眼斑双锯鱼(Amphiprion ocellaris)发育中体色花纹时序发生的色素细胞变化和控制基因表达的分析I.胚胎时期

眼斑双锯鱼(Amphiprion ocellaris)属于鲈形目、雀鲷科、双锯鱼属, 是热带珊瑚礁观赏鱼类的首选品种。不同体色和花纹与色素细胞相关基因的表达直接相关, 在其胚胎发育的过程中, 色素细胞的形态和数量一直在变化, 但不同发育时期各种色素细胞的动态变化及其控制基因表达情况等仍需明确研究。记录了眼斑双锯鱼胚胎时期色素细胞变化较为明显的6个发育时期, 观察了不同时期的体色及色素细胞的变化特点, 并利用荧光定量PCR检测了各发育时期的10个体色控制基因的表达情况。结果显示: 在胚胎发育阶段, 整个胚胎的颜色从橙黄逐渐转变为暗红、黑红、黑色到最终透明, 观察到卵黄囊表面幼体黑色素细胞到成体黑色素细胞的转变过程, 鱼体表面在翻转期出现成体黑色素细胞、在器官形成期出现红色素细胞, 眼睛及腹部在孵化期出现虹彩色素细胞。结合荧光定量PCR结果分析发现: 卵黄囊表面幼体黑色素细胞推测是外胚层中的神经嵴细胞直接发育而成, 不需要迁移过程; Ednrb、TYR、Tbx2b基因对幼体黑色素细胞的形成和发育有重要作用, Pax3、Dct、Aim1基因与成体黑色素细胞的分化、迁移、形成过程密切相关; 其中TYR基因的相对表达量在体节期出现显著增加, 说明TYR基因在胚胎发育初期就参与黑色素细胞的形成; Tbx2b基因不仅对黑色素细胞形成有影响, 对眼斑双锯鱼心脏的形成也有一定作用; Fms基因对红色素细胞形成有重要作用; Ltk基因的表达要早于虹彩色素细胞的出现; 相关性分析发现各基因之间存在相互作用。     

卵胎生许氏平鲉仔鱼与稚鱼发育形态学特征观察

本文对许氏平鲉胚后发育组织进行了较为详细观察,研究结果表明:许氏平蚰0日龄(dph)仔鱼,即未产出仔鱼,体长3.8~4.7mm,油球大而明显;初孵仔鱼体长5.0~6.5mm,已经开口,尚未摄食,体表仅分布有黑色素;2日龄仔鱼,开始摄食轮虫,鳔已充气,并出现虹彩色素;3日龄仔鱼,脑颅后方增加一丛色素,尾部放射丝出现,仔鱼摄食量大大增加;4日龄仔鱼,卵黄囊吸收殆尽,头颅后部和腹腔上缘开始分化出点状黄色素;5日龄仔鱼,胸鳍可垂直向外伸展,上下颌开启频繁;12日龄仔鱼,具有集群和趋光性;20日龄仔鱼,体前部呈黑色,在水中分散均匀,摄食旺盛;25目龄稚鱼,全长6.55~8.42mm,体前部开始出现黑色条带痕迹;30日龄稚鱼,头部眼的上缘有一横带,鱼体侧出现3条黑褐色的横条带;40日龄稚鱼,全长15.50~35.55mm,鱼体体型与成鱼相似,体侧有6~7条黑褐色的横条带;60日龄幼鱼,具有成鱼的形态,鱼体发育进入幼鱼期,并开始转入水体下层活动。通过对许氏平鲉由早期发育的形态学观察,了解其仔、稚鱼的发育规律和种属的特异性,不仅可以丰富对鱼类早期生活史的认识,而且对进一步进行许氏平鲉的生产育苗起到积极的推进作用。     

七带石斑鱼早期形态发育与仔稚幼鱼培育的观察研究

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、p H7.9~8.2、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。     

