微囊藻毒素检测方法的研究进展

随着水体富营养化日益严重,有毒蓝藻水华引起的污染事件在世界范围内频繁发生,其产生的毒素对生物有潜在的危害。所以迫切需要建立一种工作原理简单易行、分析速度快、灵敏度较高的统一的检测方法,对水体中特别是饮用水源中的微囊藻毒素进行检测。文章简要介绍了近年来发展起来的几类微囊藻毒素检测技术,以及微囊藻毒素检测技术尚要解决的问题和未来的发展方向。     

韶关市3座供水水库蓝藻种群及微囊藻毒素的季节变化

为了解供水水库的蓝藻种群和微囊藻毒素的季节变化,于2012年1—12月对韶关的苍村、瀑布和花山3座供水水库进行了采样分析。结果表明,3座水库为中营养型水库,监测蓝藻共9属(种),优势种为鱼腥藻和微囊藻,蓝藻最高丰度为5.67×107L-1;降水导致营养盐物质带入水库和水体不稳定性是蓝藻种群和优势种在夏秋两季季节变化和占优势的主要影响因子。3座水库ρ(微囊藻毒素)为0.1~0.9μg/L,最高值接近世界卫生组织对饮用水中MC-LR的指导性限制值(1μg/L)标准。微囊藻与微囊藻毒素呈显著正相关性(R=0.871,P0.01),表明产微囊藻毒素的蓝藻主要为微囊藻,当水库发生微囊藻水华时有发生微囊藻毒素的风险。     

海河流域东北部地表水水源地藻类及代谢物研究

同时对海河流域东北部6个水库型地表水水源地开展了藻类及微囊藻毒素(MC-LR和MC-RR)相关研究工作。研究结果表明:海河流域东北部6个水库型地表水水源地中水体富营养化程度有明显改善,但于桥水库、洋河水库水体仍呈富营养化状态并检出MC-LR和MC-RR。另外,富营养化程度高的水体中蓝藻占明显优势,并且微囊藻毒素浓度与其成正相关关系。虽然检出的MC-RR和MC-LR浓度均低于《生活饮用水卫生规范》标准限制,但是在蓝藻暴发期间(特别是暴发后期)应加密监测,确保微囊藻毒素浓度不会对供水人群健康造成危害。     

应用荧光定量PCR技术监测玄武湖蓝藻水华

应用荧光定量PCR技术和显微计数法对玄武湖蓝藻水华进行了长期监测,结果表明,荧光定量PCR法可同步监测蓝藻、微囊藻和有毒微囊藻的数量,及时准确反映玄武湖蓝藻水华优势种群微囊藻和有毒微囊藻的动态变化。与显微计数法相比,具有需要的样品量少、时效性强、检出下限较低、自动化程度高等优势,可有效地应用于蓝藻水华的监测。     

杭州贴沙河微囊藻毒素污染特征及健康风险评价

在2014年9月杭州贴沙河出现蓝藻异常增殖期间,利用固相萃取-液质联用法对水体中胞外微囊藻毒素(EMC)和总微囊藻毒素(TMC)进行监测。共检出以MC-LR为主的8种微囊藻毒素(MC)单体,TMC总的质量浓度为63.9 ng/L~1 090 ng/L,其中MC-LR质量浓度为31.6 ng/L~472 ng/L,毒性等效MC-LR浓度为51.8 ng/L~862 ng/L,检出的MC-LR浓度值均低于限值标准。采用USEPA推荐模型对水体中MC污染的健康风险进行评价。各批次水样中MC-LR的非致癌健康风险指数(HI)为0.03~0.39,毒性等效MC-LR的HI介于0.04和0.72之间,均低于基准值,说明贴沙河作为饮用水水源尚无明显的健康风险。     

2017年夏季巢湖水华期间浮游植物与蓝藻毒素的时空变化特征

蓝藻水华暴发过程中水华物种的演替及其与蓝藻毒素的关系，对于湖泊的风险评估具有重大意义。于2017年水华暴发较为严重的夏季（6-8月）对巢湖水体的理化参数、浮游植物和蓝藻毒素进行了调查和分析。结果表明：夏季巢湖基本上处于富营养-超富营养的状态。24个样本共鉴定出浮游植物7门72属117种，以绿藻门、蓝藻门和硅藻门为主。浮游植物的群落组成和细胞密度存在明显的时空差异性，其中6月浮游植物平均细胞密度为1.35×10⁸ cells/L，优势种属主要为微囊藻（Microcystis spp.，优势度为0.397）、水华长孢藻（Dolichospermum flos-aquae，优势度为0.195）和水华束丝藻（Aphanizomenon flos-aquae，优势度为0.181）；7月浮游植物平均细胞密度为1.31×10⁸ cells/L，优势种属主要为微囊藻（优势度为0.741）和黏伪鱼腥藻（Pseudanabaena mucicola，优势度为0.072）；8月浮游植物平均细胞密度为1.01×10⁸ cells/L，优势种属主要为微囊藻（优势度为0.646）。11种蓝藻毒素在夏季巢湖水体中均有不同程度的检出，其中以微囊藻毒素MC-LR、MC-RR和MC-YR为主，最高检出浓度分别为0.115、0.107、0.018 μg/L。此外，分析了拟柱孢藻毒素浓度与水华束丝藻细胞密度的关系，显示两者之间存在非常显著的正相关性（P<0.01），表明水华束丝藻可能是其最主要的产毒蓝藻。11种蓝藻毒素的浓度均未超过饮用水安全标准的规定，但其潜在的安全风险依然需要密切注意。     

