自然水體中若氮磷含量過高，造成優養化，會使水中藍綠菌(舊稱藍綠藻)快速增生，進而造成水體中毒素和臭味等代謝物質的增加，影響人類與環境生物的飲用安全，同時也增加用水疑慮。目前各國針對藻類滋生所產生的危害風險，主要是以水體水質污染來評估，然而對於水體周界上方的氣膠粒子所造成的危害，評估文獻仍非常少。水中藻細胞若隨氣膠粒子傳輸至空氣中，其伴隨之毒素亦可藉由呼吸途徑，造成水體周邊活動民眾的暴露風險。因此，有效量化優養化水體周界空氣中有害藻細胞與毒素濃度，為推估暴露量的基本工作。
本研究利用PS-1、PM10和PM2.5的Cyclone等三種粒徑不同的空氣採樣器，分別於成功大學校園中成功湖、台南市官田區葫蘆埤、和金門太湖湖庫周界採集氣膠粒子，並將採集到的樣本分別進行甲醇萃取、藻體溶出回收和核苷酸(DNA)萃取等，再結合酵素免疫學技術(ELISA)和定量聚合酶鏈鎖反應(qPCR)，來量化優養化水體周界空氣中的代表性產毒藻(微囊藻與柱孢藻)與毒素(微囊藻毒素及柱孢藻毒素)濃度。研究發現，在利用甲醇萃取法的毒素回收率試驗中，以100%甲醇將藻體破碎、高純氮吹乾後，再添加1%甲醇回溶，可得到70%至120%之毒素回收率；定量PCR法在回收濾紙藻個數上，則回收率較佳，並與添加細胞數具相關性，因此後續實驗中以定量PCR法進行樣本藻細胞數分析。
在三個湖庫周界的實地採樣分析結果顯示，當湖泊中存在微囊藻、柱孢藻和藻毒時，空氣採樣器能有效地將帶有柱孢藻毒、及微囊藻之氣膠採集於濾紙上，並可藉由甲醇萃取和ELISA法，量測到空氣中有0.02-0.002 ng/m3的柱孢藻毒，顯示毒素會隨著氣膠懸浮於湖泊周界的空氣中。本研究旨在建立優養化水體周界上方氣膠粒子中藻體與藻毒濃度的採樣分析方法，相較於過去著重在毒素對飲用水的影響，新的觀點指出若空氣中的氣膠藻體和藻毒濃度達到一定程度以上，同樣將對優養化水體周界活動的民眾造成暴露的風險，建議主管機關後續納入評估與考量。

Cyanobacteria blooms and cyanotoxins have posed a great threat to human beings for many years, causing liver and renal pathology, mainly through oral intake of drinking water contaminated by the toxins. However, in a few recent studies, inhalation of aerosolized cyanobacteria and cyanotoxins is considered to be another pathway for exposure to cyanotoxins. Therefore, establishing a feasible and efficient detection method for the air samples will provide a better basis to evaluate the risk of inhalation exposure to cyanotoxins.
In this study, we developed sampling and analytical procedures and collected aerosols near eutrophic reservoirs and lakes in Taiwan, including two places in Taiwan main island and one in Kinmen island by PS-1 sampler, PM10 and PM2.5 cyclones. Enzyme linked immunosorbent assay (ELISA) and quantitative polymerase chain reaction (qPCR) methods were employed to quantify the aerosolized microcystins/cylindrospermopsins and four target genes within Microcystis and Cylindrospermopsis. The results show that, when 1% methanol was used to dissolve the dried cyanotoxins, recovery efficiency is ranged from 70% to 120% for ELISA. Besides, qPCR also had a good correlation with the cell enumerations in liquid phase, and therefore, the qPCR approach was further applied in determining the copy/cell number on the sampling filters.
