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研究生:李勇廷
研究生(外文):Yong-ting Li
論文名稱:不同臭氧化生物濾床出水中有機物性質之變動
論文名稱(外文):The variations of the organic property in the effluent from the different ozonated biofilters
指導教授:賴文亮賴文亮引用關係
指導教授(外文):Wen-Liang Lai
學位類別:碩士
校院名稱:大仁科技大學
系所名稱:環境管理研究所
學門:環境保護學門
學類:環境資源學類
論文種類:學術論文
論文出版年:2008
畢業學年度:96
語文別:中文
論文頁數:122
中文關鍵詞:變性梯度膠體電泳生物濾床聚合酶鏈鎖反應
外文關鍵詞:biofilterDGGEPCR
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以0.2 µm過濾後之澄清湖原水,控制臭氧與非揮發性溶解性有機碳(O3/NPDOC)比值分別為0、1與2,再加入適量之澄清湖水之原生菌,控制其啟始菌數,之後,利用蠕動幫浦將臭氧化之原水打入預先滅菌填充玻璃珠層析管,進行約100 天之操作,研究除進行22種胺基酸及取自Aldrich公司生產腐植酸標準品進行螢光激發發射光譜圖(excitation emission fluorescent matrix, EEFM)進行全譜分析比較,另濾床在生物活性過程EEFM、胺基酸成份及含量、有機物分子量大小及濾床對NPDOC去除能力差異比較外,另濾床操作穩定後(96 days),不同臭氧化生物濾床,在不同深度水相及濾料附著生物膜之有機物性質,包括EEFM、胺基酸成份及含量、有機物分子量大小與菌相歧異度差異之比較,均是本研究之監測重點。
研究結果顯示,在人工配製之22種胺基酸之EEFM圖中,Tyrosine與Tryptophan之螢光強度值遠高於同濃度之其它胺基酸,而Tyrosine之波峰位置為280/300 nm與230/300 nm,Tryptophan 僅在280/360 nm出現波峰;而所有胺基酸出現波峰之發射波長最長為380 nm。Aldrich之EEFM圖則出現四個波峰位置,包括220/430 nm、260/460 nm、310/440 nm與480/530 nm,其發射波長最低為430 nm。
生物濾床在穩定過程(57 days),對NPDOC之去除能力,以O3/NPDOC=1最佳,其次為O3/NPDOC=2,最低者為O3/NPDOC=0之生物濾床。且三種臭氧化生物濾床出水中有機物分子量高於20,000之含量減少,但分子量小於5,000 者之含量則呈現增加。而屬似腐植質之螢光強度值,亦隨操作時間之增加而增加。生物濾床在穩定過程(57 days),對NPDOC之去除能力,以O3/NPDOC=1最佳,其次為O3/NPDOC=2,最低者為O3/NPDOC=0之生物濾床。且三種臭氧化生物濾床出水中有機物分子量高於20,000之含量減少,但分子量小於5,000 者之含量則呈現增加。
生物濾床操作穩定後(96 days),不同深度水相及濾料附著生物膜有機物之EEFM之特性發現,兩者皆出現似胺基酸及似腐植質之成份,但在螢光強度值部分,水相部分,似腐植質均高於似胺基酸部分,而附著於濾料之生物膜有機物則是似胺基酸大於似腐植質。不同深度濾床水相有機物分子量大於20,000之分佈面積百分率較附著於濾料之生物膜有機物高,而生物膜有機物主要為分子量小於5,000者。關於臭氧化生物濾床不同深度濾料附著生物膜及水相中微生物之16S rDNA的V6-V8區域之變性梯度膠泳(Denaturing Gradient Gel Electrophoresis, DGGE)圖,生物膜菌相之豐富度高於水相;而DGGE電泳圖譜條帶上的條帶數,利用Quantity One 4.6.2軟體進行非加權配對群團分析法,結果呈現生物膜菌相之相似度低於水相。
The source water, Cheng Chin Lake, filtered by 0.2 µm membrane filter , was added with different ozone dosages to make the ratios of ozone to NPDOC (non-purgeable organic carbon) be 0, 1 and 2 respectively. Then, the indigenous bacterium taken from Cheng Chin Lake was added into the ozonated source water to make sure the bacterial number close to 103 cells/mL. Later, ozonated source water with microorganisms was transferred into a 5 L bottle wrapped with aluminum foil to cut off sunlight. It was draw into a chromatography packed with glass bead using peristaltic pump. The effluent from this filter was also collected into the collection bottle, and pumped into filter again. This process is called recirculated biofilter. The operational days of three biofilter systems were about 100 days. In this study, the EEFMs (excitation emission fluorescent matrices) of 22 amino acids and humic acids purchased from Aldrich Company produce were measured and compared. The effluents of three stabilizing biofilters were monitored using parameters such as NPDOC, EEFM, amino acids, distribution of molecular weight. Of course, the differences of organic and bacterial parameter between bulk and attached bead filter were compared in different depths in varied ozonated biofilters.
