隨著工業的進步與生活水準的提升，民眾對於水源的品質與安全也越來越重視，供水單位也因此致力於水庫原水以及供水水源的管理，但由於水源區內農業的開發以及家庭污水之排放，導致大多數的水庫常有水質優養化的現象，進而衍生出藻類有害代謝物的問題。目前已知部分藍綠菌於生長過程亦會產生二次代謝物(secondary metabolites)，例如微囊藻毒素(microcystins, MCs)、柱孢藻毒素(cylindrospermopsin, CYN)與二甲基冰片(2-methylisoborneol, 2-MIB)，這些代謝物不僅造成水質之惡化，更會影響了飲用水的安全。因此了解藍綠菌及其代謝物的存在，對於淨水處理單位非常重要。
此外，為了瞭解環境水體中產毒/產臭的來源，分離與鑑定菌株是必要的步驟。當完成菌株的純化分離後，即可針對分離菌株進行外觀與生理特性等進行研究。然而，於一系列的世代培養下，雖然可以簡便地維持菌株的生命，但已有許多研究皆指出，長期的世代培養會導致菌株基本特性的改變，例如型態、生理、生化以及遺傳性質等。因此，發展簡便的長期培養技術，對於菌株分離與保存非常重要。
以目前法規規範藻類代謝物之傳統監測方法而言，須經採完水樣後，送之具有複雜儀器設備之實驗室進行分析，由水庫發現藻華開始，採樣、運送樣品、分析、回報結果等，通常至少需要2-3天的時間。若水源水質遭到污染進而超過了標準值，不僅無法於第一時間掌握水庫的狀況進而採取相關應變措施外，可能因此增加民眾健康危害之風險，甚至造成不可逆的傷害。有鑑於此，若能縮短相關檢測技術所需之時間，並進一步將其運用於現地檢測，可讓管理單位及各級淨水廠有足夠的時間提出相關的應對措施以及預警策略，降低飲用水受到污染所造成相關之危害，以提升飲用水安全的保障。
本研究應用即時定量連鎖聚合酶反應(quantitative polymerase chain reaction, qPCR)系統配合TaqMan偵測方法(qPCR-TaqMan系統)，搭配專一性引子(primer)與探針(probe)，提供快速分析水中主要藻類風險(藻類毒素與臭味)的能力，並結合顯微鏡鏡檢、酵素免疫連結法(ELISA)與氣相層析質譜儀(GC-MSD)針對台灣地區水庫水樣中的產毒/產臭基因、目標藍綠菌細胞數、藻類毒素濃度以及臭味物質濃度進行監測，並探討三者間的相關性，以期可即時掌握柱孢藻毒素對飲用水體的潛在危害，進一步保障民眾用水安全。此外，對於不同抗冷凍劑應用於藍綠菌的保存技術中，本研究結果顯示，5%的二甲基亞碸(Dimethyl sulfoxide；DMSO)對於大部分的藍綠菌有顯著的保存作用。
本研究發展的qPCR技術已成功應用於台灣及離島地區共29座水庫中，其結果顯示顯微鏡鏡檢與qPCR分析的總細胞數，以及ELISA分析的藻類毒素與qPCR分析的產毒基因數皆有良好的相關性(R2值範圍介於0.392到0.740之間)。同樣地，於GC/MS測得的2-MIB濃度與qPCR分析的產2-MIB基因數(mibC genes)也有良好的相關性(R2值範圍介於0.473到0.479之間)。
此結果顯示本技術可以快速、有效定量出藍綠菌代謝物發生潛勢，可有效地回饋分析結果給水庫管理單位，對於水源安全提供快速且有效之保障。

Lakes and reservoirs represent important drinking water sources in Taiwan and many other countries. Due to improper management of nutrients in their watersheds, many reservoirs are facing the risk of eutrophication, increasing the possibility of cyanobacterial blooms in the source waters. Some of them may produce cyanotoxins and odorants as secondary metabolites, such as microcystins (MCs), cylindrospermopsin (CYN), and 2-methylisoborneol (2-MIB). Drinking water contaminated with cyanotoxins and odorants may affect public health and consumers’ perception to public water supply. Therefore, it is very important to understand the presence of cyanobacteria and their metabolites in water supplies.
In addition, to correlate the presence of a toxin/odorant in a given reservoir to a specific cyanobacterial species, isolation of strains from each environmental sample was a necessary step. The isolated strains can then be easily screened for the production of toxins/odorants and be studied morphologically and genetically in order to assign them to a particular species. However, loss of general characteristic such as morphology, physiology, biochemistry, and genetic properties during serial sub-culturing has been evidenced in several cyanobacterial cultures. Therefore, it is important to have a reliable and simple technique for long-term preservation after acquiring or isolating cyanobacterial culture.
Currently, when water utilities are faced with cyanobacteria problems in the source water, to confirm the risk associated with cyanotoxins and odorants the standard procedures include sample collection and delivery, laboratory analysis, and reporting. The whole procedures usually take more than 24 hours. In this case, the contaminated water may have entered into public water supplies. Therefore, a rapid method to quickly provide the potential risk of cyanobacteria associated toxins and odorants would be very usefully for water utilities to justify the quality of their drinking water.
This study aimed to establish a quantitative real-time polymerase chain reaction (qPCR) to quickly determine the risk of cyanobacteria associated toxins (MCs and CYN) and odorants (2-MIB) in fresh water bodies based on the detection of cyanotoxin and odorant-producing cyanobacterial genes. Concurrently, an enzyme-linked immunosorbent assay (ELISA) and a solid-phase microextraction (SPME) concentration followed by a gas chromatograph-mass selective detector (GC-MSD) was employed to measure cyanotoxins and odorants produced by cyanobacteria, respectively. Samples were also enumerated for cyanobacteria cells with microscope. In addition, the efficacy of the various cryoprotective agents was compared for several isolates of cyanobacteria. This study highlighted that the 5 % dimethyl-sulphoxide (DMSO) for cryopreservation was shown to be a preferable choice of cryopreservative for most strains tested.
This qPCR based approaches have been successfully applied in 29 drinking-water sources in Taiwan and her offshore islands. Results show that cell numbers of Microcystis and Cylindorspermopsis enumerated with microscopy, and MCs and CYN concentrations measured with the enzyme-linked immuno-sorbent assay method, correlated well with their corresponding gene copies determined with the qPCR systems (range of coefficients of determination R2 = 0.392−0.740). There are also statistically significant correlations between mibC genes and 2-MIB concentrations (range of coefficients of determination R2 = 0.473−0.479).
The results demonstrate that the developed qPCR based approaches were able to quantify the targeted cyanobacteria and toxin/odorant producing genes. In particular, the obtained gene information correlated well with cell numbers enumerated with microscopy, toxin concentrations measured with ELISA, and 2-MIB concentration measured with GC/MS. This scheme should provide timely information to managers of reservoirs and water utilities to diagnose the potential risk associated with toxigenic cyanobacteria, cyanotoxins, and 2-MIB in drinking water lakes and reservoirs.

Abed, I.J. (2015) PCR-Based Test for the Early Warning of Both Cylindrospermopsin and Saxitoxin in Iraqi Freshwater. Journal of Al-Nahrain University 18(3), 109-114.
