中华绒螯蟹幼体及仔蟹阶段肝胰腺发育的组织学研究

通过对中华绒螯蟹幼体及仔蟹进行连续采样和组织切片,系统地研究了该阶段中华绒螯蟹肝胰腺的发育情况。结果表明,中华绒螯蟹的肝胰腺前体出现于心跳期胚胎,由卵黄囊特化而来,为1对囊状结构,此阶段首次出现肝胰腺细胞(柱状上皮细胞)。胚胎孵化后,随着幼体的发育,肝胰腺由最初的1对囊状结构逐渐分叶生长。从Ⅰ期溞状幼体至Ⅴ期溞状幼体,肝胰腺分叶数量分别为4-6-8-12-12,肝胰腺细胞高度由(12.63±4.91)μm增至(44.16±13.57)μm。肝胰腺中的细胞组成也随着幼体的发育逐渐完善,Ⅱ期溞状幼体的肝胰腺中首次出现E细胞和B细胞,Ⅲ期溞状幼体肝胰腺中首次出现F细胞,而R细胞直到Ⅱ期仔蟹才首次出现。从大眼幼体到Ⅲ期仔蟹,肝胰腺细胞高度由(44.93±18.27)μm增至(56.38±5.69)μm。     

中华绒螯蟹Timeless基因在不同发育阶段的表达谱分析

Timeless基因是重要内源性的生物钟基因之一,在生物体内周期性表达,保持昼夜节律时间并激活输出途径。本文分析了中华绒螯蟹Timeless基因在其溞状幼体Ⅰ~Ⅴ期、大眼幼体期和仔蟹Ⅰ~Ⅲ期共9个不同发育阶段以及扣蟹、成蟹(性成熟)不同组织间的mRNA表达情况。荧光定量PCR结果揭示Timeless基因在溞状幼体Ⅰ期至仔蟹Ⅲ期9个发育阶段均有表达,在溞状幼体Ⅴ期的表达量最高,显著高于其他时期(P0.05),随后在大眼幼体时期急剧下降,最后仔蟹时期又呈稳步上升趋势。在扣蟹时期的8个组织中,Timeless基因在肝胰腺和肌肉中的表达量较高,与其他组织的表达均呈显著差异(P0.05);在成蟹时期肝胰腺中的表达量最高且与其他组织中的表达水平均差异显著(P0.05)。研究结果表明,中华绒螯蟹Timeless基因在幼体发育阶段均有表达,组织表达主要在肝胰腺和肌肉中,并且在不同性别的成蟹组织间Timeless mRNA表达存在显著差异。推测该基因可能在中华绒螯蟹的生长发育、生理适应和代谢调控过程中具有重要作用。     

蟹苗运输及放养初期关键点剖析

中华绒螯蟹,俗称河蟹、大闸蟹,是我国淡水渔业的重要养殖经济品种。河蟹通过海水繁殖,第Ⅴ期溞状幼体经3~5 d发育蜕皮后即变态为大眼幼体,俗称蟹苗。大眼幼体呈龙虾型,既可游泳、又可爬行,体形扁平,胸足5对,具有大螯和口器,杂食偏肉食性,可主动捕食大型浮游动物。大眼幼体已具备较强的渗透压调节能力,经过几天的逐步淡化     

一株中华绒螯蟹幼体益生菌的筛选及其种属特性的初步研究

通过单株细菌感染中华绒螯蟹Ⅰ期溞状幼体的方法筛选出2株病原菌,它们对幼体的72 h致死率为100%;获得有益菌1株,与对照组相比提高中华绒螯蟹Ⅰ期溞状幼体的变态率近50%,该菌株有望成为中华绒螯蟹幼体的益生菌株.根据16S rRNA序列同源性分析结果,将其初步鉴定为节杆菌属的一种.     

