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研究生:張志文
研究生(外文):Zhi-Wen Zhang
論文名稱:藻相之顯微鏡照像及其萃出液之螢光光譜圖特性分析
論文名稱(外文):The Observation of Algae by Microscope and Excitation-Emission Fluoresence Matrix of Its Extraction
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:2006
畢業學年度:93
語文別:中文
論文頁數:83
中文關鍵詞:三鹵甲烷生成潛能激發發射螢光光譜螢光顯微鏡
外文關鍵詞:Epifluorescent MicroscopeExcitation-Emission Fluorescence Matrix (EEFM)Trihalomethane formation potential (THMFP)
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依環保署之相關監測資料顯示,國內湖泊、水庫優養化情況近年來越趨嚴重。由於優養化湖水中藻類會影響水廠之操作及水質之處理或處理後之水質,從研究文獻得知,大部分研究者著重於除藻技術開發,少見相關研究報告進行藻類螢光光譜特性分析。由於螢光光譜儀具有未破壞有機物性質之分析特性,國內在此之研究亦顯得相當不足,故本研究除比較不同顯微照像技術進行藻類外觀及活性觀察外,另利用螢光光譜儀進行水體藻類與溶解性有機質之螢光光譜圖(Excitation-Emission Fluoresence Matrix,EEFM)比較,藉以瞭解兩者間之關連性外,並進行湖水中優勢藻科對消毒副產物生成潛能之影響。
研究結果顯示,藻量與溫度、葉綠素a、總磷與水體濁度均呈現高度之正相關性。至於優勢藻含量百分比之變化,澄清湖 (CCL)與鳳山 (FS)湖水兩次採樣之主要優勢藻均為綠球藻科 (Chlorococcaceae)與色球藻科(Chroococcaceae),二科佔優勢藻量之90 %;牡丹 (MD)水庫第一次及第二次,雖仍以綠球藻科與色球藻科為主,但其藻量僅佔總藻量之70 %,該水庫第三及第四次採樣結果則以菱形藻科 (Nitzschiaceae)及圓篩藻科 (Coscinodiscaceae)為主要優勢藻,其含量均佔總藻量之72 %。另以螢光顯微鏡觀察發現不同藻體所發出之螢光強度及顏色有明顯之差異,在相同藻種之顆粒直鏈藻(Melosira granulate (Eherenberg) Ralfs Var. granulate)及單角盤星藻(Pediastrum simplex Meyen Var. simplex),比較有無添加Lugol''s 溶液,皆可觀察到有添加Lugol''s 溶液之藻體自發性螢光減弱及顏色改變之現象。
另CCL 與FS 湖水中各兩次採樣之優勢藻數含量百分比與不同分子量有機物碳含量百分比之變化,皆呈現一致性之變化,亦即綠球藻科增加時,水中大於5K分子量有機物碳含量百分比呈現增加的現象,而色球藻科減少時,水中小於5K 之有機碳含量百分比則展現出減少之現象。但在MD 湖水第一及第二次採樣結果,有機碳含量百分比恰與CCL及FS之結果相反,其與優勢藻圓篩藻科之減少及鼓甲亞藻科之增加相關。CCL、FS及MD水庫過濾前與過濾後之THMFP (THM formation potential)形成之差異性,濾前均高於濾後。且由THMFP除以葉綠素a或藻數之比值進行比較,顯示藻科對THMFP之形成有影響,綠球藻科、圓篩藻科、鼓甲亞藻科及菱形藻科等四藻科不利於THMFP之形成,而色球藻科、水網藻科及顫藻科則有利於THMFP之形成。
原水及其濾液與藻體萃出液之EEFM之比較發現,CCL與FS之水樣測試結果在220~230/300~310、230/330~350及270~280/330~340之EX/EM波峰位置相近,另在MD第一及二次採樣,亦出現相似波峰,顯然與湖水中綠球藻科及色球藻科之優勢藻科相關。至於MD第三及四次採樣,在210~250/300~310出現EX/EM波峰,其濾液中主要波峰以240~250/300~310最為顯著,明顯與前兩次採樣結果相異,與該次採樣優勢藻菱形藻科及圓篩藻科相關。亦即水體中有機物性質變化與藻科之變化有明顯之關聯性。
According to data collected from monitoring stations in lakes and reservoirs maintained by the Environmental Protection Administration (EPA), the problem of eutrophication has escalated in recent years. The phenomenon of eutrophication gives rise to algae proliferation. Due to its interference with the quality of treated water, and the operation of water treatment facilities, the development of technologies for the removal of algae growth has been the focus of many research papers for the past several years, both domestically as well as internationally, and less attention has been paid the study of characteristics of algae through spectral analysis.
The fluorescence meter, due to its non-destructive nature of measurement, makes it feasible to quality the difference in structural characteristics between that of suspended particles and dissolved organic matter. Due to the lack of research carried out on the study of the fluorescent characteristics of algae, and the correlation of characteristics between dissolved organic matter and algae in the eutrophic lake, the use of the fluorescent meter EEFM (Excitation-Emission Fluoresence Matrix) to compare this relationship is worth considering. In this study, in addition to the carrying out of spectral analysis, the THMFP (Trihalomethane formation potential) from different kinds of algae will also be described.