The finding in this study reveals fluorescent intensities of peak locations in EEFM for tyrosine and Tryptophan were far higher than those for other amino acids. The peak positions of Tyrosine is 280/300 nm and 230/300 nm, however, Tryptophan presents the only peak in 280/360 nm. The longer emission wavelength for 22 amino acids measured is less than 380 nm. Four peak locations were existed in humic acids, including 220/430 nm, 260/460 nm, 310/440 nm and 480/530 nm. Comparing the ability of NPDOC removal among the three stabilizing biofilters (57 days), its order from large to little was O3/NPDOC =1, O3/NPDOC =2, and O3/NPDOC =0. The variations of contents of molecular weight of organic matter form the effluents from three ozonated biofilters show the same trends. Molecular weight higher than 20,000 was decreased with operational days while molecular weight less than 5,000 was increased with operational days.
Regarding to organic properties between the bulk solutions and attached in bead filters in three stabilized biofilters (96 days), both protein-like and humic-like substances were observed from peak locations of EEFM. For the fluorescent intensities in bulk solution in different depths of three biofilters, humic-like substance is higher than protein-like; however, this result was contrast to the bead filters in different depths of three biofilters. The content of molecular weight greater than 20,000 existed in bulk solution was higher than that of bead filter. The content of molecular weight in bead filter was dominant in less than 5,000. The differences of microorganism species between the bulk solutions and attached in bead filters in three stabilized biofilters (96 days) were compared using 16S rDNA with V6-V8 of DGGE (Denaturing Gradient Gel Electrophoresis). It finds that the abundance of microorganism species attached in bead filter exceeded that of bulk solution compared in the same depth and biofilter. The similarities of microorganism species in microorganism species attached in bead filter was less than that of bulk solution applying the software of Quantity One 4.6.2 into the fingerprint of DGGE.
誌謝
摘要
Abstract
表目錄
圖目錄
壹、前言
一、研究背景
二、研究目的
貳、文獻回顧
一、螢光激發發射光譜儀 (Fluorescent spectrophotometer)應用於有機物之定性分析
(一)有機物之分類
(二)UV(ultraviolet)應用在有機物之定性分析
(三)螢光激發發射光譜圖應用在有機物之定性分析
二、影響生物濾床操作效能之因子
(一)生物濾床對BOM之去除模式
(二)空床接觸時間(Empty bed contact time, EBCT)
(三) O3/DOC之比值
(四)濾床種類
三、分子生物技術應用於淨水處理系統中微生物之鑑定
(一) 自來水處理系統菌相之變化
(二)、菌相之鑑定
参、研究架構、系統操作與參數分析
一、研究流程之規劃
二、研究系統圖
(一)生物濾床之操作
(三) 臭氧反應系統及操作
(三) 臭氧反應系統及操作
三、生物性參數分析
(一)總細菌數
(二) 菌體之死活判別
(三) 菌體之鑑定
(四) 利用PCR增殖16S rDNA片段
(五) 變性梯度膠體電泳
(六) SYBR green I 染色與膠體拍攝與分析統計
四、化學性參數分析
(一) 揮發性之溶解性有機碳(non-purgeable dissolved organic carbon)
(二)有機物分子量大小
(三) 醣類
(四) 氨基酸
(五)螢光激發與發射光譜圖
肆、結果與討論
一、不同胺基酸及腐植酸之特徵螢光激發發射光譜圖
二、生物濾床穩定過程中不同特徵螢光激發發射光譜圖、胺基酸及分子量大小之變動
二、生物濾床穩定過程中不同特徵螢光激發發射光譜圖、胺基酸及分子量大小之變動
三、生物濾床在不同深度水相及濾料菌相分佈及有機物性質之差異
(一)有機性
(二)生物性
伍、結論
陸、建議
柒、參考文獻
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