,, (58 dph),, 6 cm,, 30 dph 7.98 mm±1.461mm, [24] mm± mm 60 dph mm± 1.961mm, 島 [28] 60 dph 30 mm, ( 25 m 3, 22~24 ; 15m 3, 22~28 ) [20] (E. akaara ) [25] (E. coioides ) [29] [17] ( 2),,,, 表 2 七带石斑鱼与其他 4 种石斑鱼的相关数据对比 Tab.2 Comparison of relevant data of Epinephelus septemfasciatus to those of other groupers (d) (mm) (d) (mm) (mm) (mm) : [1],,,. [J]., 2011, 30(4): [2],,,. [J]., 2008, 39(6) : [3] Kline R J, Khan I A, Soyano K, et al. Role of folliclestimulating hormone and androgens on the sexual inversion of seven-band grouper Epinephelus septemfasciatus [J]. North Am J Aquac, 2008, 70(2) : [4] Koh I C C, Yoloi K, Tsuji M, et al. Cryopreservation of sperm from seven-band grouper, Epinephelus septemfasciatus[j]. Cryobiology, 2010, 61(3) : [5],,,. [J]., 2011, 20(1) : [6],,. [J]. ( ), 2005, 23(3): [7],,,. [J]., 2010, 7(5): [8] Teruya K, Yoseda K, Oka M, et al. Effects of photoperiod on survival, growth and feeding of seven band grouper Epinephelus septemfaciatus larvae[j]. Nippon Suisan Gakkaishi, 2008, 74(4) : [9] Kokawa Y, Takami I, Nishizawa T. A mixed infection in seven band grouper Epinephelus septemfasciatus affected with viral nervous necrosis (VNN)[J]. Aquaculture, 2008, 284(1/2/3/4) : [10],,,. 66 / 2014 / 38 / 12

10 [J]., 2010, 31(1): [11] Tanaka S, Takagi M, Miyazaki T. Histopathological studies on viral nervous necrosis of seven band grouper, Epinephelus septemfasciatus Thunberg, at the grow-out stage[j]. J Fish Dis, 2004, 27(7) : [12],,,. [J]., 2009, 30(5) : [13],,,. [J]., 2010, 17(1) : [14],,,. [J]., 2009, 28(2): [15],,,. [J]., 2010, 26(01): [16],,,. [J]., 2003, 27(4): [17],,,. [J]., 2012, 33(3): [18],,,. [J]., 2011, 32(2): [19],,,. [J]., 2012, 33(3): [20],,,. ( ) [J]., 2011, 32(4) : [21],,. [J]., 1985, 9(4): [22],. [J]. ( ), 2001, 19(2): [23],,,. [J]., 2003, 4: [24],,,. [J]., 2011, 32(5): [25],,. [J]., 2001, 10(4): [26]., [M]. :, 2007: [27]., [M]. :, 2007: [28] 島,, 塚島,. 発飼魚態態 [J]. 魚魚雑誌, 1991, 38(1): [29],,,. [J]., 2008, 27(2): Marine Sciences / Vol. 38, No. 12 /

11 Studies on the early morphological development and breeding of larval, juvenile and young fish of Epinephelus septemfasciatus ZHOU Qing-jie 1, YU Dao-de 2, GAO Tian-xiang 1, GUAN Shu-guang 2 (1. Fisheries College, Ocean University of China, Qingdao , China; 2. Marineculture Institute of Shandong Province, Qingdao , China) Received: Nov., 4, 2013 Key words: Epinephelus septemfasciatus; artificial breeding; morphological development; fry rearing Abstract: In order to develop a set of feasible artificial breeding techniques and accumulate biological data and fry rearing experience of Epinephelus septemfasciatus, the morphological characteristics of every early developmental stage and fry rearing of Epinephelus septemfasciatus were observed and studied. The time for the embryonic development is 31 h and 5 min in the water with temperature (23.5±0.5), salinity 31, ph 7.9~8.2, DO 6~8 mg/l. The embryonic development of E.septemfasciatus can be divided into five stages, namely cleavage stage, blastula stage, gastrula stage, neurula stage and organogenesis stage. The post embryonic development of E.septemfasciatus was divided into larval stage, juvenile stage as well as young fish stage, based on the features of the yolk-sac, pelvic fin spine, second dorsal fin spine, scale and body color. In the sea water with temperature 24~26, salinity 28, the pre-larvae stage developed from hatching to 2 dph. The embryon developed 3 days to the post-larvae stage. The juvenile stage developed from 31 to 57 days after hatching. After 58 days, the fish developed into the early young fish period. The extension and shrinkage of the second spine of dorsal fin and the first spine of ventral fin are the most obvious changes during early development of E.septemfasciatus. The ratio of the second spine of dorsal fin to total length attained its maximum (about 0.81) when the larvae were 28~30 days old. All these increase the difficulties and obstacles in the fry rearing. ( 本文编辑 : 谭雪静 ) 68 / 2014 / 38 / 12

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