丝鳍圆天竺鲷的繁殖习性、胚胎发育和胚后发育观察

本文以丝鳍圆天竺鲷(Sphaeramia nematoptera)为研究对象,观察记录了其繁殖习性以及胚胎发育和胚后发育的过程。研究结果表明:丝鳍圆天竺鲷的繁殖活动由雌鱼主导,在凌晨1点至3点交配,产卵受精后由雄性口孵,产卵间隔期为11～18d,无季节性。雌鱼单次产卵3126～4882粒,受精卵近圆球形,卵径(720±10)μm,内有数颗油球;在水温(27.0±0.5)℃条件下,受精卵经过卵裂期、囊胚期、原肠期、神经胚期、器官形成期到出膜需187h。丝鳍圆天竺鲷胚后发育分为仔鱼期、稚鱼期、幼鱼期和成鱼期,初孵仔鱼全长(2.8±0.1)mm,口裂高度(0.202±18)mm,孵化后第二天仔鱼开口摄食;孵化后24d鳍条形成,身体出现两条黑色条带,结束浮游阶段,进入稚鱼期;孵化后50d鳞片完全形成,体色斑纹等外形特点与成鱼相同,进入幼鱼期;孵化后360d性腺发育成熟,具备生殖能力。     

卵形鲳鲹胚后发育阶段的体色变化和鳍的分化

采集在池塘进行育苗的31天前的卵形鲳鲹Trachinotus ovatus仔稚幼鱼,对其早期发育阶段的体色变化和鳍的发生、发育进行了连续观察。在水温24.73±2.11℃、盐度20‰—24‰、pH8.0—8.2条件下,初孵仔鱼体表已具有黑色素,第2天眼点的褐色素增加,此时仔鱼未开口、眼点未有视物功能;第3天开口时,眼大而突出、布满黑色素,变为黑色的眼睛,具有视觉功能;第17天当仔鱼变态为稚鱼时,鱼体变得不透明;在第22天变态为幼鱼时,鱼体体表为褐色;30天的幼鱼体表为银白色,与成鱼的体色一致。各鳍开始分化和发育的顺序依次为胸鳍→尾鳍→背鳍→臀鳍→腹鳍。1天仔鱼胸鳍原基出现;2—3天仔鱼开始摄食,胸鳍为扇形;第17天各鳍发育基本完成,标志着稚鱼期的开始。体色的变化和鳍的形成和发育对仔稚鱼的摄食、行为、生长、变态和存活等生命活动有着至关重要的作用,也是仔稚鱼变态发育的重要特征之一,与人工育苗生产技术尤其是投饵、分池等密切相关。     

光唇鱼( Acrossocheilus fasciatus) 胚胎及仔、稚鱼的发育

采用干法授精方法获得受精卵,在人工培育条件下观察了光唇鱼胚胎及仔、稚鱼发育过程。结果表明,光唇鱼受精卵呈圆球形,沉性、弱粘性。在水温23—25℃下,胚胎发育历时46h45min,经历了胚盘形成、卵裂、囊胚、原肠胚、神经胚、器官形成和孵化等阶段。在水温24—27℃下,仔、稚鱼发育历时22d,初孵仔鱼具有较大的卵黄囊,胸鳍原基及肛门原基形成;4日龄仔鱼开口摄食,进入混合营养期;7日龄仔鱼卵黄囊消失,营外源性营养,体侧8条横斑形成,各鳍均已出现,进入晚期仔鱼阶段;14日龄仔鱼各鳍鳍条发育基本完成;16日龄仔鱼鳞片开始出现,进入稚鱼期;22日龄稚鱼全身被鳞,进入幼鱼期。     

人工繁育花鲈早期发育形态特征与仔鱼培育技术研究

对人工繁育的花鲈(Lateolabrax maculatus)早期发育阶段的形态特征及苗种培育技术进行了观察与研究。结果表明:在水温17±1℃、盐度30、pH8.0、溶解氧7.0mg/L的条件下,花鲈受精卵经过72h完成胚胎发育过程。在胚后发育阶段,根据卵黄囊体积、鳍条发育情况,将花鲈前期发育划分为仔鱼期、稚鱼期、幼鱼期;仔鱼期又可分为前期仔鱼和后期仔鱼。本文对培育过程中死亡率较高的仔鱼期进行了详细观察,发现在水温16~17℃,盐度30的海水中培育,初孵仔鱼至5日龄(day post hatching,dph)仔鱼是前期仔鱼,6~35dph为后期仔鱼。花鲈个体之间发育速度差异较大、仔鱼之间的残食现象是整个培育过程中比较明显的特征,这些特征与冬季水温较低造成的越冬困难一起限制了花鲈苗种的全人工繁育技术的建立。     

