【作者】 刘立鹤；

【导师】 陈立侨；

【作者基本信息】 华东师范大学 ， 动物学， 2005， 博士

【摘要】 中华绒螯蟹（Eriocheir sinensis H.Milne-Edwards）是我国特有的水产珍品,广泛分布于我国的咸淡水或淡水水域,其中以长江流域河蟹的品质最佳,其营养丰富、肉味鲜美、风味独特,倍受国内消费者的青睐。近年来,河蟹养殖面积的不断扩大,产量与日俱增,2004年全国总产量达到47万吨,产值数百亿元。然而,养蟹业在迅速发展的同时,也出现了一系列亟待解决的问题:养殖规模的扩大,需要更多的小杂鱼,而小杂鱼资源越来越有限,这就迫切需要杂鱼的可替代物。维持目前的养殖规模,需要大量优质的苗种,而优质苗种来源于性腺发育良好的亲体。但目前为止,人们对河蟹的性腺发育、亲体选育缺乏足够的认识。饲料产业化正在逐步形成。而营养研究,尤其是亲体营养研究极其缺乏,研究相对滞后。 现有的甲壳动物营养研究中,脂类的研究相对较丰富。已有河蟹脂质营养研究表明:脂类作为河蟹重要的能量来源和营养物质,是参与血淋巴中营养转运的主要物质,同时也是构成卵黄蛋白原和卵黄磷蛋白的重要组分,河蟹的性腺发育和繁殖对脂类有着极其重要的需求。不同脂肪源饲料可显著影响河蟹亲体的繁殖性能,以及河蟹组织、器官和卵中的脂肪酸组成。饲料中HUFA组成和含量与繁殖性能密切相关,研究已证实EPA,DHA是合成卵正常发育和增殖的物质基础,EPA与河蟹的蜕皮、生长及亲体的产卵力,DHA与细胞增殖,亲体的孵化率密切相关,而且HUFA与维生素C、E存在明显的交互作用,同时添加能显著地提高河蟹亲体的繁殖性能。 本研究的主要目的是在已有工作基础上,以脂质营养为主线,通过脂质营养对中华绒螯蟹雌体繁殖性能、卵巢发育期其生理生化组成、消化生理、脂肪合成、类固醇激素及MIH基因表达变化的研究,深入探讨了脂质营养在河蟹亲体性腺发育过程中的作用和以及其通过内分泌途径来调控生殖的可能途径。并在此基础上,进一步研究了HUFAs与V_E,β-胡萝卜素交互作用对中华绒螯蟹繁殖性能及生理生化的影响,以期为人工配合饲料中合理使用脂质,提供理论依据和数据支持。 一、脂质营养对中华绒螯蟹繁殖性能的影响 在蛋白水平40%左右的基础饲料中分别添加6%的鱼油、花生油、磷脂和猪油,投喂河蟹28周,以冰鲜作为对照组,研究了不同脂肪源饲料对中华绒螯蟹繁殖性能的影响,研究表明脂质营养对河蟹繁殖性能影响极其显著:河蟹卵巢发育过程中,冰鲜组河蟹HSI逐渐下降,GSI持续上升,Ⅲ₁～Ⅲ₂期增幅最大,Ⅲ₂期雌蟹GSI值高达9.15,显著高于同期人工饲料组,而同期血淋巴中卵黄蛋白原（Vg）的含量显著增加,这说明了Ⅲ₁～Ⅲ₂期是冰鲜组雌蟹营养转移,卵巢营养快速积累时期。与冰鲜组不同,饲料组蟹的性腺发育期物质积累明显滞后于冰鲜组,GSI峰值多出现在性腺成熟Ⅳ期,Ⅳ期磷脂、鱼油组蟹的GSI最高分别为11.50,11.25;花生油组次之（8.85）;猪油组的GSI最低,仅为7.30。投喂不同的脂质对雌蟹性腺发育的影响有更多还原

【Abstract】 In crustaceans, it is well known that lipid is a kind of indispensable nutrient because of its important physiological function on cell, tissue, body structure and supplying energy to growth, development and reproduction. Lipid and fatty acids of broodstock diet have been identified as major dietary factors that determine successful reproduction, quality of offspring, and improve fatty acid profiles of hepatopancreas and egg. At the same time, some studies on crustacean endocrine had shown the Crustacean molting and reproduction regulated by steroids hormone. Up to date there are no reports on dietary lipid sources and contents influence on reproduction performance by endocrine regulation or other methods and how to regulate this process during the ovarian maturation.The Chinese mitten-handed crab, Eriocheir sinensis, is considered to be one of the most important species for commercial production in China. Lots of studies on Chinese mitten-handed crab has been conducted, however, Studies on nutrition of Chinese-mitten crab Broodstock is still few. From former studies on lipid nutrition of Chinese-mitten crab Broodstock reviewing, lipid nutrition has special effect on ovarian maturation of the Chinese mitten-handed crab, which, especially n-3 HUFAs, significantly affects reproduction performance of the Chinese mitten-handed crab, female broodstock, improves quality of offspring, optimize fatty acid profile of hepatopancancreas and egg. Moreover, dietary HUFAs and vitamin E, C are indispensable and cooperative to egg developing of crab and must be supplemented in the broodstock crab diet.It is still a secret how reproduction is affected by lipid nutrition. So in this paper, The main aim is to further probe into the mechanism how lipid nutrient affects ovarian development and reproduction performance, and regulation metabolism of lipid. To accumulate basal information for reproductive lipid nutrition, and provide reference for breeding broodstock in seed production and formulation of broodstock diet. In addition, effects of a-tocopherol and β -carotene, highly unsaturated fatty acids, in broodstock diet on reproduction performance, non-specific immunity and biochemical profiles of Chinese mitten-handed crab, are studied on basis of former research finding.1. Effect of dietary lipid sources and contents on ovarian development and reproduction performance on female Eriocheir sinensis, broodstockA 6 month feeding and reproduction trial was studied to investigate the effect of feeding eight kinds of compound diets containing different lipid sources and contents （6% fish oil, 6% peanut oil, 6% pork lard, 6% lecithin,0%,15% fish oil,15%porklard,15%lecithin） and fresh frozen-fish as the control diet on reproduction performance of Chinese mitten-handed crab broodstock. Broodstock crab fed the diet containing pork lard and peanut oil showed poor fecundity, low hatchability. Fecundity and hatchability of female crab fed 6% pork lard were only 56.9%, 32.57% of that fed 6% fish oil respectively. Broodstock fed the diets containing Alaska fish oil showed the highest fecundity, 2.787x103 cell/g. However, egg hatchability showed no significance between diet fed containing 6% lecithin and control group.During ovarian maturation, Hepatopancreas somatic index （HSI） is continuously decreasing, while Gonad somatic index （GSI） increasing in control group, especial during Ⅲ₁^Ⅲ₂ stage). In contrast to control group, the trend of nutrients transferring from hepatopancreas and storage to ovary in the group fed with diets lagged ,most sharp peak in Ⅲ₂^Ⅳ stage, which is more evident in groups fed diet containing Pork lard and peanut oil, especially Pork lard, which increased Vg content in hemolymph so as to nutrients sufficient storage in ovary at Ⅲ₁ stage. However, at Ⅳ ovarian mature stage GSI of female crab was 11.50 for lecitin,11.25 for fish oil, 8.85 for peanut oil and 7.30 for pork lard.Further studies of dietary lipid contents （0%, 15% fish oil, 15% pork lard, 15% lecithin） on reproduction performance of Chinese mitten-handed cra更多还原