Quantitative Abundance and Concentration of Toxigenic and Odor Producing Cyanobacteria in Laguna Lake: New Discovery in the Philippines

菲律賓的新發現：量化Laguna湖中產生毒性與臭味的藍菌門之豐度與濃度

10.6937/TWC.202003/PP_68(1).0006

Ricardo F. de Leon Jr；Delia B. Senoro；Lennie Santos-Borja；Tsair-Fuh Lin；Yi-Ting Chiu；Yi-Hsuan Chen

2-methylisoborneol ； Blue green algae ； Cell-bound ； Inland water ； Taste and odor ； 2-甲基異莰醇 ； 藍藻 ； 細胞結合 ； 內陸水 ； 口感和氣味

台灣水利

68卷1期（2020 / 03 / 01）

52 - 66

英文

Laguna Lake in the Philippines is the biggest inland water source for domestic supply, with average depth of 2.5 meters, can significantly support water supply demand in the metropolis. However, the presence of cyanobacteria, known as blue-green algae, is a major threat to the lake water quality which directly affects nearby communities. Aiming to provide information for mitigation strategies to improve the water quality, the objective of this study is to quantitatively assess and monitor the harmful cyanobacteria in the water and sediment. The first ever 2-methylisoborneol (2-MIB) detection was recorded in Laguna Lake in August 2016. Collection of secondary qualitative data for the period 2012-2016 and quantitative detection and analysis of primary data from 2016 were considered for this study. To grab sampling on board, a boat was conducted with Ekman dredge and stainless water sampler for sediments and water column, respectively. Using the real time quantitative polymerase chain reaction, enzyme-linked immunosorbent assay, and gas chromatography, several species of harmful cyanobacteria were detected at various stations among the lake. The assessment of the 2012 data revealed the dominance of microcystis at cell density of 1,181,700 per liter at one of the major stations of the lake. Succeeding results from the 2016 monitored data uncovered abundance of toxigenic cyanobacteria species, high concentration of toxins such as microcystins and cylindrospermopsins, and high number of gene copies that produced taste and odor problems in the water. The cell density of microcystis monitored at the Central Bay area of the lake was 16,000,000 cells/mL while that of cylindrospermopsis was 220 cells/mL at the South Bay side. The abundance of 2-MIB in water column range from 210 to 100,000 copies/mL, while the concentrations range from 5.0 ng/L to 774.7 ng/L. This record suggests high activity of cyanobacterial bloom during the same period. The detection and quantitative analysis of the harmful blue-green algae in the lake provides a clearer picture of its water quality and implications to human health if not addressed properly. Such information may prove crucial in developing remediation strategy, appropriate water treatment system, research direction, as well as developmental cooperation among primary stakeholders within the Philippines. Concerned regulatory agencies can also benefit from the detection technology to implement measures and direct policy enhancement towards sustainable management of the lake.

菲律賓的Laguna湖是國內最大的內陸水源，其平均深度為2.5公尺，可以極大滿足大都市的用水需求。然而，被廣泛稱呼為藍綠藻的藍菌門如今大量存在於湖中，對Laguna湖的水質產生嚴重的威脅，並對周遭社區產生直接的影響。為提供有效的策略以減輕藍菌門對水質的傷害，本研究旨在量化、評估與監測湖水與沉積物中的有害藍菌門。2016年8月記錄了有史以來首次在Laguna湖檢測出的2-甲基異莰醇（2-MIB）。本文亦將2012年至2016年的次級質性資料以及2016年的定量檢測與分析的初級資料納入研究中。本研究分別使用艾克曼採泥器和不銹鋼採水器對沉積物與水柱進行採樣。通過使用即時聚合酶連鎖反應（Real-time PCR）、酵素結合免疫吸附分析法（ELISA）、氣相色譜法，我們在湖泊的數個站點檢測到了幾種有害的藍菌門。2012年資料的評估顯示，在湖泊的主要站點之一，微囊藻屬作為主要的藍菌門其細胞密度達到1,181,700 cells /L。2016年監測到的資料揭示了大量製造出毒素的藍菌門物種，高濃度的毒素如微囊藻與柱孢藻，以及其大量的複製基因所造成的口感和氣味問題。在Laguna湖的中央海灣區域，被監測到的微囊藻毒細胞密度達到16,000,000 cells/mL；在南灣區域，柱孢藻毒素則達到220 cells/mL。水柱中2-MIB豐度介於210到100,000 copies/mL，其濃度介於5.0 ng/L到774.7 ng/L之間。而這些記錄顯示藍藻水華在同一時期內高度活躍。針對湖中有害藍藻進行檢測與定量分析將有助於了解其造成的水質問題及對人類健康的隱患。這些資料對製定補救策略、適合的水處理系統、研究方向，以及菲律賓的主要利益相關者之間的發展合作至關重要。相關的監管機構可以受益於檢測技術，並實施措施與政策以推動Laguna湖的永續性管理。