鱼胚胎发育及其仔、稚、幼鱼形态特征和生活习性的初步研究

作者对鱼(Miichthys miiuy)的早期发育(从受精卵到45日龄幼鱼)形态特征及生态习性进行了描述。鱼的受精卵在水温21.4℃～22.0℃，盐度25.7条件下，经过29 h30 min仔鱼孵出。从受精卵到仔鱼出膜可分为卵裂期、囊胚期、原肠期、胚体形成期等4个主要时期，然后进入胚后发育阶段。孵出后的3日龄仔鱼已能在水体中平游，极少数个体可开口摄食轮虫。4日龄仔鱼全部开口摄食，并对光反应敏感，出现明显的集群现象。22日龄的后期仔鱼各鳍鳍条发育形成，鳞片开始出现，进入到稚鱼期。39日龄后，鱼全身披满细小的鳞片，全部分布于育苗池的底部，开始进入到幼鱼发育阶段。根据仔、稚、幼鱼形态特征和生活习性对鱼的分类进行了探讨。     

东海陆架区几种灯笼鱼后期仔鱼的研究

1975-1976年“金星”号调査船在东海陆架区先后进行了三个航次的调查,曾采到灯笼鱼科中的六个属七个种的后期仔鱼共646尾。它们是无须灯笼鱼Diogenichthys panurgus Bolin,粗鳞灯笼鱼Myctophum asperum ( Richardson),巴氏灯笼鱼Gonichthys barnesi Whitely,神鰯灯笼鱼Diaphus theta ( Eigenmann et Eigenman),多光灯笼鱼Ceratoscopelus townsendi ( Eigenmann et Eigenmann),柳氏戴芬鱼Diaphus lutkeni Brauer及七星鱼Benthosema pterota ( Alcock)。在我国海域,除七星鱼的成鱼和神鰯戴芬鱼的后期仔鱼与稚鱼有过记录外,其它各种,包括其早期变态时期的后期仔鱼皆系首次记录。 灯笼鱼科的鱼类广布于热带和亚热带水深200-500米的大洋中层水域,是大洋性重要经济鱼类和海洋哺乳动物的重要饵料的组成部分。对灯笼鱼科鱼类的后期仔鱼及稚鱼的分布和形态的研究,对进一步探讨成鱼的生态及其地理分布有着重要的意义。 本文就七种灯笼鱼的后期仔鱼的出现及其变态时期的特征进行研讨。     

鲤科经济鱼类马口鱼(Opsariichthys bidens)胚胎发育及仔稚鱼形态与生长观察研究

为了解鲤科经济鱼类马口鱼(Opsariichthys bidens)胚胎及仔稚鱼发育特点,采用人工干法授精获得受精卵,在人工孵化及培育条件下观察了马口鱼胚胎发育及仔稚鱼的形态变化与生长特性。马口鱼成熟卵及受精卵均呈圆形、沉性,卵径分别为1.00—1.20mm及1.4—1.6mm。在26—28oC水温下,胚胎发育历时37h,历经了胚盘形成期、卵裂期、囊胚期、原肠胚期、神经胚期、器官发生期、心跳期和孵出期等阶段,所需积温为968.95oC·h。初孵仔鱼全长3.74±0.04mm。在26—28oC水温下,仔鱼孵后12h,胸鳍原基形成;孵后48h,肠道形成;孵后84h,一鳔室形成;孵后96h,多数开始平游;孵后108h,卵黄囊消失,早期仔鱼发育至后期仔鱼,营外源性营养;孵后14d,两鳔室形成;孵后17d,全长12.46±0.76mm,各鳍发育完整,鳞片开始出现,进入稚鱼期;孵后23d,全长达17.09±1.08mm,外部形态特征与成鱼相近。马口鱼仔稚鱼生长方程显示异速生长特性。本研究积累了马口鱼早期生活史资料,为马口鱼人工繁殖及鱼苗培育提供理论参考。     

