中华绒螯蟹（Eriocheir sinensis）肌肉中粗脂肪含量与死亡,产 …

本文报道了中华螯蟹越冬时,越冬后不同性别、不同生活状况肌肉中粗脂肪含量及变化。越冬时、越冬后死亡的♂♀河蟹肌肉中粗脂肪的含量均不到８％,而同时越冬成活的♂♀蟹肌肉中粗脂肪含量均高于１０％。顺利越冬并能产卵的雌蟹肌肉中粗脂肪含量为４．９６％,顺利越冬并交配的♂性肌肉中粗脂肪含量为４．１８％。产卵并抱卵孵化的♀蟹肌肉中粗脂肪含量为５．８５％,产卵但♀蟹中途死亡的其肌肉中粗脂肪的含量为５．４７％。交配不     

中华绒螯蟹（Eriocheir sinensis）体内维生素C含量与死亡和产卵的关系

本文报道了中华绒螯蟹越冬后肝胰脏、肌肉和生殖腺维生素Ｃ的含量及变化。肝胰脏为中华绒螯蟹储存维生素Ｃ的器官;越冬后的雌性中华绒螯蟹,其肌肉和生殖腺的维生素Ｃ高于雄性;正常活动雌性或雄性,其肝胰脏、肌肉和生殖腺的维生素Ｃ含量均高于死亡的个体的维生素Ｃ含量;产卵且抱卵孵化的雌体,其肝脏维生素Ｃ的含量高于那些不产卵的雌体,维生素Ｃ含量的降低是越冬时或越冬后中华绒螯蟹死亡的主要原因之一。     

中华绒螯蟹（Eriocheir sinensis）体内维生素C含量与死亡和产…

本文报道了中华绒螯蟹越冬后肝胰脏、肌肉和生殖腺维生素Ｃ的含量及变化。肝胰脏为中华绒螯蟹储存维生素Ｃ的器官;越冬后的雌性中华绒螯蟹,其肌肉和生殖腺的维生素Ｃ高于雄性;正常活动雌性或雄性,其肝胰脏、肌肉和生殖腺的维生素Ｃ含量均高于死亡的个体的维生素Ｃ含量;产卵且抱卵孵化的雌体,其肝脏维生素Ｃ的含量高于那些不产卵的雌体。维生素Ｃ含量的降低是越冬时或越冬后中华绒螯蟹死亡的主要原因之一。     

中华绒螯蟹(Eriocheir sinensis)精子顶体反应的研究

分别用卵水、海水、caCl_2或NaCl水溶液对中华绒螯蟹成熟精子进行人工诱导顶体反应,结果表明:精子的生理性成熟、同种卵或Ca~(++)的存在、碱性环境以及与一定的固体接触均为精子顶体反应触发的重要条件。3月份精子诱导率最高。 电镜观察证明,中华绒螯蟹精子的顶体反应可分四个阶段:(1)辐射臂收缩;(2)顶体囊外翻;(3)顶体管前伸;(4)片层结构脱落。     

中华绒螯蟹(Eriocheir sinensis)雄性生殖系统的组织学研究

中华绒螯蟹雄性生殖系统的组织学研究表明:生精小管一侧为管壁上皮,另一侧为生发区,生殖细胞由生发区基底部同管腔增殖。输精管分为输精细管和贮精囊,管壁上皮具分泌功能,贮精囊有肌肉层。射精管壁肌肉层较厚,粘膜形成纵行皱襞。副性腺内壁为单层立方上皮。生殖系统发育有明显的季节性,8月开始发育加速,10月进入高峰,4月开始发育停滞。     

饥饿对中华绒螯蟹(Eriocheir sinensis)幼体发育的影响

对刚孵化的中华绒螯蟹第一期的蚤状幼体经不同时间的饥饿后再投喂,发现饥饿可以明显降低幼体的存活率和延长幼体的发育期。实验表明:对中华绒螯蟹第一期的蚤状幼体的饥饿时间(t)和发育期长(D)呈线性关系(D=4.6303+1.3226t r=0.970p<0.01)。对于中华绒螯蟹第一期的蚤状幼体,当起始饥饿时间超过了4d,再予以投饵,幼体均不能恢复正常的发育和蜕皮功能,得出中华绒螯蟹的不可恢复点(the point of no-return,PNR)大约为4d。通常以产生50%的幼体死亡的饥饿期即PNR₅₀,来表明幼体对饥饿的抵抗能力,实验得出中华绒螯蟹第一期的蚤状幼体的PNR₅₀大约为48h。     

