微囊藻毒素生物学功能研究进展

我国淡水水体蓝藻水华的频繁发生已成为我国目前和今后长时期内面临的重大水环境问题.微囊藻(Microcystis)因其产生对人类健康具有危害的微囊藻毒素而尤其受到重视.研究者对构成蓝藻水华的主要种类、微囊藻毒素的毒性、毒理等方面的认识逐渐明晰,但对微囊藻毒素生物学功能的了解还相对较少.本文对微囊藻毒素的产生,尤其是近年来随着技术的发展在微囊藻毒素生物学功能方面的研究开展讨论:总结了微囊藻毒素可能作为化感物质、参与光合作用、在微囊藻的越冬或复苏中可能发挥的作用以及可能参与微囊藻群体形成及维持等4个方面的生物学功能;分析了微囊藻毒素作为信号分子的研究现状;探讨了全球气候变化及实验室模拟条件下微囊藻毒素产毒与无毒株的竞争,并对微囊藻毒素未来的研究热点及组学技术可能在其研究中的应用进行了展望.     

太湖微囊藻毒素时空分布特征及与环境因子的关系

采用高效液相色谱法(HPLC)对太湖水体中溶解态微囊藻毒素(MC-LR;MC-RR)浓度进行检测,探讨微囊藻毒素浓度的时空变化及其与氮、磷、总有机碳、蓝藻生物量以及温度等湖泊环境因子之间的关系.结果表明:微囊藻毒素表底层浓度基本一致;在一天之内的变化没有明显规律;冬季微囊藻毒素浓度较高;位于竺山湾和西部沿岸地区的两个点...     

2013年洱海水华期间微囊藻毒素和浮游植物动态变化

2013年秋季,洱海再次出现大规模水华,周边几十万居民的饮用水安全引起当地政府的极大关注.为了解水华发生期间洱海水体微囊藻毒素的含量和浮游植物的种类及丰度,分别在2013年9月和10月两次在洱海采集水样,测定其中两种主要微囊藻毒素MC-LR和MC-RR含量,探讨微囊藻毒素含量对洱海周边水源地的安全影响.同时也对水华期间的浮游植物特别是蓝藻进行研究,从而分析微囊藻毒素与浮游植物种类和丰度的关系.结果表明,洱海水体中微囊藻毒素含量偏低,暂不会对人类健康产生威胁,但在洱海的水鸟体内仍然检测到微囊藻毒素的富集,显示洱海蓝藻水华对水生生物的潜在危害不容忽视.浮游植物的研究发现,洱海水华的优势种属为微囊藻(Microcystis)和暗丝藻(Psephonema aenigmaticus).9月份暗丝藻更占优势,10月份则是微囊藻更占优势.研究结果显示处于富营养化初期的洱海水体中湖湾微囊藻毒素的含量与浮游植物中产毒蓝藻的种类和丰度密切相关.     

微囊藻毒素粗提物对水环境中温和气单胞菌活的非可培养状态的影响

细菌VBNC状态(Viable but non-culturable state),又称"活的非可培养状态",是自然水体中广泛存在的细菌的一种特殊存活状态,复杂的环境因子变化是VBNC状态转换的可能原因.本研究利用接种温和气单胞菌的无菌湖水,研究铜绿微囊藻(Microcystis aeruginosa)培养液和微囊藻毒素粗提液对水环境中温和气单胞菌VBNC状态转换的影响,结果发现,温和气单胞菌在无菌湖水中同时存在VBNC状态和可培养状态;产毒微囊藻培养液和微囊藻毒素粗提液均能促进温和气单胞菌中VBNC细菌与可培养细菌之间的状态转换,说明微囊藻毒素可能是水环境中影响细菌VBNC状态转换的重要环境因素,其进一步的分子机制研究正在进行中.     

河蟹养殖池塘微囊藻水华毒性及其光合作用活性特征

苏州市吴中区一河蟹养殖池塘在2013年7月和8月发生了严重的微囊藻水华.采用单一和双重PCR扩增微囊藻毒素合成酶基因,用以检测微囊藻水华是否产毒,结果显示为阳性.同时,采用高效液相色谱测定微囊藻水华的毒性大小.结果表明:7月和8月微囊藻水华的胞内微囊藻毒素浓度分别为1.49和0.88μg/L,胞外微囊藻毒素的浓度分别为0.75和1.09μg/L.另外,采用浮游植物荧光仪Phyto-PAM测定河蟹养殖水体形成水华的微囊藻的光合作用活性.结果显示:7月和8月水华微囊藻的最大光量子产量Fv/Fm分别为0.48和0.44,实际光量子产量ΦPSII分别为0.38和0.32,表明形成水华的微囊藻有较高的生长潜力.非光化学荧光淬灭值NPQ分别为0.28和0.36.从快速光响应曲线RLC的特征参数来看,7月水华微囊藻的光合作用活性和光能利用效率高于8月.本研究结果表明,河蟹养殖池塘水体受到微囊藻水华和微囊藻毒素的污染,进而可能对河蟹食品安全构成潜在威胁.     