Al-Tebrineh, J., Gehringer, M.M., Akcaalan, R. and Neilan, B.A. (2011) A new quantitative PCR assay for the detection of hepatotoxigenic cyanobacteria. Toxicon 57(4), 546-554.
Al-Tebrineh, J., Pearson, L.A., Yasar, S.A. and Neilan, B.A. (2012) A multiplex qPCR targeting hepato- and neurotoxigenic cyanobacteria of global significance. Harmful Algae 15, 19-25.
Alster, A., Kaplan-Levy, R.N., Sukenik, A. and Zohary, T. (2010) Morphology and phylogeny of a non-toxic invasive Cylindrospermopsis raciborskii from a Mediterranean Lake. Hydrobiologia 639(1), 115-128.
Andersen, R. (1996) Maintaining cultures for biotechnology and industry. Hunter-Cevera, J. and Belt, A. (eds), pp. 29-64, Academic Press. Inc, London.
Andersen, R.A. and Kawachi, M. (2005) Traditional microalgae isolation techniques. Algal culturing techniques, 83-100.
Apha (1985) Standard methods for the examination of water and wastewater, Apha.
Apt, K.E. and Behrens, P.W. (1999) Commercial developments in microalgal biotechnology. Journal of Phycology 35(2), 215-226.
Baker, J.A., Entsch, B., Neilan, B.A. and McKay, D.B. (2002) Monitoring Changing Toxigenicity of a Cyanobacterial Bloom by Molecular Methods. Applied and Environmental Microbiology 68(12), 6070-6076.
Baker, L., Sendall, B.C., Gasser, R.B., Menjivar, T., Neilan, B.A. and Jex, A.R. (2013) Rapid, multiplex-tandem PCR assay for automated detection and differentiation of toxigenic cyanobacterial blooms. Molecular and Cellular Probes 27(5–6), 208-214.
Baker, P. and Humpage, A. (1994) Toxicity associated with commonly occurring cyanobacteria in surface waters of the Murray-Darling Basin, Australia. Marine and Freshwater Research 45(5), 773-786.
Banker, R., Carmeli, S., Hadas, O., Teltsch, B., Porat, R. and Sukenik, A. (1997) Identification of cylindrospermopsin in Aphanizomenon ovalisporum (cyanophyceae) isolated from lake kinneret, israel. Journal of Phycology 33(4), 613-616.
Baxa, D.V., Kurobe, T., Ger, K.A., Lehman, P.W. and Teh, S.J. (2010) Estimating the abundance of toxic Microcystis in the San Francisco Estuary using quantitative real-time PCR. Harmful Algae 9(3), 342-349.
Beaty, M. and Parker, B. (1990) Investigations of cryopreservation and storage of eukaryotic algae and protozoa. J. Phycol 26(5).
Benson, E.E. (2008) Cryopreservation of phytodiversity: a critical appraisal of theory & practice. Critical reviews in Plant sciences 27(3), 141-219.
Berglind, L., Johnsen, I.J., Ormerod, K. and Skulberg, O.M. (1983) Oscillatoria brevis (Kutz.) Gom. and some other especially odouriferous benthic cyanophytes in Norwegian inland waters. Water Science and Technology 15(6-7), 241-246.
Boerjesson, T.S., Stoellman, U.M. and Schnuerer, J.L. (1993) Off-odorous compounds produced by molds on oatmeal agar: Identification and relation to other growth characteristics. Journal of Agricultural and Food Chemistry 41(11), 2104-2111.
Botes, D.P., Kruger, H. and Viljoen, C.C. (1982) Isolation and characterization of four toxins from the blue-green alga, Microcystis aeruginosa. Toxicon 20(6), 945-954.
Botes, D.P., Tuinman, A.A., Wessels, P.L., Viljoen, C.C., Kruger, H., Williams, D.H., Santikarn, S., Smith, R.J. and Hammond, S.J. (1984) The structure of cyanoginosin-LA, a cyclic heptapeptide toxin from the cyanobacterium Microcystis aeruginosa. Journal of the Chemical Society, Perkin Transactions 1, 2311-2318.
Bowling, L. (1994) Occurrence and possible causes of a severe cyanobacterial bloom in Lake Cargelligo, New South Wales. Marine and Freshwater Research 45(5), 737-745.
Bowling, L., Egan, S., Holliday, J. and Honeyman, G. (2016) Did spatial and temporal variations in water quality influence cyanobacterial abundance, community composition and cell size in the Murray River, Australia during a drought-affected low-flow summer? Hydrobiologia 765(1), 359-377.
Bowling, L.C., Merrick, C., Swann, J., Green, D., Smith, G. and Neilan, B.A. (2013) Effects of hydrology and river management on the distribution, abundance and persistence of cyanobacterial blooms in the Murray River, Australia. Harmful Algae 30, 27-36.
Brock, N.L., Ravella, S.R., Schulz, S. and Dickschat, J.S. (2013) A Detailed View of 2-Methylisoborneol Biosynthesis. Angewandte Chemie International Edition 52(7), 2100-2104.
Butakova, E.A. (2013) Specific features of odor-causing compounds (geosmin and 2-methylisoborneol) as secondary metabolites of cyanobacteria. Russian Journal of Plant Physiology 60(4), 507-510.
Cañavate, J.P. and Lubian, L.M. (1995) Relationship between cooling rates, cryoprotectant concentrations and salinities in the cryopreservation of marine microalgae. Marine Biology 124(2), 325-334.
Carmichael, W.W. (1986) Algal Toxins. Advances in Botanical Research 12, 47-101.
Carmichael, W.W. (1992) Cyanobacteria secondary metabolites—the cyanotoxins. Journal of Applied Bacteriology 72(6), 445-459.
Carmichael, W.W. (2001) Health effects of toxin-producing cyanobacteria:“The CyanoHABs”. Human and ecological risk assessment: An International Journal 7(5), 1393-1407.
Castenholz, R.W. (1988) Culturing methods for cyanobacteria. Methods in enzymology 167, 68-93.
Chapman, A.D. and Schelske, C.L. (1997) Recent appearance of Cylindrospermopsis (cyanobacteiua) in five hypereutrophic Florida lakes. Journal of Phycology 33(2), 191-195.
Chiu, Y.-T., Chen, Y.-H., Wang, T.-S., Yen, H.-K. and Lin, T.-F. (2017) A qPCR-Based Tool to Diagnose the Presence of Harmful Cyanobacteria and Cyanotoxins in Drinking Water Sources. International Journal of Environmental Research and Public Health 14(5), 547.
Chiu, Y.-T., Yen, H.-K. and Lin, T.-F. (2016) An alternative method to quantify 2-MIB producing cyanobacteria in drinking water reservoirs: Method development and field applications. Environmental Research 151, 618-627.
Chorus, I. (2001) Cyanotoxins: occurrence, causes, consequences. Chorus, I. (ed), pp. 75-78, Springer Science & Business Media, Berlin.
Chow, G.C. (1960) Tests of Equality Between Sets of Coefficients in Two Linear Regressions. Econometrica 28(3), 591-605.
Christiansen, G., Fastner, J., Erhard, M., Borner, T. and Dittmann, E. (2003) Microcystin biosynthesis in Planktothrix: Genes, evolution, and manipulation. Journal of Bacteriology 185(2), 564-572.
Chu, F., Huang, X. and Wei, R. (1989) Enzyme-linked immunosorbent assay for microcystins in blue-green algal blooms. Journal-Association of Official Analytical Chemists 73(3), 451-456.
