本文選題：抗性基因　切入點(diǎn)：抗性菌　出處：《天津大學(xué)》2016年碩士論文　論文類型：學(xué)位論文

【摘要】：大量研究顯示現(xiàn)在的消毒技術(shù)不僅不能除去ARGs和ARB,反而會(huì)使水中ARGs和ARB比例增加。但ARGs發(fā)生水平轉(zhuǎn)移和傳播的原因,卻未見研究報(bào)道。所以,本課題的目的是研究UV消毒對(duì)耐藥菌和耐藥基因(質(zhì)粒)的滅活規(guī)律,以及消毒過程中損傷菌的形成規(guī)律;探討UV消毒損傷菌的性質(zhì);明確游離質(zhì)粒轉(zhuǎn)化消毒損傷菌的形成條件和影響因素,為進(jìn)一步提出一種有效控制ARGs水平轉(zhuǎn)移和傳播的消毒新工藝提供科學(xué)依據(jù)和技術(shù)支撐。研究?jī)?nèi)容如下:1.以E.coli 25922(RP4)為研究對(duì)象,觀察UV消毒對(duì)其滅活及損傷菌形成的規(guī)律,對(duì)消毒釋放耐藥質(zhì)粒轉(zhuǎn)化活性,耐藥基因轉(zhuǎn)座子及耐藥基因的影響,還對(duì)5種耐藥菌的UV消毒抗性進(jìn)行了比較。UV消毒對(duì)E.coli 25922(RP4)作用的先后順序?yàn)?耐藥菌損傷耐藥菌滅活耐藥質(zhì)粒滅活耐藥基因的轉(zhuǎn)座子損傷耐藥基因的去除。5種耐藥菌(RP4)消毒抗性由強(qiáng)到弱為:S.aureus 6538約E.faecalis 33186Salmonell 50312約E.coli 25922P.aeruginosa 27853。2.采用透射電鏡、拉曼光譜分析及生物化學(xué)的方法,觀測(cè)了UV消毒致?lián)p傷菌的形態(tài)、物質(zhì)結(jié)構(gòu)、膜通透性和相關(guān)功能的改變。發(fā)現(xiàn)UV消毒可使細(xì)菌內(nèi)容物減少,且β-半乳糖苷酶活性提高及拉曼光譜分析結(jié)果均提示消毒后細(xì)菌的膜通透性發(fā)生了改變.此外,菌體中相關(guān)抗氧化酶活性也有明顯升高。3.以E.coli K12為待測(cè)目標(biāo)菌,采用大濾膜法(MF)、多管發(fā)酵法(MTF)、酶底物法(EST)分別對(duì)臭氧損傷菌進(jìn)行檢測(cè),計(jì)算其對(duì)臭氧損傷E.coli的檢出率。結(jié)果發(fā)現(xiàn):MTF在檢測(cè)野生損傷菌方面表現(xiàn)出較高的靈敏性,其檢出率最高可達(dá)100%,EST最多可檢測(cè)4.76%損傷的E.coli,而MF檢出率為0。4.通過實(shí)驗(yàn)室模擬水處理中的消毒處理,研究了游離ARGs轉(zhuǎn)化UV消毒損傷菌的條件及其影響因素。結(jié)果發(fā)現(xiàn):損傷菌轉(zhuǎn)化頻率較對(duì)照組高1~2個(gè)數(shù)量級(jí)。環(huán)境溫度、pH值,作用時(shí)間、氨氮、CODMn、金屬離子、受體菌濃度等因素雖對(duì)轉(zhuǎn)化有影響,但實(shí)驗(yàn)組轉(zhuǎn)化頻率均比相應(yīng)對(duì)照組高。以上結(jié)果說明:UV消毒促進(jìn)了ARGs的釋放,同時(shí)形成了大量具感受態(tài)性質(zhì)的消毒損傷菌,這兩者的存在促進(jìn)了自然轉(zhuǎn)化的發(fā)生。
[Abstract]:A large number of studies have shown that the current disinfection techniques not only can not remove ARGs and ARB, but also increase the proportion of ARGs and ARB in water. However, the causes of horizontal transfer and transmission of ARGs have not been reported. The purpose of this paper is to study the inactivation of drug-resistant bacteria and resistant genes (plasmids) in UV disinfection, and the formation of damaged bacteria in the process of disinfection, and to explore the properties of UV disinfection of drug-resistant bacteria and drug-resistant genes (plasmids). The forming conditions and influencing factors of free plasmids transformed into disinfectant damaged bacteria were determined. In order to provide scientific basis and technical support for a new disinfection process to effectively control the horizontal transfer and spread of ARGs, the research contents are as follows: 1. Taking E.coli 25922RP4) as the research object, the regularity of the inactivation and the formation of damaged bacteria by UV disinfection was observed. The effects on the transformation activity, transposon and drug resistance gene of drug resistance gene by disinfection and release of drug resistance plasmids. The sequence of UV disinfection resistance to E. coli 25922 (RP4) by UV disinfection was also compared. The sequence was as follows: drug resistant bacteria damage resistant bacteria, inactivated plasmid resistant plasmids, transposon damage resistance genes, removal of 5 drug resistance genes. The sterilizing resistance of RP4 was from strong to weak to that of: S.aureus 6538, E. faecalis 33186 Salmonell 50312, E. coli 25922P.aeruginosa 27853.2. transmission electron microscopy was used. The changes of morphology, material structure, membrane permeability and related functions of the damaged bacteria caused by UV disinfection were observed by Raman spectroscopy and biochemical methods. It was found that UV disinfection could reduce bacterial contents. Moreover, the increase of 尾 -galactosidase activity and Raman spectrum analysis indicated that the membrane permeability of bacteria changed after disinfection. In addition, the activity of antioxidant enzymes in bacteria also increased significantly. 3. E.coli K12 was used as the target bacteria. The ozone damaging bacteria were detected by large filter membrane method, multi-tube fermentation method and enzyme substrate method, respectively, and the detection rate of ozone damaged E. coli was calculated. The results showed that: MTF showed a high sensitivity in the detection of wild injured bacteria. The highest detectable rate of EST can be as high as 4.76% damage E.coli, while MF detection rate is 0.4. Disinfection treatment in simulated water treatment in laboratory, The conditions and influencing factors of free ARGs transformation for UV disinfection of damaged bacteria were studied. The results showed that the transformation frequency of the injured bacteria was 1 ~ 2 orders of magnitude higher than that of the control group. The environmental temperature, pH value, time of action, ammonia nitrogen, CODMn, metal ions, etc. Although the concentration of receptor bacteria had an effect on the transformation, the transformation frequency of the experimental group was higher than that of the corresponding control group. The existence of both promotes the occurrence of natural transformation.
【學(xué)位授予單位】：天津大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2016
【分類號(hào)】：R123.6

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