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1、 密 级： 学校代码：10075 分类号： 学 号：20091059 理学硕士学位论文 类球红细菌和枯草芽孢杆菌对养殖 水体的净化作用 学位申请人：岳 强 指 导 教 师 ：康现江 教授 学 位 类 别 ：理学硕士 学 科 专 业 ：水生生物学 授 予 单 位 ：河北大学 答 辩 日 期 ：二一二年五月 Classified Index： CODE：10075 U.D.C： NO：20091059 A Dissertation for the Degree of M. Science Purification of aquaculture water with Rhodobacter spha
2、eroides and Bacillus subtilis Candidate: Yue Qiang Supervisor: Prof. Kang Xianjiang Academic Degree Applied for: Master of Science Specialty: Hydrobiology University: Hebei University Date of Accomplishment: May，2012 摘 要 I 摘 要 近年来，随着人们生活水平的提高，水产养殖在我国蓬勃发展。但是随着养殖规模 的不断扩大，养殖集约化程度不断提高，在养殖过程中残饵和排泄物的不断积累，
3、使养 殖水体污染日益严重，这成为疾病频繁发生的主要诱因，直接危害着养殖对象。因而， 如何改善养殖水质成为水产养殖成败的关键问题之一， 利用有益微生物来改善养殖水体 的微生态环境、改善水质是当前研究的热点。本文从以下 4 个方面进行了研究。 1光合细菌的鉴定。本文通过对一株从白洋淀底泥中分离出的光合细菌进行碳源 利用、氮源利用等一系列生理生化指标的检测和 16SrDNA 的同源性比对，初步确定这 株光合细菌是类球红细菌(Rhodobacter sphaeroides)。 2类球红细菌和枯草芽孢杆菌净化养殖水体的研究。以本文分离的类球红细菌和 实验室保藏的枯草芽孢杆菌为添加对象，对养殖水体净化效果
4、进行研究。首先确定类球 红细菌和枯草芽孢杆菌单独使用时最佳用量，综合考虑净水效果和成本等因素，类球红 细菌的最佳使用量为 8104 cfu/mL，芽孢杆菌的最佳使用量为 18104 cfu/mL。为了研究 类球红细菌和枯草芽孢杆菌发挥协同作用的最佳配比，以 3:1、2:1、1:1、1:2、1:3 五种 配比的组合菌进行养殖水体净化实验。结果表明 1:1 配比组合即类球红细菌使用量为 4104 cfu/mL 和芽孢杆菌使用量为 9104 cfu/mL 组合的综合净水效果最好，氨氮的去除 率达到 72%，亚硝酸氮的去除率达到 82.5%。 3类球红细菌和枯草芽孢杆菌对中华鳖养殖池水的净化效果的研究
5、。实验结果表 明，混合菌组比单一菌组的水质净化效果要好，实验结束后，混合菌组氨氮的去除率为 42.9%，亚硝酸氮的去除率为 60.8%，COD 的去除率为 31.1%。 4类球红细菌和枯草芽孢杆菌对中华锯齿米虾养殖水体净化效果的研究，实验表 明添加混合菌的中华锯齿米虾养殖水体氨氮含量、亚硝酸氮含量和 COD 含量要明显低 于单一菌种添加组和对照组，45 天养殖结束后混合菌组与对照组相比氨氮降低 74.3%， 亚硝酸氮降低 70.6%，COD 降低 56.9%。 关键词 中华锯齿米虾 中华鳖 类球红细菌 枯草芽孢杆菌 水质净化 Abstract II Abstract In recent yea
6、rs, as peoples living standards improve, aquaculture in China has rapid development, while the farming intensification improved constantly with the expansion of the farming scale, and bait and excretion are accumulated continuously in the water of aquaculture, causing increasingly serious pollution
7、in aquaculture, which lead to the disease occurred frequently in aquaculture and endanger the culture organisms directly. Therefore, how to improve the aquaculture water quality become the key issues for the success or failure of aquaculture, and the use of beneficial micro-organisms to improve the
8、micro-ecological environment and water quality of the aquaculture water become a hot topic in current research. There are four aspects followed in the paper: 1 The identification of photosynthetic bacteria. In this paper, one photosynthetic bacteria isolated from the sediment of Baiyangdian is ident
9、ified as Rhodobacter sphaeroides, based on the results of morphological, physiological and biochemical characteristics and the homological analysis of 16S rDNA sequence. 2 The purification of aquaculture water with photosynthetic bacteria and Bacillus subtilis. Photosynthetic bacteria, isolated befo
10、re, and B.subtilis, stored in lab, are studied about their purification capacity to aquaculture water. The results showed that, the optimum concentration of photosynthetic bacteria for purification is 8104 cfu/ml, and the optimum concentration of B.subtilis is 18104 cfu/ml, according to the comprehe
11、nsive analysis of water purification effect and the cost. To find the synergy ratio of R.sphaeroides and B.subtilis, experiments about water purification with five kinds of ratio combination of bacteria (3:1, 2:1, 1:1, 1:2, 1: 3) were studied, which results showed that the best ratio of R.sphaeroide
12、s and B.subtilis is 1:1 that ammonia and nitrite reduced 72% and 82.5% respectively. 3 The purification of aquaculture water of Trionyx sinensis with R.sphaeroides and B.subtilis. The results of experiments showed that, the purification effect in the aquaculture water in which there are mixed bacter
13、ia is better than that in the aquaculture water in which there is only one bacteria, and the ammonia ,the nitrite and the COD are reduced 42.9%, 60.8% and 31.1% after 8 days, respectively. 4 The purification of aquaculture water of Neocaridina denticulate sinensis with R.sphaeroides and B.subtilis.
14、The purification study showed similar results that the ammonia, the nitrite and the COD in the aquaculture water in which there are mixed bacteria is lower Abstract III obviously than that in the aquaculture water in which there is only one bacteria. After 45 days, the ammonia, the nitrite and the C
15、OD in the water in which there are mixed bacteria are reduced 74.3%, 70.6%, and 56.9%, respectively. Key words Neocaridina denticulate sinensis Trionyx sinensis Rhodobacter sphaeroides Bacillus subtilis Identification of bacteria 目 录 IV 目 录 第 1 章 绪 论1 1.1 研究背景1 1.1.1 亚硝酸氮和氨氮的危害1 1.1.2 净化水质的方法1 1.2 微
16、生态制剂2 1.2.1 微生态制剂生物菌的特征3 1.2.2 光合细菌3 1.2.3 芽孢杆菌5 1.3 研究的目的和意义6 第 2 章 光合细菌的鉴定7 引 言.7 2.1 材料与方法7 2.1.1 实验材料7 2.1.2 主要仪器7 2.1.3 实验方法8 2.2 结果12 2.2.1 细菌的生理生化鉴定12 2.2.2 细菌的 16S rDNA 测定.12 2.3 讨论14 第 3 章 类球红细菌与枯草芽孢杆菌的最佳投放量15 引 言.15 3.1 材料与方法15 3.1.1 实验材料15 3.1.2 实验方法15 3.2 结果和讨论17 3.2.1 生长曲线的测定17 目 录 V 3.2.2 最大活菌数的测定18 3.2.3 类球红细菌投放组对水质的影响19 3.2.4 芽孢杆菌投放组对水质的影响20 3.2.5 类球红细菌和枯草芽孢杆菌最佳混合比例22 第
