稻田网箱养殖辽河水系中华绒螯蟹幼蟹的个体生长

本实验在室外网箱内监测辽河水系中华绒螯蟹(Eriocheir sinensis)幼蟹阶段个体生长发育的每一次蜕壳生长情况。2013年实验选取雄蟹和雌蟹各200只,记录了每个蜕壳阶段雌蟹和雄蟹的生长,包括壳长、壳宽、总重和蜕壳间隔时长(d),并且观察雌蟹和雄蟹形态特征的变化。实验共进行111 d,幼蟹共蜕壳11次。实验结束时,雄蟹剩余34只,雌蟹剩余42只;雄蟹的特定生长率为(7.176 5±0.168 4)%/d,雌蟹的特定生长率为(7.283 3±0.174 3)%/d;雌蟹生长蜕壳过程中腹部的形态变化大,腹部由三角形变成卵圆形;雄蟹在生长蜕壳过程中螯足的增长明显较雌蟹快,并且在本实验最后一次即第11次蜕壳后螯足腹面内侧出现1小撮绒毛,外侧也出现少量绒毛,但不易被发现。     

温度和体重对中华绒螯蟹幼蟹饥饿代谢的影响

于1998年9月—12月对均重为3．64、1．69和0．52g三种规格的中华绒螯蟹幼蟹在15、20、25和30℃四种温度条件下的饥饿代谢进行了测定,以探讨体重和温度变化对代谢的影响。结果表明,耗氧率(Ro)和氨氮排泄率(Rs)都随温度升高而增大,随体重增大而减小,其关系式可分别表示为：Ro=0．1309We^0.0490T和Ro=0．4254W^-0.1999;Rs=3．3649e^0.0484T和RN=0．4400W^-2.900不同体重幼蟹在不同温度条件下的饥饿代谢率差异显,饥饿代谢率随体重的增大而减少,随温度升高而增大。同时,体重对能源物质的供能比有较大影响,三种规格的幼蟹,其代谢底物中蛋白质、碳水化合物和脂肪提供的能量比分别为6．4：41．8：518、6．7：27．8：65．5和7．3：43．8：48．9。     

湖泊放养长江水系中华绒螯蟹的性成熟规律研究

于8－11月份,结合中华绒螯蟹捕捞期,即生死洄游期的到来,通过性腺发育的组织学和细胞学观察,重点观察了长江下游安徽杨赛湖等人工放养长江水系中华绒螯蟹的性成熟规律。结果表明：①在性成熟时期,性腺发育与肝脏变化极为密切,肝脏指数逐渐下降,而性腺指数逐渐上升;②雌、雄蟹的性成熟存在规律性差异,8－9月份,雄蟹的性腺指数和性／肝指数均高于雌蟹,而10－11月份,雌蟹的性腺指数和性／肝指数均高于雄蟹;③经性腺的组织学和细胞学观察,雄蟹的性腺发育略早于雌蟹;④洄游高峰期的出现是河蟹性细胞走向成熟的外在表现。作者建议把肝脏指数7.5作为捕捞长江水系河蟹最佳时机的指标。     

不同发育阶段中华绒螯蟹对底质的喜好性研究

为探究不同发育阶段中华绒螯蟹对底质类型的喜好性, 采用单因子实验方法, 研究不同发育阶段中华绒螯蟹个体和群体对泥、泥沙(泥﹕沙=1﹕1)、中沙(0.35—0.50 mm)和细沙(0.25—0.35 mm)等4种底质的喜好性。亲蟹个体出现在各底质上的时间百分比和群体出现在各底质上的数量百分比结果均表明亲蟹对泥底质的喜好性显著高于其他三种底质(P＜0.05); 扣蟹个体和群体实验结果均表明扣蟹对泥和中沙底质的喜好性显著高于其他两种底质(P＜0.05); 仔蟹群体实验结果表明仔蟹最喜好泥底质, 其次是泥沙, 两者显著多于中沙和细沙(P＜0.05)。昼夜研究结果表明, 亲蟹个体和群体、扣蟹群体及仔蟹群体在白天和夜晚对底质的选择均具有显著性差异(P＜0.05), 但扣蟹个体在夜晚对底质的选择无显著性差异(P＞0.05)。对栖息方式的研究表明, 亲蟹无显著的埋栖行为, 扣蟹喜好埋栖在泥和中沙里面, 仔蟹喜好埋栖在泥里面, 其次是泥沙。研究揭示, 中华绒螯蟹对不同底质的喜好性具有显著差异, 且不同发育阶段中华绒螯蟹对底质的喜好及栖息方式也不尽相同, 为其生境修复和资源养护提供了参考。     