Codd, G., Steffensen, D., Burch, M. and Baker, P. (1994) Toxic blooms of cyanobacteria in Lake Alexandrina, South Australia—learning from history. Marine and Freshwater Research 45(5), 731-736.
Codd, G.A. (2000) Cyanobacterial toxins, the perception of water quality, and the prioritisation of eutrophication control. Ecological Engineering 16(1), 51-60.
Codd, G.A., Lindsay, J., Young, F.M., Morrison, L.F. and Metcalf, J.S. (2005a) Harmful Cyanobacteria. Huisman, J., Matthijs, H.C.P. and Visser, P.M. (eds), pp. 1-23, Springer Netherlands, Dordrecht.
Codd, G.A., Morrison, L.F. and Metcalf, J.S. (2005b) Cyanobacterial toxins: risk management for health protection. Toxicology and Applied Pharmacology 203(3), 264-272.
Cook, D., Newcombe, G. and Sztajnbok, P. (2001) The application of powdered activated carbon for mib and geosmin removal: predicting pac doses in four raw waters. Water Research 35(5), 1325-1333.
Dai, J., Peng, H., Chen, W., Cheng, J. and Wu, Y. (2013) Development of multiplex real-time PCR for simultaneous detection of three Potyviruses in tobacco plants. Journal of applied microbiology 114(2), 502-508.
Dawson, R.M. (1998) the toxicology of microcystins. Toxicon 36(7), 953-962.
Day, J., Benson, E., Harding, K., Knowles, B., Idowu, M., Bremner, D., Santos, L., Santos, F., Friedl, T. and Lorenz, M. (2005) Cryopreservation and conservation of microalgae: the development of a pan-European scientific and biotechnological resource (the COBRA project). CryoLetters 26(4), 231-238.
Day, J.G., Benson, E.E. and Fleck, R.A. (1999) In vitro culture and conservation of microalgae: applications for aquaculture, biotechnology and environmental research. In Vitro Cellular & Developmental Biology-Plant 35(2), 127-136.
Day, J.G. and Brand, J.J. (2005) Algal culturing techniques. Anderson, R.A. (ed), pp. 165-188, Academic Press, London.
Day, J.G., Watanabe, M.M. and Turner, M.F. (1998) Ex-situ conservation of protistan and cyanobacterial biodiversity: CCAP-NIES collaboration 1991–1997. Phycological Research 46, 77-83.
Dickschat, J.S., Nawrath, T., Thiel, V., Kunze, B., Müller, R. and Schulz, S. (2007) Biosynthesis of the Off-Flavor 2-Methylisoborneol by the Myxobacterium Nannocystis exedens. Angewandte Chemie International Edition 46(43), 8287-8290.
Dionigi, C.P., Lawlor, T.E., McFarland, J.E. and Johnsen, P.B. (1993) Evaluation of geosmin and 2-methylisoborneol on the histidine dependence of TA98 and TA100 Salmonella typhimurium tester strains. Water Research 27(11), 1615-1618.
Dittmann, E. and Börner, T. (2005) Genetic contributions to the risk assessment of microcystin in the environment. Toxicology and Applied Pharmacology 203(3), 192-200.
Dittmann, E., Fewer, D.P. and Neilan, B.A. (2013) Cyanobacterial toxins: biosynthetic routes and evolutionary roots. FEMS Microbiology Reviews 37(1), 23-43.
Dittmann, E., Neilan, B.A., Erhard, M., Von Döhren, H. and Börner, T. (1997) Insertional mutagenesis of a peptide synthetase gene that is responsible for hepatotoxin production in the cyanobacterium Microcystis aeruginosa PCC 7806. Molecular Microbiology 26(4), 779-787.
Domingos, P., Rubim, T.K., Molica, R.J.R., Azevedo, S. and Carmichael, W.W. (1999) First report of microcystin production by picoplanktonic cyanobacteria isolated from a northeast Brazilian drinking water supply. Environmental Toxicology 14(1), 31-35.
Duy, T.N., Lam, P.K., Shaw, G.R. and Connell, D.W. (2000) Reviews of Environmental Contamination and Toxicology, pp. 113-185, Springer.
Dwyer, Daniel J., Camacho, Diogo M., Kohanski, Michael A., Callura, Jarred M. and Collins, James J. (2012) Antibiotic-Induced Bacterial Cell Death Exhibits Physiological and Biochemical Hallmarks of Apoptosis. Molecular Cell 46(5), 561-572.
Elnifro, E.M., Ashshi, A.M., Cooper, R.J. and Klapper, P.E. (2000) Multiplex PCR: Optimization and Application in Diagnostic Virology. Clinical Microbiology Reviews 13(4), 559-570.
Fan, J., Rao, L., Chiu, Y.-T. and Lin, T.-F. (2016) Impact of chlorine on the cell integrity and toxin release and degradation of colonial Microcystis. Water Research 102, 394-404.
Fastner, J., Erhard, M. and von Döhren, H. (2001) Determination of Oligopeptide Diversity within a Natural Population of Microcystis spp. (Cyanobacteria) by Typing Single Colonies by Matrix-Assisted Laser Desorption Ionization–Time of Flight Mass Spectrometry. Applied and Environmental Microbiology 67(11), 5069-5076.
Fergusson, K.M. and Saint, C.P. (2003) Multiplex PCR assay for Cylindrospermopsis raciborskii and cylindrospermopsin-producing cyanobacteria. Environmental Toxicology 18(2), 120-125.
Fischer, W.J., Garthwaite, I., Miles, C.O., Ross, K.M., Aggen, J.B., Chamberlin, A.R., Towers, N.R. and Dietrich, D.R. (2001) Congener-independent immunoassay for microcystins and nodularins. Environmental Science & Technology 35(24), 4849-4856.
Fortin, N., Aranda-Rodriguez, R., Jing, H., Pick, F., Bird, D. and Greer, C.W. (2010) Detection of Microcystin-Producing Cyanobacteria in Missisquoi Bay, Quebec, Canada, Using Quantitative PCR. Applied and Environmental Microbiology 76(15), 5105-5112.
Frank, C.A.P. (2002) Microcystin-producing cyanobacteria in recreational waters in southwestern Germany. Environmental Toxicology 17(4), 361-366.
Gavrieli, Y., Sherman, Y. and Ben-Sasson, S.A. (1992) Identification of programmed cell death in situ via specific labeling of nuclear DNA fragmentation. The Journal of Cell Biology 119(3), 493-501.
Gerber, N.N. (1979) Volatile Substances from Actinomycetes: Their Role in the Odor Pollution of Water. CRC Critical Reviews in Microbiology 7(3), 191-214.
Giglio, S., Chou, W.K.W., Ikeda, H., Cane, D.E. and Monis, P.T. (2011) Biosynthesis of 2-methylisoborneol in cyanobacteria. Environmental Science & Technology 45(3), 992-998.
Griese, M., Lange, C. and Soppa, J. (2011) Ploidy in cyanobacteria. FEMS Microbiology Letters 323(2), 124-131.
Grout, B.W. (1995) Cryopreservation of plant protoplasts. Methods in Molecular Biology 38, 91-101.
Hamilton, T.L., Bryant, D.A. and Macalady, J.L. (2016) The role of biology in planetary evolution: cyanobacterial primary production in low‐oxygen Proterozoic oceans. Environmental Microbiology 18(2), 325-340.
Harada, K.-i., Ohtani, I., Iwamoto, K., Suzuki, M., Watanabe, M.F., Watanabe, M. and Terao, K. (1994) Isolation of cylindrospermopsin from a cyanobacterium Umezakia natans and its screening method. Toxicon 32(1), 73-84.
Harding, K., Day, J.G., Lorenz, M., Timmermann, H., Friedl, T., Bremner, D.H. and Benson, E.E. (2004) Introducing the concept and application of vitrification for the cryo-conservation of algae–a mini-review. Nova Hedwigia 79(1-2), 207-226.
Henegariu, O., Heerema, N.A., Dlouhy, S.R., Vance, G.H. and Vogt, P.H. (1997) Multiplex PCR: critical parameters and step-by-step protocol. BioTechniques 23(3), 504-511.
Hindson, C.M., Chevillet, J.R., Briggs, H.A., Gallichotte, E.N., Ruf, I.K., Hindson, B.J., Vessella, R.L. and Tewari, M. (2013) Absolute quantification by droplet digital PCR versus analog real-time PCR. Nat Meth 10(10), 1003-1005.
Hubalek, Z. (2003) Protectants used in the cryopreservation of microorganisms. Cryobiology 46(3), 205-229.
Huggett, J.F., Foy, C.A., Benes, V., Emslie, K., Garson, J.A., Haynes, R., Hellemans, J., Kubista, M., Mueller, R.D., Nolan, T., Pfaffl, M.W., Shipley, G.L., Vandesompele, J., Wittwer, C.T. and Bustin, S.A. (2013) The Digital MIQE Guidelines: <em>M</em>inimum <em>I</em>nformation for Publication of <em>Q</em>uantitative Digital PCR <em>E</em>xperiments. Clinical Chemistry 59(6), 892-902.
Hyeon, J.-Y., Park, C., Choi, I.-S., Holt, P.S. and Seo, K.-H. (2010) Development of multiplex real-time PCR with Internal amplification control for simultaneous detection of Salmonella and Cronobacter in powdered infant formula. International Journal of Food Microbiology 144(1), 177-181.
Izaguirre, G. (1992) A Copper-Tolerant Phormidium Species from Lake Mathews, California, That Produces 2-Methylisoborneol and Geosmin. Water Science and Technology 25(2), 217-223.
Izaguirre, G., Hwang, C.J., Krasner, S.W. and McGuire, M.J. (1982) Geosmin and 2-Methylisoborneol from Cyanobacteria in Three Water Supply Systems. Applied and Environmental Microbiology 43(3), 708-714.
Izaguirre, G., Hwang, C.J., Krasner, S.W. and McGuire, M.J. (1983) Production of 2-Methyliso-Borneol by Two Benthic Cyanophyta. Water Science and Technology 15(6-7), 211-220.
Izaguirre, G. and Taylor, W.D. (2004) A guide to geosmin- and MIB-producing cyanobacteria in the United States. Water Science and Technology 49(9), 19-24.
Jüttner, F. (1987) Volatile organic substances. The cyanobacteria, 453-469.
Jüttner, F. and Watson, S.B. (2007) Biochemical and ecological control of geosmin and 2-methylisoborneol in source waters. Applied and Environmental Microbiology 73(14), 4395-4406.
Jiang, Y., Xiao, P., Yu, G., Sano, T., Pan, Q. and Li, R. (2012) Molecular Basis and Phylogenetic Implications of Deoxycylindrospermopsin Biosynthesis in the Cyanobacterium Raphidiopsis curvata. Applied and Environmental Microbiology 78(7), 2256-2263.
Jochimsen, E.M., Carmichael, W.W., An, J., Cardo, D.M., Cookson, S.T., Holmes, C.E., Antunes, M.B., de Melo Filho, D.A., Lyra, T.M. and Barreto, V.S.T. (1998) Liver failure and death after exposure to microcystins at a hemodialysis center in Brazil. New England Journal of Medicine 338(13), 873-878.
Jones, G. (1993) On the occurrence and control and toxic cyanobacterial blooms in Australia. J Phycol 41, 400-401.
Jones, K.H. and Senft, J.A. (1985) An improved method to determine cell viability by simultaneous staining with fluorescein diacetate-propidium iodide. Journal of Histochemistry & Cytochemistry 33(1), 77-79.
Kaebernick, M., Rohrlack, T., Christoffersen, K. and Neilan, B.A. (2001) A spontaneous mutant of microcystin biosynthesis: genetic characterization and effect on Daphnia. Environmental Microbiology 3(11), 669-679.
Kakimoto, M., Ishikawa, T., Miyagi, A., Saito, K., Miyazaki, M., Asaeda, T., Yamaguchi, M., Uchimiya, H. and Kawai-Yamada, M. (2014) Culture temperature affects gene expression and metabolic pathways in the 2-methylisoborneol-producing cyanobacterium Pseudanabaena galeata. Journal of Plant Physiology 171(3–4), 292-300.
Kerr, J.F.R., Wyllie, A.H. and Currie, A.R. (1972) Apoptosis: A basic biological phenomenon with wide-ranging implications in tissue kinetics. British Journal of Cancer 26(4), 239-257.
Kim, H., Cho, E. and Park, S. (2006) Analysis of factors affecting the cryopreservation of garlic shoot tips. Journal of Biomedical Nanotechnology 2(2), 129-132.
Kiss, T., Vehovszky, Á., Hiripi, L., Kovács, A. and Vörös, L. (2002) Membrane effects of toxins isolated from a cyanobacterium, Cylindrospermopsis raciborskii, on identified molluscan neurones. Comparative Biochemistry and Physiology Part C: Toxicology & Pharmacology 131(2), 167-176.
Komatsu, M., Tsuda, M., Ōmura, S., Oikawa, H. and Ikeda, H. (2008) Identification and functional analysis of genes controlling biosynthesis of 2-methylisoborneol. Proceedings of the National Academy of Sciences of the United States of America 105(21), 7422-7427.
Kono, S., Kuwano, K., Ninomiya, M., Onishi, J. and Saga, N. (1997) Cryopreservation of Enteromorpha intestinalis (Ulvales, Chlorophyta) in liquid nitrogen. Phycologia 36(1), 76-78.
Koskenniemi, K., Lyra, C., Rajaniemi-Wacklin, P., Jokela, J. and Sivonen, K. (2007) Quantitative Real-Time PCR Detection of Toxic Nodularia Cyanobacteria in the Baltic Sea. Applied and Environmental Microbiology 73(7), 2173-2179.
Kotak, B.G., Lam, A.K.Y., Prepas, E.E., Kenefick, S.L. and Hrudey, S.E. (1995) VARIABILITY OF THE HEPATOTOXIN MICROCYSTIN-LR IN HYPEREUTROPHIC DRINKING WATER LAKES1. Journal of Phycology 31(2), 248-263.
Krasner, S.W., Hwang, C.J. and McGuire, M.J. (1983) A standard method for quantification of earthy-musty odorants in water, sediments, and algal cultures. Water Science and Technology 15(6-7), 127-138.
