Monitoring of potentially toxic cyanobacteria using an online multi-probe in drinking water sources

Toxic cyanobacteria threaten the water quality of drinking water sources across the globe. Two such water bodies in Canada (a reservoir on the Yamaska River and a bay of Lake Champlain in Québec) were monitored using a YSI 6600 V2-4 (YSI, Yellow Springs, Ohio, USA) submersible multi-probe measuring in vivo phycocyanin (PC) and chlorophyll-a (Chl-a) fluorescence, pH, dissolved oxygen, conductivity, temperature, and turbidity in parallel. The linearity of the in vivo fluorescence PC and Chl-a probe measurements were validated in the laboratory with Microcystis aeruginosa (r(2) = 0.96 and r(2) = 0.82 respectively). Under environmental conditions, in vivo PC fluorescence was strongly correlated with extracted PC (r = 0.79) while in vivo Chl-a fluorescence had a weaker relationship with extracted Chl-a (r = 0.23). Multiple regression analysis revealed significant correlations between extracted Chl-a, extracted PC and cyanobacterial biovolume and in vivo fluorescence parameters measured by the sensors (i.e. turbidity and pH). This information will help water authorities select the in vivo parameters that are the most useful indicators for monitoring cyanobacteria. Despite highly toxic cyanobacterial bloom development 10 m from the drinking water treatment plant's (DWTP) intake on several sampling dates, low in vivo PC fluorescence, cyanobacterial biovolume, and microcystin concentrations were detected in the plant's untreated water. The reservoir's hydrodynamics appear to have prevented the transport of toxins and cells into the DWTP which would have deteriorated the water quality. The multi-probe readings and toxin analyses provided critical evidence that the DWTP's untreated water was unaffected by the toxic cyanobacterial blooms present in its source water.     

Use of in vivo phycocyanin fluorescence to monitor potential microcystin-producing cyanobacterial biovolume in a drinking water source

The source water of a drinking water treatment plant prone to blooms, dominated by potential microcystin-producing cyanobacteria, was monitored for two seasons in 2007-2008. In the 2008 season, the median value for potential microcystin-producing cyanobacterial biovolume was 87% of the total phytoplankton biovolume in the untreated water of the plant. Depth profiles taken above the plant's intake identified three sampling days at high risk for the contamination of the plant's raw water with potentially toxic cyanobacteria. Chlorophyceae and Bacillariophyceae caused false positive values to be generated by the phycocyanin probe when cyanobacteria represented a small fraction of the total phytoplanktonic biovolume present. However, there was little interference with the phycocyanin probe readings by other algal species when potential microcystin-producing cyanobacteria dominated the phytoplankton of the plant's untreated water. A two-tiered method for source water monitoring, using in vivo phycocyanin fluorescence, is proposed based on (1) a significant relationship between in vivo phycocyanin fluorescence and cyanobacterial biovolume and (2) the calculated maximum potential microcystin concentration produced by dominant Microcystis sp. biovolume. This method monitors locally-generated threshold values for cyanobacterial biovolume and microcystin concentrations using in vivo phycocyanin fluorescence.     

A phycocyanin probe as a tool for monitoring cyanobacteria in freshwater bodies

In many countries, the presence of cyanobacteria in freshwater bodies used for both drinking water and recreational purposes is under increasing public health attention. Water managers are considering how to implement monitoring that leads to a minimization of the risks incurred by the users of potentially contaminated sites. To address this question, this study involved assessing the performance of a submersible probe for measuring phycocyanin-specific fluorescence as a function of cyanobacterial biomass, with the aim of applying it as a tool for surveillance management. Its advantages and limits compared to more traditional analyses are discussed. The monitoring of cyanobacteria in the water bodies of western France was carried out using a minifluorimeter specific to the fluorescence of phycocyanin, a pigment specific to cyanobacteria. The results are compared with the analyses recommended by the World Health Organisation (chlorophyll a and cell counting). This study based on nearly 800 samples shows a significant correlation between the phycocyanin content and the cyanobacterial biomass, expressed as the number of cells per mL (R2 = 0.73). This submersible probe is simple and rapid to use, making it possible to take into account horizontal and vertical heterogeneities in the proliferation growth. In this way, we are able to detect at an early stage the conditions that could potentially lead to a risk, in order to start sampling. Due to its sensitivity, this tool proves suitable for monitoring aimed at reducing the risks incurred by the users of contaminated sites and launching preventative actions. The use of the phycocyanin probe provides an effective tool to complement traditional analyses of cyanobacterial presence. It is suggested that a surveillance protocol based on phycocyanin concentration can significantly improved the accuracy of the extent of cyanobacterial bloom development in the light of spatial and temporal variabilities associated with these occurrences.     