The field sampling results show that Microcystis, Cylindrospermopsis and cylindrospermopsins were detected in the air samples collected near the studied reservoirs/lakes, with the gene copies of the two cyanobacteria ranging from below detection limit to 102 copies/m3 air and cylindrospermopsin from below detection limit to 0.02 ng/m3 air. This study demonstrates the presence of the harmful cyanobacteria and toxins near eutrophic reservoirs/lakes. However, more studies are required to better quantify the concentrations of the harmful cyanobacetria and toxins, and to link with the water quality of the reservoirs/lakes.

Backer L.C., McNeel S.V., Barber T., Kirkpatrick B.,Williams C., Irvin M., Zhou Y., Johnson T.B., Nierenberg K., Aubel M., LePrell R., Chapman A., Foss A., Corum S., Hill V., Kieszak S.M., and Cheng Y.S. Recreational exposure to microcystins during algal blooms in two California lakes. Toxicon, 55: 909-921. (2010)
Bishop C.T., Anet E.F.L. and Gorham P.R. Isolation and Identification of the Fast-Death Factor in Microcystis Aeruginosa Nrc-1. Canadian Journal of Biochemistry and Physiology, 37(3): 453-471. (1959)
Boopathi T. and Ki J.S. Impact of Environmental Factors on the Regulation of Cyanotoxin Production. Toxins, 6: 1951-1978. (2014)
Brookes J.D. and Carey C.C. Resilience to Blooms. Science, 334(6052), 46-47. (2011)
Byth S. Palm Island mystery disease. Medical Journal of Australia, 2(1): 40-42. (1980)
Cane D.E., Walsh C.T. and Khosla C. Harnessing the biosynthetic code: combinations, permutations, and mutations. Science, 282: 63-68. (1998)
Carey C.C., Ibelings B.W., Hoffmann E.P., Hamilton D.P. and Brookes J.D. Eco-physiological adaptations that favour freshwater cyanobacteria in a changing climate. Water Research, 46: 1394-1407. (2012)
Carmichael W.W., Azevedo M.F.O., An J.S., Molica R.J.R., Jochimsen E.M., Lau S., Rinehart K.L., Shaw G.R., and Eagelsham G.K. Human Fatalities from Cyanobacteria: Chemical and Biological Evidence for Cyanotoxins. Environmental Health Perspectives, 109: 663-668. (2001)
Chen H, Burke E, and Prepas E.E. Cyanobacterial toxins in fresh waters. In: Nriagu JO, editor. Encyclopedia of environmental health. Amsterdam: Elsevier; 860–71. (2011)
Cheng Y.S., Zhou Y., Irvin C.M., Kirkpatrick B., Backer L.C. Characterization of Aerosols Containing Microcystin. Marine Drugs, 136-150. (2007)
Chien A., Edgar D.B., and Trela J.M. Deoxyribonucleic acid polymerase from the extreme thermophile Thermus aquaticus. Bacteriol, 127: 1550-1557. (1976)
Christiansen G., Fastner J., Erhard M., Börner T., and Dittmann E. Microcystin biosynthesis in Planktothrix: genes, evolution and manipulation. J. Bacteriol. 185: 564-572. (2003)
Codd G.A., Bell S.G., Kaya K., Ward C.J., Beattie K.A., Metcalf J.S. Cyanobacterial toxins, exposure routes and human health. Eur. J. Phycol. 34: 405-416. (1999)
Codd G.A., Morrison L.F. and Metcalf J.S. Cyanobacterial toxins: risk management for health protection. Toxicology and Applied Pharmacology, 203: 264-272. (2005)
Dittmann E. and Börner T. Genetic contributions to the risk assessment of microcystin in the environment. Toxicol. Appl. Pharmacol. 203: 192-200. (2005)
Duy T.N., Lam P.K.S., Shaw G.R., and Connell D.W. Toxicology and risk assessment of freshwater cyanobacterial (blue-green algal) toxins in water. Reviews of Environmental Contamination and Toxicology, 163: 113-186. (2000)
Engvall E. and Perlmann P. Enzyme-linked immunosorbent assay (ELISA) quantitative assay of immunoglobulin G. Immunochemistry, 8(9): 871-874. (1971)