1. American Water Works Association=AWWA(2016).,Water Research Foundation.
2. Arguelles, E.(2016).Morphotaxonomic Account of Epilithic Microalgae and Cyanobacteria in Los Baños, Laguna (Philippines).IAMURE International Journal of Ecology and Conservation,16,22-39.
3. Baldia, SF,Conaco, MCG.,Nijishima, T,Imanishi, S.,Harada, K.(2003).Microcystin production during algal bloom occurrence in Laguna de Bay, the Philippines.Fisheries Science,69,110-116.
4. Chiu, Yi-Ting,Hung-Kai, Yen,Tsair-Fuh, Lin(2016).An Alternative Method to Quantify 2-MIB Producing Cyanobacteria in Drinking Water Reservoirs: Method Development and Field Applications.Environmental Research,151,618-627.
5. Cuvin-Aralar, M. L.,Santiago, A. E.,Gonzal, A. C.,Santiago, C. B.,Romana-Eguia, M. R.,Baldia, S. F.,Palisoc, F., Jr.(2001).Incidence and causes of mass fish kill in a shallow tropical eutrophic lake (Laguna de Bay, Philippines).9th International Conference on the Conservation and Management of Lakes. Conference proceedings
6. Cuvin-Aralar, Maria Lourdes,Fastner, Jutta,Focken, Ulfert,Becker, Klaus,Aralar, Emiliano V.(2002).Microcystins in Natural Blooms and Laboratory Cultured Microcystis Aeruginosa from Laguna de Bay.Philippines,182,179-182.
7. DAO=DENR Administrative Order(2016).DAO. 2016. “DENR Administrative Order (2016). Water Quality Guidelines and General Effluent Standards of 2016. Department of Environment and Natural. Republic of the Philippines.”.
8. Dequito, AQD.,Trailfalgar, RFM,Saclauso, CA,Briones, ZFA,Yap, EES(2015).Biodegration of 2-methylisoborneol (2-MIB) and geosmin by bacteria isolated from aquaculture pond water and sediment.ELBA Bioflux,7(2),87-93.
9. Ding, Z,Xia, W,Zheng, H,Chen, X,Yin, L(2014).Analysis of Five Earthy-Musty Odorants in Environmental Water by HS-SPME/GC-MS.International Journal of Analytical Chemistry
10. Endo, I,Johnson, B,Skeehan, E.(2017).,Institute for Global Environmental Strategies.
11. Fan, JJ,Ho, L,Hobson, P,Brookes, J(2013).Evalauting the Effectiveness of Copper Sulphate, Chlorine, Potassium Permanganate, Hydrogen Peroxide and Ozone on Cyanobacteria Cell Integrity.Water Research,47,5153-5164.
12. Graham,Jennifer L,Loftin, Keith A,Ziegler, Andrew C,Meyer, Michael T(2008).,未出版
13. Herrera, EC,Nadaoka, K,Blanco, AC,Hernandez, EC(2014).Hydrodynamic Investigation of Shallow Tropical Lake Environment (Laguna Lake, Philippines) and Associated Implications for Eutrophic Vulnerability.ASEAN Eng. J.,1(48)
14. Jaraula, Caroline Marie B,Siringan, Fernando P.,Klingel, Ralf,Sato, Hiroshi,Yokoyama, Yusuke(2014).Records and Causes of Holocene Salinity Shifts in Laguna de Bay, Philippines.Quaternary International,349,207-220.
15. Jia, J,Shi, W,Chen, Q,Lauridsen, TL(2017).Spatial and Temporal Variations Reveal the Response of Zooplankton to Cyanobacteria.Harmful Algae,64,63-73.
16. Li, Q,Shang, L,Gao, T,Zhang, L,Ou, T,Huang, G,Chen, C,Li, C(2014).Use of Principal Component Scores in Multiple Linear Regression Models for Simulation of Chlorophyll-a and Phytoplankton Abundance at a Karst Deep Reservoir, Southwest of China.Acta Ecologica Sinica,34,72-78.
17. Lin, TF,Chang, DW,Lien, SK,Tseng, YS,Chiu, YT,Wang, YS(2009).Effect of Chlorination on the Cell Integrity of Two Noxious Cyanobacteria and Their Releases of Odorants.Journal of Water Supply Research and Technology,58,539-551.
18. Lin, Tsair-fuh,Liu, Chia-ling,Yang, Feng-chen,Hung, Hsu-wen(2003).Effect of Residual Chlorine on the Analysis of Geosmin , 2-MIB and MTBE in Drinking Water Using the SPME Technique.Water Res.,37(1),21-26.
19. LLDA(2013).LLDA. 2013. “Laguna de Bay: 2013 Ecosystem Health Report Card: Restore the Ecological Balance of the Lake. A Joint Implementation of Laguna Lake Development Authority (LLDA) and the Partnerships in Environmental Manage.” Manila, Philippines..
