石磺繁殖生物学的实验研究

用实验室小型试验、贝类育苗场生产性试验和自然群体实地观察的方法，结合养殖池塘内石磺（Onchidium sp.）繁殖习性跟踪观察，2年内研究了沪浙地区石磺的繁殖季节，繁殖方式，繁殖力，受精卵的发育孵化，水温和盐度对胚胎发育的影响，胚胎停止发育现象，幼虫发育及变态前形态变化，对石磺繁殖生物学作了比较系统的实验研究。结果显示石磺雌雄同体，雄性先熟，异体交配，体长5 cm体重10 g以上的个体为繁殖的主要群体，其生物学最小型为体长3.1 cm和体重3.5 g；交配期多在气温22 ℃以上的5月中下旬至9月下旬，6～8月为繁殖盛期。产卵前经过5～10 h的求爱和1～5 h的交配过程，交配后15 d左右产出受精卵，多在大潮日至小潮日期间3～5 d内的晚间产卵；繁殖期具有6个明显的产卵高峰。卵群表面积15～30 cm²，每平方厘米卵群含卵子2 650±300个，每个卵群含卵子4.4～8.0万粒。水里与潮湿环境中卵群孵化率没有明显差异；26～35 ℃的水温下，卵群孵化时间10～14 d，水温23 ℃以下石磺胚胎出现停止发育的现象。适合卵群孵化的海水盐度为6～20，孵化出膜2～3 d后的面盘幼虫开始摄食单胞藻。自由生活面盘幼虫经23 d培育后出现明显的革质膜；变态后的幼体可能存在不断蜕去革质膜的过程，面盘幼虫的变态过程和变态条件仍有待研究。     

大竹蛏生产性人工繁育试验

2007年对大竹蛏进行了生产性人工繁育试验。对亲本不同批次催产效果、不同规格亲本产卵差异、孵化密度进行了比较,研究了D形幼虫发育与温度的关系以及幼虫发育各阶段生长规律等,结果显示,大竹蛏属分批成熟、分批产卵的潜居性贝类,不同批次个体平均产卵量(36~118)×104cell,个体总产卵量294×104cell。壳长8~10 cm的亲贝产卵量大,催产效果好。受精卵适宜的孵化密度为5~10 cell/mL。在水温21~24.5℃条件下,D形幼虫经8~5 d生长,发育变态为附着稚贝。D形幼虫壳长日生长33.33±4.22μm,1 mm前的稚贝壳长日生长108.33±22.57μm,1~10 mm的稚贝平均旬生长量2.12 mm,10.4~30.6 mm的苗种平均旬生长量为2.02 mm。当水温下降到15℃后,大竹蛏苗种生长滞缓。2007年培育出壳长11.52±0.27 mm的稚贝2 165×104cfu,平均成活率12.1%。     

钝缀锦蛤繁殖周期和胚胎发育

2013年8月至2014年7月,采用组织学和实验生态学方法对钝缀锦蛤(Tapes conspersus)的性腺发育、生殖周期、肥满度、胚胎和幼虫发育进行了研究。结果显示,钝缀锦蛤性腺发育以1年为1个周期,性腺发育过程可分为增殖期、生长期、成熟期、排放期和休止期5个时期;繁殖期在每年10月至翌年3月,繁殖盛期为3月、10月和11月(水温20.3~24.6℃),分批产卵。8月肥满度最高,为33.43%;4月最低,为18.31%。钝缀锦蛤卵径为70~80μm;在水温24~26℃、盐度28~32条件下,受精卵经16 h左右发育为D形幼虫;初孵D形幼虫大小为90~110μm,浮游幼虫经7~8 d培育进入附着变态期,此时壳长为210~230μm,再经4~5 d发育变态为稚贝。本研究为钝缀锦蛤的人工繁育及种质资源保护和利用提供了基础资料。     