Fergusson K.M. and Saint C.P. Multiplex PCR assay for Cylindrospermopsis raciborskii and cylindrospermopsin-producing cyanobacteria. Environ Toxicol, 18(2): 120-125. (2003)
Fleming L.E., Backer L.C., and Baden D.G. Overview of aerosolized Florida red tide toxins: exposures and effects. Environmental Health Perspectives, 113: 618-620. (2005b)
Fleming L.E., Bean J.A., Kirkpatrick B., Chung Y.S., Pierce R., Naar J., Nierenberg K., Backer L.C., Wanner A., Reich A., Zhou Y., Watkins S., Henry M., Zaias J., AbrahamW.M., Benson J., Cassedy A., Hollenbeck J., Kirkpatrick G., Clarke T. and Baden D.G. Exposure and effect assessment of aerosolized red tide toxins (brevetoxins) and asthma. Environmental Health Perspectives, 117: 1095-1100. (2009)
Fleming L.E., Kirkpatrick B., Backer L.C., Bean J.A., Wanner A., Reich A., Dalpra D., Zaias J., Cheng Y.S., Pierce R., Naar J., Abraham W.M. and Baden D.G. Aerosolized red-tide toxins (brevetoxins) and asthma. Chest, 131: 187-194. (2007)
Fleming L.E., Kirkpatrick B., Backer L.C., Bean J.A., Wanner A., Dalpra D., Tamer R., Zaias J., Cheng Y.S., Pierce R., Naar J., Abraham W., Clark R., Zhou Y., Henry M.S., Johnson D., Van de Bogart G., Bossart G.D., Harrington M., and Baden D.G. Initial evaluation of the effects of aerosolized Florida red tide toxins (brevetoxins) in persons with asthma. Environmental Health Perspectives, 113: 650-657. (2005a)
Funari E. and Testai E. Human health risk assessment related to cyanotoxins exposure. Critical Reviews in Toxicology, 38: 97-125. (2008)
Griffiths D.J. and Saker M.L. The Palm Island mystery disease 20 years on: A review of research on the cyanotoxin cylindrospermopsin. Environmental Toxicology, 18(2): 78-93. (2003)
Harada K., Ogawa K., Kimura Y., Murata H. and Suzuki M. Microcystins from Anabaena flos-aquae NRC 525-17. Chemical Research in Toxicology, 4: 535-540. (1991)
Hummert C., Dahlmann J., Reichelt M., and Luckas M. Analytical techniques for monitoring harmful cyanobacteria in lakes. Lakes and Reservoirs: Research and Management, 6: 159-168. (2001)
Innis M.A. and Gelfand D.H. Optimization of PCRs. In: Innis MA, Gelfand DH, Sninsky JJ, White TJ, editors. PCR protocol. San Diego: Academic Press, pp. 3-12. (1990)
Karlin S. and Mrazek J. What drives codon choices in human genes. Journal of Molecular Biology, 262(4): 459-472. (1996)
Mackintosh C. and Klumpp S. Tautomycin from the bacterium Streptomyces verticillatus. Another potent and specific inhibitor of protein phosphatases 1 and 2A. FEBS Lett. 277: 137-140. (1990)
Maizels M. and Budde W.L. A LC/MS Method for the Determination of Cyanobacteria Toxins in Water. Analysis Chemistry, 76: 1342-1351. (2004)
Michinaka A., Yen H.K., Chiu Y.T., Tsao H.W. and Lin T.F. Rapid on-site multiplex assays for total and toxigenic Microcystis using real-time PCR with microwave cell disruption, Water Sci. Technol., 66: 1246-1252. (2012)
Mihali T.K., Kellmann R., Muenchhoff J., Barrow K.D. and Neilan B.A. Characterization of the gene cluster responsible for cylindrospermopsin biosynthesis. Applied and Environmental Microbiology, 74(3), 716-722. (2008)
Mikalsen B., Boison G., Skulberg O.M., Fastner J., Davies W., Gabrielsen T.M., Rudi K. and Jakobsen K.S. Natural variation in the microcystin synthetase operon mcyABC and impact on microcystin production in Microcystis strains. Bacteriol, 185(9): 2774-2785. (2003)
Moffitt M.C. and Neilan B.A. On the presence of peptide synthetase and polyketide synthase genes in the cyanobacterial genus Nodularia. FEMS Microbiol. Lett. 196: 207-214. (2001)