20. LLDA(2016).LLDA. 2016. “2016-2026 Laguna de Bay Basin Master Plan: 2016 and Beyond Towards Climate-Resiliency and Sustainable Development.”.
21. LLDA. 2012. “2009 to 2012 Annual Water Quality Report on the Laguna de Bay and Its Tributary Rivers.”
22. Markou, Giorgos,Georgakakis, Dimitris(2011).Cultivation of Filamentous Cyanobacteria (Blue-Green Algae) in Agro-Industrial Wastes and Wastewaters : A Review.Applied Energy,88(10),3389-3401.
23. Pahila, JG,Yap, EE S.,Traifalgar, RFM,Saclauso, CA(2014).Establishment of sensory threshold levels of geosmin and 2-methylisoborneol for Filipinos.Aquaculture, Aquarium, Conservation & Legislation International Journal of the Bioflux Society,7(2),94-102.
24. Ramanan, Rishiram,Kim, Byung-hyuk,Cho, Dae-hyun,Oh, Hee-mock,Kim, Hee-sik(2016).Algae – Bacteria Interactions : Evolution, Ecology and Emerging Applications.Biotechnology Advances,34(1),14-29.
25. Rosales, CAM,Rollon, AP(2011).Factors Affecting the Occurrence and Growth of Toxic Cyanobacteria Microcystis Aeruginosa in Laguna De Bay, Philippines.Journal of Environmental Science and Technology,5,1109-1116.
26. Ruan, E. D.,Aalhus, J. L.,Summerfelt, S. T.,Davidson, J.,Swift, B.,Juárez, M.(2013).Determination of off-flavor compounds, 2-methylisoborneol and geosmin, in salmon fillets using stir bar sorptive extraction-thermal desorption coupled with gas chromatography-mass spectrometry.Journal of Chromatography A,1321,133-136.
27. Santos-Borja, Adelina(2017).Laguna de Bay, Philippines: A Laboratory for Multiple-Use Resource Management.International Workshop on Sustainable Soil and Groundwater Protection and Remediation,Tainan, Taiwan:
28. Senoro, Delia B.,Maravillas, Sharon L.,Ghafari, Nima,Rivera, Clarissa C.,Quiambao, Erwin C.,Lorenzo, Maria Carmina M.(2016).Modeling of the Residue Transport of Lambda Cyhalothrin, Cypermethrin, Malathion and Endosulfan in Three Different Environmental Compartments in the Philippines.Sustainable Environment Research Elsevier,1-9.
29. Su, M.,Yu, J.,Zhang, J.,Chen, H.,An, W.,Vogt, R. D.,Yang, M.(2015).MIB-producing cyanobacteria (Planktothrix sp.) in a drinking water reservoir: Distribution and odor producing potential.Water Research,68,444-453.
30. Su, Ming,Yu, Jianwei,Pan, Shenling,An, Wei,Yang, Min(2014).Spatial and Temporal Variations of Two Cyanobacteria in the Mesotrophic Miyun Reservoir, China.Journal of Environmental Sciences,26,289-298.
31. Suurnäkki, Suvi,Gomez-Saez, Gonzalo V.,Rantala-Ylinen, Anne,Jokela, Jouni,Fewer, David P.,Sivonen, Kaarina(2015).Identification of Geosmin and 2-Methylisoborneol in Cyanobacteria and Molecular Detection Methods for the Producers of These Compounds.Water Research,68(9),56-66.
32. Wang, Z,Song, G,Shao, J,Tan, W,Li, Y(2016).Establishment and Field Applications of Real-Time PCR Methods for the Quantification of Potential MIB-Producing Cyanobacteria in Aquatic Systems.Journal of Applied Phycology,28,325-333.
33. WAVES(2016).WAVES. 2016. “WAVES Executive Summary Philippines May 2016: Laguna de Bay Ecosystem Accounts.".
34. Wynne, Timothy T,Stumpf, Richard P(2015).Spatial and Temporal Patterns in the Seasonal Distribution of Toxic Cyanobacteria in Western Lake Erie from 2002-2014.Toxins,7,1649-1663.
35. Yajima, H,Choi, J(2013).Changes in Phytoplankton Biomass due to Diversion of an Inflow into the Urayama Reservoir.Ecological Engineering,58,180-191.
36. Zamyadi, A,Ho, L,Newcombe, G,Daly, RI,Burch, M,Baker, P,Prevost, M(2010).Release and Oxidation of Cell-Bound Saxitoxins during Chlorination of Anabaena Circinalis Cells.Environmental Science and Technology,44,9055-9061.
37. Zhang, J.,Li, L.,Qiu, L.,Wang, X.,Meng, X.,You, Y.,Yu, J.,Ma, W.(2017).Effects of temperature and DIC on the odor production of 2-methylisoborneol-producing Anabaena spiroides & Planktothrix spp.Proceedings of the 2017 IWA Symposium of Lake and Reservoir Management,Shanghai, China: