藍綠藻(cyanobacteria)以及其所產生之毒素與臭味物質已被證實為飲用水中影響人體生命安全之主要因素之一。為能有效偵測並去除水中藍綠藻及其代謝物質，本研究首先針對快速偵測藻藍蛋白之胞內螢光偵測法進行探討。在該研究中，藍藻種類、藻體生長期、水中色素(chlorophyll-a)、濁度以及細胞型態(團粒現象)皆可影響螢光偵測法測定水中藍綠藻濃度之結果。在色素與濁度的測試上，頂脊藻(Chodatella sp.)所含有之葉綠素與高嶺土所形成之濁度被應用在研究中進行螢光偵測法之干擾測試，結果顯示色素與濁度的存在將大幅影響藻藍蛋白螢光測定之準確度，干擾程度呈等比例上升。在細胞型態的影響上，藉由Microcystis aeruginosa PCC 7005於實驗室培養時會形成不同團粒大小之特性，進行團粒型藻體對螢光偵測法之干擾測試，其結果利用一理論模式進行模擬，並將該模式應用於台灣數座水庫中，成功修正藻體團粒對於藻藍蛋白螢光偵測法之干擾。
在胞內螢光偵測法可有效應用於水中藻類濃度之偵測後，利用氧化劑進行水體氧化，藉以抑制藻體數量，使其不至形成藻華。然而，以往的殺藻劑多以硫酸銅、氯等化學物質為主，雖可有效破壞藻體，但也造成代謝物包括毒素、臭味物質的釋出，而氧化後的水體中亦可能殘留或形成其他對人體有害之化學物質。因此，本研究針對新型殺藻劑進行開發，藉由過氧化氫經由擬太陽光燈管之照射後，轉化形成之氫氧自由基進行藻體破壞。研究中針對可產生臭味物質geosmin之捲曲魚腥藻 (Anabaena circinalis)進行測試，分別測試藻體在黑暗與不同光照強度下，過氧化氫/氫氧自由基對細胞及其所釋放的代謝物質之破壞程度。過氧化氫濃度為10 ~ 60 mg/L，而氧化時間則為三個小時內，光照強度控制在0 ~ 100 W/m2。其結果顯示，光照強度可影響過氧化氫轉換為氫氧自由基之速率，而氫氧自由基對魚腥體細胞破壞之動力參數為4.7 * 109 M-1 S-1。研究中利用動力模擬之質量平衡方程式，藉由細胞破裂、代謝物釋出及自由基降解代謝物之結果，模擬並成功的預測代謝物質geosmin在系統中釋出並降解之現象。本研究也成功應用了流式細胞儀搭配螢光染劑的方式，有效並快速的偵測細胞在氧化過後的殘餘活性。藉由本研究之結果，成功的證實了過氧化氫/氫氧自由基可有效的針對藍綠藻細胞進行破壞，並同時降解藻體釋出之代謝物質。
在新型殺藻劑的開發上，氮參雜的二氧化鈦結合可見光之光觸媒技術應用，已成功地利用澳大利亞水質中心培養之Anabaena circinalis 318CR與Microcystis aeruginosa 338 進行初步的藻體破壞測試。研究中利用掃描式電子顯微鏡針對改良之二氧化鈦表面進行觀察，並應用X-ray photoelectron spectroscopy (XPS)與Visible-UV証實二氧化鈦基本性質已由於氮原子之參雜，而使其對於光之吸收由紫外光光譜紅移至可見光區段，且其能隙由理論值之3.2eV降低為2.99eV，顯示該材料可經由可見光之照射進而造成電子躍遷，進而產生氫氧自由基。研究中應用改良型二氧化鈦對於A. circinalis及M. aeruginosa 去除效率之最佳化進行測試，發現N/Ti 比例為20%、二氧化鈦濃度20 mg/L與10mg/L之過氧化氫對於藻體有最大的破壞效果；於40分鐘的接觸時間後，藻體完整性將低於5%。研究中應用2’,7’-dichlorodihydrofluorescein diacetate (DCFH-DA) 能與自由基反應進而產生螢光物質之特性，證實藻體內氫氧自由基之濃度可在氧化60分鐘後於A. circinalis及M. aeruginosa分別增高3.2倍與2.5倍。研究中亦將改良型二氧化鈦固定於石英板，並將其放置於氧化槽中，藉以將二氧化鈦與水體分離。結果發現，由於藻體與氧化劑的接觸情況改變，自由基無法完全破壞藻體生長機能，氧化後未完全破壞的藻體細胞將於五天後重新生長，且A. circinalis具有比M. aeruginosa更強的再生能力。
氧化劑的添加並非去除藻體活性的唯一方式。藻類代謝物質β-cyclocitral (木頭味，β-檸檬醛)被應用於測試其對於不同藻類的細胞活性之影響。測試的藻種分別為銅綠微囊藻Microcystis aeruginosa PCC 7005 、PCC 7820以及谷皮菱形藻Nitzschia palea，差別在於產生β-cyclocitral之能力為PCC 7820 > PCC 7005 > N. palea。研究中利用掃描式電子顯微鏡觀察與β-cyclocitral 接觸後的藻體表面，發現細胞隨著暴露時間的多寡而有不同程度的細胞破損。流式細胞儀的應用則觀察到細胞完整性下降，且藻細胞的葉綠素亦有降低之情形。藻體在抵抗β-cyclocitral破壞的強度則與其產生β-cyclocitral的能力成正比關係，β-cyclocitral約5 ~ 10 mg/L在同樣的時間下可破壞15 ~ 20%的微囊藻細胞，相對於菱形藻僅需0.1 ~ 0.5 mg/L的劑量即可達到同樣的效果。一階動力方程式的應用顯示細胞破裂的程度與β-cyclocitral之劑量有相關性，此數據亦可瞭解β-cyclocitral影響微囊藻及菱形藻細胞活性所需之劑量與時間。本研究利用β-cyclocitral對於非該物質產生者之細胞活性影響結果顯示，藻體代謝物除可造成人體健康風險外，亦有可能是其在面對物種競爭上維持其能持續生存的物質之一。

The effect of calibrated range, algal growth phase, chlorophyll-a, turbidity and colony structure, on measurement of phycocyanin by in-vivo fluoroscopy (IVF) was investigated. Microcystis aeruginosa PCC 7820, Anabaena circinalis and Planktothricoides raciborskii were used to investigate variation in phycocyanin content in the different cyanobacteria and growth phases. The green alga, Chodatella sp., and Kaolin particles were used as the sources of chlorophyll-a and turbidity respectively to examine how turbidity can impact on phycocyanin measurements. Another cyanobacterium, Microcystis aeruginosa PCC 7005, which forms large colonies, was used to investigate the relationships between colony size and phycocyanin levels measured using IVF. Experimental results showed that chlorophyll-a, turbidity, and colonial cyanobacteria significantly interfered with the measurement of phycocyanin fluorescence. Models were developed to compensate for the effect of chlorophyll-a, turbidity and colony size on the measurement. The models were successfully used to correct phycocyanin probe data collected from several reservoirs in Taiwan to produce cell counts with good correlation between measurements made using the phycocyanin probe and microscopy.