Phytoplankton and water quality in a Mediterranean drinking-water reservoir (Marathonas Reservoir, Greece)

Phytoplankton and water quality of Marathonas drinking-water Reservoir were examined for the first time. During the study period (July-September 2007), phytoplankton composition was indicative of eutrophic conditions although phytoplankton biovolume was low (max. 2.7 mm3 l?1). Phytoplankton was dominated by cyanobacteria and diatoms, whereas desmids and dinoflagellates contributed with lower biovolume values. Changing flushing rate in the reservoir (up to 0.7% of reservoir's water volume per day) driven by water withdrawal and occurring in pulses for a period of 15-25 days was associated with phytoplankton dynamics. Under flushing pulses: (1) biovolume was low and (2) both 'good' quality species and the tolerant to flushing 'nuisance' cyanobacterium Microcystis aeruginosa dominated. According to the Water Framework Directive, the metrics of phytoplankton biovolume and cyanobacterial percentage (%) contribution indicated a moderate ecological water quality. In addition, the total biovolume of cyanobacteria as well as the dominance of the known toxin-producing M. aeruginosa in the reservoir's phytoplankton indicated a potential hazard for human health according to the World Health Organization.     

YSI6600V2水质多参数仪现场快速法与实验室分光光度法测定湖泊藻类叶绿素的比较

通过将YSI6600V2水质多参数仪测得的叶绿素a值与实验室分光光度法叶绿素a测定值进行比较,分析了YSI6600V2多参数水质监测仪测定叶绿素a的准确性与局限性。结果表明,YSI6600V2测定值多数偏低,且相关性较差。将YSI6600V2测得的蓝藻密度样本分为2类后,相关性得到显著提升。提出了利用蓝藻密度对样本进行分类后,分别进行叶绿素a校正的方法,并说明了此种校正方法的局限性。     

Spatiotemporal distribution pattern of cyanobacteria community and its relationship with the environmental factors in Hongze Lake,China

Hongze Lake, located in the east route of the South-to-North Water Diversion Project (SNWDP), is a potential drinking water source for the residents along this water diversion project. Based on a monthly sampling at 11 stations in three regions of Hongze Lake, the spatiotemporal distribution pattern of cyanobacteria community was comprehensively investigated from March 2011 to February 2013. A total of 23 cyanobacterial species which belong to 16 genera were identified, and Microcystis was the most predominant cyanobacterial genus mainly composed of Microcystis wesenbergii in Hongze Lake. The cyanobacterial abundance ranged from 0 to 2.6?×?10⁷ cells/L, and the average cyanobacteria abundance of Northern region was significantly higher than those of Western region and Eastern region in the 2-year study. The total cyanobacteria abundance and the Microcystis abundance both took on a similar seasonal regularity in the three regions. The results of correlation analysis indicated that Microcystis abundance was correlated with water temperature, chemical oxygen demand (COD)_Mn, nitrate (NO₃-N), and total nitrogen (TN)/total phosphorus (TP) mass ratio, among which water temperature had the highest correlation coefficient. In summer, cyanobacteria blooms may take place under suitable environmental conditions at some special areas in Hongze Lake, especially where the concurrence of slow water exchange and steady wind direction exists.     