National Rivers Authority. Toxic blue-green algae. London: National Rivers Authority. (1990)
Neilan B.A., Jacobs D., DelDot T., Blackall L.L., Hawkins P.R., Cox P.T. and Goodman A.E. rRNA sequences and evolutionary relationships among toxic and nontoxic cyanobacteria of the genus Microcystis. Int. J. Syst. Bacteriol. 47: 693-697. (1997)
Newcombe G, editor. International guidance manual for the management of toxic cyanobacteria. London: IWA Publishing. (2012)
Nishizawa T., Asayama M. and Shirai M. Cyclic heptapeptide microcystin biosynthesis requires the glutamate racemase gene. Microbiology, 147(5): 1235-1241. (2001)
Norris R.L., Eaglesham G.K., Pierens G., Shaw G.R., Smith M.J., Chiswell R.K., Seawright A.A. and Moore M.R. Deoxycylindrospermopsin, an analog of cylindrospermopsin from Cylindrospermopsis raciborskii. Environ. Toxicol, 14: 163-165. (1999)
Pearson L.A., Barrow K.D. and Neilan B. A. Characterization of the 2-hydroxy-acid dehydrogenase mcyI, encoded within the microcystin biosynthesis gene cluster of Microcystis aeruginosa PCC7806. Journal of Biological Chemistry, 282(7): 4681-4692. (2007)
Pearson L.A., Hisbergues M., Börner T., Dittmann E. and Neilan B. A. Inactivation of an ABC transporter gene, mcyH, results in loss of microcystin production in the cyanobacterium Microcystis aeruginosa PCC7806. Appl. Environ. Microbiol 70: 6370-6378. (2004)
Philipp R. and Bates A.J. Health-risks assessment of dinghy sailing in Avon and exposure to cyanobacteria (Blue-green algae). J. Inst. Water Environ. Manage, 6: 613-617. (1992)
Percudani R., Pavest A. and Ottonello S. Transfer RNA gene redundancy and translational selection in saccharomyces cerevisiae. Journal of Molecular Biology, 268(2): 322-330. (1997)
Puddick J., Prinsep M.R., Wood, S.A., Cary S.C., Hamiton D.P. and Holland P.T. Further Characterization of Glycine-Containing Microcystins from the McMurdo Dry Valleys of Antarctica. Toxins, 7: 493-515. (2015)
Pyo D. and Shin H. Extraction and analysis of microcystins RR and LR in cyanobacteria using a cyano cartridge. Journal of Biochemical and Biophysical Methods, 51: 103-109. (2002)
Rao P.V.L., Gupta N., Bhaskar A.S.B. and Jayaraj R. Toxins and bioactive compounds from cyanobacteria and their implications on human health. Journal of Environmental Biology, 3: 215-224. (2002)
Rasmussen J.P., Giglio S., Monis P.T., Campbell R.J. and Saint, C.P. Development and field testing of a real-time PCR assay for cylindrospermopsin-producing cyanobacteria. Appl Microbiol, 104(5): 1503-1515. (2008)
Rinta-Kanto J.M., Ouellette A.J.A., Boyer G.L., Twiss M.R., Bridgeman T.B. and Wilhelm S.W. Quantification of toxic Microcystis spp. during the 2003 and 2004 blooms in western Lake Erie using quantitative real-time PCR. Environmental Science & Technology, 39(11): 4198-4205. (2005)
Rippka R. Isolation and purification of cyanobacteria. Methods in enzymology, 167. 3. (1988)
Rouhiainen L., Vakkilainen T., Siemer B.L., Buikema W., Haselkorn R. and Sivonen K. Genes coding for hepatotoxic heptapeptides (microcystins) in the cyanobacterium Anabaena strain 90. Appl. Environ. Microbiol. 70: 686-692. (2004)
Saiki R.K., Scharf S., Faloona F., Mullis K.B., Horn G.T., Erlich H.A. and Arnheim N. Enzymatic amplification of beta-globin genomic sequences and restriction site analysis for diagnosis of sickle cell anemia. Science 230: 1350-1354. (1985)
