發(fā)布時(shí)間：2018-07-15 22:12

【摘要】：目前我國(guó)正大力推廣再生水的應(yīng)用，保障再生水的水質(zhì)安全是關(guān)鍵問題。病原菌在消毒壓力下會(huì)進(jìn)入活性但不可培養(yǎng)（VBNC）狀態(tài)，無法用培養(yǎng)法檢測(cè)出，但仍保留原有的毒力因子和致病性，可在消毒壓力解除后復(fù)活繁殖而導(dǎo)致再生水回用的微生物健康風(fēng)險(xiǎn)。本研究針對(duì)VBNC病原菌復(fù)活風(fēng)險(xiǎn)，建立了逆轉(zhuǎn)錄活性和細(xì)胞膜完整性的檢測(cè)方法體系，深入分析再生水系統(tǒng)中VBNC病原菌的存在去除特性和復(fù)活特性，探究VBNC病原菌復(fù)活的關(guān)鍵影響因子；應(yīng)用焦磷酸測(cè)序技術(shù)，研究病原菌的群落結(jié)構(gòu)變化；評(píng)估再生水市政雜用的微生物健康風(fēng)險(xiǎn)。在此基礎(chǔ)上提出控制VBNC病原菌復(fù)活的方法。 本研究結(jié)果表明，進(jìn)水中可培養(yǎng)大腸桿菌、糞腸球菌、沙門氏菌、分枝桿菌的濃度水平為3~5LogCFU/100mL。經(jīng)過深度處理工藝，出水中處于VBNC狀態(tài)的大腸桿菌、糞腸球菌、沙門氏菌濃度水平為1.5~3.8Log CE/100mL，占各自總活性菌含量的40~100%；分枝桿菌對(duì)消毒劑的抗性較強(qiáng)，出水中VBNC狀態(tài)分枝桿菌濃度為1Log CE/100mL，占總活性菌的20%。膜處理工藝對(duì)可培養(yǎng)菌與活性菌的去除量相近約為1Log。消毒工藝對(duì)培養(yǎng)菌的去除量可達(dá)2~3Log，對(duì)活性菌的去除量1Log，對(duì)分枝桿菌作用不顯著。 再生水貯存輸送過程中，處于VBNC狀態(tài)的病原菌大量復(fù)活，四種細(xì)菌的復(fù)活率達(dá)0.3~2.9Log。余氯與微生物濃度水平存在相關(guān)性。有機(jī)物和營(yíng)養(yǎng)元素會(huì)促進(jìn)微生物在管網(wǎng)中的復(fù)活，但其變化與微生物的復(fù)活不存在顯著相關(guān)性。 采用454焦磷酸測(cè)序分析再生水生產(chǎn)、貯存和輸送過程中群落結(jié)構(gòu)的變化，結(jié)果表明經(jīng)過消毒工藝、貯存和輸送，微生物群落結(jié)構(gòu)發(fā)生顯著變化。管網(wǎng)末端，Proteobacteria是主要的門類，包含大量病原菌Burkholderiales。采用典范對(duì)應(yīng)分析（CCA），考察理化因子對(duì)生物群落的影響，發(fā)現(xiàn)TN、TP、NO3-N和TOC是最重要的影響因素。 對(duì)再生水用作市政雜用的微生物健康風(fēng)險(xiǎn)進(jìn)行評(píng)價(jià)，，得出年健康風(fēng)險(xiǎn)值與病原菌濃度水平的相關(guān)曲線。再生水出廠中，VBNC病原菌含有的潛在風(fēng)險(xiǎn)值比可培養(yǎng)病原菌高2個(gè)數(shù)量級(jí)。經(jīng)貯存輸送后，該風(fēng)險(xiǎn)值成為顯性風(fēng)險(xiǎn)值，影響再生水回用的微生物健康風(fēng)險(xiǎn)。
[Abstract]:At present, our country is popularizing the application of reclaimed water. It is a key problem to ensure the safety of reclaimed water quality. The pathogen can enter into the active but unculturable (VBNC) state under disinfection pressure, which can not be detected by culture method, but it still retains the original virulence factor and pathogenicity. Microbial health risks that can be reactivated to reproduce after disinfection pressure is released, leading to reuse of reclaimed water. In this study, the reverse transcription activity and cell membrane integrity of VBNC pathogens were analyzed by reverse transcriptase assay, and the removal and reactivation characteristics of VBNC pathogens were analyzed. To explore the key factors influencing the revival of VBNC pathogens; to study the community structure changes of pathogens by pyrosequencing technology; and to evaluate the microbial health risk of municipal mixed use of reclaimed water. On this basis, the method of controlling the resurrection of VBNC pathogens was put forward. The results showed that the concentration of Escherichia coli, Enterococcus faecalis, Salmonella and Mycobacterium in the influent was 3 5 LogCFU / 100 mL. After advanced treatment, the concentration levels of Escherichia coli, Enterococcus faecalis and Salmonella in VBNC in the effluent were 1.5 ~ 3.8Log CE / 100 mL, accounting for 40,100 of the total active bacteria, and the resistance of Mycobacterium to disinfectants was stronger. The concentration of Mycobacterium in VBNC state was 1 Log CE / 100 mL, accounting for 20% of the total active bacteria. The removal of culturable bacteria and active bacteria by membrane treatment process was similar to that of 1Log. The removal amount of culture bacteria was 2 ~ 3Log. the removal amount of active bacteria was 1Log. the effect on Mycobacterium was not significant. During the storage and transportation of reclaimed water, the pathogenic bacteria in the state of VBNC were resurrected in large numbers, and the resurrection rate of the four bacteria reached 0.3n2.9Log. Residual chlorine was correlated with microbial concentration. Organic matter and nutrient elements can promote the resurrection of microorganisms in the pipe network, but there is no significant correlation between the changes and the resurrection of microorganisms. 454 pyrosequencing was used to analyze the changes of community structure during the production, storage and transportation of reclaimed water. The results showed that the microbial community structure changed significantly after disinfection, storage and transportation. Proteobacteria at the end of pipe network is the main category, including a large number of pathogenic bacteria Burkholderia. Canonical correspondence analysis (CCA) was used to investigate the effects of physicochemical factors on biota. It was found that TNT TPNO3-N and TOC were the most important factors. The microorganism health risk of reclaimed water used as municipal miscellaneous was evaluated and the correlation curve between annual health risk value and pathogen concentration level was obtained. The potential risk value of VBNC pathogens in reclaimed water was two orders of magnitude higher than that of culturable pathogens. After storage and transportation, the risk value becomes dominant risk value, which affects the microbial health risk of reclaimed water reuse.
【學(xué)位授予單位】：清華大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2013
【分類號(hào)】：R123;X824

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本文編號(hào)：2125543