Management of toxic cyanobacteria for drinking water production of Ain Zada Dam

Blooms of toxic cyanobacteria in Algerian reservoirs represent a potential health problem, mainly from drinking water that supplies the local population of Ain Zada (Bordj Bou Arreridj). The objective of this study is to monitor, detect, and identify the existence of cyanobacteria and microcystins during blooming times. Samples were taken in 2013 from eight stations. The results show that three potentially toxic cyanobacterial genera with the species Planktothrix agardhii were dominant. Cyanobacterial biomass, phycocyanin (PC) concentrations, and microcystin (MC) concentrations were high in the surface layer and at 14 m depth; these values were also high in the treated water. On 11 May 2013, MC concentrations were 6.3 μg/L in MC-LR equivalent in the drinking water. This study shows for the first time the presence of cyanotoxins in raw and treated waters, highlighting that regular monitoring of cyanobacteria and cyanotoxins must be undertaken to avoid potential health problems.     

YSI 6600传感器在太湖蓝藻预警工作中的应用

通过对太湖饮用水源地2个取水口水体中的蓝藻密度(CBD)和叶绿素(Chl)分别进行 YSI 6600-v2水质多参数仪和实验室显微镜检分析,探讨了CBD和Chl在预警工作中的应用.结果表明,在CBD与Chl相关性显著时期内,藻类群落结构相对稳定;CBD和Chl相关性不显著时期内,藻类群落结构变化较大.ρ(CBD)/ρ(...     

A comparative assessment of molecular biological and direct microscopic techniques for assessing aquatic systems

Molecular techniques have clear value for community characterization; however, almost all previous datasets are based upon non-molecular measurements and it is hard to compare "old" data with "new" data because few correlations have been made. Therefore, the purpose of this evaluation was to simultaneously use molecular and non-molecular methods within the same sampling program to determine how data compare. Three methods were used for characterizing microbial populations in Lake 260 (L260) at Experimental Lakes Area (Ontario, Canada) during a whole-lake exposure study. Methods included whole-cell microscopic counts (for bacteria, cyanobacteria, algae and zooplankton), chlorophyll a, and small sub-unit (ssu)-rRNA hybridization using EUB-338, EUB-785, CYAN-785-a/b, EUCA-1379 and UNIV-1390 gene probes. Strong correlations were found between the EUB-785 probe signal and "bacteria minus cyanobacteria" direct counts, and the EUB-338 probe signal and "bacteria plus cyanobacteria" counts. Furthermore, the difference in probe signal between EUB-338 and EUB-785 (a presumptive signature for cyanobacteria and plastids) correlated with cyanobacterial direct counts and also with chlorophyll a. However, EUCA-1379 probe signal did not correlate with algae counts, and UNIV-1390 probe signal only correlated with total bacteria counts. Results suggest that, although ssu-rRNA methods are fast, reproducible, and specifically detect "viable" organisms, their use may be limited to non-eukaryotic populations unless new probes are developed that are more specific.     

Temporal distribution of cyanobacteria in the coast of a shallow temperate estuary (Río de la Plata): some implications for its monitoring

The aim of this study was to analyze the temporal distribution of phytoplanktonic cyanobacteria in a site located in the freshwater tidal zone near the extraction point for the drinking water supply. Samples were taken considering three timescales as follows: hours, days, and weeks, during the period of highest development of cyanobacteria. The phytoplankton density, microcystin concentration (LR, RR, YR), and chlorophyll-a were related to meteorological variables (wind and temperature), tidal high, and physical-chemical variables (nutrients, pH, conductivity, light penetration). The results obtained in this study showed that the variables that primarily modulate the temporal distribution of cyanobacteria were temperature, pH, light penetration, conductivity, and nutrients (particularly NO₃ ^? and NH₄ ⁺), while the winds and tide had a secondary effect, only evidenced at an hourly scale. Therefore, this timescale would be the most suitable for monitoring cyanobacterial populations, when the amount of cyanobacterial cells exceeds the alert I level proposed by the World Health Organization. This recommendation is particularly important for the water intake zones in Río de la Plata, which are vulnerable to the damage generated by cyanobacteria on the water quality.     

