评价水产动物生长性能的生理生化指标和分子生物学指标

以奥利亚罗非鱼(Oreochromis aureus)为实验对象，设计了3种不同的摄食类型，分别是鲜活饵料组、饥饿3周后饱食投喂组和人工饲料组。鲜活饵料组投喂冰冻赤子爱胜蚓，利用蚯蚓体内丰富的营养成分和活性物质，以期获得奥利亚罗非鱼良好的生长状况；饥饿后饱食组是指饥饿3周后，以人工饲料饱食投喂2周，用于研究饥饿与补偿生长获得快速生长时血液理化指标的变化情况；人工饲料组作为对照组。纯淡水条件下养殖，水温25±2℃。测定了奥利亚罗非鱼在3种摄食类型饲喂下某些血液生理生化指标变化的情况，并将指标变化情况与增重率做相关性分析，试图找出能够反映奥利亚罗非鱼生长性能的血液生理生化指标。 研究结果表明，奥利亚罗非鱼在饥饿3周后获得了补偿生长，补偿生长时的增重率和特定生长率显著高于人工饲料组(P<0.05)，高于鲜活饵料组，但差别不显著；相关性分析研究表明血清总蛋白、胆固醇、四碘甲状腺原氨酸（T4）与增重率极显著相关(P<0.01)，血红蛋白显著相关(P<0.05)，红细胞、白细胞、碱性磷酸酶高度相关(相关系数为0.580、0.551和0.557)，因此，建议血清总蛋白、胆固醇和血红蛋白可作为能够反映罗非鱼生长性能的新指标。 根据序列设计引物，PCR反应条件：变性温度：95 ℃，3 min；退火温度：57℃，20 sec；延伸温度：72℃，5 min，共36个循环，从牙鲆、黑鲪和鲈鱼中克隆出胰岛素样生长因子（IGF-Ⅰ）部分序列，首次证实了IGF-Ⅰ在3种海水鱼中的存在。 利用蛋氨酸与ZnSO4•7H2O，在pH 5.5、80℃下，反应1小时，采用蛋氨酸与硫酸锌2：1的配料比，合成出了产物蛋氨酸螯合锌，蛋氨酸螯合锌外观白色，粉状，室温下微溶于水，不溶于乙醇，并用原子吸收光谱法测定其含锌量为15%，螯合率为88.2%。

Abstract: Three feeding categories were devised to feed tilapia (Oreochromis aureus): fresh earthworm, starvation for 3 weeks and re-feeding, formulated feed. The fresh earthworm group was fed with ice earthworm, and the earthworm contained rich nutrition gradients and active materials, the tilapia were expected to grow fast and well. The tilapia in compensatory group were starving for three weeks then fed with formulated feed for two weeks, and were used to study the change of blood biochemical indices during the period of starving and re-feeding. The formulated group was control group, and the tilapia were fed formulated feed bought from Qingdao aquaculture feed company. The tilapia were about 12 cm long, and about 90 g weight, and were divided to three feeding group, each group was triplicate, 18 tilapia each were raised in a tank, and its cubage was 200 L. The tilapia were raised in fresh water, and the temperature of water was 25±2 ℃, pH value was 8.0±0.5, three meals a day, 08:00, 14:00, 19:00, cleaned the leftover feed 1h after feeding, and exchange the 1/3 tank water. The experiment had investigated the changes of blood biochemical indices under different feed categories. Studies were carried out on the relationships between growth performance and blood indices, trying to find out which induces can exactly reflect growth performance. The result showed that; The tilapia acquired a compensatory growth, when the tilapia were during the period of compensatory growth, the weight gain ratio and specific growth rate was significantly higher than that in formulated group (p<0.05), and higher than that in fresh earthworm group, but not significantly. The fresh earthworm group and formulated group was significantly higher than that of compensatory group in total weight gain and specific growth rate (p<0.05). The fresh earthworm group was higher than that of the formulated group in total weight gain and specific growth rate, but not significantly. Glucose, triacylglyerol, the red blood cell count and hemoglobin dropped significantly after starving for one week, but then was stable in the following starving weeks; Total serum protein concentration did not drop significantly. When re-feeding for one week, the biochemical indices went up obviously. The serum total protein concentration，cholesterol and T4 was significantly correlated to the growth performance (p<0.01), the correlation coefficient was 0.872，0.803 and 0.876, separately; The correlated coefficient of hemoglobin was 0.615, and was significantly correlated to the growth performance (p<0.05); the red blood cell count, white blood cell count and AKP was high correlated to the growth performance, the correlation coefficient was 0.580，0.551 and 0.557, separately. Triacyglycerol and phosphorus was correlated to the growth performance, and the correlated coefficient was 0.366 and 0.231.The serum calcium was minus correlated to the growth performance, and the correlated coefficient was －0.121. From the study, the authors suggested the serum total protein, cholesterol, and hemoglobin be the new indices which could reflect growth performance. According to the sequence, we designed primers, and we cloned the part of the IGF-Ⅰgene sequence at the following conditions: denature temperature at 95 ℃，3 min; annealing temperature at 72℃，5 min, and 36 cycles from the Pacific fluke, Japanese sea bass, slickhead, and confirmed that IGF-Ⅰgene was in this three kinds of fish. Using Methioninum and Zinc sulfate heptahydrate as reagent, they were mixed together at pH 5.5、80℃ for 1 h, and their ratio was 2:1, finally we got the production Methionine chelate Zinc.The Methionine chelate Zinc is white, flouder, and it is hard to be dissolved in water at room temperate, and can not be dissolved in ethanol, it was detected by atomic absorption spectrometry, the Zinc was 15% percent, and its chlate ratio is 88.2%.