大西洋庸鲽Hippoglossus hippoglossus和大西洋鲑Salmo salar配子质量研究

IOCAS-IR > 海洋环流与波动重点实验室

	大西洋庸鲽Hippoglossus hippoglossus和大西洋鲑Salmo salar配子质量研究
其他题名	The Game Quality Research about Atlantic Halibut Hippoglossus hippoglossus and Atlantic Salmon Salmo salar
	丁福红
学位类型	博士
导师	李军
	2008-06-06
学位授予单位	中国科学院海洋研究所
学位授予地点	海洋研究所
学位专业	海洋生物学
关键词	大西洋庸鲽大西洋鲑配子脂肪酸冷冻保存
摘要	种质问题是养殖健康发展的基础。在鱼类养殖中，卵子和精子的质量直接关系到受精、胚胎发育，仔稚鱼发育以及幼鱼生长等一系列过程。本论文针对大西洋庸鲽和大西洋鲑的配子质量进行研究。研究内容涉及大西洋庸鲽精子冷冻保存方法；促性腺激素释放激素类似物（GnRHa）使用对其精子冷冻保存效果、以及脂肪酸组成的影响；野生和驯养大西洋鲑卵子在脂肪酸、类胡萝卜素、矿物盐方面的差异比较。精子冷冻保存通过提高对精子的利用效率，进而对于种质改良，推进鱼类养殖科研和生产具有重要意义。本实验建立了大西洋庸鲽精子大容量冷冻保存方法。八种抗冻剂冷冻保存实验结果表明：10% 及15% DMSO配以 HBSS 或KS 的抗冻剂组合冷冻保存效果最佳，4 mL体积冷冻保存可获得与1.6 mL同样的保存效果。在繁殖季节后期注射GnRHa激素缓释剂，可获得质量稳定的大西洋庸鲽精液，将激素注射方法与精子冷冻保存方法相结合对于提高雄鱼利用率，扩大生产规模具有重要实用价值。本项研究分三个时间采集注射GnRHa激素后的雄鱼精子以及同期未注射激素的雄鱼精子，对所有精子样品使用同样的方法进行冷冻保存，检测冷冻保存后解冻精子的受精率与活力。结果表明，激素注射与否对于冷冻保存后精子的受精率和活力无显著影响，两类冷冻精液均达到鲜精水平。实验结果还表明，注射激素14天后的精子的密度显著的降低。说明GnRHa激素的使用可以显著降低精子密度，但不会影响精子的冷冻保存效果。本相研究同时对注射GnRHa 缓释激素和未注射GnRHa 缓释激素的大西洋庸鲽精液脂肪酸成分进行分析，以检测该激素使用对精子生化组分的影响。结果表明激素的使用对在DHA (22:6n-3,二十二碳六烯酸)、EPA(20:5n-3,二十碳五烯酸)、AA(20:4n-6，花生四烯酸)等重要脂肪酸，不饱和脂肪酸、饱和脂肪酸以及n-3、n-6等重要种类的脂肪酸总量及其比例没有显著影响。精液脂肪酸中DHA含量最高，约占25%；PUFA约为44%。作为世界性的重要养殖品种，野生和驯养大西洋鲑在形态、生化组成以及遗传等方面表现出的差异被广泛关注。本论文，对野生和驯养大西洋鲑受精卵关键生化成分进行分析，通过与野生受精卵比较阐明驯养受精卵的质量状况，为亲鱼营养需求提供指导依据。本实验中野生配子和驯养配子的受精率没有显著差异，但重要脂肪酸组成、类胡萝卜素以及矿物盐含量都存在多方面显著差异。两类受精卵脂肪酸中含量最高的依次为18:1n-9（油酸）、DHA（二十二碳六烯酸）、16:0（棕榈酸）、EPA（二十碳五烯酸）。野生受精卵的单不饱和脂肪酸总量显著高于驯养受精卵，而多不饱和脂肪酸（PUFA）比例显著低于驯养的受精卵。在主要必需不饱和脂肪酸（EFA）中，DHA和EPA在野生受精卵中的比例高于驯养受精卵，AA（花生四烯酸）低于驯养受精卵。野生受精卵虾青素（Ax）的含量低于驯养受精卵而鸡油菌素（Cx）含量高于驯养受精卵。野生受精卵中多种矿物盐的含量（铝、铜、铁、硒和锌）含量显著高于驯养的受精卵。差别最大的为铜。诸多方面的差异表明，野生亲鱼与驯养亲鱼产出的卵子确实存在显著差异，因此关注亲鱼的营养极为重要。
其他摘要	Game quality control as a basic issue for the development of fish aquaculture is intimately related to fertilization, embryo and larval developments. The main topic of this thesis is about quality determination of sperm and eggs from Atlantic halibut and Atlantic salmon. Development of Atlantic halibut (Hippoglossus hippoglossus) aquaculture will be enhanced with cryopreservation of halibut sperm by ensuring a reliable supply of sperm of desired quality and quantity. To assist in its commercial application, the cryopreservation of large volumes of halibut sperm was investigated. In this research, by testing eight kinds of extenders, 10% or15% DMSO combined with HBSS or KS were proved to be optimal for the cryopreservation of Atlantic halibut sperm, an optimized protocol for the cryopreservation of Atlantic halibut sperm using a volume as large as 4 mL has been established by inspecting fertilization rate and motility of post-thawed sperm. During the broodstock breeding, the hormone of GnRHa is generally used to enhance the milt product during the end of spawning season. Combining the hormone using and cryopreservation will dramatically increase the efficiency of sperms application. In this research, the effect hormone implant on sperm cryopreservation was detected by comparing with the milt from no-implanted males. No significant difference was shown between the two groups, both implanted and no-implanted milt got the same fertilization rates and motility as fresh milt after cryopreservation. To test the effect of hormone implant on sperm composition, fatty acids analyzing of halibut sperm were conducted. The results showed that the essential fatty acids ( DHA, EPA, AA )and the ratio between important fatty acids series(n-3, n-6, PUFA, SAT) were not affected by the hormone implantation. DHA was the mainly fatty acid in halibut sperm with percentage of 25%.The PUFA content was around 44%. The changes between wild and captive Atlantic salmon has been noticed in parts of morphology, physical composition and genealogy. In this research, the quality of embryos of wild and captive salmon were compared to indicate their difference and to provide evidence for improving broodstock breeding conditions. The results showed that the fertilization rates of the two kinds of eggs has no significant difference, while the compositions of fatty acids ,carotenoids and mineral were different in many aspects. In those embryos, the most high percentage of fatty acids were 18:1n-9、22:6n-3、16:0、20:5n-3.PUFA was higher in captive embryos than in wild embryos, while MONO was lower in captive embryos. EPA 20:5n-3 and DHA 22:6n-3 were higher in wild embryos. Astoxanthin was lower in wild embryos with the contract result of canthaxanthin. The contents of several kinds of minerals ,like（aluminum,copper,iron,selenium, zinc）were higher in wild embryos ,especially copper. These kinds of difference between wild and captive embryos indicated that more attention should be paid on the nutrition condition of broodstocks to decrease the gaps and improve the eggs quality for culturing.
页数	86
语种	中文
文献类型	学位论文
条目标识符	http://ir.qdio.ac.cn/handle/337002/787
专题	海洋环流与波动重点实验室
作者单位	中国科学院海洋研究所
推荐引用方式 GB/T 7714	丁福红. 大西洋庸鲽Hippoglossus hippoglossus和大西洋鲑Salmo salar配子质量研究[D]. 海洋研究所. 中国科学院海洋研究所,2008.