Kurmayer, R., Christiansen, G. and Chorus, I. (2003) The Abundance of Microcystin-Producing Genotypes Correlates Positively with Colony Size in Microcystis sp. and Determines Its Microcystin Net Production in Lake Wannsee. Applied and Environmental Microbiology 69(2), 787-795.
Kurmayer, R., Dittmann, E., Fastner, J. and Chorus, I. (2002) Diversity of microcystin genes within a population of the toxic cyanobacterium Microcystis spp. in Lake Wannsee (Berlin, Germany). Microbial Ecology 43(1), 107-118.
Kurmayer, R. and Kutzenberger, T. (2003) Application of real-time PCR for quantification of microcystin genotypes in a population of the toxic cyanobacterium Microcystis sp. Applied and Environmental Microbiology 69(11), 6723-6730.
Lawton, L.A. and Codd, G.A. (1991) Cyanobacterial (Blue-Green Algal) Toxins and their Significance in UK and European Waters. Water and Environment Journal 5(4), 460-465.
Lee, L.G., Connell, C.R. and Bloch, W. (1993) Allelic discrimination by nick-translation PCR with fluorgenic probes. Nucleic Acids Research 21(16), 3761-3766.
Li, R., Carmichael, W.W., Brittain, S., Eaglesham, G.K., Shaw, G.R., Liu, Y. and Watanabe, M.M. (2001a) First report of the cyanotoxins cylindrospermopsin and deoxycylindrospermopsin from Raphidiopsis curvata (cyanobacteria). Journal of Phycology 37(6), 1121-1126.
Li, R., Carmichael, W.W., Brittain, S., Eaglesham, G.K., Shaw, G.R., Mahakhant, A., Noparatnaraporn, N., Yongmanitchai, W., Kaya, K. and Watanabe, M.M. (2001b) Isolation and identification of the cyanotoxin cylindrospermopsin and deoxy-cylindrospermopsin from a Thailand strain of Cylindrospermopsis raciborskii (Cyanobacteria). Toxicon 39(7), 973-980.
Li, Z., Hobson, P., An, W., Burch, M.D., House, J. and Yang, M. (2012) Earthy odor compounds production and loss in three cyanobacterial cultures. Water Research 46(16), 5165-5173.
Lin, T.-F., Liu, C.-L., Yang, F.-C. and Hung, H.-W. (2003) Effect of residual chlorine on the analysis of geosmin, 2-MIB and MTBE in drinking water using the SPME technique. Water Research 37(1), 21-26.
Lin, T.-F., Wong, J.-Y. and Kao, H.-P. (2002) Correlation of musty odor and 2-MIB in two drinking water treatment plants in South Taiwan. Science of The Total Environment 289(1–3), 225-235.
Lin, T., Watson, S., Devesa, R., Bruchet, A., Burlingame, G., Dietrich, A. and Suffet, M. (2012) Off-flavours in the aquatic environment: A global issue. Global Trends & Challenges in Water Science, Research and Management–A compendium of hot topics and features from IWA Specialist Groups. International Water Association, London, UK, 58-63.
Lorenz, M., Friedl, T. and Day, J.G. (2005) Algal culturing techniques. Anderson, R.A. (ed), pp. 145-156, Academic Press, London.
Lourey, M.J., Thompson, P.A., McLaughlin, M.J., Bonham, P. and Feng, M. (2013) Primary production and phytoplankton community structure during a winter shelf-scale phytoplankton bloom off Western Australia. Marine Biology 160(2), 355-369.
Luukkainen, R., Sivonen, K., Namikoshi, M., Fardig, M., Rinehart, K.L. and Niemela, S.I. (1993) Isolation and identification of 8 microcysitns from 13 Oscillatoria agardhii strains and structure of a new microcystin. Applied and Environmental Microbiology 59(7), 2204-2209.
Lysáková, M., Kitner, M. and Poulíčková, A. (2007) The epipelic algae at fishponds of Central and Northern Moravia (the Czech Republic). Fottea 7, 69-75.
Marbun, Y.R., Yen, H.-K., Lin, T.-F., Lin, H.-L. and Michinaka, A. (2012) Rapid on-site monitoring of cylindrospermopsin-producers in reservoirs using quantitative PCR.
Mattheis, J.P. and Roberts, R.G. (1992) Identification of geosmin as a volatile metabolite of Penicillium expansum. Applied and Environmental Microbiology 58(9), 3170-3172.
Mez, K., Hanselmann, K. and Preisig, H.R. (1998) Environmental conditions in high mountain lakes containing toxic benthic cyanobacteria. Hydrobiologia 368(1), 1-15.
Michinaka, A., Yen, H.K., Chiu, Y.T., Tsao, H.W. and Lin, T.F. (2012) Rapid on-site multiplex assays for total and toxigenic Microcystis using real-time PCR with microwave cell disruption. Water Science and Technology 66(6), 1247-1252.
Mihali, T.K., Kellmann, R., Muenchhoff, J., Barrow, K.D. and Neilan, B.A. (2008) Characterization of the Gene Cluster Responsible for Cylindrospermopsin Biosynthesis. Applied and Environmental Microbiology 74(3), 716-722.
Mikalsen, B., Boison, G., Skulberg, O.M., Fastner, J., Davies, W., Gabrielsen, T.M., Rudi, K. and Jakobsen, K.S. (2003) Natural variation in the microcystin synthetase operon mcyABC and impact on microcystin production in Microcystis strains. Journal of Bacteriology 185(9), 2774-2785.
Misson, B., Sabart, M., Amblard, C. and Latour, D. (2012) Benthic survival of Microcystis: Long-term viability and ability to transcribe microcystin genes. Harmful Algae 13, 20-25.
Mohamed, Z.A. and Al Shehri, A.M. (2009) Microcystin-producing blooms of Anabaenopsis arnoldi in a potable mountain lake in Saudi Arabia. Fems Microbiology Ecology 69(1), 98-105.
Montgomery, D.C., Peck, E.A. and Vining, G.G. (2015) Introduction to linear regression analysis, John Wiley & Sons.
Mori, F., Erata, M. and Watanabe, M.M. (2002) Cryopreservation of cyanobacteria and green algae in the NIES-Collection. Microbiol. Cult. Coll 18(1), 45-55.
Morris, G. (1978) Cryopreservation of 250 strains of Chlorococcales by the method of two-step cooling. British Phycological Journal 13(1), 15-24.
Mur, L.R. (1983) Some aspects of the ecophysiology of cyanobacteria. Annales de l'Institut Pasteur / Microbiologie 134(1), 61-72.
Mur, L.R., Skulberg, O.M. and Utkilen, H. (1999) Toxic Cyanobacteria in Water: A guide to their public health consequences, monitoring and management. Chorus, I. and Bartram, J. (eds), E & FN Spon, London.
Neilan, B., Saker, M., Fastner, J., Törökné, A. and Burns, B. (2003) Phylogeography of the invasive cyanobacterium Cylindrospermopsis raciborskii. Molecular Ecology 12(1), 133-140.
Newcombe, G., House, J., Ho, L., Baker, P. and Burch, M. (2010) Management strategies for cyanobacteria (Blue-Green Algae): A guide for water utilities. Water Quality Research Australia (WQRA), Reserach Report 74.
Ngwa, F.F., Madramootoo, C.A. and Jabaji, S. (2013) Development and application of a multiplex qPCR technique to detect multiple microcystin-producing cyanobacterial genera in a Canadian freshwater lake. Journal of Applied Phycology 26(4), 1675-1687.