中华绒螯蟹幼体的栖息生境选择与行为特征

为探究中华绒螯蟹在早期发育阶段对不同栖息生境的选择偏好及其行为特征，实验以长江口近岸微生境类型为基础，在室内建立了藨草、泥底、铁板沙、砾石等4种模拟生境，选取中华绒螯蟹大眼幼体、Ⅰ、Ⅲ、Ⅴ期仔蟹分别开展了对不同生境选择的行为观察和定量分析。结果显示，中华绒螯蟹大眼幼体、Ⅰ、Ⅲ、Ⅴ期仔蟹在藨草生境中分布比例最高，显著高于其他三种生境。随着仔蟹的生长发育，Ⅲ、Ⅴ期仔蟹在泥底和铁板沙生境中占比增大。在藨草生境中，中华绒螯蟹幼体具有游泳、附着藨草、表栖、埋栖等4种行为，大眼幼体和Ⅰ期仔蟹主要表现为附着藨草行为，Ⅲ期仔蟹的表栖、埋栖行为显著增加。幼体在藨草生境中的蜕壳率显著高于泥生境，而死亡率显著低于泥生境。研究表明，中华绒螯蟹幼体偏好栖息于有利于其生存、发育的藨草等植被生境中，这可能与藨草等植被生境为中华绒螯蟹幼体提供了适宜的庇护场所有关。本研究结果指出了植被生境对中华绒螯蟹幼体阶段的重要性，丰富了早期生活史资料，并为其资源保护和栖息地生态修复提供了基础参考。     

对中华绒螯蟹生态育苗池中桡足类控制的初步研究

比较了蛛虫速灭、鱼虫恨和强力混杀3种渔药对桡足类和中华绒螯蟹溞状幼体的急性毒性。试验结果表明,蛛虫速灭对桡足类的96h半致死添加量为0.011mL/m3,鱼虫恨为0.036mL/m3,强力混杀精为0.014mL/m3,蛛虫速灭对桡足类的杀灭能力最强;对中华绒螯蟹溞状幼体Ⅰ期的急性毒性鱼虫恨最大,蛛虫速灭最小。蛛虫速灭在0.18～0.20mL/m3的剂量下,对溞状幼体毒性低,幼体能正常变态,可用于蟹苗池桡足类的控制。强力混杀精对桡足类和中华绒螯蟹溞状幼体的毒性很接近,鱼虫恨对中华绒螯蟹溞状幼体的毒性大于对桡足类的毒性,两者均不适宜在中华绒螯蟹生态池控制桡足类。     

中华绒螯蟹人工育苗中的几个技术问题探讨

对中华绒螯蟹人工育苗中亲蟹的准备,Ｓｈｏｕ状幼体Ⅰ期到Ⅱ期的变态及Ｓｈｏｕ状幼体Ⅴ期到大眼幼体的变态等关键环节中应注意的问题进行了探讨。     

环境条件对中华绒螯蟹大眼幼体蜕壳生长的影响

试验结果表明,黑暗条件下大眼幼体蜕壳数量少、成活率低;光照可刺激并促进大眼幼体的蜕壳与生长,但长时间光照会影响Ⅰ期到Ⅳ期幼蟹的蜕壳生长;泥土和隐蔽物作为栖息环境有利于提高中华绒螯蟹幼体的成活率;苗种质量是提高中华绒螯蟹大眼幼体到Ⅳ期幼蟹成活率的关键。     

中华绒螯蟹Ⅰ期幼蟹上岸死亡原因初探

中华绒螯蟹Ⅰ期幼蟹上岸死亡原因初探李晓晴贾学文黄俊章（营口市老边区水产局关键词：Ⅰ期幼蟹上岸死亡原因生产中常有河蟹大眼幼体蜕皮变态为Ⅰ期幼蟹,约４天后上岸死亡的现象。死亡样体亦无病原生物感染或寄生。经解剖发现,胃中无食物,死亡时多为步足撑地,呈脱壳状...     

基于COⅠ序列绒螯蟹属DNA条形码和遗传多样性研究

对绒螯蟹属的中华绒螯蟹如东和七里海群体、日本绒螯蟹、合浦绒螯蟹及狭颚绒螯蟹共80条线粒体COⅠ片段进行扩增和测序,并与GenBank中绒螯蟹属的台湾绒螯蟹2条和近方蟹属的绒毛近方蟹19条COⅠ基因序列进行联配分析。结果显示,101条序列包含44种单倍型,序列组成表现明显的碱基偏倚性。中华绒螯蟹如东、七里海群体与日本绒螯蟹间的遗传距离分别为1.210%和1.078%,明显低于COⅠ基因DNA条形码鉴别种的遗传距离为2%的阈值,表明中华绒螯蟹和日本绒螯蟹为同一物种;而合浦绒螯蟹与中华绒螯蟹如东和七里海群体及与日本绒螯蟹的遗传距离分别为4.823%、5.101%以及5.011%,明显大于2%的鉴别阈值,说明合浦绒螯蟹为独立的种。以绒毛近方蟹为外群,基于群体内及群体间的遗传距离构建的邻接树显示,中华绒螯蟹与日本绒螯蟹聚在一起,合浦绒螯蟹则聚成单系。本文测序的5个群体除狭颚绒螯蟹外,其余均具有遗传多样性,单倍型多样性为0.593±0.144~0.779±0.068,核苷酸多样性为0.00156~0.01336;此外,中华绒螯蟹如东群体与日本绒螯蟹、合浦绒螯蟹和中华绒螯蟹七里海群体分别共享单倍型H1、H2和H3,说明这些蟹类可能有种质资源混杂或是遗传污染的现象。     

Ontogeny of feeding apparatus and foregut of mud crab Scylla serrata Forsskål larvae

The development of the feeding apparatus of the mud crab Scylla serrata larvae was studied using electron microscopy for mandibles and light microscopy for other paired mouthparts and the foregut. The six paired mouthparts, which consisted of the mandibles, maxillules, maxillae, first maxillipeds, second maxillipeds and third maxillipeds, were dissected from specimens representing each larval stage. The first five paired appendages were already present in newly hatched larvae while third maxillipeds appeared only at the megalopa stage. Mandibles displayed complex incisor and molar processes at early zoeal stages, which became simple in morphology at megalopa. Mandibular palp buds were observed at the zoea 5 stage and these became fully developed as three‐segmented mandibular palps at the megalopa stage. Endopods of other paired mouthparts exhibited increased number of setae and size as the individual metamorphosed from zoeal stages to megalopa and crab instar. The foregut appeared as a continuous cavity at zoea 1 where the cardiopyloric valve was indistinct while the filter gland was clearly identifiable. Zoea 2 and succeeding zoeal stages exhibited a setose foregut; the gastric mill and its lateral and median teeth were prominent at zoea 3 stage. The significance of these morphological changes is discussed in terms of its implication in larval feeding management.     

中华绒螯蟹(Eriocheir sinensis)种苗土池培育技术研究Ⅰ-Ⅰ期溞状幼体的适宜密度

在６口海水土池（计１．９ｈｍ２）中分别按下列密度（万／ｍ３）布苗：３,４,５,３,４和０．２７,比较研究了中华绒螯蟹（Ｅｒｉ－ｏｃｈｅｉｒｓｉｎｅｎｓｉｓ）Ⅰ期氵蚤状幼体（Ｚ１）的密度对幼体变态率和成活率的影响。结果表明,Ｚ１的密度为４万／ｍ３时,大眼幼体的变态率（大眼幼体密度占Ｚ５密度的％;２１．６％）和成活率（大眼幼体的密度占Ｚ１密度的％;２．４％）及产量（４２ｋｇ／ｈｍ２）最高。     

Effects of Starvation and Feeding on Lipid Class and Fatty Acid Profile of Late Stage Mud Crab,Scylla serrata,larvae

Lipid class and fatty acid (FA) analysis were conducted on newly molted, fed, and starved zoea V and megalopa of the mud crab, Scylla serrata (S. serrata). Larvae starved for 4 d showed a substantial decrease in total FA content, from 49.67 μg/mg to 13.94 μg/mg ash‐free dry weight (AFDW) at the zoea V stage, and from 38.47 μg/mg to 10.40 μg/mg AFDW at the megalopa stage. This depletion indicates that S. serrata larvae effectively utilize stored lipid reserves for energy during periods of food deprivation. Megalopa subjected to longer starvation periods, however, did not utilize lipid as the major energy source after day 4, suggesting increased reliance on protein catabolism during prolonged starvation. At both larvae stages the major FAs were 18:1n‐9, 16:0, 20:5n‐3 (eicosapentaenoic acids, EPA), 18:3n‐3 (linolenic acid, LNA), 18:0 and 22:6n‐3 (docosahexaenoic acid, DHA) and this FA profile persisted in both fed and starved larvae. The highly unsaturated fatty acids (HUFA), EPA, DHA, and arachidonic acid (20:4n‐6, AA) were not conserved in tissue during starvation, indicating that HUFA requirements might be lower for S. serrata larvae than shown for other crustaceans. Similarly, a high level of LNA in newly molted zoea V and megalopa were rapidly depleted in unfed larvae, indicating that this FA had an important role as an energy reserve. Throughout the study, FAs from the polar lipid fraction dominated larvae tissues, while FAs from the neutral lipid constituted the largest accessible energy reserve during starvation (depleted from 23.05 to 1.23 μg/mg AFDW in zoea V, and from 19.00 to 1.27 μg/mg AFDW in megalopa). The results of this study provide new insight into lipid utilization of S. serrata larvae during development, an important step toward development of formulated diets for use in mud crab hatcheries.     

The effect of dietary n-3 HUFA levels and DHA/EPA ratios on growth, survival and osmotic stress tolerance of Chinese mitten crab Eriocheir sinensis larvae

The effect of varying levels of dietary n-3 highly unsaturated fatty acid (HUFA) and docosahexaenoic acid/eicosapentaenoic acid (DHA/EPA) ratios on growth, survival and osmotic stress tolerance of Eriocheir sinensis zoea larvae was studied in two separate experiments. In experiment I, larvae were fed rotifers and Artemia enriched with ICES emulsions with 0, 30 and 50% total n-3 HUFA levels but with the same DHA/EPA ratio of 0.6. In experiment II, larvae were fed different combinations of enriched rotifers and Artemia, in which, rotifers were enriched with emulsions containing 30% total n-3 HUFA, but different DHA/EPA ratio of 0.6, 2 and 4; while Artemia were enriched with the same emulsions, but DHA/EPA ratio of 0.6 and 4. In both experiments, un-enriched rotifers cultured on baker's yeast and newly-hatched Artemia nauplii were used as control diets. Larvae were fed rotifers at zoea 1 and zoea 2 stages; upon reaching zoea 3 stage, Artemia was introduced.Experiment I revealed no significant effect of prey enrichment on the survival of megalopa among treatments, but higher total n-3 HUFA levels significantly enhanced larval development (larval stage index, LSI) and resulted in higher individual dry body weight of megalopa. Furthermore higher dietary n-3 HUFA levels also resulted in better tolerance to salinity stress. Experiment II indicated that at the same total n-3 HUFA level, larvae continuously receiving a low dietary DHA/EPA ratio had significantly lower survival at the megalopa stage and inferior individual body weight at the megalopa stage, but no negative effect was observed on larval development (LSI). The ability to endure salinity stress of zoea 3, zoea 5 and megalopa fed diets with higher DHA/EPA ratio was also improved.     

梭子蟹全人工工厂化育苗技术

2000年笔者在日照市水产研究所利用1450m^3水体进行梭子蟹全人工工厂化育苗生产，共出Ⅱ-Ⅲ期幼蟹512．7万只，创造了出苗量为3500只／m^3，总产为150多万元的直接经济效益，同时对幼体培育的密度进行了初步试验，结果表明，从Z1至M期幼体培育密度10－15万/m^3为宜，M期以后密度为0．8－1．5万/m^3为宜。     

三疣梭子蟹卵巢发育过程中雌二醇的免疫定位和变化

雌二醇(estradiol,E_2)是甲壳动物体内重要的性类固醇激素,对其卵巢发育和卵黄发生起着十分重要的调控作用。本实验采用免疫组织化学方法系统研究了三疣梭子蟹(Portunus trituberculatus)卵巢发育过程中E_2在卵巢、肝胰腺、胸神经节、脑神经节和大颚器中的分布及变化。结果表明:(1)蟹卵巢中E_2的免疫阳性主要分布于滤泡细胞和卵巢发育中后期的卵母细胞细胞质(II-V期),卵巢发育过程中卵母细胞细胞质和细胞核中的E_2免疫阳性均为先上升后下降的趋势,最大免疫阳性分别出现在IV期和II-III期,滤泡细胞中始终存在较强的E_2免疫阳性;(2)肝胰腺中的E_2免疫阳性主要分布于F细胞及R细胞细胞核中,R细胞细胞核中的E_2免疫阳性从III期开始显著下降,F细胞中始终存在较强的E_2免疫阳性;(3)卵巢发育(I-V期)的神经组织,E_2主要分布于胸神经团的神经细胞和神经髓质以及脑神经节神经细胞细胞核内,其中脑神经节神经细胞细胞核中始终保持着强免疫阳性,卵巢发育早期(I-II期)胸神经团中的神经髓质中为E_2中等免疫阳性,神经细胞中一直为弱免疫阳性;(4)就大颚器而言,E_2强免疫阳性始终存在于大颚器腺细胞的核仁以及细胞核周围的细胞质内。以上结果表明,E_2在三疣梭子蟹卵巢、肝胰腺、神经组织和大颚器中广泛分布,且免疫阳性与卵巢发育具有一定的相关性,E_2可能通过作用于多个靶器官来调控三疣梭子蟹的卵巢发育。     

Effect of starvation during late megalopa stage of Mithraculus forceps (Brachyura: Majidae) on larval duration, synchronism of metamorphosis, survival to juvenile, and newly metamorphosed juvenile size

Larval resistance to temporary starvation is considered a key factor for successful development in the wild. Subjecting larvae to temporary starvation during early and/or late development is occasionally used in larviculture to reduce production costs.Mithraculus forceps is a popular species in the marine aquarium industry for their ability to control nuisance algae in aquarium tanks; a larval culture methodology was previously proposed in order to avoid collection from the wild. In an attempt to reduce production/feeding costs of M. forceps larval culture (two zoea stages and a megalopa), larvae (megalopa stage) were starved after 7 and 8 days post-hatch (DPH); starvation treatments were compared with those of the control treatment where larvae were fed continuously. No differences were found in survival to juvenile, metamorphosis synchronism or larval duration between the treatments, which suggest that at least by day 7 DPH, megalopae have already achieved the point-of-reserve-saturation (PRS) and are able to successfully metamorphose to crab stage without feeding (facultative lecithotrophic); newly metamorphosed juveniles are slightly smaller (particularly the ones starved after 7DPH). This flexible way of development might be very advantageous in the wild and might allow the reduction of costs of larval culture in captivity.     

育苗水中添加微生物对河蟹苗成活率的影响

在中华绒螯蟹的育苗过程中，采用添加微生物的方法调节育苗水体的水质，结果表明：与对照池相比，试验池中的氨氮明显下降，溶氧量增加，pH值变化不大；河蟹各期幼体的成活率提高，从SouⅠ期幼体至大眼幼体，试验池的成活率为13.9%，对照池仅8.1%。因此，在河蟹的育苗生产上，利用添加有益微生物的水质循环装置，能有效地提高育苗产量，节约成本。     

两品系褶皱臂尾轮虫种群增长及投喂河蟹苗效果的研究

采用实验生态学方法对两品系褶皱臂尾轮虫(内蒙品系和盘锦品系)的种群增长和投喂蟹苗的效果进行了比较研究。结果表明,两品系轮虫的种群增长差异不显著,两者平均瞬时增长率分别为0.42 d-1和0.40 d-1。在投喂蟹苗的效果方面,褶皱臂尾轮虫内蒙品系好于盘锦品系。用内蒙品系轮虫投喂的蟹苗,幼体变态至大眼幼体的存活率为71.65%,用盘锦品系轮虫投喂的蟹苗,变态至大眼幼体的存活率为62.50%。     

The effect of poly‐β‐hydroxybutyrate on the performance of Chinese mitten crab (Eriocheir sinensis Milne‐Edwards) zoea larvae

Two experiments were conducted regarding the effect of poly‐β‐hydroxybutyrate (PHB) on the survival and development of Chinese mitten crab (Eriocheir sinensis Milne‐Edwards) larvae. Different PHB delivery approaches (particulate, enriched in filter feeding prey, or a combination of both) and feeding levels were applied to crab larvae from the zoea 2 (Z2) up to the megalopa stage. Bio‐encapsulation into rotifers and Artemia at a PHB dose of 100 mg L^?1 seemed an efficient approach to deliver PHB to the larvae and resulted in a significantly enhanced survival, development rate and osmotic tolerance. The results indicate that PHB should only be applied as a feed additive rather than as a food source, and that the availability of an adequate amount of nutrients seems to be of major importance for the PHB to induce these beneficial effects.