城市河道污染水体生物修复技术研究进展

概述了蓝藻毒素的结构、特性以及蓝藻毒素的危害，综述了近年来国内城市河道污染水体生物修复技术的研究和应用现状，并评述了水体生物修复技术存在的问题以及污染水体生物修复的发展方向。     

宁波市饮用水源地爆发蓝藻水华时微囊藻毒素的污染分析

通过固相萃取净化,建立了超高效液相色谱-串联四极杆质谱法测定水中微囊藻毒素MC-LR和MC-RR的方法,并用该方法监测了宁波市两个饮用水源地爆发蓝藻水华时水体中的微囊藻毒素.监测结果表明,姚江水体中的微囊藻毒素污染水平低,MC-LR和MC-RR没有被检出;梅湖水库水体中MC-RR也未被检出,采取措施后,虽MC-LR有时被检出,但最高时也低于国家标准值(1.0μg/L).     

太湖梅梁湾水源水中微囊藻毒素浓度的变化

对太湖梅梁湾水源水中的总藻毒素TMC[(TMC-LR) (TMC-RR)]和胞外藻毒素EMC[(EMC-LR) (EMC-RR)]进行了跟踪检测.结果表明,水体中TMC-RR、TMC-LR、EMC-RR、EMC-LR质量浓度平均分别为1.819 μg/L、1.090 μg/L、0.491 μg/L和0.077 μg/L,无锡市的主要水源地水质已受到微囊藻毒素的污染.提出,应加强水源地水体中微囊藻毒素浓度的监测,确保饮用水的安全.     

微囊藻毒素-LR污染饮水的健康风险及其机制

水体富营养化导致藻类繁殖和水华暴发从而使得藻类释放大量藻毒素，已经演变成为严重的公共卫生问题之一。微囊藻毒素通过饮水严重危害动物和人类的健康安全，最主要的靶器官是肝脏，从而造成肝细胞功能缺陷，比如黄疸和磷酸化活性改变。这些毒性效应主要表现为对磷酸酶的抑制和蛋白磷酸酶活性的调节。综上，微囊藻毒素会造成组织结构损伤、凋亡，即使在低浓度下，也会影响细胞周期，导致肿瘤发生。     

Quantitative assessment of aerosolized cyanobacterial toxins at two New Zealand lakes

The cyanobacterial toxins, nodularin and microcystin, are highly efficient inhibitors of cellular protein phosphatases. Toxicity primarily evolves following ingestion of cyanobacterial material or toxins and results in liver and renal pathology. Ingestion is the main route of exposure in the World Health Organizations current risk assessment of nodularin and microcystins. Nasally applied microcystin appears to have a 10-fold higher availability and toxicity than orally ingested toxins, suggesting that aerosolized toxins could represent a major risk for human populations close to lakes with cyanobacterial blooms. In this study, nodularin and microcystin levels in aerosols were assessed using high and low volume air samplers for 4, 12 and 24 h periods at lakes Forsyth and Rotorua (South Island, New Zealand). These lakes were experiencing blooms of Nodularia spumigena and Microcystis sp., respectively. Using the high volume samplers up to 16.2 pg m(-3) of nodularin and 1.8 pg m(-3) of microcystins were detected in the air. Aerosolized nodularin and microcystins do not appear to represent an acute or chronic hazard to humans. The latter was concluded based on calculations using average human air intakes, the highest nodularin or microcystin concentrations measured in the air in this study, and assuming inhalatory toxicities comparable to toxicological data obtained following intraperitoneal applications in mice. However, as the toxin concentrations in the air were calculated over extended sampling periods, peak values may be underestimated. Aerosolized toxins should be considered when developing risk assessments particularly for lakeside populations and recreational users where inhalation of cyanotoxins may be a secondary exposure source to a primary oral exposure.     