中华绒螯蟹一龄幼蟹(Eriocheir sinensis)越冬前后脂肪酸和4种类胡萝卜素含量的变化

实验以越冬前后的中华绒螯蟹一龄幼蟹作为研究对象,利用高效气相色谱法分别检测一龄幼蟹越冬前后肝胰腺及头胸甲中脂肪酸含量变化、利用液相二级质谱法分别测定一龄幼蟹越冬前后肝胰腺、头胸甲和血淋巴中4种类胡萝卜素含量变化。结果表明:肝胰腺中的饱和脂肪酸(SFA) C20:0、单不饱和脂肪酸(MUFA) C18:1n9作为主要能量脂肪酸被动用,其含量越冬后显著下降(p0.05),多不饱和脂肪酸(PUFA)中的C18:3n3、C22:6n3越冬后含量显著升高(p0.05)。头胸甲的饱和脂肪酸(SFA) C16:0、C18:0,单不饱和脂肪酸(MUFA)中的C18:1n7作为主要能量脂肪酸被动用,越冬后含量下降显著(p0.05)。多不饱和脂肪酸(PUFA) C18:2n6、C20:3n6越冬后含量显著升高(p0.05)。越冬后头胸甲中4种类胡萝卜素含量均极显著下降(p0.01)。其中,血淋巴中虾青素、叶黄素含量极显著升高(p0.05),β-胡萝卜素含量极显著下降(p0.05)。肝胰腺中虾青素与叶黄素、β-胡萝卜素变化趋势与血淋巴的一致,角黄素含量显著下降(p0.05)。越冬期间气温、水温的短暂升高可能使一龄幼蟹短暂摄食。越冬前经育肥的扣蟹越冬后能量物质无显著变化,越冬后根据肝胰腺颜色来判断一龄幼蟹质量的方法不科学。     

中华绒螯蟹(Eriocheir japonica sinensis)线粒体ND5基因序列变异分析

不同水系河蟹已发现严重混杂,水产业中急需一种快速鉴别不同水系的分子标记。基于研究13条长约853bp的线粒体ND5基因序列分析得出,其平均碱基组成为40.2%A,30.9%T,9.2%G,19.7%C,共定义了8个单元型,单元型多样性(h)为0.885;序列间存在16个变异位点,核苷酸多样性(π)为0.004;平均遗传距离为0.004。其中单元型EJSⅠ和EJSⅡ占据了53.8%的个体数量,推测可能是较为原始类型;两单元型在NJ和UPG-MA系统发生树中分居于不同的分支,可能蕴涵着各自进化历史。通过研究表明,ND5序列的变异水平将为采用PCR-RFLP或位点特异性PCR等分子标记分辨不同水系河蟹起着重要作用。     

Development and characterization of new microsatellite loci from Chinese mitten crab (Eriocheir sinensis)

A set of new microsatellite loci were isolated from Chinese mitten crab and characterized in two Chinese mitten crab populations. The number of the alleles ranged from 3 to 8 and observed and expected heterozygosity varied from 0.0870 to 0.9565 and 0.3092 to 0.8367 in each population, respectively. Loci HLJEsin1, HLJEsin4, HLJEsin30 in CLN population and HLJEsin6 in CJS population presented significant deviation from Hardy–Weinberg equilibrium (PHWE < 0.001). These markers will be useful for population studies.     

Microsatellite analysis of genetic diversity and population structure of Chinese mitten crab （Eriocheir sinensis）

Chinese mitten crab (Eriocheir sinensis) has higher commercial value as food source than any other species of Eriocheir in China.To evaluate the germplasm resources and characterize the genetic diversity and population structure of the crabs in different water systems,two stocks and two farming populations were assessed with 25 polymorphic microsallite loci available in public GenBank.Basic statistics showed that the average observed heterozygosity (Ho) amongst populations ranged from 0.5789 to 0.6824.However,a remarkable presence of inbreeding and heterozygote deficiencies were observed.To analyze population structure,pairwise FST coefficients explained only ～10.3% variability from the subdivision of mitten crab populations,the remaining variability stems from the subdivision within subpopulations.Although the four populations had slight differentiation,different allelic frequencies resulted in distinct population structures.Two stocks and one farming population were clustered together to the phylogenetic branch of Yangtze crab,with an approximate membership of 95%.Whereas,another fanning population was clustered singly to the phylogenetic branch of the Liaohe crab,with a membership of 97.1%.The tests for individual admixture showed that Yangtze crab had probably been contaminated with individuals from other water systems.Genetic relationships between populations also supported the conclusion that Yangtze crab and Liaohe crab had different gene pools in spite of the origins of the same species.     