Use of three monitoring approaches to manage a major <Emphasis Type="Italic">Chrysosporum ovalisporum</Emphasis> bloom in the Murray River,Australia, 2016

An unusual bloom of Chrysosporum ovalisporum (basionym Aphanizomenon ovalisporum) occurred for the first time in the Murray River and distributary rivers in New South Wales, Australia, from mid-February to early June 2016. At its greatest extent, it contaminated a combined river length of ca. 2360 km. Chrysosporum ovalisporum usually comprised >99% of the total bloom biovolume at most locations sampled, which at times exceeded 40 mm³ l^?1. The origins of the bloom were most likely reservoirs on the upper Murray River, with cyanobacterial-infested water released from them contaminating the river systems downstream. An integrated approach using three analytical methods: (1) identification and enumeration by microscopy, (2) multiplex quantitative polymerase chain reaction (qPCR), and (3) toxin analysis, was used to obtain data for the assessment of risk to water users and management of the bloom. qPCR indicated some cyrA and stxA genes responsible for cylindrospermopsin and saxitoxin biosynthesis respectively were present, but mostly below the level of quantification. No mcyE genes for microcystin biosynthesis were detected. Toxin analysis also revealed that cylindrospermopsin, saxitoxin and microcystin were all below detection. Lack of measurable toxicity in a species usually considered a cylindrospermopsin producer elsewhere meant the possibility of relaxing management guidelines; however, high (Red) alerts needed to be maintained due to risk to water users from other biohazards potentially produced by the cyanobacteria such as contact irritants. A three-tiered monitoring strategy is suggested for monitoring cyanobacterial blooms to provide enhanced data for bloom management.     

淡水水体中微囊藻毒素的监测技术

随着水体富营养化的日益加剧 ,有毒蓝藻水华在世界范围内频繁发生。有毒蓝藻产生的毒素对人类和动物有潜在的危害 ,迫切要求对水体中特别是饮用水源中的蓝藻毒素进行监测。微囊藻毒素是蓝藻产生的一类环状七肽 ,可以抑制蛋白磷酸酶 1和 2 A。微量微囊藻毒素还促进肿瘤的发生。简要介绍了近年来发展起来的几类微囊藻毒素监测技术 ,以期引起有关部门的重视     

Detection and quantification of major toxigenic Microcystis genotypes in Moo-Tan reservoir and associated water treatment plant

Two molecular methods, denaturing gradient gel electrophoresis (DGGE) and quantitative real-time polymerase chain reaction (qPCR) with the Universal ProbeLibrary (UPL) probe, were developed and used for the characterization and quantification of several microcystin producers in Moo-Tan Reservoir (MTR), Taiwan and its associated water treatment plant (Shih-Men Water Treatment Plant, SMWTP). Internal transcribed spacer (ITS) sequence, a highly diversified region between the 16S rRNA and 23S rRNA genes, was used to further identify the isolated strains from MTR and also used in DGGE for the detection of the specific DNA fragments and biomarkers for 11 strains observed in MTR. These ITS-DGGE biomarkers were successfully applied to monitor the community changes of potential toxigenic Microcystis sp. over a period of five years. Two highly specific primers were combined with UPL probes to measure microcystins synthesis gene (mcyB) and phycocyanin intergenic spacer region (cpcB) concentrations in water samples. The copy concentrations of UPL-mcyB and UPL-cpcB correlated well with MC-RR concentrations/water temperature and Microcystis sp. cell numbers in the water samples, respectively. For SMWTP, toxin concentrations were low, but the DGGE bands clearly demonstrated the presence of potential microcystin producers in both water treatment plants and finished water samples. It was demonstrated that toxigenic Microcystis sp. may penetrate through the treatment processes and pose a potential risk to human health in the drinking water systems.     