脉红螺繁殖生物学的研究

本文研究了脉红螺繁殖方式,繁殖季节,繁殖力,孵化,幼虫发育及变态和稚,幼螺发育过程,阐明了脉红螺在山东沿海每年只有一个繁殖期。繁殖期内亲螺有多次交尾现象,平均每年雌螺产卵２．１次,产卵袋６５５．９个,每个卵袋平均含有受精卵１１４９粒,个体繁殖力７５．４万粒。     

方格星虫多次产卵培育及幼体发育效果研究

为研究方格星虫多次人工培育的繁育性能,采用室内产卵培育和滩涂恢复养殖相结合的方法,在繁殖期内对同一批方格星虫进行了3次人工培育产卵试验,对孵出的幼体进行人工培育。结果表明:于6月、7月和8月进行的3次人工培育,雌虫平均产卵3.11×10⁴、2.70×10⁴和1.75×10⁴粒,平均受精率为(84.4±2.8)%、(87.6±1.9)%和(83.4±2.4)%；海球幼体体长日均增长49 μm、65 μm和46 μm,变态成活率(27.3±6.3)%、(32.2±4.6)%和(37.8±5.9)%；随着培育产卵次数增多,亲虫的死亡率升高,相对个体产卵量减少；海球幼体变态率升高,且差异极显著(P<0.01)；变态后稚虫的生长速度与后期海球幼体的大小成正相关。     

背瘤丽蚌钩介幼虫寄生变态发育和有效积温的研究

选用黄颡鱼(Pelteobagrus fulvidraco)作为寄主鱼,研究了背瘤丽蚌(Lamprotula leai)钩介幼虫寄生变态发育的过程、生物学零度和有效积温等重要生物学特征。结果显示,背瘤丽蚌钩介幼虫在黄颡鱼的鳃丝、鳍条或须上均能完成变态发育。在水温(25±1)℃时,寄生期为8 d。钩介幼虫腹缘先与寄主鱼体表组织接触,寄生后5 h,大部分被鱼体组织包裹;第2天,内幼虫足丝消失,钩介幼虫全部被包在包囊中;随着钩介幼虫寄生变态发育的进行,其壳颜色逐渐加深;第7天,变成黑色不透明,并有少量钩介幼虫变态为稚蚌;第8天,稚蚌脱落达到高峰期;第9天,稚蚌全部脱落。在寄生阶段,钩介幼虫壳长、壳高和铰合部长未发生显著变化。钩介幼虫寄生变态发育的生物学零度为12.8℃,有效积温为100.2℃.d。     

水温对石磺胚胎发育的影响

通过实验室研究，对石磺（Onchidium sp．）胚胎发育过程进行了详细观察，比较了不同水温下石磺胚胎发育的速度，探讨了膜内面盘幼虫孵化出膜的主要条件。结果表明：石磺的胚胎发育过程类似于肺螺亚纲其他贝类，行体内受精，刚产出的卵为受精卵，细胞未进行分裂，卵裂从卵产出体外开始，分别经过卵裂期，囊胚期，原肠期，膜内担轮幼虫期，膜内面盘幼虫期，经过孵化期孵化出膜为自由生活的面盘幼虫期。对各阶段幼体的大小、形状、内部器官等特征以及发育变态过程进行了详细的显微观察和描述。26℃，28℃，30℃3个水温组中，从受精卵到膜内面盘幼虫孵化出膜分别需要300h，290h，288h；发现影响幼虫孵化出膜的主要因素是水温和海水的刺激作用；繁殖盛期卵群工厂化水泥池自然水温孵化时间为10～14d。并报道了水温低于23℃时石磺胚胎停止发育现象。     

糙海参胚胎和幼体发育的形态观察

为提高糙海参育苗技术,研究描述了糙海参从受精卵发育到稚参的形态变化,在显微镜下测定了受精卵、胚胎和幼体的大小,确定了糙海参胚胎发育的过程。结果表明,通过利用阴干、流水刺激法对成熟亲参进行人工催产,得到大量的受精卵,其受精率为90%以上。糙海参胚胎和幼体发育可分为受精卵、卵裂期、囊胚期、旋转囊胚期、原肠期、初耳状幼体、中耳状幼体、大耳状幼体、樽形幼体和稚参等阶段;在平均水温29℃,平均盐度34条件下,糙海参受精卵经3 h发育形成囊胚,4 h进入旋转囊胚期,5 h进入原肠期,19 h完成胚胎发育变态为耳状幼体;并经过7 d的生长与发育进入樽形幼体;第15天变态为稚参。观察发现,多精入卵现象则会导致胚胎发育不正常,最终使胚胎发育停止并死亡。在糙海参幼体发育过程中,大耳状幼体期幼虫臂的大小及其球状体的形成均可作为判断其幼体发育健康状况的重要指标:幼虫臂越大,球状体出现率越高,其幼虫的变态率和成活率也越高。对比发现,位于糙海参尾部的突出结构——尾突,为仿刺参和新西兰刺参所没有,这一结构差异同时导致了其骨片位置也不同;并且,糙海参胚胎和幼体与其他种类的海参在发育时间上存在一定差异。     

日本米虾胚胎发育及离体孵化

本研究采用自制离体孵化装置，对日本米虾(Caridina japonica)不同发育期胚胎进行离体孵化研究，结果显示，水温为25.5℃时，日本米虾受精卵孵化大约需要25 d，发育积温为637.5℃。胚胎发育历经受精卵、卵裂期、囊胚期、原肠胚期、前无节幼体期、后无幼体期、前溞状幼体期和膜内溞状幼体期8个时期。各期离体胚胎均能孵化出幼体，膜内溞状幼体期离体胚胎孵化率最高，为(80.7±2.4)%，非离体孵化的对照组为(79.1±4.9)%，二者差异不显著；卵裂期离体胚胎孵化率最低，为(28.2±2.6)%，与对照组差异显著。各组离体胚胎所孵化出的Ⅰ期(ZⅠ)和Ⅱ期溞状幼体(ZⅡ)的变态率无显著差异。温度对日本米虾前溞状幼体期胚胎离体孵化影响显著，在15.0℃~32.5℃范围内，随水温升高孵化时间逐渐缩短，15.0℃时，前溞状幼体离体孵化时间为(436.8±124.8) h，32.5℃时缩短至(228.0±88.8) h，但温度高于29.0℃时，孵化出的幼体变态率开始下降。本研究可为日本米虾繁殖生物学及甲壳动物胚胎离体孵化技术研究提供参考。     

长臀性腺发育及其组织结构观察

采用常规石蜡组织切片和长期数据统计观察分析,对长臀精卵巢发育和周年变化特点以及其繁殖习性进行了探讨。依据精卵巢外形以及组织学特征,将长臀性腺发育分为6个时期,其中Ⅰ期性腺只出现在一龄以内的个体,Ⅱ期性腺将持续一年左右时间,经过Ⅲ期、Ⅳ期约一年的发育,长臀进入Ⅴ期发育至成熟并首次产卵排精。性成熟个体精卵巢发育只有后五期。长臀约3龄达性成熟,繁殖期是4—6月,5月为繁殖高峰期。长臀个体性腺发育不同步,但为一次产卵型。     

Embryonic and larval development of black skirt tetra (Gymnocorymbus ternetzi,Boulenger, 1895) under laboratory conditions

The embryonic and larval development of black skirt tetra, Gymnocorymbus ternetzi, are described under controlled laboratory conditions. In addition, major histomorphological changes and the allometric growth patterns during larval development have been described. The laboratory‐reared broodstock, that is 1 year of age, were spawned. Hatching occurred 20–21 h after spawning at 24 ± 0.5°C. The cleavage was finished in 2 h and the early blastula stage occurred at 2:04 hours after spawning. The gastrulation started at 3:20 hours and 30% epiboly was observed at 3:34 hours after spawning. Eight‐somite stage was observed at 08:33 hours. And embryonic developmental stage was completed at 21 h after spawning. The newly hatched larvae were 1442 ± 14.3 μm in mean total length (TL). The mouth opened at 3 days after hatching (DAH). The yolk sac had been totally absorbed and the larvae started to swim actively within 3–4 days. Notochord flexion began at 11 DAH. The metamorphosis was completed and the larvae transformed into juveniles at 32 DAH. In this paper, the full developmental sequence from egg to juvenile of G. ternetzi is described for the first time.     