Phylogenetic analysis of intestinal bacteria in the Chinese mitten crab (Eriocheir sinensis)

AIMS: To identify the dominant intestinal bacteria in the Chinese mitten crab, and to investigate the differences in the intestinal bacteria between pond-raised and wild crabs. METHODS AND RESULTS: The diversity of intestinal bacteria in the Chinese mitten crabs was investigated by denaturing gradient gel electrophoresis (DGGE) fingerprinting, 16S rRNA gene clone library analysis and real-time quantitative PCR. The principal component analysis of DGGE profiles indicated that substantial intersubject variations existed in intestinal bacteria in pond-raised crab. The sequencing of 16S rRNA genes revealed that 90-95% of the phylotypes in the clone libraries were affiliated with Proteobacteria and Bacteroidetes. Some genera were identified as unique in wild crabs and in pond-raised crabs, whereas Bacteroidetes was found to be common in all sampled crab groups. Real-time quantitative PCR indicated that the abundance of Bacteroides and the total bacterial load were approximately four-to-10 times higher in pond-raised crabs than in wild crabs. A significant portion of the phylotypes shared low similarity with previously sequenced organisms, indicating that the bacteria in the gut of Chinese mitten crabs are yet to be described. CONCLUSIONS: The intestinal bacteria of pond-raised crabs showed higher intersubject variation, total diversity and abundance than that observed in wild crabs. The high proportion of the clones of Proteobacteria and Bacteroidetes in the clone library is an indication that these bacteria may be the dominant population in the gut of the Chinese mitten crab. SIGNIFICANCE AND IMPACT OF THE STUDY: This study demonstrated obvious differences in the intestinal bacterial composition of pond-raised crabs and wild crabs. This knowledge will increase our understanding of the effects of aquaculture operations on bacterial community composition in the crab gut and provide necessary data for the development of probiotic products for crab cultivation.     

饥饿胁迫对中华绒螯蟹(Eriocheir sinensis)仔蟹的影响

在26.2 ～28.4℃水温条件下,研究了饥饿对中华绒螯蟹(Eriocheir sinensis)仔蟹的形态、行为、存活及体重损失的影响,同时确定了仔Ⅱ的营养饱和储存点(PRS)和不可恢复点(PNR).结果表明:饥饿胁迫下中华绒螯蟹仔Ⅰ、仔Ⅱ、仔Ⅲ的初次死亡时间(T1)分别为8.0、14.0和20.3 d,50％死亡时间(Ts0)分别为11.4、16.0和25.5 d,100％死亡时间(T100)分别为15.0、22.0和32.3 d,耐饥饿能力为仔Ⅲ＞仔Ⅱ＞仔Ⅰ;饥饿期间中华绒螯蟹体内水分含量持续升高,干重量降低显著,干重损失速率也随时间延长逐渐减小;先饱食后饥饿的给饵模式中仔Ⅱ蜕皮率随初始饱食时间延长而升高,50％个体完成蜕皮所需饱食时间(PRS50)为2.10d,各处理组仔Ⅱ的蜕皮周期与持续饱食组均无显著差异(P＞0.05),但只有饱食3d以上才能达到持续饱食饲养蟹的增重率;先饥饿后饱食的给饵模式中,仔Ⅱ的蜕皮率随着初始饥饿时间的延长而下降,其中仔Ⅱ50％不能蜕皮的初始饥饿时间(PNR50)和100％不能蜕皮的初始饥饿时间(PNR100)分别为10 d和14 d,并且蜕皮周期相对延长,延长时间约等于初始饥饿时间,不存在额外的摄食时间来弥补饥饿期间损失的能量,各处理组蜕皮后仔蟹与对照组体重无显著差异(P＞0.05).     