池养条件下中华绒螯蟹的若干生物学特征

开展中华绒螯蟹(Eriocheirsinensis)生物学研究,对指导养殖具有重要意义.1993年6月20日购于上海市崇明县的仔蟹放养于湖北省保安湖渔场的一个试验塘(2330m2),每天投喂小杂鱼和螺蚌肉.在试验期间,河蟹成活率为18.6%.协方差分析表明,性别对各月体重与壳长、体重与壳宽及壳宽与壳长回归方程的截距和斜率均无显著影响(p＞0.05);但是,两性数据混合后的不同月份间回归方程的截距却有显著差异(p＜0.01).在每次取样时,雄蟹与雌蟹的壳长、壳宽及体重均不存在显著差异(p＞0.05),这表明雄蟹与雌蟹可能具有相同的生长率.在各次取样时统计的性比(雄∶雌)均无异于理论值1∶1(X2检验,p＞0.10).在九月中旬发现大部分雌蟹开始青春期蜕壳,在11月下旬雌蟹性成熟比例为88%,性成熟雌蟹壳长变幅为26.2-49.8mm,均值为34.1mm.     

高温季节中华绒螯蟹的人工育苗

中华绒螯蟹(Eriocheir sinensis H.Milne-Edwards),俗称河蟹(Mitten crab),是一种甲壳动物,它含低脂肪高蛋白,特别美味可口,近年一直是较昂贵的水产品之一。河蟹价格居高不下的主要原因是河蟹养殖业受到诸多因素的制约,其中重要因素之一是蟹苗的来源。由于水利工程、环境污染、狂捕乱捞等原因,天然蟹苗越来越少,濒临灭绝。绝大部分人工育苗利用天然海水,在海边进行,河蟹氵蚤状幼体发育至大眼幼体的最适温度是20℃-26℃1-3,育苗水体加温容易,降温难。冬春季节过后,随着气温升高,海水中有害生物大量繁殖,蟹苗繁殖受到了限制。       

长江口中华绒螯蟹放流亲蟹的繁殖力评估

在长江口中华绒螯蟹亲蟹生殖洄游期间, 通过研究中华绒螯蟹放流群体和自然群体繁殖力随壳宽的变化规律, 比较放流群体和自然群体繁殖力的差异, 从而评估人工增殖放流亲蟹的繁殖力。结果显示, 随着壳宽的增大, 中华绒螯蟹放流群体和自然群体的繁殖力都显著增加(P<0.05)。在相同壳宽范围内, 放流群体和自然群体的繁殖力之间没有显著差异(P>0.05)。回归分析显示, 放流群体繁殖力(F)与壳宽(CW)呈幂函数关系: F=3.979CW6.208(R2=0.822);自然群体F与CW呈幂函数关系: F=1.696CW6.636(R2=0.673)。协方差分析显示, 放流群体与自然群体F与CW的两条曲线在显著性为0.05时拟合较好。研究结果表明, 放流群体与自然群体的繁殖力与壳宽之间无显著性差异, 推断放流亲蟹能够适应长江口天然水域环境, 并与自然群体的繁殖力水平相当。     

维生素C对中华绒螯蟹雄性生殖的影响

采用不同Vc的人工配合饲料 ,研究了Vc对中华绒螯蟹雄性生殖的影响。实验分为 5组 ,其添加量分别为 0mg 1 0 0g饲料 (实验 1组 )、2 0 0mg 1 0 0g饲料 (实验 2组 )、40 0mg 1 0 0g饲料 (实验 3组 )、60 0mg 1 0 0g饲料 (实验 4组 )和 80 0mg 1 0 0g饲料 (实验 5组 ) ,实验为期 5 0d。结果如下 :1 )肝胰腺、精巢和副性腺中Vc的积累量以实验 3组最高 ,分别为 2 2 61 μg mgprot、1 0 2 1 7μg mgprot、7 661 μg mgprot;2 )肝体指数和性腺指数以实验 3组最高 ,分别为 0 0 5 2 3和 0 0 2 68;3 )增重率同样以实验 3组最高 ,实验 1组最低 ,分别为 3 1 3 3 %和 2 1 2 %;4)睾酮含量以实验 2组最高 ,为 1 2 5 7ng ml,但实验 3组的睾酮含量与实验 2组接近 ;5 )SOD酶活性与其Vc含量呈负相关 ,且精巢和副性腺的酶活性高于肝胰腺。结果表明 :适量添加Vc可促进雄蟹肝胰腺、精巢和副性腺对Vc的积累 ,同时可促进精巢的发育及睾酮的分泌 ,而过量添加可能会对精巢的发育产生不良影响 ;河蟹体内的抗氧化系统之间存在着相互的协调和平衡 ,生殖期性腺对其自身的抗氧化保护需求要高于其它组织     

中华绒螯蟹亲蟹的饥饿代谢研究

1998年 10～ 12月 ,对体重为 5 4 .5 9(± 2 .37) g的亲蟹在 2 0 (± 0 .5 )℃温度条件下进行饥饿实验 ,研究了饥饿对中华绒螯蟹亲蟹代谢的影响 .结果表明 ,经过 30d的饥饿处理 ,亲蟹的耗氧率降低为摄食状态的 5 0 .0 % ;CO2 排出率减小为摄食状态的 6 3.4 % ;NH3 N排泄率减小为 5 9.1% .耗氧率和CO2 排出率、NH3 N排泄率的变化差别较大 ,耗氧率的减小可分为 4个阶段 ,而CO2 排出率和NH3 N排泄率的减小仅可分 3个阶段 .与此同时 ,亲蟹的标准代谢水平同样受到饥饿的影响 ,饥饿 30d后 ,由 4 .4 5J·g-1·h-1减小为 2 .36J·g-1·h-1;在饥饿亲蟹的代谢中 ,脂肪消耗最多 ,其次是碳水化合物 .     

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.     

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.     

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).     

Characterization of microsatellite loci for the Chinese mitten crab,Eriocheir sinensis

The Chinese mitten crab, Eriocheir sinensis is an important human food source in Asia and causes considerable ecological and economical damage as a recent invader of North America and Europe. Here we report the isolation and characterization of 12 highly polymorphic microsatellite loci for E. sinensis. The number of observed alleles per locus ranged from 7 to 40 and the observed heterozygosity from 0.50 to 0.95. These markers should prove a useful tool to investigate the colonization process and its evolutionary implications.     

Effects of atrazine on osmoregulation in the Chinese mitten crab,Eriocheir sinensis

This study integrates results from acute contamination with atrazine of isolated perfused gills, and from in vivo chronic contamination of euryhaline Chinese mitten crabs, Eriocheir sinensis, acclimated to freshwater. Atrazine 1 mg/l in contact with the basolateral membrane (IN) increases the transepithelial potential difference (TEP) from -20.8 +/- 4.9 to -29.7 +/- 3.8 mV in isolated perfused posterior gills (P < 0.01). This effect is only partially explained by a modification of Na(+) and Cl(-) active influxes. No TEP modification is detected when atrazine is added (OUT) indicating that molecular mechanisms located on the basolateral membrane are likely to be the only ones affected. Another explanation would be that cuticular barrier prevents atrazine penetration into the gill. Haemolymph osmolarity, Na(+) and Cl(-) concentrations of crabs living in freshwater contaminated with atrazine 1 mg/l during 14 days are not significantly modified. We conclude that although atrazine can disturb osmoregulatory mechanisms of isolated gills, this pollutant would be of minor importance in affecting osmoregulatory capacities of the Chinese mitten crab in natural conditions.     

The immune responses in Chinese mitten crab Eriocheir sinensis challenged with double-stranded RNA

Double-stranded RNA (dsRNA) is a virus-associated molecular pattern which induces antiviral innate immune responses and RNA interference (RNAi) in mammals. In invertebrates, RNAi phenomenon has been widely studied, but dsRNA-induced innate immune response is seldom reported. In the present study, two different dsRNAs specific for green fluorescent protein (GFP) and the putative D1 protein of photosystem II (NoPSD) from Nannochloropsis oculata, were employed to challenge Chinese mitten crab Eriocheir sinensis. The temporal changes of phenoloxidase (PO), acid phosphatase (ACP), superoxide dismutase (SOD) and malondialdehyde (MDA) content, as well as the mRNA expression of some immune-related genes were examined in order to estimate the effect of dsRNAs on the innate immunity of E. sinensis. The activities of PO, ACP and SOD significantly increased after dsRNA treatment, whereas malondialdehyde (MDA) content did not change significantly. Among the examined genes, only the mRNA expression of EsALF, an antibacterial peptide in E. sinensis, was significantly up-regulated (about 5 fold, P < 0.05) at 12 h after dsRNA treatment, while no significant expression changes were observed among the other immune genes. The increase of PO, ACP and SOD activities, and mRNA expression level of EsALF after dsRNA stimulation indicate that phenoloxidase, hydrolytic enzyme, antioxidation and EsALF were involved in dsRNA-induced innate immunity, suggesting that broad-spectrum immune responses could be induced by dsRNA in E. sinensis.     

Acute tolerance and metabolic responses of Chinese mitten crab (Eriocheir sinensis) juveniles to ambient nitrite

The lethal concentration of nitrite to the Chinese mitten crab Eriocheir sinensis was tested by exposing the animals to 17.78, 23.71, 31.62, 42.17, and 56.23 mg NaNO2 L(-1) at 20 degrees C for 24, 48, 72, and 96 h. The corresponding LC50 value for each time exposure was 43.87 (38.70-51.70), 40.24 (34.88-46.01), 38.87 (33.72-46.01) and 38.87 (33.72-46.01) mg NaNO2 L(-1) or 29.25 (25.80-34.47), 26.83 (23.25-30.67), 25.91(22.48-30.67), 25.91(22.48-30.67) mg NO2-N L(-1), respectively. The physiological response of the crab to nitrite toxicity was further investigated by exposing the crab to 0, 10, 20, 30 and 40 mg NaNO2 L(-1) for 2 d. The changes of nitrogenous compounds in haemolymph, oxyhemocyanin and metabolism were measured at 3, 6, 24 and 48 h upon exposure. Haemolymph nitrite was significantly enhanced by the increase of nitrite from 10 to 40 mg NaNO2 L(-1) during the 2-day exposure. The concentrations of nitrate, urea and glutamate in haemolymph increased concomitantly with the exposing time and ambient nitrite levels, suggesting that the formation of nitrate, urea and glutamine may be the possible end products of nitrite detoxification in crabs. The diffusion of nitrite caused a reduction of oxyhemocyanin, resulting to hypoxia in tissues. Under a hypoxia condition, crabs increased energy demand for metabolism as indicated by the elevated levels of glucose and lactate in haemolymph. Our data showed that ambient nitrite could affect oxygen carrying capacity through oxyhemocyanin reduction and the increase of energy catabolism in crabs. This study suggests that nitrite could be detoxified through the pathway of nitrate, urea and glutamine formation in crabs.