本文選題：富營(yíng)養(yǎng)化 + 生物反應(yīng)器　； 參考：《浙江師范大學(xué)》2016年碩士論文

【摘要】：由于水體中氮、磷等營(yíng)養(yǎng)物質(zhì)的過多進(jìn)入,水體富營(yíng)養(yǎng)化開始成為最常見的水污染現(xiàn)象,為人類的生活和生產(chǎn)活動(dòng)帶來嚴(yán)重危害。水體富營(yíng)養(yǎng)化的修復(fù)與治理己成為國(guó)內(nèi)外學(xué)者廣泛關(guān)注的研究課題之一。目前,吸附材料和微生物是富營(yíng)養(yǎng)化水體治理中較受關(guān)注的部分。但吸附材料的功能易受溫度、pH、磷濃度等的影響；參與實(shí)際應(yīng)用的微生物只占總量的極少數(shù)；特別是對(duì)隨季節(jié)而發(fā)生變化的富營(yíng)養(yǎng)化水體治理的詳細(xì)研究也較少見有報(bào)道。本研究中,運(yùn)用新材料構(gòu)建生物膜反應(yīng)器處理富營(yíng)養(yǎng)化水。一方面,進(jìn)行水體主要指標(biāo)(總磷：TP、總氮：TN、葉綠素a:Chl-a)的測(cè)定,驗(yàn)證生物反應(yīng)器對(duì)富營(yíng)養(yǎng)水的凈化能力；另一方面,利用MPN (most probable number,最大可能數(shù)法)培養(yǎng)體系和含有Rpf (resuscitation promoting factor,復(fù)蘇促進(jìn)因子)的培養(yǎng)上清液,分離富集處理系統(tǒng)中不同時(shí)期的優(yōu)勢(shì)菌群,確定反應(yīng)器中是否存在VBNC (viable but non-culturable,活的但非可培養(yǎng))狀態(tài)細(xì)菌及其不同處理時(shí)期水環(huán)境中細(xì)菌的組成,同時(shí)運(yùn)用PCR-DGGE探索解析Rpf對(duì)處理系統(tǒng)中VBNC狀態(tài)細(xì)菌的影響；對(duì)分離菌株進(jìn)行全基因組提取、PCR擴(kuò)增、16SrRNA基因測(cè)序、比對(duì)、系統(tǒng)發(fā)育樹構(gòu)建以及主要機(jī)能探索。取得的主要研究成果如下：1.新材料生物膜反應(yīng)器對(duì)富營(yíng)養(yǎng)化水有較好的水質(zhì)凈化處理能力,可以有效降低富營(yíng)養(yǎng)化水中的TP、TN和Chl-a。TP.TN及Chl-a平均去除率分別為95.9%,92.4%,95.8%,其中一些常見因素如溫度、pH、磷起始濃度等的變化對(duì)總磷的去除效果影響較小。2.MPN培養(yǎng)體系實(shí)驗(yàn)結(jié)果表明：每1.0 g濾料中,實(shí)驗(yàn)組(添加活性Rpf)細(xì)菌總數(shù)(2.9×109-2.1×1010)比對(duì)照組(添加失活Rpf)細(xì)菌總數(shù)(5.3×107-2.4×109)大,Rpf效果值(VR)為8.7-54.7。以上數(shù)據(jù)表明：新材料能有效富集水體中的微生物；富營(yíng)養(yǎng)化水處理系統(tǒng)中存在對(duì)Rpf敏感的優(yōu)勢(shì)菌群。分離菌株的16SrRNA基因比對(duì)結(jié)果和系統(tǒng)進(jìn)化關(guān)系顯示：生物處理系統(tǒng)的優(yōu)勢(shì)VBNC細(xì)菌主要來自Bacillus、Burkholderia、Enterobacter、Pseudomonas和Brevibacillus等屬；分離菌株與已知近緣標(biāo)準(zhǔn)菌株16S rRNA基因序列的相似性為97.6-100.0%。3.以取自生物反應(yīng)器的樣品構(gòu)建的MPN培養(yǎng)體系為模板進(jìn)行全基因組提取、PCR擴(kuò)增、DGGE電泳解析、目的基因序列鑒定與比對(duì)。DGGE圖譜表明,整體上實(shí)驗(yàn)組比對(duì)照組的菌種多樣性高,證實(shí)富營(yíng)養(yǎng)化反應(yīng)器中存在對(duì)Rpf敏感的VBNC菌群。4.菌群變遷觀測(cè)結(jié)果表明：處理系統(tǒng)中的微生物種類在不同階段存在一定差異。反應(yīng)器的每個(gè)階段都存在具有降解有機(jī)物和反硝化聚磷功能的Burkholderia cepacia和Bacillus cereus菌株；每個(gè)階段擔(dān)任溶藻功能的細(xì)菌則來自不同的屬：如Bacillus、Brevibacillus、Ochrobactrum和Enterobacter等屬；具有潛在脫氮功能的細(xì)菌則來自于Bacillus、Enterobacter和Pseudomonas等屬。新材料生物膜反應(yīng)器運(yùn)行近2年,可以有效凈化富營(yíng)養(yǎng)化水,對(duì)格柵、初沉池等進(jìn)行適當(dāng)調(diào)整,有望能起到更好的處理效果。此外,新材料因其強(qiáng)吸附能力,可成為功能菌群的良好載體,在實(shí)際生產(chǎn)中有望得到推廣使用。根據(jù)富營(yíng)養(yǎng)水處理反應(yīng)器中存在VBNC菌,及利用Rpf對(duì)復(fù)蘇活性化VBNC菌環(huán)境功能的研究事實(shí),可以推測(cè),不同的污水處理系統(tǒng)中存在VBNC菌群,利用Rpf對(duì)VBNC細(xì)菌進(jìn)行復(fù)蘇活化的實(shí)際應(yīng)用研究,將能為改進(jìn)污水處理工藝和設(shè)備及其提高生物處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.
【學(xué)位授予單位】：浙江師范大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2016
【分類號(hào)】：X703;X52

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