李芝珊臺灣大學：環境衛生研究所陳佳郁Chen, Jia-YuhJia-YuhChen2007-11-282018-06-302007-11-282018-06-302005http://ntur.lib.ntu.edu.tw//handle/246246/59781飲用水與民眾健康息息相關，為了防止水媒疾病（water-born diseases）的傳播，消毒已成為飲用水處理中不可或缺的過程。加氯消毒因成本低、操作簡單、效果好，且是唯一有能力提供餘量的方法，因此為目前最普遍使用的消毒方式。臭氧因具有超強氧化力、反應速率快，且無副產物等優點，成為目前最具潛力的消毒方法之一。本研究的主旨便是評估臭氧與氯對水中微生物之殺菌效率。 本研究包括二部分：第一部分以培養方法評估水中臭氧與加氯消毒之劑量效應關係，探討各項參數（菌種、菌濃度、臭氧濃度、暴露時間）對殺菌效率之影響，並討論美國環境保護署水中臭氧與加氯消毒標準下，不同微生物的殺菌效率；第二部分為應用非培養方法（Non-culture Method）－螢光染色法搭配流式細胞儀（Flow Cytometry, FCM）評估水中臭氧與加氯消毒效率，且同時與傳統培養方法（Culture Method）比較。 培養方法結果顯示，對於臭氧與氯的抵抗力：枯草桿菌內孢子＞青黴菌孢子＞酵母菌＞大腸桿菌；對臭氧殺菌效率之影響：臭氧濃度＞菌種差異＞暴露時間＞菌濃度；對加氯消毒效率之影響：菌種差異＞餘氯濃度＞暴露時間＞菌濃度；在美國環境保護署水中臭氧與加氯消毒標準下，對於細胞型態微生物來說，足以達到良好的殺菌效果；但對於孢子型態微生物的殺菌效率卻不及99.9％。 非培養方法結果顯示，比起傳統培養方法，以螢光染劑搭配流式細胞儀評估水中臭氧與加氯消毒效率時，可在殺菌機制與過程上提供更多的資訊，例如微生物在殺菌過程中總濃度的變化、細胞膜的受損程度或變化等，為一快速又十分有用的評估方法。此外，研究結果顯示消毒過程中，當微生物失去在培養基上的生長能力時，仍能保有一定程度的活性，因此使用傳統培養方法確實會高估殺菌效率。For preventing the spread of water-born diseases, disinfection has already become an essential part in drinking water treatment plant. Chlorine disinfection is the most popular disinfection method because of its cheap, simple, and efficient properties. On the other hand, Ozone is a very strong and fast-reacting oxidant without byproduct, as considered as a potential disinfection method. Our current study was to evaluate the inactivation efficiency of ozone and chlorine on microorganisms in water. By culture and non-culture（flow cytometry with fluorochrome, FCM/FL）methods, the influences of microorganism species, microorganism concentrations, ozone and chlorine concentrations, and exposure time on inactivation efficiencies of ozone and chlorine were evaluated. By culture method, it was found that the resistant abilities of tested microorganisms to both ozone and chlorine were as follows：endospore of Bacillus subtilis＞Penicillium citrinum spores＞Candida famata＞Escherichia coli. For ozone disinfection, the importance of the evaluated parameters were ozone concentration > species > exposure time > microbial concentrations. For chlorine disinfection, the importance was species, chlorine concentration, exposure time and microbial concentrations. Under the USEPA standards, the suggested doses could not provide efficient disinfection for spore type microorganisms. It is known that microorganism could be still viable when they lose their ability to grow on culture medium. In comparison with non-culture method, it was clearly demonstrated that inactivation efficiencies of ozone and chlorine on microorganisms were overestimated by culture method.摘 要..................................................I ABSTRACT...............................................II 總 目 錄............................................III 表 目 錄..............................................V 圖 目 錄.............................................VI 第一章 前 言...........................................1 1.1 研究緣起............................................1 1.2 研究目的............................................2 第二章 文獻回顧.........................................3 2.1 水中臭氧消毒........................................3 2.1.1 臭氧特性..........................................3 2.1.2 臭氧生成基本原理..................................3 2.1.3 臭氧消毒機制......................................5 2.1.4 影響臭氧消毒效率的因子............................6 2.2 加氯消毒...........................................11 2.2.1 氯的特性.........................................11 2.2.2 氯在水中的反應...................................11 2.2.3 加氯消毒機制.....................................12 2.2.4 影響加氯消毒效率的因子...........................12 2.3 微生物之分析.......................................14 2.3.1 培養方法（Culture Method）.......................14 2.3.2 非培養方法（Non–culture Methods）...............14 2.3.3 螢光顯微鏡（Epifluorescence Microscopy，EFM）....15 2.3.4 流式細胞儀（Flow Cytometry，FCM）................15 2.3.5 螢光染色法.......................................16 2.3.6 應用非培養方法於水中臭氧及加氯消毒之研究.........18 第三章 材料與方法......................................26 3.1驗儀器與材料........................................26 3.1.1 實驗儀器.........................................26 3.1.2 實驗材料.........................................27 3.1.3 實驗菌種之製備...................................28 3.2 實驗方法與步驟.....................................31 3.2.1 臭氧對水中微生物殺菌效率之評估...................31 3.2.2 氯對水中微生物殺菌效率之評估.....................33 3.2.3 應用螢光染色法搭配流式細胞儀於水中臭氧與加氯消毒效率評估...................................................35 3.3 統計分析...........................................39 第四章 結果與討論......................................45 4.1 臭氧對水中微生物殺菌效率之結果.....................45 4.1.1 臭氧對大腸桿菌殺菌效率之結果.....................45 4.1.2 臭氧對酵母菌殺菌效率之結果.......................47 4.1.3 臭氧對青黴菌孢子殺菌效率之結果...................49 4.1.4 臭氧對枯草桿菌內孢子殺菌效率之結果...............52 4.1.5 臭氧對水中微生物殺菌效率之綜合比較...............55 4.1.6 美國環保署水中臭氧消毒標準下之殺菌效率...........58 4.2 氯對水中微生物殺菌效率之結果.......................60 4.2.1 氯對大腸桿菌殺菌效率之結果.......................60 4.2.2 氯對酵母菌殺菌效率之結果.........................61 4.2.3 氯對青黴菌孢子殺菌效率之結果.....................64 4.2.4 氯對枯草桿菌內孢子殺菌效率之結果.................67 4.2.5 氯對水中微生物殺菌效率之綜合比較.................69 4.2.6 美國環保署加氯消毒標準下之殺菌效率...............71 4.3 應用螢光染劑搭配流式細胞儀於水中臭氧與加氯消毒效率評估之結果.................................................73 4.3.1臭氧殺菌效率評估之結果............................73 4.3.2氯對大腸桿菌殺菌效率評估之結果....................78 第五章 結論與建議....................................122 5.1 結論..............................................122 5.2建議...............................................123 參考文獻..............................................1242319064 bytesapplication/pdfen-US臭氧氯大腸桿菌枯草桿菌內孢子酵母菌青黴菌孢子ozonechlorineinactivationEscherichia coliBacillus subtilis sporesPenicillium citrinumCandida famata var. flarerithesishttp://ntur.lib.ntu.edu.tw/bitstream/246246/59781/1/ntu-94-R92844001-1.pdf