可口革囊星虫生殖周期的观察

通过对可口革囊星虫体腔生殖细胞的发育和形态特征的周年观察,逐月统计体腔卵母细胞的数量,研究其变化趋势。结果表明,可口革囊星虫雌雄异体,性比为1∶1,无可见的生殖腺结构,原生殖细胞在体腔液中发育。体腔内全年都有生殖细胞存在,排卵期与产卵期出现重叠现象,卵母细胞分批成熟,分批产卵。根据卵母细胞的形态特点和存在方式可分为5个发育阶段:细胞增殖期、细胞质生长期、胶质膜形成期、成熟前期和成熟期。雄性生殖细胞在成熟之前,以细胞团的形式存在于体腔液中,其发生和产出过程,在时间上与卵母细胞同歩。成熟的生殖细胞经肾管排出、体外完成受精。宁德地区可口革囊星虫的生殖季节为5~9月,其中7~8月为繁殖盛期。体腔卵母细胞的长径与短径呈线性关系:Y=1.14X-0.7316(R2=0.9711)。     

Morphological, physiological and biochemical parameters characterizing the over-ripening of rainbow trout eggs

In rainbow trout (Oncorhynchus mykiss) freshly ovulated eggs and over-ripened eggs which had been retained in the coelomic cavity for 7, 14 and 21 days were investigated in aspects of morphology, physiology and biochemistry. Egg viability was significantly reduced from 85.9±16.4% in freshly ovulated eggs to 25.1±21.9% in over-ripened eggs which had been retained in the coelomic cavity for 21 days. Further during over-ripening in the ovarian fluid the pH significantly decreased, while the levels of proteins, of esterified and non esterified fatty acids and the activities of aspartate aminotransferase and acid phosphatase significantly increased. Also egg parameters changed: the wet weight of the unhardened eggs increased, the weight increase during hardening and the levels of esterified and of non esterified fatty acids significantly decreased. In freshly ovulated eggs the yolk consisted of a homogenous mass and the perivitelline space was small, but in over-ripened eggs the yolk was non homogenous with numerous vesicular inclusions and the perivitelline space was enlarged. When freshly ovulated eggs were incubated in water the cortical reaction was detectable within 5 min, in over-ripened eggs hardly no extrusion of cortical vesicles was visible and the width of the perivitelline space was very irregular.For the investigated freshly ovulated and over-ripened samples the egg viability significantly correlated with ovarian fluid parameters (pH, protein, non esterified fatty acids, esterified fatty acids, aspartate aminotransferase, acid phosphatase) and egg parameters (weight increase during hardening, weight of the hardened eggs).     

金乌贼和曼氏无针乌贼胚胎发育及其盐度耐受能力的比较研究

采集金乌贼(Sepia esculenta)及曼氏无针乌贼(Sepiella maindroni)受精卵,观察了金乌贼胚胎发育过程,比较分析了2种乌贼受精卵和仔乌的形态特征;分别研究了金乌贼受精卵在盐度20,25,30(对照)、33,36及曼氏无针乌贼受精卵在盐度15,20,25,30(对照)、36条件下的孵化率,分析了2种乌贼受精卵在192 h盐度突变过程中卵液渗透压和受精卵Na~+/K~+-ATPase活力的变化,比较了其在相同盐度20,25,30,36条件下的孵化率及耐受盐度突变的生理适应过程.结果表明,金乌贼胚胎发育第7天器官开始分化,第14天器官开始形成,盐度30、水温22～24℃时,其孵化时间为20～21 d;盐度30处理组,2种乌贼受精卵的孵化率均显著高于其他盐度处理组(P<0.01);在192 h盐度突变过程中,2种乌贼受精卵卵液渗透压随环境盐度的变化而变化,金乌贼受精卵卵液渗透压与环境渗透压相等,而曼氏无针乌贼受精卵卵液渗透压比环境渗透压平均高出60 mOsm/kg,2种乌贼受精卵均未检测到Na~+/K~+-ATPase活性.比较发现,虽然2种乌贼受精卵调节渗透压能力均较弱,但曼氏无针乌贼受精卵比金乌贼能更好地适应盐度变化.本研究旨在探明两种乌贼受精卵对盐度的适应性,为开展规模化人工繁育与资源增殖提供基础依据.     