Association between biotic and abiotic parameters and the occurrence of cyanobacteria in a Brazilian reservoir

The water reservoir of Vargem das Flores, Brazil, has been subject to eutrophication problems. The occurrence of cyanobacterial blooms in this water body has raised concerns regarding the presence of toxic metabolites. In order to understand the factors that promote the proliferation and distribution of cyanobacteria in the reservoir, physicochemical parameters, species composition, and toxicity of the reservoir water column were analyzed at monthly intervals over a 3-year period (2004-2007) using a range of analytical techniques. The results showed that cyanobacteria were absent in the dry period, returning after the onset of stratification. A succession pattern was observed within this group. The Aphanocapsa genus was dominant in the last rainy period of the study (October 2006 to March 2007). However toxicity was only detected when the genera Microcystis and Sphaerocavum were present (October 2005). The principal components analysis showed negative correlation between cyanobacteria and orthophosphate. The change of dominant cyanobacteria along the 3 years of study and the final prevalence of nontoxic genera in the reservoir is an evidence of water quality improvement in response to the management techniques implemented in the reservoir basin.     

Seasonal dynamics of freshwater pathogens as measured by microarray at Lake Sapanca,a drinking water source in the north-eastern part of Turkey

Monitoring drinking water quality is an important public health issue. Two objectives from the 4 years, six nations, EU Project μAqua were to develop hierarchically specific probes to detect and quantify pathogens in drinking water using a PCR-free microarray platform and to design a standardised water sampling program from different sources in Europe to obtain sufficient material for downstream analysis. Our phylochip contains barcodes (probes) that specifically identify freshwater pathogens that are human health risks in a taxonomic hierarchical fashion such that if species is present, the entire taxonomic hierarchy (genus, family, order, phylum, kingdom) leading to it must also be present, which avoids false positives. Molecular tools are more rapid, accurate and reliable than traditional methods, which means faster mitigation strategies with less harm to humans and the community. We present microarray results for the presence of freshwater pathogens from a Turkish lake used drinking water and inferred cyanobacterial cell equivalents from samples concentrated from 40 into 1 L in 45 min using hollow fibre filters. In two companion studies from the same samples, cyanobacterial toxins were analysed using chemical methods and those dates with highest toxin values also had highest cell equivalents as inferred from this microarray study.     

Monitoring toxic cyanobacteria and cyanotoxins (microcystins and cylindrospermopsins) in four recreational reservoirs (Khon Kaen, Thailand)

The toxic cyanobacterial communities of four recreational reservoirs (Bueng Kaen Nakhon, Bueng Thung Sang, Bueng Nong Khot, and Bueng See Than) in Amphur Muang, Khon Kaen Province, Thailand, were investigated. Water samples were collected via monthly sampling from June to October 2011 for the study on the diversity and density of toxic cyanobacteria and toxin quantification. The main toxic cyanobacteria present in these reservoirs were Aphanocapsa sp., Cylindrospermopsis sp., Leptolyngbya sp., Limnothrix sp., Microcystis sp., Oscillatoria sp., Planktolyngbya sp., Planktotrix sp., and Pseudanabaena sp. The dominant bloom-forming genera in the water samples from Bueng Nong Khot and Bueng See Than were Microcystis sp. and Cylindrospermopsis sp., respectively. Enzyme-linked immunosorbent assays specific for cyanotoxins were performed to detect and quantify microcystins and cylindrospermopsins, with the highest average microcystins content (0.913 μgL^?1) being found in the sample collected from Bueng Nong Khot and the highest average cylindrospermopsins content (0.463 μgL^?1) being found in the sample collected from Bueng See Than. The application of 16S rRNA analyses to cyanobacterial isolates BKN2, BNK1, BNK2, and BST1 indicated that these isolates are most closely related to Limnothrix planctonica (JQ004026) (98 % similarity), Leptolyngbya sp. (FM177494) (99 % similarity), Microcystis aeruginosa (DQ887510) (99 % similarity), and Limnothrix redekei (FM177493) (99 % similarity), respectively.     