Nishizawa, T., Asayama, M., Fujii, K., Harada, K. and Shirai, M. (1999) Genetic analysis of the peptide synthetase genes for a cyclic heptapeptide microcystin in Microcystis spp. Journal of Biochemistry 126(3), 520-529.
Ohtani, I., Moore, R.E. and Runnegar, M.T.C. (1992) Cylindrospermopsin: a potent hepatotoxin from the blue-green alga Cylindrospermopsis raciborskii. Journal of the American Chemical Society 114(20), 7941-7942.
Oldfield, E. and Lin, F.-Y. (2012) Terpene Biosynthesis: Modularity Rules. Angewandte Chemie International Edition 51(5), 1124-1137.
Oliver, R.L. (1994) Floating and sinking in gas-vacuolate cyanobacteria. Journal of Phycology 30(2), 161-173.
Otsuka, S., Suda, S., Li, R., Watanabe, M., Oyaizu, H., Matsumoto, S. and Watanabe, M.M. (1999) Phylogenetic relationships between toxic and non-toxic strains of the genus Microcystis based on 16S to 23S internal transcribed spacer sequence. FEMS Microbiology Letters 172(1), 15-21.
Ouahid, Y. and del Campo, F.F. (2009) Typing of toxinogenic Microcystis from environmental samples by multiplex PCR. Applied Microbiology and Biotechnology 85(2), 405-412.
Ouahid, Y., Perez-Silva, G. and del Campo, F.F. (2005) Identification of potentially toxic environmental Microcystis by individual and multiple PCR amplification of specific microcystin synthetase gene regions. Environmental Toxicology 20(3), 235-242.
Ouellette, A.J.A., Handy, S.M. and Wilhelm, S.W. (2006) Toxic microcystis is widespread in Lake Erie: PCR detection of toxin genes and molecular characterization of associated cyanobacterial communities. Microbial Ecology 51(2), 154-165.
Padisák, J. (1997) Cylindrospermopsis raciborskii (Woloszynska) Seenayya et Subba Raju, an expanding, highly adaptive cyanobacterium: worldwide distribution and review of its ecology. Archiv Für Hydrobiologie Supplementband Monographische Beitrage 107(4), 563-593.
Paerl, H.W., Fulton, R.S., Moisander, P.H. and Dyble, J. (2001) Harmful Freshwater Algal Blooms, With an Emphasis on Cyanobacteria. TheScientificWorldJOURNAL 1.
Park, H.-D., Iwami, C., Watanabe, M.F., Harada, K.-I., Okino, T. and Hayashi, H. (1998) Temporal variabilities of the concentrations of intra- and extracellular microcystin and toxic Microcystis species in a hypertrophic lake, Lake Suwa, Japan (1991–1994). Environmental Toxicology and Water Quality 13(1), 61-72.
Park, H.-K. (2006) Long-term preservation of bloom-forming cyanobacteria by cryopreservation. Algae 21(1), 125-131.
Park, S.Y., Kim, E.Y., Cui, X.S., Tae, J.C., Lee, W.D., Kim, N.H., Park, S.P. and Lim, J.H. (2006) Increase in DNA fragmentation and apoptosis-related gene expression in frozen-thawed bovine blastocysts. Zygote 14(02), 125-131.
Pearson, L.A. and Neilan, B.A. (2008) The molecular genetics of cyanobacterial toxicity as a basis for monitoring water quality and public health risk. Current Opinion in Biotechnology 19(3), 281-288.
Pereira, P. and rio Tenreiro, R. (2010) Multiplex PCR for detection of microcystins-producing cyanobacteria from freshwater samples.
Pollak, F.C. and Berger, R.G. (1996) Geosmin and Related Volatiles in Bioreactor-Cultured Streptomyces citreus CBS 109.60. Applied and Environmental Microbiology 62(4), 1295-1299.
Poulíčková, A., Hašler, P., Lysáková, M. and Spears, B. (2008) The ecology of freshwater epipelic algae: an update. Phycologia 47(5), 437-450.
Pouria, S., de Andrade, A., Barbosa, J., Cavalcanti, R.L., Barreto, V.T.S., Ward, C.J., Preiser, W., Poon, G.K., Neild, G.H. and Codd, G.A. (1998) Fatal microcystin intoxication in haemodialysis unit in Caruaru, Brazil. The Lancet 352(9121), 21-26.
Preußel, K., Stüken, A., Wiedner, C., Chorus, I. and Fastner, J. (2006) First report on cylindrospermopsin producing Aphanizomenon flos-aquae (Cyanobacteria) isolated from two German lakes. Toxicon 47(2), 156-162.
Quesada, A., Moreno, E., Carrasco, D., Paniagua, T., Wormer, L., Hoyos, C.d. and Sukenik, A. (2006) Toxicity of Aphanizomenon ovalisporum (Cyanobacteria) in a Spanish water reservoir. European Journal of Phycology 41(1), 39-45.
Rücker, J., Stüken, A., Nixdorf, B., Fastner, J., Chorus, I. and Wiedner, C. (2007) Concentrations of particulate and dissolved cylindrospermopsin in 21 Aphanizomenon-dominated temperate lakes. Toxicon 50(6), 800-809.
Rai, A.N. and Bergman, B. (2002) Creation of new nitrogen-fixing cyanobacterial associations, pp. 65-68, JSTOR.
Rantala, A., Rajaniemi-Wacklin, P., Lyra, C., Lepistö, L., Rintala, J., Mankiewicz-Boczek, J. and Sivonen, K. (2006) Detection of Microcystin-Producing Cyanobacteria in Finnish Lakes with Genus-Specific Microcystin Synthetase Gene E (mcyE) PCR and Associations with Environmental Factors. Applied and Environmental Microbiology 72(9), 6101-6110.
Rasmussen, B., Fletcher, I.R., Brocks, J.J. and Kilburn, M.R. (2008a) Reassessing the first appearance of eukaryotes and cyanobacteria. Nature 455(7216), 1101-1104.
Rasmussen, J.P., Giglio, S., Monis, P.T., Campbell, R.J. and Saint, C.P. (2008b) Development and field testing of a real-time PCR assay for cylindrospermopsin-producing cyanobacteria. Journal of applied microbiology 104(5), 1503-1515.
Reed, B.M. (2008) Plant cryopreservation: a practical guide, Springer.
Rhodes, L., Holland, P., Adamson, J., Selwood, A. and McNabb, P. (2002) Mass culture of New Zealand isolates of Pseudo-nitzschia australis for production of a new isomer of domoic acid. Harmful Algae, 125-127.
Rhodes, L., Smith, J., Tervit, R., Roberts, R., Adamson, J., Adams, S. and Decker, M. (2006) Cryopreservation of economically valuable marine micro-algae in the classes Bacillariophyceae, Chlorophyceae, Cyanophyceae, Dinophyceae, Haptophyceae, Prasinophyceae, and Rhodophyceae. Cryobiology 52(1), 152-156.
Rinehart, K.L., Harada, K., Namikoshi, M., Chen, C., Harvis, C.A., Munro, M.H., Blunt, J.W., Mulligan, P.E. and Beasley, V.R. (1988) Nodularin, microcystin, and the configuration of Adda. Journal of the American Chemical Society 110(25), 8557-8558.
