臺灣博碩士論文加值系統

柱孢藻（Cylindrospermopsis raciborskii（Woloszynska） Seenaya et Subba Raju）是一種淡水藍綠藻，在金門的一些水庫裡有大量滋長的情況，並形成藻華而影響水庫之水質。本研究針對榮湖、金沙、山西及太湖水庫進行為期一年的現場生態調查，針對水庫環境與柱孢藻作相關分析，並研究柱孢藻分離株之生理特性，來探討此藻種優勢生長的原因。水庫現場調查結果發現，四個水庫中皆有柱孢藻優勢的情況，而水庫水質皆介於優養化到超優養化間，且庫水透明度低、導電度高、總磷及總氮高、略帶鹽度及含有機污染物。在柱孢藻分離株之生理研究發現，高濃度的硝酸氮、中高水溫（24~32℃）、低濃度銨氮(＜4 mM )及尿素，有利於柱孢藻生長，且此藻對鹽度有耐受性（~ 4

A filamentous freshwater cyanobacterium, Cylindrospermopsis raciborksii (Woloszynska) Seenaya et Subba Raju, was to be found dominant in some freshwater reservoirs. The water quality of the reservoirs in Kinmen was deteriorated due to the blooms of this organism. Physio-chemical analyses of water were carried out monthly in four reservoirs viz., Rong-lake, Jin-Sha, Shan-Xi and Tai-lake over the period from September, 2003 to September, 2004. Physio-chemical factors affecting the abundance of C. raciborskii in the reservoirs were analyzed. The growth physiology of C. raciborskii was studied with an isolate of this species. The results indicated that all of four reservoirs showed dominance of C. raciborskii, associated with low transparency, high electrical conductivity,low salinity, moderate organic pollutants and higher concentrations of both TN and TP. All of four water reservoirs were in eutrophic or hypertrophic condition. The isolate of C. raciborskii preferred high concentration of nitrate, low concentration of urea and moderate water temperature varying from 24

一、前言……………………………………………………………………………..1二、材料與方法…………………………………………………………………....7
2.0 研究地點……………………………………………………………………7
2.1 水庫水樣的月調查…………………………………………………………7
2.1.1 水樣的物理化學分析………………………………………………...7
2.1.2氨氮（NH4+-N）濃度的測定…………………………………………8
2.1.3硝酸氨(NO3--N)濃度的測定………………………………………......8
2.1.4亞硝酸氨(NO2--N)濃度的測定……………………………...………...8
2.1.5磷酸鹽(PO43--P)濃度的測定……………………………………..…....8
2.1.6總氮（TN）測定……………………………………………………...9
2.1.7總磷（TP）測定………………………………………………………..9
2.1.8 可溶性有機物（DOM）測定………………………………..………...9
2.1.9葉綠素a濃度的測定方法……………………………………………10
2.1.10水庫水質優養化程度評估………………………………………….10
2.2 水庫浮游藻類採樣與分析………………………………………………..11
2.2.1 定性片的製作與計數……………………………………………….11
2.2.2 定量片的製作與計數……………………………………………….12
2.2.3藻類體積分析………………………………………………………..12
2.3柱孢藻的分離與培養……………………………………………………...12
2.3.1 單株藻種分離……………………………………………………….12
2.3.2 培養條件…………………………………………………………….13
2.3.3 其他測試藻種的培養……………………………………………….14
2.4 柱孢藻的生理生長研究…………………………………………………..14
2.4.1 生長速率的計算…………………………………………………….14
2.4.2鹽度…………………………………………………………………..15
2.4.3最佳生長溫度………………………………………………………...15
2.4.4光合作用……………………………………………………………...15
2.4.5柱孢藻在不同氮源下之生長………………………………………...16
2.4.6 柱孢藻在不同氨氮(NH4+) 濃度下之生長……………………........16
2.4.7柱孢藻在不同尿素（（NH2）2 CO）濃度下之生長………………..16
2.4.8柱孢藻在不同硝酸氮（NO3-）濃度下之生長………………………17
2.4.9柱孢藻在不同磷酸鹽（KH2PO4）濃度下之生長………………….17
2.4.10 柱孢藻的C：N：P元素分析……………………………………...17
2.5 柱孢藻之藻毒研究………………………………………………………..17
2.5.1水庫水的毒性研究…………………………………………………..17
2.5.2 柱孢藻對六種藻的生長影響……………………………………….18
2.5.3 柱孢藻對浮游藻類的生長影響…………………………………….18
2.5.4 柱孢藻萃取物質對浮游動物的抑制測試………………………….19
2.6 用PCR分析柱孢藻產生毒素之可能性………………………………….21
2.6.1柱孢藻DNA的萃取…………………………………………………21
2.6.2 PCR條件……………………………………………………………..21
2.6.3瓊脂醣膠電泳……………...................................................................22
2.7 相關係數分析…………………………………………………………...…22