An Experimental Study on the Gait Patterns and Kinematics of Chinese Mitten Crabs

Despite the many studies on eight-legged animals and the importance of their mechanics of terrestrial locomotion, the mechanical energy of crabs in voluntary locomotion on uneven, unpredictable terrain surfaces has received little attention thus far. In this paper, motion video images of Chinese mitten crab (Eriocheir sinensis Milne-Edwards) locomotion on five types of terrains were recorded using a high-speed three-dimensional (3D) recording video system. The typical variables of locomotion such as gait patterns, duty factor, mechanical energy of the mass center, mass-specific rate of the total mechanical power of the mass center, and percentage recovery, were analyzed. Results show that the Chinese mitten crab uses random gaits instead of the alternating tetrapod gait with the increasing terrain roughness. The duty factors of the rows of the leading legs are greater for all terrains than those of the rows of the trailing legs. On smooth terrain, the duty factors of the rows of the trailing legs are greater than that on rough terrains. Kinematic measurements and calculations reveal that similar to mammals, birds, and arthropods, the Chinese mitten crab uses two fundamental gaits to save mechanical energy: the inverted pendulum gait and the bouncing gait. The bouncing gait is the main pattern of mechanical energy conservation. The low probability of injury and energy expenditure due to adaptations to various terrains induce the Chinese mitten crab to modify the mass-specific rate of the total mechanical power of the mass center. The statistical results of percentage recovery also reveal that the Chinese mitten crab has lower energy recovery efficiency over rough terrains compared with smooth terrains.     

Reproductive function of Selenoprotein M in Chinese mitten crabs (Eriocheir sinesis)

Selenoproteins are present in all major forms of life, including eukaryotes, bacteria and archaea. In eukaryotic animals, selenoproteins often function as antioxidants, but rare or absent in other phyla, such as plants and fungi (except for the green alga Chlamydomonas). Selenoprotein M (SelM) is a selenocysteine containing protein with redox activity, which is involved in the antioxidant response. However, information remains limited about SelM physiology and function in marine invertebrates, particularly in crustaceans. Hence, we investigated the reproductive functionality of SelM in the Chinese mitten crab (Eriocheir sinensis), which is a commercially important yet disease vulnerable aquaculture species. The full-length SelM cDNA (928bp) strand was cloned by using PCR, based on an initial expressed sequence tag (EST) that was isolated from a hepatopancreatic cDNA library. The SelM cDNA contained a 390bp open reading frame (ORF) that encoded a putative 129 amino acid (aa) protein. SelM mRNA expression in E. sinensis was (a) tissue-specific, with the highest expression observed in the hepatopancreas, testis, ovaries and intestines. Based on this information, we then detected the different stages of tissue expression for SelM in the testis, ovary, and male crab hepatopancreas and hemolymph, and the enzyme activity of SelM in the testis. Overall, SelM was isolated successfully from the Chinese mitten crab, and its involvement in the regulation of reproduction during the period of rapid development in E. sinensis was confirmed.     

Symbiotic Bacteria in Gills and Guts of Chinese Mitten Crab (Eriocheir sinensis) Differ from the Free-Living Bacteria in Water

Aquatic animals have a close relationship with water, but differences in their symbiotic bacteria and the bacterial composition in water remains unclear. Wild or domestic Chinese mitten crabs (Eriocheir sinensis) and the water in which they live were collected from four sampling sites in Jiangsu and Shanghai, China. Bacterial composition in water, gills or guts of E. sinensis, were compared by high-throughput sequencing using 16S rRNA genes. Analysis of >660,000 sequences indicated that bacterial diversity was higher in water than in gills or guts. Tenericutes and Proteobacteria were dominant phyla in guts, while Actinobacteria, Proteobacteria and Bacteroidetes were dominant in gills and water. Non-metric multidimensional scaling analysis indicated that microbiota from gills, guts or water clearly separated into three groups, suggesting that crabs harbor a more specific microbial community than the water in which they live. The dominant OTUs in crab gut were related to Mycoplasmataceae, which were low in abundance in gills, showing that, like mammals, crabs have body-site specific microbiota. OTUs related to Ilumatobacter and Albimonas, which are commonly present in sediment and seawater, were dominant in gills but almost absent from the sampled water. Considering E. sinensis are bottom-dwelling crustacean and they mate in saline water or seawater, behavior and life cycle of crabs may play an important role in shaping the symbiotic bacterial pattern. This study revealed the relationship between the symbiotic bacteria of Chinese mitten crab and their habitat, affording information on the assembly factors of commensal bacteria in aquatic animals.