Fertility of rainbow trout (Salmo gairdneri) gametes: Gamete viability in artificial media

A series of experiments was performed to test the effect of potassium ions (K⁺) on the storage of rainbow trout (Salmo gairdneri) sperm cells and eggs. Storage of eggs for up to 10 min in saline solutions, extender with K⁺ concentrations of ≤3.4 mM and coelomic fluid did not affect fertility. Fish Extender #6 had a detrimental effect on eggs within 15 s of application and reduced fertility to near zero within 10 min. Variation in the K⁺ concentration indicated that high (≥6.8 mM) concentrations decreased egg fertility while low (≤3.4 mM) concentrations had no effect on fertility. Insemination of eggs before the addition of Fish Extender #6 allowed storage for 14 h. Semen diluted (1:10) in coelomic fluid maintained fertility for 45 s, whereas semen diluted (1:1) in coelomic fluid maintained fertility for 60 min. Fish Extender #6 at K⁺ concentrations between 0 mM and 68 mM had no effect on fertility of semen diluted 1:2.5 after 1 h of storage.     

可口革囊星虫体液细胞组成及营养成分研究

利用显微镜观察可口革囊星虫体液的有形成分是由血细胞、颗粒细胞、生殖细胞和桑椹胚细胞组成。利用氨基酸自动分析仪、液相色谱研究了体液的无形成分有18种常见氨基酸,氨基酸总量为2.34 mg/m l。8种必需氨基酸为0.8 mg/m l,占氨基酸总量的34.19%,呈鲜味游离氨基酸和呈甘味游离氨基酸的总含量为1.06 mg/m l,占体液氨基酸总量的45.3%。含有18种脂肪酸,总含量为0.69 mg/m l。其中,饱和脂肪酸含量为0.22 mg/m l,占脂肪酸总量的32.88%;不饱和脂肪酸含量为0.47 mg/m l,约占脂肪酸总量的68.12%,其中单不饱和脂肪总含量为0.22 mg/m l,占脂肪酸总量的32.88%;多不饱和脂肪酸总含量为0.252 mg/m l,约占脂肪酸总量的36.52%。     

Further evidence for the presence of Renibacterium salmoninarum in salmonid eggs and for the failure of povidone-iodine to reduce the intra-ovum infection rate in water-hardened eggs

Abstract. Unfertilized, water-hardened eggs, obtained from a coho salmon, Oncor-hynchus kisutch (Walbaum), with coelomic fluid containing large enough numbers of the kidney disease bacterium, Renibacterium salmoninarum , to make the fluid cloudy (∼4 × 10⁹ cells/ml), were examined by cultural and histological methods to determine (a) whether an earlier finding of the pathogen in coho eggs could be corroborated, and (b) whether iodine (500 mg/l, in the form of povidone-iodine) was as effective at ridding them of the intra-ovum pathogen as it was at freeing them of the extra-ovum pathogen. The results confirmed that the pathogen does indeed occur within the egg (11–6%-15-l % of the eggs were infected in this manner), and suggested a location in the yolk for the pathogen. The findings also indicated that while iodine was very effective in killing the pathogen on the surface of the eggs it was completely ineffective in reducing the prevalence of the intra-ovum infections. The results point out the need for a method for treating eggs internally and confirm the inadvisability of using eggs from salmonids with cloudy coelomic fluid.     

密斯特黄彩鲶人工繁殖与胚胎发育的研究

从海南省山溪中捕捞收集到野生密斯特黄彩鲶,在池塘养殖条件下培育到性成熟,于2008年4月至10月,3次对其进行了人工繁殖和胚胎发育的系统研究。结果表明,密斯特黄彩鲶受精卵呈圆球形,透明,略带淡黄色。卵径平均1．0mm,沉性,有黏性,属黏性卵。黄彩鲶胚胎发育过程分为7期,包括受精卵阶段、卵裂期、囊胚期、原肠胚期、神经胚期、器官形成期、孵出期,历时约25h,胚胎发育总积温约为693．7h·℃。     

A Lactobacillus sp. from diseased female rainbow trout, Salmo gairdneri Richardson, in Newfoundland, Canada

Abstract. Post-spawning mortalities of 2- to 3-year old female rainbow trout, Salmo gairdneri Richardson, at a Newfoundland hatchery were studied. The fish were found to have a mixed bacterial infection involving primarily a Lactobacillus sp. (a Gram-positive, chain-forming coccobacillus) and to a lesser degree Enterobacteriaceae, Aeromonas hydrophila and Pseudomonas fluorescens. The bacteria were isolated from the visceral organs, kidney, and ascitic fluid present in the coelomic cavity. The fish had extensive degeneration and necrosis of the liver, spleen and kidney, and sloughing of intestinal epithelium, but whether these were caused directly by the bacterial infection was not established because the fish appeared to have received substantial stress from the mechanical stripping and from the retention of dead, unshed eggs in the coelomic cavity. Tissue sections suggested that substantial growth of the Lactobacillus sp. was occurring within these unshed eggs. The present study describes this unusual observation and compares the Lactobacillus sp. to two other Lactobacillus isolates from cultured trout.     