三、結果
3.1 水庫水質之理化特性狀況……………………………………………......23
3.1.1 水溫………………………………………………………………….23
3.1.2 透明度……………………………………………………………….23
3.1.3 導電度……………………………………………………………….23
3.1.4 酸鹼值……………………………………………………………….23
3.1.5 溶氧………………………………………………………………….23
3.1.6 氨氮………………………………………………………………….24
3.1.7 硝酸氮……………………………………………………………….24
3.1.8 溶解磷……………………………………………………………….24
3.1.9 總磷………………………………………………………………….24
3.1.10 有機氮……………………………………………………………...24
3.1.11 TN:TP 比值………………………………………………………25
3.1.12 有機碳………………………………………………………….......25
3.1.13 鹽度………………………………………………………………...25
3.2 水庫水質優養化評估…………………………………………………...…25
3.2.1 單項水質優養化評估…………………………………………….....25
3.2.2 卡爾森優養指數評估……………………………………………….26
3.3 柱孢藻的優勢度…………………………………………………………...26
3.3.1 柱孢藻的形態……………………………………………………….26
3.3.2 四個水庫的藻類密度……………………………………………….27
3.3.3 四個水庫藻類組成………………………………………………….27
3.3.4 柱孢藻的優勢……………………………………………………….28
3.3.5 藻乾重與葉綠素a間的關係………………………………………..28
3.4 柱孢藻生長優勢與環境因子的相關性…………………………………..29
3.5 柱孢藻之生長條件測試…………………………………………………..29
3.5.1 溫度條件與光合作用測試結果…………………………………….29
3.5.2 鹽度條件測試……………………………………………………….29
3.5.3 對於氮鹽的需求…………………………………………………….30
3.5.4 對於磷鹽的需求…………………………………………………….30
3.5.5 柱孢藻之C:N:P元素組成…………………………………………..31
3.6 柱孢藻大量存在對水庫中藻類與浮游動物的生長抑制現象…………..31
3.6.1水庫水的毒性測試…………………………………………………...31
3.6.2 柱孢藻對其他藻類的抑制生長作用……………………………….31
3.6.3 柱孢藻對水庫藻種之抑制………………………………………….32
3.6.4 柱孢藻對浮游動物的抑制作用…………………………………….32
3.7 PCR分析Cylindrospermopsin之可能性……………………….............32
四、討論……………………………………………………………………………33
五、參考文獻………………………………………………………………………41
六、圖表……………………………………………………………………………50
七、附錄…………………………………………………………………………….82

參考文獻
小島貞男著，張清源譯，1984，翡翠水庫水質預測與污染防治對策，台北翡翠水庫管理局。
台灣日研設計顧問有限公司，1998，水質保護及資訊管理服務建議書。
林襟江、盧誠吉、洪銘堅、陳時仁，1985，德基水庫層化及優養化之研究，經濟部水資源統一規劃委員會。
馮纘華，1982，Lake Eutrophication and Its Control，工業污染防治第四期，115-165頁。
曾怡禎、陳是瑩，1984，澄清湖生態的研究，第一屆給水技術研討會論文集，175-195頁。
江殷儒，2002，鯉魚潭中葡萄藻代謝物質之生物毒性，國立臺灣大學植物學研究所碩士論文，113頁。
吳俊宗，2003，藻類與金沙地區自來水水質關係之探討，金門縣自來水廠，75頁。
莊西進、許永面，2002，金門國家公園環境長期監測，15-17頁，金門國家公園管理處。
陳章波，2000，金門國家公園生態環境監測架構之建立，11-15頁，內政部營建署金門國家公園管理處委託研究報告。
郭城孟、陳尊賢 2002，金門國家公園土壤調查分析及植生適應性研究,1-98頁，內政部營建署金門國家公園管理處委託研究報告。
詹智全，2001，國內水庫優養化評估因子及藻類指標間之相關性分析研究，國立中興大學環境工程學系碩士論文，7-19頁。
蔡財興，1998，花蓮鯉魚潭水質優養化之研究，國立東華大學資然資源管理研究所碩士論文，3-7頁。

Arfi, R. and Bouvy, M. 1995. Size, composition and distribution of particles related to wind-induced resuspension in a shallow tropical lagoon. J. Plankton Res. 17: 557-574.