白刺参，浅青刺参，深青刺参和紫刺参色素组成及黑色素含量的比较分析

Sea cucumber, Apostichopus japonicus(Selenka), is a commercially important marine species in China. Among the differently colored varieties sold in China, white and purple sea cucumbers have the greatest appeal to consumers. Identification of the pigments that may contribute to the formation of different color morphs of sea cucumbers will provide a scientific basis for improving the cultivability of desirable color morphs. In this study,sea cucumbers were divided into four categories according to their body color: white, light green, dark green, and purple. The pigment composition and contents in the four groups were analyzed by high performance liquid chromatography(HPLC). The results show that the pigment contents differed significantly among the white, lightgreen, dark-green, and purple sea cucumbers, and there were fewer types of pigments in white sea cucumber than in the other color morphs. The only pigments detected in white sea cucumbers were guanine and pteroic acid.Guanine and pteroic acid are structural colors, and they were also detected in light-green, dark-green, and purple sea cucumbers. Every pigment detected, except for pteroic acid, was present at a higher concentration in purple morphs than in the other color morphs. The biological color pigments melanin, astaxanthin, β-carotene, and lutein were detected in light-green, dark-green, and purple sea cucumbers. While progesterone and lycopene,which are also biological color pigments, were not detected in any of the color morphs. Melanin was the major pigment contributing to body color, and its concentration increased with deepening color of the sea cucumber body. Transmission electron microscopy analyses revealed that white sea cucumbers had the fewest epidermal melanocytes in the body wall, and their melanocytes contained fewer melanosomes as well as non-pigmented pre-melanosomes. Sea cucumbers with deeper body colors contained more melanin granules. In the body wall of dark-green and purple sea cucumbers, melanin granules were secreted out of the cell. The results of this study provide evidence for the main factors responsible for differences in coloration among white, light-green, darkgreen, and purple sea cucumbers, and also provide the foundation for further research on the formation of body color in sea cucumber, A. japonicus.     

Embryonic exposure to environmentally relevant concentrations of PCB126 affect prey capture ability of Fundulus heteroclitus larvae

Early life stages from a marine fish species, Fundulus heteroclitus, were exposed to sublethal doses of 3,3',4,4',5 pentachlorobiphenyl (PCB126) to evaluate its effects on ecologically relevant responses: growth and behavior. A few hours after fertilisation, eggs were treated topically with PCB126 (2.5-50 pg egg?1). Four days post-hatching (dph), morphological changes (body length and malformations), spontaneous locomotor activity (active swimming speed, rate of travel, % inactivity), prey capture ability (Artemia franciscana nauplii) and whole body EROD activity were evaluated in larvae. Untreated larvae collected at 0.5, 1, 2, 3, 4 dph were also examined. PCB126 did not increase the mortality or malformation rates. Body length and spontaneous locomotor activity were altered only in larvae treated with the highest dose. Treatment with PCB126 caused a dose-responsive reduction in prey capture ability (rate of decline in the number of Artemia) and induction of EROD activity. The lowest observed effective dose for both of these responses was 5.0 pg PCB126 egg?1 or 5.0 TCDD-toxic equivalents pg g?1 egg, using a TCDD-toxic equivalent factor of 0.005 and an egg mass of 5 mg. Prey capture efficiency (number of Artemia captured per feeding strike) was reduced at ≥ 10.0 pg egg?1. In untreated developing larvae, prey capture ability and efficiency increased as post-hatching development progressed and EROD activity remained low. The pattern of behavioral responses observed in PCB126-exposed Fundulus larvae differed from that observed in less-developed larvae indicating that other mechanisms than retarded development were involved. Behavioral dysfunction was a more sensitive response to PCB126 than morphological alterations and it occurred at environmentally relevant concentrations.     

褐菖鲉(Sebastiscus marmoratus)受精卵以及仔鱼期氨基酸与脂肪酸变化研究

为研究褐菖鲉(Sebastiscus marmoratus)发育早期氨基酸和脂肪酸的组成及变化规律,采用GC/MS法等方法分析了褐菖鲉的受精卵、初产仔鱼、前仔鱼期和后仔鱼期四个阶段的氨基酸和脂肪酸组成特点及其含量变化。结果表明:(1)总氨基酸含量从受精卵至初产仔鱼显著下降、前仔鱼期又显著回升、至后仔鱼期再次呈显著下降(P0.05);(2)总游离氨基酸量以初产仔鱼最高,是受精卵的10倍,各期的游离氨基酸占总氨基酸比值为1.4%—20.0%;(3)受精卵中以DHA、C16:0两者实际含量最高,分别为104.88 mg/g和68.74 mg/g,C18:1n-9和EPA次之,分别为41.23 mg/g和27.23 mg/g;(4)褐菖鲉内源性营养阶段被选择性消耗的主要脂肪酸依次为C16:0C18:1EPADHAARA,就脂肪酸的利用率而言MUFASFAPUFA;(5)褐菖鲉仔鱼在开口后的外源营养阶段对脂肪酸的利用率为PUFASFAMUFA,其中DHA相对EPA和ARA被选择性消耗。研究表明,褐菖鲉受精卵的游离氨基酸/总氨基酸比值符合海水鱼类沉性卵的特征,褐菖鲉在早期发育不同阶段对脂肪酸消耗具有不同的选择性,C16:1、C16:0、C18:0和C18:1是褐菖鲉早期发育的主要能源物质,在褐菖鲉开口饵料中添加一定水平的异亮氨酸、亮氨酸、酪氨酸等氨基酸及高度不饱和脂肪酸DHA等是十分必要的。     

黄海鲱鱼的人工孵化及胚胎发育的初步观察

对鲱科鱼类个体发育及早期形态特征的研究,国外早已进行过,尤以苏联和日本的资料最多。     

Growth characteristics,pigmentation, and the identificaton of whitebait (galaxias spp., Salmonoidea)

Definitions are given for stages in the development of five species of Galaxias, family Galaxiidae, which have marine larvae.