仿洄游培育美洲鲥亲本的性腺发育

为解决美洲鲥(Alosa sapidissima)人工培育亲本性腺发育不佳的问题,开展了美洲鲥亲本仿洄游的培育,研究观察了美洲鲥亲本卵巢和精巢不同发育阶段(II^V期)的形态特征及相关参数,确定了仿洄游培育美洲鲥亲本性腺初次发育的特征。结果显示:在美洲鲥仿洄游培育过程中,在卵巢、精巢发育前期(II期),雌雄亲本生长迅速;而性腺发育到III期后,雌雄亲本的生长滞缓,雌鱼亲本甚至出现负生长。仿洄游培育美洲鲥性腺初次发育(II^V期)过程中,雌雄亲本均有较高的肥满度,分别为1.55~2.07和1.56~1.71;卵巢发育Ⅴ期雌鱼亲本的肥满度(2.07)显著高于在卵巢II^IV期亲本(1.55~1.67);而雄鱼亲本肥满度在精巢发育各阶段(II^V期)均无显著性差异。仿洄游培育美洲鲥性腺发育良好,雌雄亲本的性腺指数均较高,性腺发育Ⅴ期雌雄亲本的性腺指数(GSI)分别为(26.10±6.46)(18.88~33.62)和(5.18±0.68)(4.49~6.11)。研究表明,仿洄游培育的水温(10.8~30.8℃)和盐度(1~15)的周年变化符合美洲鲥亲本性腺发育的要求。     

Studies on the broodstock production and larval rearing of Coral demoiselle Neopomacentrus nemurus (Bleeker 1857)

Steps taken for broodstock development of Neopomacentrus nemurus, Coral demoiselle (Bleeker, 1857) in a hatchery, its spawning and development of egg to the juvenile stage are described. Among the three treatments tried, only the trial using Parvocalanus crassirostris nauplii as the first diet helped in the development of larvae. Larval and post‐larval growth was studied for a period of 65 days post hatch. The larva measured 2–2.4 mm in total length at the time of hatching. The mouth size of larva at the time of hatching measured 237.92 µm. By the 5th day, the width of the larval body had significantly increased. All the fins were conjoined, except for the caudal fin which had begun its appearance on the 2nd day itself. The digestive system was functional by the 5th day and the copepod remains were seen in the digestive tract. The larva metamorphosed by the 15th day with fully developed fins and fin rays. On the 20th day, the larval body had begun to display signs of fin pigmentation. The larva assumed adult pigmentation by the 35th day. By the 65th day, the larva grew to a total length of about 27 mm and weighed about 0.56 g.     

Importance of broodstock holding temperature on fecundity and egg quality in three groups of photo‐manipulated Atlantic cod broodstock

We conducted a study to evaluate the effects of holding water temperature on the fecundity and egg quality of photo‐manipulated Atlantic cod broodstock. Adult cod Gadus morhua were distributed among several 25‐m³ tanks. Three separate photoperiods were used for each group to obtain three spawning per year (in May, July and December). While photoperiod was controlled, the temperature was not controlled and followed the ambient seasonal pattern. Due to the uncontrolled seasonal temperature used, each spawning group experienced different temperatures at a particular gonadal developmental stage. Otherwise, all groups were treated the same. Results showed that the May and July spawning groups released significantly more eggs per kg of fish (four and three times more respectively) than the December spawning group. The egg fertilization success and proportion of normal eggs were also higher in the May and July groups. Possible reason for the difference may be that the May and July groups experienced lower temperatures (4–7°C) during oocyte maturation and ovulation than the December group (7–9°C). Our results show the importance of using lower/natural temperatures during oocyte maturation and ovulation to obtain good quality eggs in photo‐manipulated cod broodstock.     