Blomqvist, P., Pettersson, A. and Hyenstrand, P. 1994. Ammonium-nitrogen: A key regulatory factor causing dominance of non-nitrogen-fixing cyanobacteria in aquatic systems. Arch. Hydrobiol. 132: 141-164.

Borics, G., Grigorszky, I., Szabo, S. and Padisak, J. 2000. Phytoplankton associations in a small hypertrophic fishpond in East Hungary during a change from bottom-up to top-down control. Hydrobiologia 424: 1-3.

Bormans, M., Sherman, B.S. and Webster, I.T. 1999. Is buoyancy regulation in cyanobacteria an adaptation to exploit separation of light and nutrients? Mar. Freshwat. Res. 50: 897-906.

Bouvy, M., Molica, R., De Oliveira, S., Marinho, M. and Beker, B. 1999. Dynamics of a toxic cyanobacterial bloom (Cylindrospermopsis raciborskii) in a shallow reservoir in the semi-arid region of northeast Brazil. Aquat. Microb. Ecol. 20: 285-297.

Bouvy, M., Falcao, D., Marinho, M., Pagano, M. and Moura, A. 2000. Occurrence of Cylindrospermopsis (Cyanobacteria) in 39 Brazilian tropical reservoirs during the 1998 drought. Aquat. Microb. Ecol. 23: 13-27.

Branco, C.W.C. and Senna, P.A.C. 1994. Factors influencing the development of Cylindrospermopsis raciborskii and Microcystis aeruginosa in the Paranoa Reservoir, Brasilia, Brazil. Algol. Stud. 75: 85-96.

Branco, C.W.C. and Senna, P.A.C. 1996. Phytoplankton composition, community structure and seasonal changes in a tropical reservoir (Paranoa Reservoir, Brazil). Algol. Stud. 114: 69-84.

Briand, J., Leboulanger, C., Humbert, J., Bernard, C. and Dufour, P. 2004. Cylindrospermopsis raciborskii (Cyanobacteria) invasion at mid-latitudes: Selection, wide physiological tolerance, or global warming? J. Phycol. 40: 231-238.

Briand, J.F., Robillot, C., Quiblier-Lloberas, C., Humbert, J.F., Coute, A. and Bernard, C. 2002. Environmental context of Cylindrospermopsis raciborskii (Cyanobacteria) blooms in a shallow pond in France. Water Res. 36: 3183-3192.

Caraco, N., and Miller, R. 1998. Effects of CO2 on competition between a cyanobacterium and eukaryotic phytoplankton. Can. J. Fish. Aquat. Sci. 55, 54-62.

Carlson, R.E. 1977. A trophic state index for lake. Limnol.Oceanogr. 22: 361-369.

Chapman, A.D. and Schelske, C.L. 1997. Recent appearance of Cylindrospermopsis (Cyanobacteria) in five hypereutrophic Florida lakes. J. Phycol. 33: 191-195.

Couderchet, M. and Vernet, G. (2003). Pigments as biomarkers of exposure to the vineyard herbicide flazasulfuron in freshwater algae. Ecotoxicol. Environ. Saf. 55: 271-277.

Das, S., Mandal, B. and Mandal, L. 1991. Effect of growth and subsequent decomposition of blue-green algae on the transformation of iron and manganese in submerged soils. Plant Soil. 138: 75-84.

Dokulil, M.T. and Mayer, J. 1996. Population dynamics and photosynthetic rates of a Cylindrospermopsis - Limnothrix association in a highly eutrophic urban lake, Alte Donau, Vienna, Austria. Algol. Stud. 83: 179-195.

Dokulil, M.T. and Teubner, K. 2000. Cyanobacterial dominance in lakes. Hydrobiologia 438: 1-3.

Elser, J.J. (1992). Phytoplankton dynamics and the role of grazers in Castle Lake, California. Ecology 73: 887-902.

Fabbro, L.D. and Duivenvoorden, L.J. 1996. Profile of a bloom of the cyanobacterium Cylindrospermopsis raciborskii (Woloszynska) Seenaya and Subba Raju in the Fitzroy River in tropical Central Queensland. Mar. Freshwat. Res. 47: 685-694.

Fergusson, K.M. and Saint, C.P. 2003. Multiplex PCR assay for Cylindrospermopsis raciborskii and cylindrospermopsin-producing cyanobacteria. Environ. Toxicol. 18: 120-125.

Fistarol, G..O., Legrand,C. and Gran