Remote sensing as a tool for monitoring water quality parameters for Mediterranean Lakes of European Union water framework directive (WFD) and as a system of surveillance of cyanobacterial harmful algae blooms (SCyanoHABs)

Remote sensing has been used from the 1980s to study inland water quality. However, it was not until the beginning of the twenty-first century that CHRIS (an experimental multi-angle sensor with good spectral and spatial resolutions) and MERIS (with good temporal and spectral resolutions) started to acquire imagery with very good resolutions, which allowed to develop a reliable imagery acquisition system so as to consider remote sensing as an inland water management tool. This paper presents the methodology developed, from the field data acquisition with which to build a freshwater spectral library and the study of different atmospheric correction systems for CHRIS mode 2 and MERIS images, to the development of algorithms to determine chlorophyll-a and phycocyanin concentrations and bloom sites. All these algorithms allow determining water eutrophic and ecological states, apart from generating surveillance maps of toxic cyanobacteria with the main objective of Assessment of the Water Quality as it was used for Monitoring Ecological Water Quality in smallest Mediterranean Reservoirs integrated in the Intercalibration Exercise of European Union Water Framework Directive (WFD). We keep on using it to monitor the Ecological Quality Ratio (EQR) in Spain inland water.     

Bioaccumulation and toxicity of arsenic in cyanobacteria cultures separated from a eutrophic reservoir

The bioaccumulation and toxicity of arsenate (arsenic (As)(V)) was studied using three cultures of cyanobacterial species—Oscillatoria tenuisa, Anabaena affinis, and Microcystis aeruginosa—that were isolated from a eutrophic reservoir. The As(V) uptake depended on the cyanobacterial species, the growth phase of the cyanobacteria, the duration of exposure, and the initial concentration of As(V). The specific growth rates of the three cultures immediately following the logarithmic phase were 0.033–0.041 L/day when the initial concentration of As(V) was 50 mg/L. These rates were 2.3–3.6 times less than those in the original culture medium without As(V). The rate of intake of As(V) in the logarithmic phase cultures greatly exceeded that in the stationary cultures. The accumulation of As(V) by the three cultures increased rapidly within 1 week from the initial value of 3.23?×?10^?2–5.40?×?10^?2 to 5.06?×?10^?1–6.73?×?10^?1 ng/cell in the logarithmic phase. The effective concentrations (EC₅₀) of As(V) for inhibiting the growth of the three cyanobacterial species growth of at 72 h followed the order Oscillatoria tenuisa (3.8 mg/L)?>?A. affinis (2.6 mg/L)?>?M. aeruginosa (1.2 mg/L). The cyanobacterial species that was most sensitive to As(V) was M. aeruginosa. Preliminary results from SEM-map studies suggest most of the As(V) in Microcystis aeruginosa accumulated in the cytoplasm (intercellular), while in O. tenuisa and A. affinis, a large proportion of As(V) bound to the cell wall (extracellular). These differences were understood with reference to the variation among the metabolic properties and morphological characteristics of the cyanobacterial species.     

Temporal variation in density and diversity of cyanobacteria and cyanotoxins in lakes at Nagpur (Maharashtra State), India

Toxic cyanobacteria (TCB) are known worldwide for the adverse impacts on humans and animals. Species composition and the seasonal variation of TCB in water bodies depend on interactions between physical and chemical factors. The present investigation delineates temporal variations in physico-chemical water quality parameters, viz. nutrients and density, diversity, and distribution of toxic cyanobacteria and cyanotoxins in Lake Ambazari (21°7′52″N, 79°2′22″E) and Lake Phutala (21°9′18″N, 79°2′37″E) at Nagpur (Maharashtra State), India. These lakes are important sources of recreational activities and fisheries. Toxic cyanobacterial diversity comprised Anabaena, Oscillatoria, Lyngbya, Phormidium, and Microcystis, a well-known toxic cyanobacterial genus, as dominant. Chlorophyll-a concentrations in the lakes ranged from 1.44 to 71.74 mg/m³. A positive correlation of Microcystis biomass existed with orthophosphate-P (p?<?0.05) and nitrate-N (p?>?0.05). Identification and quantification of microcystin variants were carried out by high performance liquid chromatography equipped with photodiode array detector. Among all the tested toxin variants, microcystin-RR (arginine–arginine) was consistently recorded and exhibited a positive correlation (p?<?0.05) with Microcystis in both the water bodies. Microcystis bloom formation was remarkable between post-monsoon and summer. Besides nutrient concentrations governing bloom formation, the allelopathic role of microcystins needs to be established.