【作者】 肖志猛；

【导师】 陈昌福； 杨季芳；

【作者基本信息】 华中农业大学 ， 水产养殖， 2006， 硕士

【摘要】 本文于2004年6月到2005年12月对宁波象山港养殖区域浮游细菌、浮游病毒、以及蛭弧菌生物量分布进行采样调查，并比较了几种染色法用于计数细菌生物量的优缺点；同时本文还对免疫多糖对黄鳝的免疫增强作用进行了研究，结果如下： 1．用DAPI染色法对纯培养的菌株0503和0501进行染色并计数，结果显示，DAPI染色法计数结果与细胞计数板法和平板涂布法结果非常接近，DAPI染色的细菌背景突出，计数方便。同时运用这几种方法对宁波象山港养殖区海水细菌总量进行计数，结果显示，海水、塘水、塘泥Zobell培养基计数结果分别为3.75×10~4cfu／ml、3.2×10~4cfu／ml、1.15×10~5cfu／g；DAPI染色计数结果分别为1.44×10~7cells／ml、1.16×10~7cells／ml、5.56×10~7cells／g；两种计数结果存在数量级上的差异。从DAPI染色效果来看，虽然海水中细菌大小不一，但是均可形成鲜明的亮点而从背景中突出，因此，检出度更高。 2．对宁波象山港养殖区四个采样点浮游病毒和浮游细菌生物量进行采样调查，结果显示，秋季浮游细菌生物量在10~5～10~7cells／ml数量级之间，而浮游病毒生物量无论池塘还是海湾均在10~5cells／ml数量级；冬季浮游细菌生物量在10~5～10~6cells／ml数量级之间，而浮游病毒生物量无论池塘还是海湾均为10~4cells／ml数量级。与以往报道相比，浮游病毒丰度并不算高。 3．通过以海水养殖动物病原菌为靶细菌在象山港养殖水域成功诱导分离到各种蛭弧菌，开展了蛭弧茵、弧菌、异养细菌季节生态分布特征研究。研究结果表明，不同种类或同种不同菌株宿主其所对应的敏感蛭弧菌数量呈现明显差异，副溶血弧菌敏感蛭弧菌检出率(数值)最高。同一宿主检测出的敏感蛭弧菌在近岸海水、池塘水及底泥中的分布呈现差异，其分布特征与异养细菌、弧菌的分布特征呈正相关。近岸海水中异养细菌和弧菌总数均高于塘内海水；虾塘表泥中异氧细菌和弧菌总数均高于近岸海水和池塘海水中细菌数量；夏季各种细菌数量最高，冬季最低。蛭弧菌的最适生长温度为25～30℃。 4．在饲料中添加不同剂量的免疫多糖(酵母细胞壁)，连续投喂经注射接种嗜水气单胞菌(Aeromonas hydrophila)脂多糖(Lipopolysaccharide，LPS)的黄鳝(Monopterus albus)28d后，通过测定供试黄鳝的增重量、血清中凝集抗体效价、溶菌酶活性、谷丙转氨酶活性(SGPT)、血清总蛋白含量、白细胞吞噬活性以及活菌攻毒后的免疫保护率(RPS)，探讨了将LPS作为免疫原、免疫多糖作为免疫刺激更多还原

【Abstract】 In this paper, we investigated the distribution of bacterioplankton, virioplankton, bdellovibrio and we also compared 3 methods in which we can calculate the biomass of the bacterioplankton; also we discussed the enhancement of immune response in ricefleld eel (Monopterus albus) inoculated with lipopolysaccharide of Aeromonas hydrophila UsingYeast Cell Wall, the results revealed that:1. We used DAPI method to calculate the biomass of strain of 0503 and 0501, the results revealed that the calculating method of DAPI and cell counting chamber was almost the same. DAPI method was very clear in which the bateria can easily distincted from the background. We also used DAPI method to calculate the biomass of bacterioplankton in aquaculture area, the results revealed that using culture medium method the biomass of bacterioplankton in sea water, pond and sediment was 3.75 X 10~4cfu/ml、3.2×10~4 cfu/ml、1.15×10~5 cfu /g respectively; DAPI method was 1.44× 10~7cells/ml、1.16×10~7cells/ml、5.56×10~7cells/g respectively. the 2 method in calculating have great difference. DAPI method is very bright and clear, we can easily distinct the bactiria from the backgroud.2. In Autum and Winter we investigated the the biomass of bacterioplankton and virioplankton in aquaculture area of Ningboxiangshan bay. The result showed that the biomass of bacterioplankton in autum was 10~5～10~6cells/ml in the 4 sampling locations and virioplankton was 10~5 cells/ml; In winter the biomass of bacterioplankton was about 10~5～10~6cells/ml and the biomass of virioplankton was about 10~4cells/ml. Comparing with the other report the biomass of virioplankton is not very high.3. In this study, we investigated the distribution of heterotrophic bacterium, Bdellovibrios and vibrios in different seasons in Ningboxiangshan bay by the different host of pathogenic bacterium. The results showed that the number of sensitive Bdellovibrios and vibrios was very different in the same or different host, and the detection rate of haemolyticus Bdellovibrios was most highest. An obtained similar on distributional characteristics of Bdellovibrios and vibrios in the same sensitive host in seawater alongshore, pond water and sediment revealed a presence of strong association.更多还原