有毒铜绿微囊藻对鱼和溞的毒性

用有毒铜绿微囊藻细胞及其从毒细胞中提取的毒素喂养和腹腔注射鱼类,结果表明:(1)微囊藻毒素MCYST-LR对所有实验过的鱼有毒害作用(LD₅₀为110-500μg/kg).注射毒素24h后,鱼肝电镜超微结构观察均表现出肝损伤.(2)在受控条件下,当用有毒铜绿微囊藻作为食物喂养日鲢和罗非鱼时,鱼类能在以毒藻为唯一食物的环境中生活20d以上.白鲢和罗非鱼粪便的毒性比喂食藻类的毒性分别降低约2-4.5倍,其肝的超微结构现察仍有轻度损伤.(3)在培养液中加入15μg/mLMCYST-LR和用实验室纯培养的铜绿微囊藻作为食物培养枝角类时,均可使它们在48h死亡.以无毒微囊藻培养时则生长不良亦不怀卵,表明它不是枝角类合适的食物.     

Fe(Ⅲ)改性土壤用于微囊藻细胞及微囊藻毒素的去除

蓝藻水华及其释放的蓝藻毒素给我国很多地区的饮用水安全带来较大威胁.尽管去除蓝藻和净化藻毒素的技术目前已有较多报道,但能同时对二者起作用的技术方法还较少,发展能够同时去除蓝藻和蓝藻毒素的技术显得十分必要.根据前期Fe(Ⅲ)化合物去除微囊藻毒素的工作基础,选取4种不同的土壤对其进行Fe(Ⅲ)改性,并应用于微囊藻毒素的去除.结果表明,黏土含量高低决定土壤本身对毒素的吸附能力强弱;土壤经过离子改性后,其毒素吸附能力与其阳离子交换量大小直接相关,且毒素去除能力较未改性前有显著提高,其中效率提升最高的一种土壤离子改性后其吸附能力增加了约148倍.离子改性土壤对微囊藻细胞同样具有絮凝沉降功能,沉降能力同样随土壤对Fe(Ⅲ)负载能力的提高而上升,使用剂量提高时微囊藻沉降平衡时间明显缩短.在野外蓝藻水华去除实验中,蓝藻细胞去除可以保持与室内实验相近的高去除率,但蓝藻毒素的去除效率有一定的下降,可能与天然水体中其它杂质的竞争作用有关.     

利用real-time RT-PCR研究大型蚤对铜绿微囊藻毒素合成基因转录水平影响

近年来关于浮游动物与微囊藻相互作用的研究逐渐被关注.其中有的研究认为浮游动物能够诱导产毒细胞毒素含量的变化.微囊藻毒素是由微囊藻毒素合成基因编码翻译的,目前关于浮游动物对微囊藻毒素合成基因相对表达的影响并无报道,本文首次通过实时定量逆转录PCR方法研究铜绿微囊藻PCC7806产毒相关基因mcyB和mcyD在大型蚤胁迫下...     

巢湖微囊藻毒素异构体组成的时空分布特征及影响因子

有害蓝藻释放微囊藻毒素（MCs）,严重威胁饮用水源地用水安全.为了解巢湖MCs污染状况及其异构体组成对水质的影响,于2012年夏季（8月）和秋季（11月）,2013年冬季（2月）和春季（5月）进行采样分析,研究了巢湖水体中胞内微囊藻毒素（IMCs）和胞外微囊藻毒素（EMCs）异构体的时空分布及其与环境因子的关系.结果发现,IMCs和EMCs的平均浓度变化范围分别为0.12~6.45 μg/L和0.69~1.92 μg/L.在3种常见的异构体中,MC-RR和MC-LR比例较高,MC-YR最低,MC-RR和MC-LR是巢湖水体中MCs的主要异构体类型.IMCs和EMCs的异构体浓度及其比例呈现不同的时空分布特征.微囊藻生物量、水温、总磷浓度是影响IMCs和EMCs异构体浓度及其组成变化的关键环境因子.本研究表明巢湖富营养化严重的西湖区夏季能合成更多的MC-RR异构体,而秋、冬季节偏向于释放生理毒性更强的MC-LR异构体.了解MCs异构体组成变化及其关键影响因素,有助于预测预警水体MCs污染状况和评估饮用水源地MCs风险.     