低盐胁迫对刺参 4 个盐度调节相关基因表达丰度的影响

从刺参(Apostichopus japonicus)低盐转录组数据库中选取与应激(热休克蛋白70基因)和离子传递(甘氨酸转运蛋白基因、锌转运蛋白基因、神经乙酰胆碱受体基因)相关的4个差异表达基因,利用qRT-PCR技术分析这4个基因在不同组织中的表达水平及低盐对其表达丰度的影响.结果表明甘氨酸转运蛋白基因在刺参呼吸树中表达水平最高,肠次之,体腔液表达较低;锌指蛋白基因和神经乙酰胆碱受体基因均在体腔液中表达最高,肠次之,在呼吸树中不表达;热休克蛋白70基因在体腔液中表达量最高,其次是呼吸树和肠组织.低盐胁迫下这4种基因的表达量均随着胁迫时间的延长呈波动性增减,其中甘氨酸转运蛋白在体腔液中的表达低于正常表达水平,在胁迫后3h时达到最低表达量.神经乙酰胆碱受体基因除在体腔液中48 h出现明显的上调外,在体腔液其他时间点和肠组织中处于下调表达状态.低盐胁迫下这4个基因表达丰度的变化,说明这些基因或作为功能蛋白直接参与机体的代谢调节,或作为调控蛋白调节胁迫功能蛋白的表达和活性来提高刺参对低盐胁迫的耐受能力.研究结果可为刺参盐度调节适应机制的研究奠定基础.     

扇贝精子及卵子的受精生物学特性

采用扫描电子显微镜和普通光学显微镜观察方法,结合全光谱扫描分析方法,对栉孔扇贝和虾夷扇贝的精子及卵子的受精生物学特性进行了详细的观察,包括精卵产出后形态学变化、卵水的特性、精子顶体反应、卵子皮层反应以及精子分级分离组分对卵子的作用5个方面.结果表明,这两种扇贝精卵生物学特性没有本质的区别,扇贝精子排入海水中10min以内,外形没有太大的变化,但30min以后约有1/4的精子出现头部膨大变成圆球形的现象,受精能力明显下降;扇贝卵子产入海水中1h以内受精能力没有变化,但2h以后,约1/3的卵子出现卵裂现象,未见极体的排放,且卵裂多停止在2细胞期,少数达到4细胞期,基本属于均等卵裂,并失去受精能力;滴入卵水中的精子极易发生自溶解体,未解体的精子发生凝集,10min之后卵水中基本检测不到完整的精子,卵水和钙离子载体A23187均能诱导精子发生顶体反应;精子分级分离组分分别为精浆、精子头部和精子尾部,这三部分除了少数精子头部能够附着在卵子表面外,其他都不具备激活卵子的作用.对扇贝精子和卵子的生物学特性的研究将为解释不同种扇贝的精卵相互识别并受精的现象提供基础资料.     

福建省东山岛西埔湾港养斑鰶的繁殖生物学

本文研究了福建省东山岛西埔湾港养斑鰶性腺周期发育、成熟系数周年变化、个体生殖力以及雌雄同体。港养斑鰶能在半封闭式湾内发育成熟而且自然产卵。其卵细胞发育分为6个时相,精巢发育分为6期。卵膜由透明带、外放射带和内放射带组成,内有孔道结构。第4时相中期卵母细胞的内放射带有7～8层纤维状片层。产卵后鞘膜细胞和颗粒细胞分裂增殖变成游走的吞噬细胞。西埔湾港养斑鰶Ⅰ龄鱼初次性成熟,生殖季节从12月下旬至翌年3月下旬,生殖盛期为2月,雌鱼成熟系数高达17.88％。个体绝对生殖力为7575～72621粒,平均 35310粒,系属于分批产卵类型,第1批生殖力大于第2批。     

Swelling of wolffish, Anarhichas lupus L., eggs and prevention of their adhesiveness

Wolffish, Anarhichas lupus L. has internal fertilization. The eggs are released into water before the beginning of cleavage and stick together in clutches. To develop methods for separating eggs for subsequent incubation, changes during egg swelling and egg shell hardening were observed. Uninseminated eggs did not swell in ovarian fluid, but a perivitelline space formed in sea water because of a partial cortical reaction. In inseminated eggs kept in ovarian fluid, a cortical reaction took place and swelling was caused by elevation of the yolk membrane. Shrinkage of the yolk and absorption of water were observed after inseminated eggs were placed in sea water. Eggs swelled in ovarian fluid diluted by less than 30% sea water showed abnormal cleavage during subsequent development. An effective method for preventing egg stickiness after fertilization was to distribute them in trays with stagnant sea water for at least 5-6 h. Milk was effective for loss of egg adhesiveness at 50% concentration, but some eggs cleaved abnormally and subsequent mortality was high. The influences of ovarian fluid and milk on initial egg development with respect to peculiarities of egg swelling are discussed.     