Schembri M.A., Neilan B.A. and Saint C.P. Identification of genes implicated in toxin production in the cyanobacterium Cylindrospermopsis raciborskii. Environmental Toxicology, 16(5): 413-421. (2001)
Shalev‐Alon G., Sukenik A., Livnah O., Schwarz R. and Kaplan A. A novel gene encoding amidinotransferase in the cylindrospermopsin producing cyanobacterium Aphanizomenon ovalisporum. FEMS microbiology letters, 209(1), 87-91. (2002)
Sielaff H., Dittmann E., Tandeau De Marsac N., Bouchier C., Von Döhren H., Börner T. and Schwecke T. The mcyF gene of the microcystin biosynthetic gene cluster from Microcystis aeruginosa encodes an aspartate racemase. Biochemical Journal, 373(3): 909-916. (2003)
Smith C.J. and Osborn A.M. Advantages and limitations of quantitative PCR (Q-PCR)-based approaches in microbial ecology. Microbiology Ecology, 67(1): 6-20. (2008)
Stewart I., Schluter P. and Shaw G. Cyanobacterial lipopolysaccharides and human health. Review of Environmental Health, 5: 7. (2006)
Thompson A.S., Rhodes J.C. and Pettman I. Natural Environmental Research Council Culture Collection of Algae and Protozoa-Catalogue of Strains; Freshwater Biology Association, Ambleside, UK; p. 22. (1988)
Tillett D., Dittman E., Erhard M., von Döhren H., Börner T. and Neilan B. A. Structural organization of microcystin biosynthesis in Microcystis aeruginosa PCC7806: an integrated peptide: polyketide synthetase system. Chem. Biol., 7: 253-264. (2000)
Turner P.C., Gammie A.J., Hollinrake K., Codd G.A. Pneumonia associated with contact with cyanobacteria. Brit. Med. J., 300: 1440-1441. (1990)
Tsuji T., Watanuki T., Kondo F., Ishikawa N., Watanabe M.F., Suzuki S., Nakazawa H., Suzuki M., Uchida H., and Harada K.I. Stability of microcystins from cyanobacteria--II. Effect of UV light on decomposition and isomerization. Toxicon, 33: 1619-1631. (1995)
Harada K.I., Tsuji T., Watanabe M.F., and Kondo F. Stability of microcystins from cyanobacteria-III, effect of pH and temperature. Phycologia, 35: 83-88. (1996)
Toxicologyguide Website, http://www.toxicologyguide.com/944-molecular-techniques/
USEPA. Creating a Cyanotoxin Target List for the Unregulated Contaminant Monitoring Rule. (2001)
USEPA. Drinking Water Health Advisory for the Cyanobacterial Microcystin Toxins. (2015a)
USEPA. Health Effects Support Document for the Cyanobacterial Toxin Microcystins. (2015b)
USEPA. Health Effects Support Document for the Cyanobacterial Toxin Cylindrospermopsin. (2015c)
Whipple G.C. (revised by Fair G.M. and Whipple M.C.) The Microscopy of drinking water. John Wiley and Sons Inc., pp. xix + 586, 19 plates. (1927)
WHO. Management of Cyanobacteria in Drinking-water Supplies: Information for regulators and water suppliers. (2015)
WHO. Toxic Cyanobacteria in Water: A guide to their public health consequences, monitoring and management- Laboratory analysis of cyanotoxins. (1999)
Wilson K.M., Schembri M.A., Baker P.D. and Saint, C.P. Molecular characterization of the toxic cyanobacterium Cylindrospermopsis raciborskii and design of a species-specific PCR. Applied and Environmental Microbiology, 66(1): 332-338. (2000)
Wimmer K.M., Strangman W.K. and Wright J.L.C. 7-Deoxy-desulfo-cylindrospermopsin and 7-deoxy-desulfo-12-acetylcylindrospermopsin: Two new cylindrospermopsin analogs isolated from a Thai strain of Cylindrospermopsis raciborskii. Harmful Algae, 37: 203-206. (2014)
Westerhoff P. and Mash H. Dissolved Organic Nitrogen in Drinking Water Supplies: A Review. Journal of Water Supply Research and Technology-Aqua 2002. 51, 415-448.