Embryonic development of Ganga river prawn Macrobrachium gangeticum (Bate)

Macrobrachium gangeticum (Bate) are commercially valuable prawns that are broadly distributed in the Ganga river that flows from northern to eastern India. We studied embryonic development from cleavage (blastomeres), segmentation, formation of optic vesicle, eye pigment development to larva formation, as these data are largely unknown in this species. We conducted this study using mature M. gangeticum broodstock placed in freshwater tanks. Just after spawning, females were kept in separate aerated tanks, where fecundity was estimated from the total number of eggs. Thereafter, egg samples from berried females were used to study their embryonic stages. The correlation coefficient between size and prawn weight, egg weight, egg number, body weight and egg mass of different sizes of prawns was analysed. Our results show that an increase in prawn size led to an increase in egg weight and egg number while decreases in body weight:egg mass ratio was noticed. Further results indicate that an increase in prawn weight had a positive impact on egg weight and egg number. The embryo development stages were used in this study as a tool to evaluate the development process of the prawn in a hatchery environment.     

Maturing, Spawning and Egg Collecting of the White Shrimp Penaeus vannamei Boone in a Recirculating System

A specially designed recirculating system was used to maintain P. vannamei broodstock for maturation, mating and spawning in the same tank. Two groups of different size shrimp were stocked in this system in Phase I (8 May to 5 September 1989) and Phase II (5 September to 5 December 1989). Results showed that this recirculating system was able to maintain good water quality keeping broodstock maturing, mating and spawning with high stocking density (10 shrimp/m²) and low mortality (6%). Seventy-eight percent of spawned eggs were collected by a submerged egg collector. Mating success was very poor (3.4%) in Phase I even though there was normal spawning frequency. Mating success and nauplii production were much improved in Phase II. During Phase II 104 male and female broodstock housed in a 6 m³ tank produced 1.6 ± 10⁶ nauplii (with 70% mating success) in three months. This study illustrated that broodstock size was a critical factor of successful mating and fertilization for P. vunnamei , and also, reduction of handling stress and lowered disturbance of the shrimp greatly enhanced the mating success.     

太平洋鳕(Gadus macrocephalus)亲鱼驯化培育与早期发育特征

采捕山东威海外海黄海海域的太平洋鳕(Gadus macrocephalus)亲鱼进行驯化和培育,在人工条件下成功驯化存活野生亲鱼49尾,经短期促熟培育后,通过人工授精方式获得了多批次受精卵.对胚胎和早期仔鱼发育过程进行了观察,详细描述了从受精卵到早期仔鱼各发育时期的形态特征.结果显示,太平洋鳕成熟卵子为沉性卵,圆球形,卵径为0.9-1.1 mm,无油球.胚胎发育分为5个时期,分别为卵裂期、囊胚期、原肠胚期、神经胚期和器官形成期.在水温为9-10℃、盐度为27-29的海水中孵化,受精卵历时312 h 30 min孵化出膜.初孵仔鱼全长为(3.85±0.12) mm,6日龄仔鱼开口,肛门与外界相通,进入混合营养期.8日龄仔鱼卵黄囊消耗殆尽,开始进入外源性营养阶段.仔鱼开口饵料为轮虫(Rotifer),12日龄开始摄食卤虫(Artemia saline)无节幼体.6日龄仔鱼鳔原基形成,16日龄鳔充气成为亮泡状.12日龄仔鱼形成肠道第1个生理弯曲,22日龄仔鱼第2个肠道生理弯曲形成.研究结果可为太平洋鳕亲鱼驯化培育和苗种培育提供基础资料.     

