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学校代码:研究生学号10264M100101041上 海 海 洋 大 学硕士学位论文 题 目:公子小丑鱼(Amphiprion ocellaris)胚胎发育的形态学观察及盐度对幼鱼生长的影响英文题目:Morphological observation of embryonic development and effects of salinity on growth of False clownfish Amphiprion ocellaris专 业:水产养殖研究方向:海水观赏鱼繁殖与发育姓 名:鞠晨曦指导教师:蔡生力 李云 二O一三 年 四 月 九 日 上海海洋大学学位论文原创性声明本人郑重声明:我恪守学术道德,崇尚严谨学风。所呈交的学位论文,是本人在导师的指导下,独立进行研究工作所取得的成果。除文中已经明确注明和引用的内容外,本论文不包含任何其他个人或集体已经发表或撰写过的作品及成果的内容。论文为本人亲自撰写,我对所写的内容负责,并完全意识到本声明的法律结果由本人承担。 学位论文作者签名: 日期: 年 月 日上海海洋大学学位论文版权使用授权书学位论文作者完全了解学校有关保留、使用学位论文的规定,同意学校保留并向国家有关部门或机构送交论文的复印件和电子版,允许论文被查阅或借阅。本人授权上海海洋大学可以将本学位论文的全部或部分内容编入有关数据库进行检索,可以采用影印、缩印或扫描等复制手段保存和汇编本学位论文。 保密 ,在 年解密后适用本版权书。本学位论文属于 不保密 学位论文作者签名: 指导教师签名:日期: 年 月 日 日期: 年 月 日3公子小丑鱼(Amphiprion ocellaris)胚胎发育的形态学观察及盐度对幼鱼生长的影响摘 要公子小丑鱼,学名为眼斑双锯鱼,海葵鱼属(Amphiprion),雀鲷科(Pomacentri- dae),鲈形目(Perciformes)。公子小丑鱼是最常见的海洋观赏鱼类,雌雄同体,雄性先熟,分布于西太平洋礁岩海域。公子小丑鱼以其鲜艳的色彩、漂亮的花纹及可爱的形象而深受水族爱好者的喜爱。挑选合适的野生的公子小丑鱼个体在实验室条件下进行配对,调控适宜的外界环境因子并进行营养强化使其产卵,产卵以后进行人工孵化。仔鱼在孵出的第2d即开口食用由小球藻(Chlorella sp)强化的褶皱臂尾轮虫(Brachionus plicatilis),全长4.24.4mm,为仔鱼期,46d变态为稚鱼,全长5.15.4mm;开始投喂刚孵出的卤虫无节幼体,10d以后开始投喂经小球藻强化的卤虫(Artemia sp),1622d 进入幼鱼期,全长10.012.4 mm,3545d开始投喂自制人工配合饵料。 在实验室水温261、盐度281的条件下,对公子小丑鱼胚胎发育过程中的形态学变化进行了观察研究。结果表明,公子小丑鱼受精卵属附着性卵,呈长椭圆形,长径2.0mm0.1mm,短径0.95mm0.1mm;其卵裂方式为端黄卵盘状卵裂;胚胎发育可分为10个主要阶段:卵裂前期、卵裂期、囊胚期、原肠期、神经胚期、体节期、翻转期、血管形成期、器官形成期和孵化期。发育过程中胚胎的颜色逐渐加深,呈现由橙黄色到棕黄色的变化;在26水温条件下,整个胚胎发育过程需200h左右。在水温261的条件下,研究了盐度对公子小丑鱼幼鱼生长、摄食、饵料利用率和生长激素及其受体mRNA水平的影响。第42d和70d的试验结果都表明, 公子小丑鱼幼鱼在各盐度实验组成活率无显著差异; 当盐度为25和30时生长最快,盐度20时次之,盐度15和35时较慢;与之相应的饵料转换效率也是在盐度25和30时最高,盐度20、35、15依次降低;相对体重增长率也是在盐度25和30时最高,盐度20、35、15依次降低。第70d时取样进行分子生物学实验,结果表明,公子小丑鱼脑垂体GHmRNA水平在盐度15时最高,盐度25次之,盐度20、30、35时较低;肝脏GHR1mRNA水平在盐度20、25、30时最高,盐度35次之,盐度15时最低;肝脏GHR2mRNA水平在盐度25时最高,盐度20时次之,盐度30、35、15时较低。公子小丑鱼幼鱼GHR1和GHR2mRNA表达量与其生长呈正相关。说明公子小丑鱼幼鱼最适宜的生长盐度为25到30。关键词:公子小丑鱼,胚胎发育,形态学,盐度,成活率,生长,生长激素,生长激素受体Morphological Observation of Embryonic Development and Effects of Salinity on Growth of False clownfish Amphiprion ocellarisABSTACTS False clownfish is placed in Amphiprion ocellaris, Pomacentridae, Perciformes. It is a kind of common marine ornamental fish, hermaphrodite and cross-fertilization and widespread in the sea of south China, Philippines and the western Pacific. False clownfish is a popular fish for ornamental fish enthusiasts for the bright colore, beautiful pattern and lovely figure. The fish was collected from the wild as broodstock, and cultured to mature and spawned in artificial condition. The larval fish could be fed rotifer strengthened by unicellular algae chlorella spp. in the next day after hatching and developed into juvenile at 46 days after hatching, newly hatched Artemia nauplii were supplied, from the 10th day Artemia nauplii strengthen by chlorella spp. were supplied, then developed into parr 1622 days after hatching. The parrs could be fed by artificially made compound feed at 35 45 days after hatching.The study was carried out to investigate the incubation period and the changes of morphological character during the embryonic development of Amphiprion ocellaris at water temperature (261),salinity(281)in laboratory condition. It was found that the eggs belonged to the telolecithal and were subject to discoidal cleavage;The adhesive fertilized eggs, which were ellipsoidal, were 2.0mm0.1 mm long and 0.95mm0.1mm in diameter;The process of embryonic development could be divided into ten stages: pre-cleavage stage, cleavage stage, blastula stage, gastrula stage, neurula stage, somite stage, turnover stage, angiogenesis stage, organ formation stage and hatching stage. The color of the embryo changed gradually during the development procession, from orange-yellow to claybank. The whole incubation period is about 200h at water temperature 26. The effects of salinity on growth, feeding, food conversion rate (FCR), growth hormone and receptor mRNA leve of young Amphiprion ocellaris were studied. The results of the 42th and 70th day showed that there is no significant effect on the survive rate for young Amphiprion ocellaris at each salinity levels. The young Amphiprion ocellaris had a highest growth at the 25 and 30 salinity levels, higher growth at 20 and low growth at 15 and 35. The feed conversion ratio is also higher at the 25 and 30 salinity levels and then reduced along the 20, 35, 15 salinity levels. The growth rate of relative body weight is conformed to this law. The molecular biology study began at the 70th day, the result showed that the highest level of pituitary GHmRNA was at salinity 15 and higher at 35, the lowest level was at salinity 20, 30, 35. The highest level of hepar (liver) GHR1mRNA was at salinity 20, 25, 30 and higher at 35,
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