Wood S.A. and Dietrich D.R. Quantitative assessment of aerosolized cyanobacterial toxins at two New Zealand lakes. Environmental Monitoring, 13: 1617-1624. (2011)
Yen H.K., Lin T.F. and Liao P.C. Simultaneous detection of nine cyanotoxins in drinking water using dual solid-phase extraction and liquid chromatography- mass spectrometry. Toxicon, 58: 209-218. (2011)
Yen H.K., Lin T.F. and Tseng I.C. Detection and quantification of major toxigenic Microcystis genotypes in Moo-Tan reservoir and associated water treatment plant. Environ. Monit, 14 (2): 687-696. (2012)
李貞慧、吳美惠、張嬉麗、廖福全、黃瑞聰、吳美炷、林彥宏、許惠佳，水公司水庫原水及其淨水場原清水中微囊藻毒之調查研究，第五屆海峽兩岸水質安全技術與管理研討會，澳門，10月28-29日 (2009)
李聖哲，台灣中部沿海地區之重金屬污染來源分析，碩士論文，國立成功大學環境工程研究所 (2011)
林財富、吳哲宏、顏宏愷，水庫有害藻類及微生物監測應變與檢測技術研究經濟部水利署 (2013)
林財富等，水源產毒藻類與有害微生物監控與管理技術之研發與應用，經濟部水利署 (2010)
林健智，微囊藻產毒基因檢測技術開發與微囊藻毒素釋放生理研究，國立台灣大學漁業科學研究所碩士論文 (2007)
洪文宗、翁英明、牟麗娥、黃壬瑰、李秓萍、劉素妙、潘復華、蕭美琪、趙春美、梁淑婷，螢光光度計應用在水庫環境連續監測探討，環保署環境檢驗所，中壢 (2008)
動植物防疫檢疫局官網，第八章-聚合酶鏈鎖反應(Polymerase chain reaction, 簡稱PCR)，行政院農委會
游漢威、莊蕙萍、董珮伶、簡義杰、黃良銘、林財富，SybrGreen-qPCR法定量氯烯類污染區域中關鍵微生物菌群與其功能性基因，中華民國環境工程學會土壤與地下水研討會，中原大學，11月13-14日 (2015)
黃鑫、魏曉璐、馮悅、張阿梅、夏雪山、劉麗，微囊藻毒素生物合成基因及其功能研究進展，《生命科學研究》，第18卷第5期：第445頁-第452頁 (2014)
廖大修與陳靜瑩，第九章-質譜儀的技術原理介紹及蛋白質鑑定，醫藥基因生物技術教學資源中心主編，蛋白質體學：第100頁-第109頁 (2004)
廖寶琦，第十一章-介質輔助雷射脫附離子化質譜法，醫藥基因生物技術教學資源中心主編，蛋白質體學：第127頁-第138頁 (2004a)
廖寶琦，第十二章-電噴灑離子化質譜法，醫藥基因生物技術教學資源中心主編，蛋白質體學：第139頁-第150頁 (2004b)
環境檢驗所，水中微囊藻毒及節球藻毒篩檢方法－盤式或條式直接競爭型酵素免疫分析法，NIEA E510.50B，行政院環境保護署 (2011)
環境檢驗所，放流水標準，行政院環境保護署 (2006)
環境檢驗所，空氣中粒狀污染物檢測法－高量採樣法，NIEA A102.12A，行政院環境保護署 (2006)
環境檢驗所，飲用水標準，行政院環境保護署 (2004)