There is an indication that algal population in three eutropic lakes had a positive correlation with temperature, presence of chlorophyll, level of total phosphorus and turbidity. The percentage of the dominant algae, Chlorococcaceae and Chroococcaceae were determined to be 90 % in CCL and FS reservoirs. In MD reservoir, the value was decreased to 70 % in the first and second sampling, but Nitzschiaceae and Coscinodiscaceae made up 72%, and were found to be the dominant algal species in the third and fourth sampling.
Also, different autofluorescent colors and intensity were observed when different kinds of algae were placed under an epifluoresent microscope. Melosira granulate (Eherenberg) Ralfs Var. granulate and Pediastrum simplex Meyen Var. simplex as an example, it was revealed that the intensity of algal autofluorescence and coloration upon the addition of Lugol''s solution was weakened as compared with the adding of no Lugol''s solution.
Two samples each were taken from CCL Lake and FS Lake, and it was found that the percentages of carbon content of different MWCO (molecular weight cut-off) in dissolved organic matter (DOM) were also varied as the percentage of algal species were changed. This suggests that the increase in Chlorococcaceae will lead to an increase in the percentage of carbon content of dissolved organic matter which the MWCOs is higher than 5K. Furthermore, it reveals that a decrease in Chroococcaceae will lead to a decrease in the percentage of carbon content of dissolved organic matter (DOM) which the MWCOs is less than 5K. However, during the first and second sampling at MD reservoir, the variation in the percentage of carbon content of dissolved organic matter were opposite from those from CCL and FS reservoirs. This effect may be attributed to the decrease in the percentage of Coscinodiscaceae among the total algal population, and an increase of Cosmarium in total algal population.
The THMFP value in the water source as obtained from CCL, FS and MD reservoirs, as compared with that of the filtered water source, indicates that the former value was larger than latter. Furthermore, Chlorococcaceae, Coscinodiscaceae, Cosmarium and Nitzschiaceae do not favorably form THMFP. On the other hand, Chroococcaceae, Hydrodictyaceae, and Oscillatoriaceae could form well THMFP.
The peak values of EEFM of the water source, filtrates and extracted algae obtained from CCL and FS reservoirs were always found to be within the range of excitation/emission wavelength (EX (nm)/EM(nm)), 220~230/300~310,230/330~350, and 270~280/330~340. However, the peak value of excitation/emission wavelength of 240~250/300~310 can be found in the third and fourth water samples taken from MD reservoir. This may be that the dominant algal species, Chlorococcaceae and Chroococcaceae, which appeared in CCL and FS reservoirs in March and April, were different from Nitzschiaceae and Coscinodiscaceae, which appeared in MD reservoir in November and December.
誌 謝
摘 要 I
ABSTRACT III
目 錄 VI
表 錄 X
圖 錄 XI
壹、前言 1
一、研究緣起 1
二、研究目的 1
貳、文獻回顧 3
一、優養化 3
(一)優養化之成因及分類 3
(二)歷年來國內水庫優養化及藻種變動 4
二、藻類衍生問題 7
(一)臭、味及色度 7
(二)毒素 9
(三)湖水水質之變化 9
(四)影響淨水廠之操作 9
1.濾床阻塞及污泥特性改變 9
2.影響氧化劑或混凝劑加量及造成自然有機物質之變動 10
3.消毒副產物(DBP, disinfection by-product)前質 12
三、螢光光譜儀運用於有機物之定性分析 13
參、研究規劃與實驗分析方法 17
一、研究流程之規劃 17
二、採樣計劃 19
三、實驗設備與分析方法 19
(一)有機物分子量大小之決定 19
1.腐植酸之配製 19
2.原水不同分子量有機物分離設備 19
(1)薄膜系統介紹操作 19
(2)薄膜系統操作 19
(3)薄膜前處理及清洗 20
(二)基本參數分析 21
1.pH值及導電度值 21
2.透視度 21
3.濁度 21
4.鹼度 22
5.氨氮 23
6.總凱氏氮 23
7.正磷酸鹽 24
8.總磷 24
9.硝酸鹽氮、亞硝酸鹽氮及溴離子含量分析(使用IC) 24
10.葉綠素a 25
(三)有機參數分析 26
1.UV254 26
2.溶解性有機碳(DOC)含量分析 26
3.三鹵甲烷生成潛能(THMFP)試驗 27
(1)採樣前處理 27
(2)採樣及分析 27
4.螢光光譜(EEFM)之操作參數及使用流程 30
(四)藻類之計數與觀察 31
1.前處理 31
2.掃描式電子顯微鏡之樣本觀察 33
肆、結果與討論 35
一、優氧化湖水中藻種之變動及其與水質參數之相關性分析 35
二、螢光顯微鏡應用於藻體外觀之判別 41
三、優養化湖水中溶解性有機物性質及藻體對三鹵甲烷生成潛能之影響 48
四、湖水中溶解性有機物及藻體萃出物之EEFM圖譜之比較 56
伍、結論 70
陸、建議 72
柒、參考文獻 73
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