Spatial and temporal variability in the relationship between cyanobacterial biomass and microcystins

The increasing incidence of toxic cyanobacterial blooms, together with the difficulties to reliably predict cyanobacterial toxin (e.g. microcystins) concentration, has created the need to assess the predictive ability and variability of the cyanobacterial biomass–microcystin relationship, which is currently used to assess the risk to human and ecosystems health. To achieve this aim, we assessed the relationship between cyanobacterial biomass and microcystin concentration on a spatiotemporal scale by quantifying the concentration of cyanobacterial biomass and microcystin in eight lakes over 9 months. On both a temporal and spatial scale, the variability of microcystin concentration exceeded that of cyanobacterial biomass by up to four times. The relationship between cyanobacterial biomass and microcystin was weak and site specific. The variability of cyanobacterial biomass only explained 25 % of the variability in total microcystin concentration and 7 % of the variability of cellular microcystin concentration. Although a significant correlation does not always imply real cause, the results of multiple linear regression analysis suggest that the variability of cyanobacterial biomass and cellular microcystin concentration is influenced by salinity and total phosphorus, respectively. The weak cyanobacterial biomass–microcystin relationship, coupled with the fact that microcystin was present in concentrations exceeding the WHO drinking water guidelines (1 μg?L^?1) in most of the collected samples, emphasizes the high risk of error connected to the traditional indirect microcystin risk assessment method.     

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.     

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.     

Detection of microcystins in Pamvotis lake water and assessment of cyanobacterial bloom toxicity

Lake Pamvotis is a shallow, eutrophic Mediterranean lake with ecological significance. This paper deals with the evaluation of cyanobacterial toxicity in Lake Pamvotis. ELISA and HPLC revealed the presence of significant amounts of MCYST-LR. Danio rerio bioassay confirmed the toxic nature of the bloom. Cyanobacterial extracts had adverse toxic effects on development of D. rerio. Also, it was shown that cyanobacterial extracts containing environmentally detected concentrations of MCYST can cause reduced survival rate of fish species. The results clearly indicate that cyanobacterial blooms in Lake Pamvotis may be regarded as human and fish health hazard. Continuous monitoring of the lake is suggested, in order to prevent future possible intoxications.     

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.     

Development of a new risk-based framework to guide investment in water quality monitoring

An innovative framework for optimising investments in water quality monitoring has been developed for use by water and environmental agencies. By utilising historical data, investigating the accuracy of monitoring methods and considering the risk tolerance of the management agency, this new methodology calculates optimum water quality monitoring frequencies for individual water bodies. Such information can be applied to water quality constituents of concern in both engineered and natural water bodies and will guide the investment of monitoring resources. Here we present both the development of the framework itself and a proof of concept by applying it to the occurrence of hazardous cyanobacterial blooms in freshwater lakes. This application to existing data demonstrates the robustness of the approach and the capacity of the framework to optimise the allocation of both monitoring and mitigation resources. When applied to cyanobacterial blooms in the Swan Coastal Plain of Western Australia, we determined that optimising the monitoring regime at individual lakes could greatly alter the overall monitoring schedule for the region, rendering it more risk averse without increasing the amount of monitoring resources required. For water resources with high-density temporal data related to constituents of concern, a similar reduction in risk may be observed by applying the framework.     

Identification of microcystins contamination in surface water samples from the Three Gorges Reservoir, China

Physicochemical and biological parameters related to water quality and microcystins (MCs) contamination in aquatic environment of the Three Gorges Reservoir were investigated in August 2004 and January 2005. A solid-phase extraction method and an HPLC equipped with photodiode array were used for MC-LR detection. A quantitative analysis showed the total MC-LR concentrations of water samples ranged from non-detectable to 0.57 μg L?1 among the seven sampling sites. The highest MC-LR concentration was found at sampling site G (Wushan), which was followed by F (Kaixian), E (Wanzhou), D (Fuling), C (Cuntan), and A (Daxigou). The correlation analysis showed the MC-LR concentration was positively correlated with chlorophyll-a concentration. This result suggests that MC concentration in water can be indirectly estimated by analyzing the chlorophyll-a concentration. Overall, the results of this study suggest that more importance should be placed on monitoring of MC contamination and water quality in the Three Gorges Reservoir to ensure drinking water safety and reduce the potential exposure of people to these health hazards.     

水中微囊藻毒素的超高效液相色谱-串联质谱法研究

建立测定水体中微囊藻毒素(MCYST)的超高效液相色谱-串联质谱分析方法.水样通过固相萃取富集净化,采用超高效液相色谱-串联质谱测定,6min内完成3种MCYST的分离及检测.MCYST-LR、MCYST-RR、MCYST-YR检出限分别为3.5、2.5、5.0ng/L,回收率为85.6％～107.9％,为水质微囊藻毒素监测提供了一种快速、准确、灵敏的分析方法.