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人工湿地异养硝化细菌的分离、鉴定及其硝化性能研究
Alternative TitleSTUDIES ON THE IDENTIFICATION AND NITRATION OF HETEROTROPHIC NITRIFIER BACTERIAL STRAINS IN THE CONSTRUCTED WETLAND
王成林
Subtype硕士
Thesis Advisor吴振斌
2008-06-19
Degree Grantor中国科学院水生生物研究所
Place of Conferral水生生物研究所
Keyword人工湿地 异养硝化细菌 16s Rdna 分离 鉴定
Abstract异养硝化菌与传统的脱氮微生物相比,具有生长速率快、脱氮效率高、对环境条件要求低等优点,而现阶段国内有关异养硝化菌的研究特别是其在人工湿地系统脱氮方面的研究还比较少。本文以下行流-上行流复合水流方式的构建湿地—复合垂直流构建湿地系统为研究对象,采用将传统方法与现代分子生物学手段相结合的新型异养硝化细菌分离筛选方法,从复合垂直流人工湿地表层基质中,成功分离出了两株异养硝化细菌,编号为H-1,H-2,通过生理生化实验,biolog菌种鉴定,16S rDNA 序列分析,以及四因素三水平的正交试验,进行最佳硝化效果研究。 主要研究结果如下: 1 . 本文采用选择性培养基进行了异养硝化菌株的筛选,共得到了2株具有较高消化活性的细菌,编号为H-1,H-2,通过biolog实验,得到其碳源利用图谱,证明了其异养性。对其生长特性进行了研究,得到其生长曲线,最适生长碳、氮源、温度和pH。 2 .对菌株H-1,H-2,进行DNA提取和16S rDNA PCR扩增,16S rDNA 的序列分析结果显示,菌株H-1,H-2与产碱杆菌属(Alcaligenes)粪产碱杆菌(A.faecalis)有98%相似性,所以认为从人工湿地中分离的两株异养硝化细菌都属于产碱杆菌属粪产碱菌种。 3 . 通过四因素三水平的正交试验,结果显示:当温度为30 oC,pH为7.5,接种量为107 CFU,溶氧2.25mg/L时,该菌株亚硝化反应效果最佳,影响亚硝化反应效果的因素顺序为:溶氧>温度>pH>接种量,温度和溶氧影响极显著,pH和接种量影响相对不显著。当温度为20℃,pH为4.5,接种量为107CFU,溶氧3.76mg/L,硝化反应效果最佳,影响硝化反应效果的因素顺序为:温度> pH >接种量>溶氧;pH和溶氧影响极显著,温度和接种量相对不显著。
Other AbstractCompared with traditional bacteria, heterotrophic nitration bacteria had the advantage of higher growing rate higher efficiency and lower requirement to the environment. However, researches on the heterotrophic nitration bacteria especially in water treatment were few at present. In this paper, the heterotrophic nitration bacteria in the surface layers of the integrated vertical flow constructed wetland were studied by traditional and molecular biology methods and two heterotrophic nitration bacteria, named by H-1, H-2, were isolated. The optimal nitrification effects were studied through physiology and biochemistry experiments, biolog experiments, 16S rDNA phylogenetic analysis and orthogonal experiments. The maim results were summarized as follows: 1. The heterotrophic nitration bacteria were screened by selective incubation media and two strains of heterotrophic nitrifier which had higher nitrification potential, named by H-1, H-2, were isolated from the surface layers of the integrated vertical flow constructed wetland. The heterotrophic characteristics were proven by C fountain in the biolog and their bacterial growth curve, the optimal C/N fountain, temperature and pH were also obtained. 2. The phylogenetic analysis based on 16S rDNA of H-1、H-2 suggested that the bacterial strain were 98% homology at the nucleotide level compared with 16SrDNA of Alcaligenes faecalis. It was thought that bacterial strains H-1and H-2 were possibly Alcaligenes A. faecalis. 3. Orthogonal experiments of four factors and three levels were designed to screen out the optimum nitration condition. The results indicated that the nitrosification effect was optimal when the temperature, pH, inoculation quantities and dissolved oxygen were 300 C, 7.5, 107 CFU and 2.25mg/L, respectively, with the sorting order being dissolved oxygen> temperature>pH> inoculation quantities; The influences of temperature and dissolved oxygen on nitrosification effect were extremely significant, while the influences of pH and inoculation quantities were quite slight. Referring to the nitrification effect, the optimal condition was obtained when the temperature, pH, inoculation quantities and dissolved oxygen were 200 C, 4.5, 107 CFU and 3.76mg/L, respectively, with the sorting order being dissolved temperature > pH > inoculation quantities > dissolved oxygen. The influences of pH and dissolved oxygen on nitrification effect were highly significant, while the influences of temperature and inoculation quantities on nitrification effect were relatively slight.
Pages60
Language中文
Document Type学位论文
Identifierhttp://ir.ihb.ac.cn/handle/342005/12376
Collection学位论文
Recommended Citation
GB/T 7714
王成林. 人工湿地异养硝化细菌的分离、鉴定及其硝化性能研究[D]. 水生生物研究所. 中国科学院水生生物研究所,2008.
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