新材料生物膜反应器净化富营养水及其功能菌群解析

发布时间：2018-06-06 08:11

本文选题：富营养化 + 生物反应器　；参考：《浙江师范大学》2016年硕士论文

【摘要】：由于水体中氮、磷等营养物质的过多进入,水体富营养化开始成为最常见的水污染现象,为人类的生活和生产活动带来严重危害。水体富营养化的修复与治理己成为国内外学者广泛关注的研究课题之一。目前,吸附材料和微生物是富营养化水体治理中较受关注的部分。但吸附材料的功能易受温度、pH、磷浓度等的影响；参与实际应用的微生物只占总量的极少数；特别是对随季节而发生变化的富营养化水体治理的详细研究也较少见有报道。本研究中,运用新材料构建生物膜反应器处理富营养化水。一方面,进行水体主要指标(总磷：TP、总氮：TN、叶绿素a:Chl-a)的测定,验证生物反应器对富营养水的净化能力；另一方面,利用MPN (most probable number,最大可能数法)培养体系和含有Rpf (resuscitation promoting factor,复苏促进因子)的培养上清液,分离富集处理系统中不同时期的优势菌群,确定反应器中是否存在VBNC (viable but non-culturable,活的但非可培养)状态细菌及其不同处理时期水环境中细菌的组成,同时运用PCR-DGGE探索解析Rpf对处理系统中VBNC状态细菌的影响；对分离菌株进行全基因组提取、PCR扩增、16SrRNA基因测序、比对、系统发育树构建以及主要机能探索。取得的主要研究成果如下：1.新材料生物膜反应器对富营养化水有较好的水质净化处理能力,可以有效降低富营养化水中的TP、TN和Chl-a。TP.TN及Chl-a平均去除率分别为95.9%,92.4%,95.8%,其中一些常见因素如温度、pH、磷起始浓度等的变化对总磷的去除效果影响较小。2.MPN培养体系实验结果表明：每1.0 g滤料中,实验组(添加活性Rpf)细菌总数(2.9×109-2.1×1010)比对照组(添加失活Rpf)细菌总数(5.3×107-2.4×109)大,Rpf效果值(VR)为8.7-54.7。以上数据表明：新材料能有效富集水体中的微生物；富营养化水处理系统中存在对Rpf敏感的优势菌群。分离菌株的16SrRNA基因比对结果和系统进化关系显示：生物处理系统的优势VBNC细菌主要来自Bacillus、Burkholderia、Enterobacter、Pseudomonas和Brevibacillus等属；分离菌株与已知近缘标准菌株16S rRNA基因序列的相似性为97.6-100.0%。3.以取自生物反应器的样品构建的MPN培养体系为模板进行全基因组提取、PCR扩增、DGGE电泳解析、目的基因序列鉴定与比对。DGGE图谱表明,整体上实验组比对照组的菌种多样性高,证实富营养化反应器中存在对Rpf敏感的VBNC菌群。4.菌群变迁观测结果表明：处理系统中的微生物种类在不同阶段存在一定差异。反应器的每个阶段都存在具有降解有机物和反硝化聚磷功能的Burkholderia cepacia和Bacillus cereus菌株；每个阶段担任溶藻功能的细菌则来自不同的属：如Bacillus、Brevibacillus、Ochrobactrum和Enterobacter等属；具有潜在脱氮功能的细菌则来自于Bacillus、Enterobacter和Pseudomonas等属。新材料生物膜反应器运行近2年,可以有效净化富营养化水,对格栅、初沉池等进行适当调整,有望能起到更好的处理效果。此外,新材料因其强吸附能力,可成为功能菌群的良好载体,在实际生产中有望得到推广使用。根据富营养水处理反应器中存在VBNC菌,及利用Rpf对复苏活性化VBNC菌环境功能的研究事实,可以推测,不同的污水处理系统中存在VBNC菌群,利用Rpf对VBNC细菌进行复苏活化的实际应用研究,将能为改进污水处理工艺和设备及其提高生物处理效果提供新的研究思路和方法。
[Abstract]:Due to the excessive entry of nitrogen, phosphorus and other nutrients in the water body, the eutrophication of water body has become the most common water pollution phenomenon, causing serious harm to human life and production activities. The restoration and treatment of water eutrophication has become one of the research topics of extensive concern at home and abroad. At present, the adsorption material and microorganism are rich camps. But the function of the adsorbed material is easily affected by temperature, pH, phosphorus concentration, and so on. The microorganism involved in the actual application is only a few of the total amount; in particular, the detailed study on the regulation of eutrophic water with the seasonal variation is seldom reported. In this study, the construction of new materials is used. Biofilm reactor was used to treat eutrophic water. On the one hand, the main indexes of water body (total phosphorus: TP, total nitrogen: TN, chlorophyll a:Chl-a) were measured to verify the purification ability of bioreactor to eutrophic water; on the other hand, MPN (most probable number, maximum possible number method) culture system and Rpf (resuscitation promoting facto) were used. R, resuscitation promoting factor) culture supernatant, separating and enriching the dominant bacteria in different periods of the system, determining whether there is VBNC (viable but non-culturable, live but non culturable) state bacteria and the formation of bacteria in the water environment at different treatment periods, and the PCR-DGGE is used to explore the Rpf to the VBN in the treatment system. The effect of C state bacteria; complete genome extraction, PCR amplification, 16SrRNA gene