本研究探討藍綠菌細胞受到氧化劑破壞時細胞破損的程度與其胞內代謝物釋出之相關性，實驗以次氯酸鈉作為主要氧化劑，部分實驗使用高錳酸鉀，並以水中兩種常見產毒之藍綠菌，包括微囊藻(Microcystis aeruginosa)及柱孢藻(Cylindrospermopsis raciborskii) 做為研究對象。所使用之微囊藻具有產高濃度β-cyclocitral與微囊藻毒素(Microcystin, MC-LR)的能力，柱孢藻會產生柱孢藻毒素(Cylindrospermopsin, CYN)。實驗前將其培養於經數道過濾之金門太湖原水與ASM培養基混合之合成水。研究中應用兩種螢光染劑FDA(fluorescein diacetate)與SYTOX Green，並搭配兩種偵測儀器-流式細胞儀(flow cytometer, FCM)及螢光顯微鏡(epifluorescence microscope, EFM)來偵測藍綠菌細胞被氧化劑所破壞的程度，並相互比較，另外以應用掃描式電子顯微鏡(scanning electron microscope, SEM)對細胞進行高放大倍率的觀察，以便觀察細胞表面的變化與狀態。在代謝物分析部分，研究中以β-cyclocitral、微囊藻毒素(Microcystin, MC-LR)與柱孢藻毒素(Cylindrospermopsin, CYN)分別做為微囊藻與柱孢藻代謝物的代表，並應用固相微萃取法(solid phase micro-extraction, SPME)搭配氣相層析質譜儀(gas chromatograph/mass spectrometry detector, GC/MSD)對β-cyclocitral進行分析；以固相萃取法(solid phase extraction, SPE)配合高壓液相層析質譜儀(Liquid Chromatography/Mass Spectrometry, LC/MS) 以及酵素連結免疫吸附法(Enzyme-Linked ImmunoSorbent Assay, ELISA)來分析藻類毒素的濃度。研究結果發現氯對於微囊藻與柱孢藻細胞的氧化力相差不大；整體來說，以氯劑量2 mg/L對微囊藻與柱孢藻進行氧化，接觸時間10分鐘後就會有90%以上的細胞被氯所破壞，若是藻數不超過2,000,000 cells/mL時，幾乎反應都會在5分鐘內完成大部分的反應，而柱孢藻在大致相同的藻數時，其氯消耗量在5～30分鐘時的反應會比微囊藻來的多。在微囊藻代謝物β-cyclocitral方面，發現細胞被破壞仍不會釋出到水體，其原因在於催化其產生的β胡蘿蔔素加氧脢(β-carotene oxygenase)被氯所抑制的關係，而毒素方面在500,000 cells/mL以下的微囊藻，以5 mg/L的氯作用下就可以達到警戒值(1ppb)以下，而在高藻數100萬時這個劑量是不夠的；在柱孢藻毒素方面，即使是約600,000 cells/mL藻數，5 mg/L仍然不夠。另外在高錳酸鉀方面，其在流式細胞儀觀察細胞的完整性較為困難，其原因在於會有其他顆粒的產生，而比較氯與高錳酸鉀對微囊藻細胞的破壞，可以發現高錳酸鉀氧化後細胞的完整性較高，且藻體有凝聚的效果。

The effect of oxidation on the cell integrity and metabolite release for two nauseous cyanobacteria, Microcystis aeruginosa and Cylindrospermopsis raciborskii, is investigated. Two oxidants were used in the experiments, where sodium hypochloride was used mainly and potassium permanganate was used in certain cases. The two cyanobacteria were grown in the laboratory with filtrated water from Tai-Lake, Kinmen, and with addition of ASM algae growth medium. A fluorescence technique, combining fluorescein diacetate (FDA) and SYTOX Green with either flow cytometer or epifluorescence microscope, was successfully developed for the determination of cell integrity. A solid-phase microextraction (SPME) concentration followed by a gas chromatograph (GC) and mass spectrometric detector (MSD) was employed to measure a metabolite for Microcystis, β-cyclocitral . , while both solid-phase extraction (SPE) with liquid chromatography/mass spectrometry (LC/MS) and enzyme-linked immunosorbent assay (ELISA) was used to detect two toxins from the two cyanobacteria. A series of oxidation experiments of cyanobacteria-laden water was conducted at different cell concentrations and different oxidant dosages. During the experiments, chlorine concentration, cell integrity, metabolite concentration, and other water quality parameters were monitored at different time. The effect of chlorine on cell integrity for both Microcystis and Cylindrospermopsis was similar, being both very fragile to chlorine. At an initial free chlorine concentration of 2 mg/L, 80->90% of cells of both cyanobacteria were ruptured within 2-5 minutes, causing immediate release of metabolites from the cells. If the cells number was lower than 2,000,000 cells/mL, the reaction between chlorine and cells finished within 5 min.
For microcystis metabolite, β-cyclocitral concentration did not increase even after Microcystis cells lysed. This may be attributed to that the enzymes responsible for β-cyclocitral production, β-carotene, is inhibited by chlorine. About the toxin, when the cell concentration of Microcystis is lower than 500,000 cells/mL, 5mg/L of free chlorine is enough to destroy MC-LR present in the water. However, when the cells concentrations were larger than 1,000,000 cells/mL, more chlorine is needed to destruct MC-LR. For Cylindrospermopsis, however, when the cell concentration is at about 600,000 cells/mL, 5mg/L chlorine can only destruct 75% of CYN.For the oxidation of Microcystis using permanganate, it is more difficult to detect cell integrity using FCM, as there were some particle generation caused by permanganation. Compared the data with those from chlorination, the cells after permanganatation were ruptured to a lesser extent. However, more experiments are needed to confirm this.

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