盐度对瘤背石磺不同部位Na /K -ATP酶活性、围心腔液和腹腔液渗透压及离子含量的影响

为了研究盐度对瘤背石磺不同部位Na+/K+-ATP酶活性、围心腔液和腹腔液渗透压及离子含量(Na+、K+和Cl-)的影响,选取健康的瘤背石磺[湿重(10.24±2.16)g]600只,在不同盐度条件下(5、15、25、35和45)采用模拟生态环境的方式饲养30 d后测定各项指标.结果显示:(1)瘤背石磺腹腔液和围心腔液的盐度和渗透压均随着环境盐度的升高而增高,腹腔液的盐度和渗透压的变动幅度分别为25～ 43和514 ～ 796 mOsm/kg,其增高幅度均大于围心腔液(分别为23 ～ 28和502～ 726 mOsm/kg);(2)围心腔液中Na+、Cl-和K+含量随环境盐度的变化具有一致性,均呈先上升后下降的趋势,且各离子浓度均在盐度25时达到最大值,分别为295.50、322.50和5.15 mmol/L,但盐度对Na+和Cl-含量有显著影响,对K+含量无显著影响.(3)瘤背石磺不同部位的Na +/K+-ATP酶活性具有组织特异性,贲门胃、肠、表皮、肌肉和围心腔膜等部位的Na +/K+-ATP酶活性较高,同时,盐度激活不同部位的Na +/K+-ATP酶活性具有显著差异性,肌肉和贲门胃的Na +/K+-ATP酶在低盐度时活性较高,而肠、肾及围心腔膜等在高盐度时Na+/K+-ATP酶被激活,酶活性较高.研究表明:盐度能显著影响瘤背石磺腹腔液和围心腔液的渗透压和离子浓度以及各器官的Na+/K+-ATP酶活性,贲门胃、肠和肌肉等均是其离子转运和渗透调节的主要部位,具有较强的渗透压调节能力.     

Breeding and larval rearing of the camouflage grouper Epinephelus polyphekadion (Bleeker) in the hypersaline waters of the Red Sea coast of Saudi Arabia

This is the first report on the successful year-round natural spawning and larval rearing of Epinephelus polyphekadion (Bleeker) in captivity and under hypersaline water conditions of 42-43%₀ salinity in the Red Sea coast of Saudi Arabia. Although the fish spawned naturally once or twice a year during 1992-94 culture period, incorporation of cod-liver oil in the broodstock diet during the 1995 culture period enabled the fish to spawn continuously for 2-3 days in each month during March, April, May and August. The egg fertilization and hatching rates also increased during the 1995 spawning period. The egg fertilization rate varied from 90 to 100% with a mean of 96.5 ± 3.38%. The egg hatching rate varied from 70 to 95% with a mean of 83.1 ± 10.12%. The fertilized egg diameter averaged 757.3 ± 37.36 μm. There was a linear relation between the fertilized egg size and the egg hatching rate. The increase in the hatching rate relevant to the egg size was statistically significant (P < 0.01). The egg development time until hatching lasted for 19 h at 29°C. The newly hatched larval size ranged from 1.55-1.71 mm with a mean of 1.65 ± 0.052 mm in total length. The larval growth was slow in the early stages and the growth curve until metamorphosis showed a curvilinear pattern. Wide variations in larval size, range 22-47 mm with a mean of 33.40 ± 7.01 mm, were observed during the metamorphosis stage at day 50. No significant difference (P > 0.05) in growth and survival was observed between the larvae reared using white and grey coloured tanks. The larval survival up to metamorphosis was 1.6-4.7% with a mean of 2.98 ± 1.56% in the grey coloured tanks and 1.6-1.9% with a mean of 1.73 ± 0.16% in the white tanks. The results demonstrated the possibility of breeding E. polyphekadion under captive culture conditions. However, methods to improve the larval survival have to be pursued further for commercial farming of this species.