Larvae of Galaxias maculates attenuates and G. brevipinnis are large, and those of G. fasciatus relatively small as they enter fresh water from the sea. Development as whitebait involves shrinkage by about 25% in total length; the head length, considered separately, shrinks by about 15%. The ratio of standard length to head length alters from the larval to the adult value before positive growth recommences. These features are probably similar for G. postvectis and G. argenteus

Minimum observed pigmentation is described for the late larvae of G m. attenuates, G. brevipinnis, and G. fasciatus and for early whitebait of these and of G. postvectis and G. argenteus Subsequent development of melanophores and of colour pattern is described and figured. Large melano‐phores along the lateral line distinguish early whitebait of G. m. attenuates, but no distinguishing feature of pigmentation has been found in other species until the juvenile pattern is apparent. This pattern is characteristic for each species, and it persists in adult G. m. attenuates. G. argenteus has distinct juvenile and adult patterns, the latter being developed in a second, superficial, layer of pigment cells. Adult G. postvectis develop a distinctive fin colour pattern unlike other New ‐Zealand Galaxias; the superficial pigment layer is without definite pattern. In adult G. brevipinnis this layer resembles the persistent juvenile pattern and the two layers combined give a reticulate appearance. In G. fasciatus the juvenile pattern persists and develops as the fish grow; the superficial pigment layer is present in adults but is not usually apparent.

Until now positive identification of whitebait has depended on colour pattern. Recently recorded identifications of unpigmented whitebait are shown to be misleading. A key is given for the identification of early and late whitebait using absolute measurements as an important character.     

长牡蛎(Crassostrea gigas)两个Dmrt家族基因的时空表达

本文以长牡蛎27个幼虫发育阶段个体以及成体的5个组织织作为实验材料,采用实时荧光定量PCR技术对其Dmrt家族中的2个基因(CgDsx和CgDmrtA2)的表达模式以及在性别决定中的作用进行了研究。结果表明,长牡蛎CgDsx基因在胚胎发育初期有大量表达,从囊胚期到担轮幼虫初期表达量最高,之后表达量开始降低,在D形幼虫后一直维持在极低的表达水平,此后仅在成体的雄性性腺中具有高度表达。由此可见,CgDsx可能也对早期胚胎发育起一定调控作用,同时参与了性别决定。CgDmrtA2在长牡蛎所有组织中均有表达,各组织间表达差异不显著,在D形幼虫至壳顶后期表达量较高,说明它参与了胚胎中后期的发育过程,可能与神经的形成相关,但是其具体功能还需要进一步研究。     

日粮中添加叶黄素与角黄素对瓦氏黄颡鱼(Pelteobagrus vachelli Richardson)体色影响研究

采用传统养殖试验方法研究了日粮中添加叶黄素与角黄素对瓦氏黄颡鱼(Pelteobagrus vachelli Richardson)体色的影响。实验设计了11组等蛋白质(42.6%)、等能量(18.5MJ/kg)的实用试验日粮,在试验日粮中分别添加0、25、50、100、200mg/kg叶黄素和25、50、100mg/kg叶黄素与25、50mg/kg角黄素,对照组为野生瓦氏黄颡鱼。结果表明,实验结束时(60d),试验3、8、9日粮组(添加50mg/kg叶黄素、50mg/kg叶黄素+25mg/kg角黄素、50mg/kg叶黄素+25mg/kg角黄素)鱼体体色与对照组鱼(野生瓦氏黄颡鱼)体色相似,试验3、8、9组鱼皮肤中叶黄素含量与对照组鱼差异不显著(P0.05)。本实验认为,在日粮中添加叶黄素对瓦氏黄颡鱼的着色效果明显较角黄素好,是养殖瓦氏黄颡鱼饲料的适宜添加色素。在瓦氏黄颡鱼的饲料中叶黄素的建议添加量为50mg/kg。