Rinta-Kanto, J.M., Ouellette, A.J.A., Boyer, G.L., Twiss, M.R., Bridgeman, T.B. and Wilhelm, S.W. (2005) 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.
Rippka, R. (1988) Methods in Enzymology, pp. 3-27, Academic Press.
Romo, S. and Bécares, E. (1992) Preservation of filamentous cyanobacteria cultures (Pseudanabaena galeata) Böcher and Geitlerinema amphibium (Ag. ex Gom.) Anagn. under low temperatures. Journal of Microbiological Methods 16(2), 85-89.
Runnegar, M.T., Kong, S.-M., Zhong, Y.-Z. and Lu, S.C. (1995) Inhibition of reduced glutathione synthesis by cyanobacterial alkaloid cylindrospermopsin in cultured rat hepatocytes. Biochemical Pharmacology 49(2), 219-225.
Runnegar, M.T., Kong, S.M., Zhong, Y.Z., Ge, J.L. and Lu, S.C. (1994) The Role of Glutathione in the Toxicity of a Novel Cyanobacterial Alkaloid Cylindrospermopsin in Cultured Rat Hepatocytes. Biochemical and Biophysical Research Communications 201(1), 235-241.
Saker, M., Thomas, A. and Norton, J. (1999) Cattle mortality attributed to the toxic cyanobacterium Cylindrospermopsis raciborskii in an outback region of north Queensland. Environmental Toxicology 14(1), 179-182.
Schöller, C.E.G., Gürtler, H., Pedersen, R., Molin, S. and Wilkins, K. (2002) Volatile Metabolites from Actinomycetes. Journal of Agricultural and Food Chemistry 50(9), 2615-2621.
Schembri, M.A., Neilan, B.A. and Saint, C.P. (2001) Identification of genes implicated in toxin production in the cyanobacterium Cylindrospermopsis raciborskii. Environmental Toxicology 16(5), 413-421.
Sevilla, E., Martin-Luna, B., Vela, L., Bes, M.T., Peleato, M.L. and Fillat, M.F. (2010) Microcystin-LR synthesis as response to nitrogen: transcriptional analysis of the mcyD gene in Microcystis aeruginosa PCC7806. Ecotoxicology 19(7), 1167-1173.
Shaw, G.R., Sukenik, A., Livne, A., Chiswell, R.K., Smith, M.J., Seawright, A.A., Norris, R.L., Eaglesham, G.K. and Moore, M.R. (1999) Blooms of the cylindrospermopsin containing cyanobacterium, Aphanizomenon ovalisporum (Forti), in newly constructed lakes, Queensland, Australia. Environmental Toxicology 14(1), 167-177.
Silva, P.G., Ferrari, S.G. and Silva, H.J. (2007) Preservation methods of Tolypothrix tenuis for use as a cyanobacterial fertilizer. Journal of Applied Phycology 19(3), 239-246.
Sint, D., Raso, L. and Traugott, M. (2012) Advances in multiplex PCR: balancing primer efficiencies and improving detection success. Methods in Ecology and Evolution 3(5), 898-905.
Sivonen, K. and Börner, T. (2008) Bioactive compounds produced by cyanobacteria. The cyanobacteria: molecular biology, genomics and evolution, 159-197.
Sivonen, K. and Jones, G. (1999) Toxic cyanobacteria in water: A guide to their public health consequences, monitoring and management. Chorus, I. and Bartram, J. (eds), pp. 44-111, E & FN Spon, London.
Sivonen, K., Namikoshi, M., Evans, W.R., Carmichael, W.W., Sun, F., Rouhiainen, L., Luukkainen, R. and Rinehart, K.L. (1992a) Isolation and characterization of a variety of microcystins fron 7 strains of the cyanobacterial genus Anabaena. Applied and Environmental Microbiology 58(8), 2495-2500.
Sivonen, K., Namikoshi, M., Evans, W.R., Fardig, M., Carmichael, W.W. and Rinehart, K.L. (1992b) 3 New microcystins, cylic heptapeptide hepatotoxins, from Nostoc sp. strain 152. Chemical Research in Toxicology 5(4), 464-469.
Socol, C.T., Vlaic, A., Cosier, V. and Criste, A. (2008) Assesment of cryopreservation systems influence on the survavial of E. coli recomvinant strains. Scientific Papers Animal Science and Biotechnologies 41(1), 138-142.
Sommer, U., Gliwicz, Z.M., Lampert, W. and Duncan, A. (1986) The PEG-model of seasonal succession of planktonic events in fresh waters. Arch. Hydrobiol 106(4), 433-471.
Spoof, L., Berg, K.A., Rapala, J., Lahti, K., Lepistö, L., Metcalf, J.S., Codd, G.A. and Meriluoto, J. (2006) First observation of cylindrospermopsin in Anabaena lapponica isolated from the boreal environment (Finland). Environmental Toxicology 21(6), 552-560.
Srinivasan, R. and Sorial, G.A. (2011) Treatment of taste and odor causing compounds 2-methyl isoborneol and geosmin in drinking water: A critical review. Journal of Environmental Sciences 23(1), 1-13.
Stitz, L., Kinnear, S. and Fabbro, L. (2013) A role for aeration and intake depth in managing toxic Cylindrospermopsis: A comparison between off-stream and riverine environments in the Fitzroy Basin, Australia. Lakes & Reservoirs: Research & Management 18(2), 179-196.
Sun, D., Yu, J., An, W., Yang, M., Chen, G. and Zhang, S. (2013) Identification of causative compounds and microorganisms for musty odor occurrence in the Huangpu River, China. Journal of Environmental Sciences 25(3), 460-465.
Suurnäkki, S., Gomez-Saez, G.V., Rantala-Ylinen, A., Jokela, J., Fewer, D.P. and Sivonen, K. (2015) Identification of geosmin and 2-methylisoborneol in cyanobacteria and molecular detection methods for the producers of these compounds. Water Research 68, 56-66.
Takano, M., Sado, J.-I., Ogawa, T. and Terui, G. (1973) Freezing and freeze-drying of Spirulina platensis. Cryobiology 10(5), 440-444.
Taton, A., Grubisic, S., Brambilla, E., De Wit, R. and Wilmotte, A. (2003) Cyanobacterial diversity in natural and artificial microbial mats of Lake Fryxell (McMurdo Dry Valleys, Antarctica): A morphological and molecular approach. Applied and Environmental Microbiology 69(9), 5157-5169.
Taylor, R. and Fletcher, R.L. (1998) Cryopreservation of eukaryotic algae – a review of methodologies. Journal of Applied Phycology 10(5), 481-501.
Te, S.H., Chen, E.Y. and Gin, K.Y.-H. (2015) Comparison of quantitative PCR and droplet digital PCR multiplex assays for two genera of bloom-forming cyanobacteria, Cylindrospermopsis and Microcystis. Applied and Environmental Microbiology 81(15), 5203-5211.
Theiss, W.C., Carmichael, W.W., Wyman, J. and Bruner, R. (1988) Blood pressure and hepatocellular effects of the cyclic heptapeptide toxin produced by the freshwater cyanobacterium (blue - green alga) Microcystis aeruginosa strain PCC-7820. Toxicon 26(7), 603-613.