Feeding Ration,Genetics, and Reproductive Traits in Female Coho Salmon: Is Bigger Better?

We assessed the effects of different feeding regimes on reproductive traits in coho salmon, Oncorhynchus kisutch. Ration treatments subjected fish to different energy levels during oocyte proliferation (Year 1) and oocyte maturation (Year 2). Fish from 10 full‐sibling families were fed at either the recommended feeding rate or double the recommended ration during the first year. At the beginning of the second year, each ration group was split again into recommended and double ration tanks for a total of four dietary treatments. At the end of the second year, 365 females from the four treatment groups were spawned and body weight, egg weight, and egg number were measured. Overall, the fish fed the double ration during both years grew the fastest (50.1% larger than controls), had the highest egg number (46.3% larger), and had the largest eggs (9.6% larger), although similar increases were observed feeding at the double ration level during the second year alone. Ration modification produced significant changes in some reproductive traits, especially egg number, while family effects had a greater influence on relative fecundity (eggs/g female body weight) and egg weight. In fact, family effects were significant for all of the traits. Results indicate that reproductive traits can be manipulated through changes in ration and/or via genetic selection, either directly or indirectly.     

Biochemical changes associated with overripening of the eggs of rainbow trout Salmo gairdneri Richardson

This study of the biochemical composition of mature unfertilised eggs of rainbow trout was undertaken to investigate normal intraspecific variation in egg composition between parent females, and to describe the biochemical changes which occur after ovulation during overripening in the parental abdominal cavity. The following aspects of egg composition were measured: wet and dry weight, chorion weight, free and bound lipid, protein phosphorus, lipid phosphorus, precipitable protein, calcium, and iron. All these were determined on single eggs, and six eggs were examined from each female. Ripe eggs, defined as those stripped within 3 days of ovulation, were obtained from 7 females. Overripe eggs, defined as those stripped 30 or more days after ovulation were obtained from 8 females. Results were expressed in absolute terms (mg or μg of component per egg) and relative terms (percentage of egg dry weight). In ripe eggs, there was significant variation between females in all these aspects of egg composition when expressed absolutely, and in all except lipid phosphorus when expressed relatively. In overripe eggs, there was significant variation in all these aspects between females when egg composition was expressed either absolutely or relatively. Comparison of ripe and overripe eggs showed that overripening was accompanied by the following significant changes expressed absolutely: increase in water content, free lipid, iron, calcium; decrease in bound lipid, precipitable protein, protein phosphorus, lipid phosphorus, total lipid and chorion weight; when expressed relatively there were increases in water content, free lipid, iron, calcium, total lipid and chorion weight, and decreases in bound lipid, precipitable protein, protein phosphorus and lipid phosphorus.     

维生素E、C和HUFA交互作用对中华绒螯蟹生殖性能的影响

通过投喂添加或不添加VE、VC和HUFA的四组实验饲料,经198d饲养实验,以雌蟹的性腺系数、产卵力、孵化率、各组织超氧化物歧化酶(SOD)活性和过氧化产物丙二醛(MDA)含量等为指标,研究这两类营养素交互作用对其生殖性能的影响。结果表明,VE、VC和HUFA对雌蟹生殖性能有显著的影响(P<0.05):在产卵力方面,1组为3790个卵细胞·g-1体重,极显著高于2组(为2317个卵细胞·g-1体重)和3组(为2129个卵细胞·g-1体重)(P<0.01),显著高于4组(为3050个卵细胞·g-1体重)和5组(对照组)(为3010个卵细胞·g-1体重)(P<0.05);而孵化率方面,1组(86.17%)极显著高于2组(36.06%)、3组(29.18%)和4组(25.73%)(P<0.01),也显著高于5组(71.12%)(P<0.05)。1组和2组雌蟹性腺中SOD活性分别为40.09Nu·mL-1和39.87Nu·mL-1,相应地,MDA含量分别为6.87nmol·mL-1和3.33nmol·mL-1;而3组和4组雌蟹,尽管其性腺中SOD活性较高,分别为79.21Nu·mL-1和85.2Nu·mL-1,但由于饲料中抗氧化性VE、VC的缺乏,导致脂质过氧化发生,其MDA含量显著增加(P<0.05),分别为12.65nmol·mL-1和25.18nmol·mL-1。4组雌蟹的产卵力较大,但因VE、VC的缺乏发生脂质过氧化,导致所产的卵质低下,从而影响了卵子的孵化率。结果表明,VE、VC和HUFA都是河蟹保持良好的生殖性能的必需营     