The application of free radical became a novel conception to control cyanobacterial bloom and their metabolites at the same time. In this study, effects of the oxidation by hydroxyl radical transformed from hydrogen peroxide on the cell lysis, release and decay of the metabolite from a geosmin producer-Anabaena circinalis were investigated. The toxicity of hydroxyl radical was researched to develop a potential tool for limiting and controlling the cyanobacterial blooms. The cyanobacterium Anabaena circinalis was tested under exposure to H2O2/OH radical in the dark and at the various irradiances. H2O2 was decomposed at rates depending on the intensity of illumination. A. circinalis was affected by H2O2 at 10 ~ 60 mg/L within 3 hr and the toxicity for algal cells was increased by light enhanced because of the release rate of OH radical. A pseudo first-order rate model was used to simulate the kinetics of cell rupture during the oxidative experiments, the value were 4.7 * 109 M-1 S-1 at the light intensity of 0 ~ 100 W/m2. An equation of dynamical system was applied to predict the simulation of the cell lysis, release and degradation of geosmin using the hydroxyl radical transformed from hydrogen peroxide combined with the sunlight-like. Geosmin was found released into water immediately after the cell lysis, and the cell integrity measurement using cell staining techniques correlated closely with geosmin release into solution.
Effects of visible-light photocatalysis using N-doped TiO2/H2O2 on the cell rupture and metabolite release from cyanobacteria Anabaena circinalis and Microcystis aeruginosa are investigated. A scanning electron microscopy coupled with an X-ray photoelectron spectroscopy (XPS) was employed to evidence that the crystal lattices of TiO2 were successfully modified with N-atoms, while a UV-Visible spectrometer was used to observe the shift of absorption wavelength for the modified photocatalyst. The doping process successfully shifted the absorption wavelength from 387.5 to 399 nm and decreased the gap band from 3.2 to 2.99 eV. A series of experiments with different doses of N-TiO2 and H2O2 were conducted to determine the effect on the studied cyanobacteria. Experimental results show that A. circinalis was more resistant than M. aeruginosa to the free radicals produced by N-doped TiO2. Combined doses of N- TiO2 at 20mg/L (N/Ti ratio 20%) and H2O2 at 10 mg/L had the host destruction capacity for the tested cells in this study, with cell viability decreased to be less than 5% after 40 min of exposure. Measurement of reactive oxygen species (ROS) inside of cyanobacteria with a fluorescent dye 2’,7’-dichlorodihydrofluorescein diacetate (DCFH-DA) indicated that the levels of ROS increased by 3.2 and 2.5 times in A. circinalis and M. aeruginosa after 60 min of oxidation. To mimic field application, cyanobacteria-laden water was allowed to flow through a column installed with glass plates coated with N-doped TiO2. The results showed that cells remain integral for the system without the photocatalyst. However, with the presence of the photocatalyst, cyanobacteria cells were readily ruptured and causing release of metabolites.
The algaecide doesn’t necessarily come from those chemicals which have the oxidative capability. Finally, the effect of an algal metabolite, β-cyclocitral, on the cell integrity of two cyanobacteria and a diatom was investigated. The cyanobacteria, Microcystis aeruginosa PCC 7005 and PCC 7820, and the diatom, Nitzschia palea, were exposed to various concentrations of β-cyclocitral. Scanning electron microscopy (SEM) results indicate that the cells of tested species were greatly altered after being exposed to β-cyclocitral. A flow cytometer coupled with the SYTOX stain and chlorophyll-a auto-fluorescence was used to quantify the effect of β-cyclocitral on cell integrity for the tested cyanobacteria and diatom. Kinetic experiments show that about 5-10 mg L-1 of β-cyclocitral for the two M. aeruginosa strains and a much lower concentration, 0.1-0.5 mg L-1, for N. palea were needed to cause 15-20% of cells to rupture. When the β-cyclocitral concentration was increased to 200-1000 mg L-1 for M. aeruginosa and 5-10 mg L-1 mg L-1 for N. palea, almost all the cells ruptured between 8-24 hr. A first-order kinetic model is able to describe the data of cell integrity over time. The extracted rate constant values well correlate with the applied β-cyclocitral dosages. The obtained kinetic parameters may be used to estimate β-cyclocitral dosage and contact time required for the control of cyanobacteria and diatoms in water bodies.

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