臺灣博碩士論文加值系統

近年來由於環保意識抬頭、水資源匱乏及水庫開發不易，廢水回收利用已成為相當重要之議題。若有害物質或致病性微生物仍存在於回收利用的廢水當中，則可能對於人們的健康及環境造成更大的傷害。因此，廢水中各種有害物質及致病性微生物是否能在廢水處理廠中有效被處理，對於廢水回收利用是相當重要的。
本研究乃針對17個國內各類型廢水處理廠，採集共41個水樣，以聚合酵素連鎖反應法(PCR)進行退伍軍人桿菌分析。根據PCR分析後之定序結果，總計共有10處廢水處理廠（58.8%）之25個水樣檢測出退伍軍人桿菌（61.0%），包含六種不同型別之退伍軍人菌—Uncultured Legionella spp.、Legionella pneumophila str. Paris、Legionella lytica、Legionella pneumophila serogroup 6、Legionella anisa以及Legionella donaldsonii，其中有2處廢水廠檢測出致病率最高的嗜肺性退伍軍人桿菌(Legionella pneumophila)。使用MPN-PCR方法推估廢水水體之退伍軍人桿菌原始濃度大多都落於6.0 × 104 CFU / L至1.9 × 105 CFU / L之間。而14個廢水處理廠之空氣樣本均無發現退伍軍人桿菌之存在。
根據實驗結果顯示，部分廢水處理廠因為處理設施之清潔，及排放前添加次氯酸鈉，而有效控制退伍軍人桿菌。紫外光使退伍軍人桿菌失活速度極快，照射劑量大於2.2 mWs / cm2時即可使失活率達到90％。蛋白質分解酵素在CT值大於330000（mg / L • min）時，即可使退伍軍人桿菌之失活率達到90％。次氯酸鈉使退伍軍人桿菌失活能力佳，在CT值大於98（mg / L • min）時，即可使退伍軍人桿菌失活率達到90％。
廢水處理廠之廢水中的確含有退伍軍人桿菌，而如何有效控制廢水中之退伍軍人桿菌，以降低從業人員及廢水再回收利用之感染風險，是未來廢水處理相當重要的課題。

Wastewater reclaiming and recycling is quite important , because of the shortage of water resource and reservoir of resuming in environmental consciousness in recent years. If harmful substance or the pathogenic microorganisms public existed in the reclaimed wastewater, it may cause great injury to people's health and environment. Whether harmful substances and pathogenic microorganisms in the wastewater can be dealt with and disinfected by the wastewater treatment plant, it is quite important for reclaiming and recycling wastewater.
This research analyzed 41 wastewater samples of 17 wastewater treatment plants in Taiwan by polymerase chain reaction (PCR). Twenty five wastewater samples (61%) of 10 wastewater treatment plants (58.8%) are detected positive for Legionella by PCR and DNA sequencing, including six kinds of different Legionella spp. - Uncultured Legionella spp., Legionella pneumophila str. Paris, Legionella lytica, Legionella pneumophila serogroup 6, Legionella anisa and Legionella donaldsonii. highly infectious Legionella pneumophila were detected in 2 wastewater treatment plants. The Legionella in the detected raw wastewater are ranged from 6.0 × 104 CFU/L to 1.9 × 105 CFU/L by MPN-PCR estimation. All of 14 air samples collected from wastewater treatment plants were not detected Legionella. Legionella in some wastewater treatment plants are effectively controlled by cleaned treatment procedures or adding HOCl to the discharged wastewater. Ultraviolet (UV) Light is an effective method. It achieved more than 90% deactivations of Legionella while exposing at 2.2 mWs/cm2. The Protease disinfection made Legionella achieved more than 90% deactivations while CT values more then 330000 mg/L•min. It is also effective that Legionella disinfected by HOCl. The HOCl disinfection made Legionella achieved more than 90% deactivations while CT values more then 98 mg/L•min.
Legionella was widely detected in wastewater so that it is very important to control the Legionella in wastewater, in order to reduce the infection risk of people in wastewater and reclaimed wastewater.