sequencing, comparison, phylogenetic tree construction and main functional exploration. The main research results are as follows: 1. new material biofilm reactor has good water quality purification ability for eutrophic water and can effectively reduce eutrophication. The average removal rates of TP, TN, Chl-a.TP.TN and Chl-a in water were 95.9%, 92.4% and 95.8% respectively. Some of the common factors, such as temperature, pH, and phosphorus initial concentration, affected the removal efficiency of total phosphorus in the.2.MPN culture system, and the results showed that the total number of bacteria (2.9 x 109-2.1 x 1010) in the experimental group (adding active Rpf) was compared with the.2.MPN culture system. The total number of bacteria (adding inactivated Rpf) was large (5.3 x 107-2.4 x 109), and the Rpf effect value (VR) above 8.7-54.7. indicated that the new material could effectively enrich the microorganisms in the water body; the eutrophic water treatment system had the dominant colony of Rpf sensitive bacteria. The relationship between the 16SrRNA gene comparison result and the phylogenetic relationship of the isolated strains showed that the biological sites were in the biological place. The dominant VBNC bacteria of the system are mainly from Bacillus, Burkholderia, Enterobacter, Pseudomonas and Brevibacillus, and the similarity between the isolated strain and the known genetic sequence of the known standard strain 16S rRNA is the whole genome extraction and PCR amplification of the MPN culture system constructed from the samples from the bioreactor. DGGE electrophoresis analysis, the target gene sequence identification and comparison.DGGE map showed that the whole experimental group was more diverse than the control group. The results showed that there was a change in the.4. bacteria group of the VBNC bacteria sensitive to the Rpf in the eutrophic reactor. The results showed that the microbial species in the treatment system were different in different stages. There are Burkholderia cepacia and Bacillus cereus strains with degrading organic and denitrifying phosphorus accumulation at each stage; bacteria that act as algae lysing from each stage are derived from different genera, such as Bacillus, Brevibacillus, Ochrobactrum and Enterobacter, and the bacteria with potential for denitrification come from Bacillus, Ente. The new material biofilm reactor, such as robacter and Pseudomonas, can effectively purify eutrophic water for nearly 2 years and adjust the grid and initial sink properly. It is expected to have better treatment effect. In addition, the new material can be a good carrier of functional bacteria because of its strong adsorption capacity. It is expected to be popularized in practical production. According to the existence of VBNC bacteria in the eutrophic water treatment reactor and the fact that using Rpf to resuscitate the environmental function of the active VBNC bacteria, we can speculate that there is a group of VBNC bacteria in different sewage treatment systems, and the practical application of Rpf for the recovery and activation of VBNC bacteria will be able to improve the wastewater treatment process and equipment and improve the biology. The treatment effect provides new research ideas and methods.
【学位授予单位】：浙江师范大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：X703;X52

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