Tillett, D., Dittmann, E., Erhard, M., von Döhren, H., Börner, T. and Neilan, B.A. (2000) Structural organization of microcystin biosynthesis in Microcystis aeruginosa PCC7806: an integrated peptide–polyketide synthetase system. Chemistry & Biology 7(10), 753-764.
Tillett, D., Parker, D.L. and Neilan, B.A. (2001) Detection of toxigenicity by a probe for the microcystin synthetase A gene (mcyA) of the cyanobacterial genus Microcystis: Comparison of toxicities with 16S rRNA and phycocyaninoperon (phycocyanin intergenic spacer) phylogenies. Applied and Environmental Microbiology 67(6), 2810-2818.
Tsao, H.-W., Michinaka, A., Yen, H.-K., Giglio, S., Hobson, P., Monis, P. and Lin, T.-F. (2014) Monitoring of geosmin producing Anabaena circinalis using quantitative PCR. Water Research 49, 416-425.
Tung, S.-C., Lin, T.-F., Yang, F.-C. and Liu, C.-L. (2008) Seasonal change and correlation with environmental parameters for 2-MIB in Feng-Shen Reservoir, Taiwan. Environmental Monitoring and Assessment 145(1-3), 407-416.
Tzovenis, I., Triantaphyllidis, G., Naihong, X., Chatzinikolaou, E., Papadopoulou, K., Xouri, G. and Tafas, T. (2004) Cryopreservation of marine microalgae and potential toxicity of cryoprotectants to the primary steps of the aquacultural food chain. Aquaculture 230(1), 457-473.
Vaitomaa, J., Rantala, A., Halinen, K., Rouhiainen, L., Tallberg, P., Mokelke, L. and Sivonen, K. (2003) Quantitative real-time PCR for determination of microcystin synthetase E copy numbers for Microcystis and Anabaena in lakes. Applied and Environmental Microbiology 69(12), 7289-7297.
Valério, E., Chambel, L., Paulino, S., Faria, N., Pereira, P. and Tenreiro, R. (2010) Multiplex PCR for detection of microcystins-producing cyanobacteria from freshwater samples. Environmental Toxicology 25(3), 251-260.
Wang, L., Li, Y. and Mustapha, A. (2007) Rapid and simultaneous quantitation of Escherichia coli O157: H7, Salmonella, and Shigella in ground beef by multiplex real-time PCR and immunomagnetic separation. Journal of Food Protection® 70(6), 1366-1372.
Wang, Z., Song, G., Shao, J., Tan, W., Li, Y. and Li, R. (2015a) Establishment and field applications of real-time PCR methods for the quantification of potential MIB-producing cyanobacteria in aquatic systems. Journal of Applied Phycology, 1-9.
Wang, Z., Xiao, P., Song, G., Li, Y. and Li, R. (2015b) Isolation and characterization of a new reported cyanobacterium Leptolyngbya bijugata coproducing odorous geosmin and 2-methylisoborneol. Environmental Science and Pollution Research 22(16), 12133-12140.
Wang, Z., Xu, Y., Shao, J., Wang, J. and Li, R. (2011) Genes Associated with 2-Methylisoborneol Biosynthesis in Cyanobacteria: Isolation, Characterization, and Expression in Response to Light. PLoS ONE 6(4), e18665.
Watson, S.B. (2003) Cyanobacterial and eukaryotic algal odour compounds: signals or by-products? A review of their biological activity. Phycologia 42(4), 332-350.
Watson, S.B. (2004) Aquatic taste and odor: a primary signal of drinking-water integrity. Journal of toxicology and environmental health. Part A 67(20-22), 1779-1795.
Watson, S.B., Ridal, J. and Boyer, G.L. (2008) Taste and odour and cyanobacterial toxins: impairment, prediction, and management in the Great Lakes. Canadian Journal of Fisheries and Aquatic Sciences 65(8), 1779-1796.
White, S.H., Duivenvoorden, L.J. and Fabbro, L.D. (2005) Impacts of a toxic Microcystis bloom on the Macroinvertebrate Fauna of Lake Elphinstone, Central Queensland, Australia. Hydrobiologia 548(1), 117-126.
Whitton, B.A. (2012) Ecology of cyanobacteria II: Their Diversity in Space and Time, Springer Dordrecht Heidelberg New York London.
WHO (2004) Guidelines for drinking-water quality, World Health Organization.
Wiedner, C., Chorus, I. and Fastner, J. (2001) The waterbodies surveyed for cyanotoxins in Germany. Cyanotoxins—Occurrence, Causes, Consequences. Springer, Berlin, 6-21.
Wilson, A.E., Sarnelle, O. and Tillmanns, A.R. (2006) Effects of cyanobacterial toxicity and morphology on the population growth of freshwater zooplankton: meta-analyses of laboratory experiments. Limnol. Oceanogr 51(4), 1915-1924.
Wilson, K.M., Schembri, M.A., Baker, P.D. and Saint, C.P. (2000) Molecular Characterization of the Toxic Cyanobacterium Cylindrospermopsis raciborskii and Design of a Species-Specific PCR. Applied and Environmental Microbiology 66(1), 332-338.
Wood, S., Williams, S.T. and White, W.R. (2001) Microbes as a source of earthy flavours in potable water - a review. International Biodeterioration & Biodegradation 48(1–4), 26-40.
Wood, S.A., Rhodes, L.L., Adams, S.L., Adamson, J.E., Smith, K.F., Smith, J.F., Tervit, H.R. and Cary, S.C. (2008) Maintenance of cyanotoxin production by cryopreserved cyanobacteria in the New Zealand culture collection. New Zealand Journal of Marine and Freshwater Research 42(3), 277-283.
Wu, J.-T. and Jüttner, F. (1988) Differential partitioning of geosmin and 2-methylisoborneol between cellular constituents in Oscillatoria tenuis. Archives of Microbiology 150(6), 580-583.
Yen, H.-K., Lin, T.-F. and Tseng, I.-C. (2012) Detection and quantification of major toxigenic Microcystis genotypes in Moo-Tan reservoir and associated water treatment plant. Journal of Environmental Monitoring 14(2), 687-696.
Yen, H., Lin, T., Tseng, I. and Su, Y. (2006) Cyanobacteria toxins and toxin producers in nine drinking water reservoirs in Taiwan. Water Science and Technology: Water Supply 6(2), 161-167.
Yoshida, M., Yoshida, T., Takashima, Y., Hosoda, N. and Hiroishi, S. (2007) Dynamics of microcystin-producing and non-microcystin-producing Microcystis populations is correlated with nitrate concentration in a Japanese lake. FEMS Microbiology Letters 266(1), 49-53.
Zhang, W., Lou, I., Ung, W.K., Kong, Y. and Mok, K.M. (2014) Application of PCR and real-time PCR for monitoring cyanobacteria, Microcystis spp. and Cylindrospermopsis raciborskii in Macau freshwater reservoir. Frontiers of Earth Science 8(2), 291-301.
Zohary, T. and Robarts, R.D. (1989) Diurnal mixed layers and the long-term dominance of Microcystis aeruginosa. Journal of Plankton Research 11(1), 25-48.
Zurawell, R.W., Chen, H., Burke, J.M. and Prepas, E.E. (2005) Hepatotoxic cyanobacteria: a review of the biological importance of microcystins in freshwater environments. Journal of Toxicology and Environmental Health, Part B 8(1), 1-37.