第一章 前言 .1
1.1 研究背景 1
1.2研究目標 2
1.3研究架構 2
第二章 文獻回顧 4
2.1退伍軍人桿菌之發現 4
2.2退伍軍人桿菌生存與培養環境 5
2.2.1退伍軍人桿菌之生存環境 5
2.2.2環境水體之前處理與退伍軍人桿菌培養方法 8
2.3退伍軍人桿菌之感染與案例 11
2.3.1退伍軍人桿菌之感染與診斷 11
2.3.2國外感染案例 14
2.3.3國內感染案例 16
2.4廢水處理廠之微生物感染風險 17
2.4.1一般微生物之感染風險 17
2.4.2退伍軍人桿菌之感染風險 18
2.5退伍軍人桿菌檢測方法之比較 20
2.6 退伍軍人桿菌之定量分析法―MPN-PCR 21
2.7退伍軍人桿菌之消毒方法 24
2.7.1紫外光 27
2.7.2次氯酸鈉 29
第三章 研究方法與材料 31
3.1研究流程 31
3.2實驗藥品 32
3.3實驗設備 33
3.4退伍軍人桿菌正控制組 35
3.4.1正控制組來源 35
3.4.2菌種保存與活化 35
3.4.3計算菌液濃度 36
3.5樣本採集 39
3.5.1採樣所需設備及注意事項 39
3.5.2採樣地點之挑選 41
3.5.3水體樣本採集 42
3.5.3空氣樣本採集 43
3.6指標性微生物檢測 44
3.7物理性指標檢測 45
3.7.1水溫 45
3.7.2 pH值 45
3.7.3導電度 45
3.7.4溶氧量 46
3.7.5濁度 46
3.8 水樣濃縮 46
3.9 DNA萃取 47
3.10聚合酵素連鎖反應法 48
3.10.1引子設計與序列 48
3.10.2反應溶液配置與反應條件 50
3.10.2.1 PCR 50
3.10.2.2 Second –PCR與Seminested -PCR 51
3.11聚合酵素連鎖反應法之結果檢驗 52
3.12退伍軍人桿菌分型 52
3.12.1 DNA純化 52
3.12.2 DNA定序及型別鑑定 54
3.12.3退伍軍人桿菌之分型 54
3.13退伍軍人桿菌定量分析 55
3.13.1 MPN-PCR定量實驗方法 55
3.13.2 MPN-PCR結果與培養結果對照 55
3.13.3廢水水樣之定量 56
3.14退伍軍人桿菌失活分析 57
3.14.1紫外光失活效能評估 57
3.14.2蛋白質分解酵素失活效能評估 58
3.14.3次氯酸鈉失活效能評估 58
第四章 結果與討論 61
4.1廢水處理廠中退伍軍人桿菌之相關分析 61
4.1.1廢水水體中退伍軍人桿菌之檢驗結果 61
4.1.2空氣樣本中退伍軍人桿菌之檢驗結果 63
4.1.3退伍軍人桿菌型別之比較 65
4.1.4廢水處理設施之清潔與添加次氯酸鈉 70
4.2廢水中微生物及物理指標之分析 72
4.2.1台灣地區廢水中微生物及物理指標之概況 72
4.2.2各類型廢水處理廠之微生物及物理指標分析 74
4.2.3各指標與退伍軍人桿菌之相關性分析 76
4.3 MPN-PCR定量分析 78
4.3.1 MPN與CFU對照公式建立 78
4.3.2環境樣本之MPN-PCR結果 79
4.4退伍軍人桿菌失活分析 81
4.4.1紫外光失活效能分析 81
4.4.2蛋白質分解酵素失活效能分析 84
4.4.3次氯酸鈉失活效能分析 87
第五章 結論 91
第六章 建議 93
參考文獻 94

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