小体鲟胚胎发育特征观察

观察了水温(18±1)℃条件下,小体鲟(Acipenser ruthenus Linnaeus)的胚胎发育特征。结果显示:成熟小体鲟卵圆球形、黑色、不透明,为粘性卵,卵径2.2～2.8 mm。水温(18±1)℃条件下,小体鲟受精卵历时110 h孵化出膜,所需总积温为2023～2308℃·h。小体鲟受精卵的卵裂方式为特殊的辐射状卵裂;原肠中期时动物极细胞覆盖胚胎三分之二;心跳期,心脏呈C型;尾端接近心脏时,眼囊内已经有色素沉着。根据胚胎发育过程的形态特征,可将小体鲟胚胎发育过程分为受精卵、卵裂、囊胚、原肠、神经胚、器官发生和出膜7个连续阶段,34个时期。另外,本研究还将小体鲟胚胎发育和鲟形目其他鱼类胚胎发育特征进行了对比分析。     

亚硒酸钠对刺参免疫反应中体腔液酶活力的影响

为了合理评价硒对水产养殖刺参的作用,实验检测和观察了室内模拟亚硒酸钠处理及病菌感染条件下刺参体腔液免疫酶的变化、组织病理变化和相对保护率。结果表明,随着亚硒酸钠处理强度的增加,刺参体腔液中超氧物岐化酶(SOD)、谷胱甘肽过氧化物酶(GP-X)、溶菌酶(LYZ)、酚氧化酶(PO)及碱性磷酸酶(ALP)活性升高;在同时感染病菌的情况下,SOD、GP-X及LYZ活性表现为先升高后下降的趋势,PO和ALP活性则持续上升,但随亚硒酸钠浓度升高,ALP活性增加的幅度减少,而较高亚硒酸钠浓度时累计发病率上升,相对保护率下降。研究结果表明,适宜浓度的亚硒酸钠可增强刺参对病菌感染的免疫力,而病菌感染可以加剧亚硒酸钠胁迫对刺参造成的免疫功能损伤,所以适度利用硒可减轻刺参养殖病害的发生。     

Seasonal shift in spawning of Atlantic cod (Gadus morhua L.) by photoperiod manipulation: egg quality in relation to temperature and intensive larval rearing

Commercial intensive fry production of Atlantic cod will be dependent on production of viable eggs independent of season. This can only be done by manipulation of maturation by photoperiod, but little is known about potential effects on egg characteristics and larval viability. In two cod broodstocks, maturation was successfully advanced or delayed 6 months compared with normal spawning season (March–April) by manipulation of photoperiod. The advanced broodstock spawned both in spring and autumn the same year. In two of the spawning tanks during autumn, ambient temperature was reduced after reaching 13.7°C during the first half of the spawning period. Egg quality and viability were monitored, and several egg batches were incubated, hatched and start‐fed for examination of growth and survival. Temperatures above 9.6°C resulted in significant reductions in fertilization and normal egg development. Concurrently, fractions of dead and unfertilized eggs increased with elevated temperature. Actual relative fecundity was not affected by temperature. Egg characteristics improved when temperature was controlled and lowered below 9.6°C. Occurrence of irregular spawners suggests that handling of broodstock fish should be avoided during maturation and spawning. Cod larvae originating from eggs of the advanced or delayed broodstocks were successfully reared beyond metamorphosis. Survival was 9.0–46.6% and 29.3% in green and clear water respectively. Survival correlated with both initial and average feeding conditions, but growth rate did not correlate with either of survival and feeding conditions. Specific growth rates (8.3–13.6% day⁻¹) is comparable with other intensive rearing trials with cod, but were lower than reported from nature‐like systems.     

莫桑比克罗非鱼幼鱼的性腺发育与分化

本文介绍了莫桑比克罗非鱼幼鱼性腺的发育和分化的观察结果。用组织学方法观察，莫桑比克罗非鱼幼鱼的性腺发育，可以明显地区分为三个阶段，即：性腺的原始阶段，性腺分化前期阶段，性腺分化完成阶段。作者认为，对莫桑比克罗非鱼的诱变试验，应在前两阶段进行；当幼鱼体长达到15mm以后，性腺分化已基本完成，性激素的处理不可能取得性诱变的明显效果。