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研究生： 葉桂寶
Ip, Gunadi
論文名稱： 使用β環檸檬醛控制並抑制微囊藻的生長
Control and Growth Inhibition of Microcystis using Beta-cyclocitral
指導教授： 林財富
Lin, Tsair-Fuh
學位類別： 碩士
Master
系所名稱： 工學院 - 環境工程學系
Department of Environmental Engineering
論文出版年： 2018
畢業學年度： 106
語文別： 英文
論文頁數： 94
外文關鍵詞： beta-cyclocitral, Microcystis aeruginosa, mechanical method, Cylindrospermopsis raciborskii, degradation kinetics
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    • Microcystis aeruginosa has been known to produce beta-cyclocitral when the cells ruptured. The chemical was known to cause rupture of cyanobacteria and algae. In this study, effect of beta-cyclocitral on the cell integrity and growth inhibition of two cyanobacteria, Microcystis aeruginosa and Cylindrospermopsis raciborskii were studied. A flowcytometer coupled with SYTOX stain was used to quantify the cell integrity of the tested cyanobacteria, while an enzyme immunosorbent assay was used to determine the cyanotoxins produced by the two cyanobacteria. Kinetic experiments show that 1 mg L-1 of beta-cyclocitral was able to cause 50% and 15% cell rupture of the tested Microcystis and Cylindrospermopsis respectively. A first-order rate model was able to describe the data of cell integrity for the first 60 minutes of rupture. Four mechanical methods, including sonication, bead mill, freeze method and sonication enhanced with bead mill were tested for the production of beta-cyclocitral from Microcystis cells. The results show that 5 freezing cycles of the freeze method is able to produce beta cyclocitral up to 15 µg L-1 with 92% efficiency and 84.3% recovery, respectively. Furthermore, beta-cyclocitral was produced from sonication of Microcystis cell and then applied for inhibition of M. aeruginosa and C. raciborskii. The experimental results may provide information of production and application of the cyanobactericide, beta-cyclocitral on the control of cyanobacteria in water bodies.

    Table of Contents ABSTRACT I ACKNOWLEDGEMENT II Table of Contents IV List of Figure IX List of Table XIII CHAPTER 1 INTRODUCTION 1 1.1 Background 1 1.2 Objectives of study 3 CHAPTER 2 LITERATURE REVIEW 4 2.1 Freshwater Algae 4 2.2 Size and Diversity of Structure of Algae and cyanobacteria 4 2.3 Cyanotoxins 9 2.4 Gas Vacuoles 11 2.5 Microcystis aeruginosa 13 2.5.1 Beta-cyclocitral 14 2.5.2. Microcystins 15 2.6 Cylindrospermopsis raciborskii 17 2.6.1 Cylindrospermopsin 19 2.7 Cell Disruption Methodology 21 2.7.1 Mechanical strength of cell walls 22 2.7.2 Contamination of products (beta-cyclocitral) 22 2.7.3 Selection of Disruption method 23 2.8 Sonication 23 2.8.1 Horns 24 2.8.2 Baths 25 2.9 Bead Mill 26 2.10. Freeze Drying (Lyophillization) 27 CHAPTER 3 MATERIAL AND METHODS 29 3.1 The Approach of this Research 29 3.2 Materials and Methods 31 3.2.1 Cyanobacteria 31 3.2.2 Cell Enumeration 31 3.2.2.1 M. aeruginosa PCC7820 cell enumeration 31 3.2.2.2 Cylindrospermopsis cell enumeration 32 3.2.3 Preliminary Oxidation Process 33 3.2.3.1 Sample Preparation 33 3.2.3.2 Sampling 34 3.2.3.3 Inhibition Test 34 3.2.3 Mechanical Methods for Cell Disruption 35 3.2.3.1 Pretreatment of Sample 35 3.2.3.2 Sonification 35 3.2.3.3 Bead Mill Method 36 3.2.3.4 Freeze Shock Method 36 3.2.3.5 Bead Mill and Sonication Method 37 3.2.4 Cyanotoxins 37 3.2.4.1 Pretreatment of samples 37 3.2.4.2 Measurement of Cyanotoxins 38 3.2.5 Application Method 38 3.2.5.1 Beta Cyclocitral extraction (Run One) 38 3.2.5.2 Inhibition test (Run One) 39 CHAPTER 4 RESULTS AND DISCUSSION 40 4.1 Preliminary Research 40 4.1.1 Cell Lysis 40 4.1.1.1 Run One 40 4.1.1.2 Run Two 45 4.1.2 Inhibition Test 47 4.1.2.1 Run One 47 4.1.2.2 Run Two 50 4.1.3 Beta-cyclocitral concentration 53 4.1.3.1 Run One 53 4.1.3.2 Run Two 54 4.1.4 Cyanotoxins Concentration 55 4.1.4.1 Run One 55 4.1.4.2 Run Two 58 4.2 Mechanical Method for M. aeruginosa cell disruption 60 4.2.1 Sonication 60 4.2.1.1 Run One 61 4.2.1.2 Run Two 62 4.2.2 Bead Mill 65 4.2.2.1 Run One 66 4.2.2.2 Run Two 67 4.2.3 Freeze Method (Lyophylization) 69 4.2.3.1 Run One 70 4.2.3.2 Run Two 71 4.2.4 Bead Mill and Sonication 75 4.3 Applications 77 4.3.1 Selection of Mechanical method 77 4.3.2 Beta cyclocitral production 77 4.3.3 Inhibition Tests 78 4.3.3.1 Run one 78 4.3.3.2 Run Two 80 CHAPTER 5 CONCLUSIONS 82 5.1 Conclusion 82 5.2 Suggestion 84 REFERENCES 85 APPENDIX A – Microcystis aeruginosa PCC7820 media 92 APPENDIX B –Cylindrospermopsis sp. media 93

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