汾河(太原市景区段)微囊藻的分子多样性及产毒能力

对太原市汾河景区采集到的微囊藻进行分离纯化,得到7株微囊藻藻株,运用ITS、PC-IGS和gyrB基因序列构建系统树来研究其分子多样性.结果表明,太原市汾河景区微囊藻具有一定的分子多样性.采用mcyA基因和ELISA检测两种方法,即全细胞PCR测定微囊藻毒素合成酶基因A( mcyA),对这些藻株的微囊藻毒素进行测定,检...     

On the diet of Nile tilapia in two eutrophic tropical lakes containing toxin producing cyanobacteria

Due to the production of toxins, cyanobacteria may adversely affect economically important fish such as Nile tilapia Oreochromis niloticus in tropical lakes. We studied the diet composition and factors affecting the diet of Nile tilapia in two tropical lakes where cyanotoxins were present. Particle-bound microcystins were present in all analysed water samples, ranging in concentration from 0.00012 to 1.11 and from 0.006 to 0.254 μg L⁻¹ in Murchison Bay in Lake Victoria and Lake Mburo, respectively. Detritus and phytoplankton were the main dietary components of the Nile tilapia, with phytoplankton contributing to over 30% by volume of stomach contents. The cyanobacteria Microcystis spp., which are also the most likely source of microcystins in the lakes, accounted for more than 80% of ingested phytoplankton. Microcystis spp. were also the most abundant cyanobacteria in both lakes (>60%). We found no significant relationship between the contribution of phytoplankton in Nile tilapia diet and the concentration of microcystins in the water but we found a close association between water transparency and the contribution of insects to Nile tilapia diets in Murchison Bay. Our results further show that none of the other measured environmental variables was a good predictor of diet items in Nile tilapia. Adult Nile tilapia in our study lakes, rely heavily on filter feeding, particularly under conditions of low water transparency, trapping detritus and phytoplankton cells especially colonies. They can ingest more mobile prey like insects and insect larvae when the water transparency and visibility increases.     

Development of a Capillary Zone Electrophoretic Method for the Rapid Separation and Detection of Hepatotoxic Microcystins

Analysis of trace amounts of various hepatotoxic microcystins in marine and freshwater samples is very important since these toxins, especially microcystin-LR, have been demonstrated to have tumour-promoting activity. In this study, instead of measuring the total amount of microcystins, we developed a capillary zone electrophoretic method for the separation and detection of individual toxin standards. No additives were used for enhancement of resolution. This technique is characterized by a high separation efficiency, short analysis time and small sample volume. In order to improve the detection sensitivity, a laser-induced fluorescence detector was used, and the labelling of microcystins was accomplished through a two-step procedure. First, the microcystin standards were converted into cysteine conjugates, followed by derivatization with Fluorescein 5-Isothiocyanate (FITC). After derivatization, the FITC-labelled microcystins were directly injected, separated and detected in 8 min. This method was shown to be a promising technique for sensitive and rapid analysis of individual microcystin toxins.     

Detection of Cyanobacterial Toxins in Water Supply Sources and Tap Water in Some Russian Cities: Searching Producers and Testing Removal Methods

Water Resources - For the first time, data are given on cyanobacterial toxins entering tap water, supplied to the population of some cities in Yaroslavl oblast. The detection of microcystins with...     

Cyanobacterial toxins in Italian freshwaters

This study focuses on the occurrence and distribution of cyanobacterial toxins from 1989 to 2006 in several Italian lakes of different characteristics and human uses, the latter including drinking water abstraction and recreation. Phytoplankton and LC/MS/MS toxin analyses were performed on surface water samples collected from 28 lakes. The most widespread species associated with toxin production belonged to the genera Microcystis, Planktothrix and Anabaena. Extracellular concentrations varied from non-detectable values up to 226.16 ng/mL for microcystins (sum of all variants), to 126 ng/mL for total cylindrospermopsin, and to 100 μg/g (dry weight) for anatoxin-a.The toxin concentrations in the lake waters did not always correlate with the cyanobacteria cell densities. This implies a need for control studies including toxin detection in water together with microscopic cell evaluations, in order to avoid possible toxin underestimates.     

Concentrations and Seasonal Trends of Extracellular Microcystins in Freshwaters of the Czech Republic – Results of the National Monitoring Program

The first outcomes of the national monitoring program on cyanobacteria and their toxins in the Czech Republic are reported. The concentrations of extracellular microcystins (MCs, analyzed by ELISA) in 206 water samples collected from 94 localities have been investigated. Observed MC concentrations were relatively high in comparison with the literature data, although weather conditions during the covered season (year 2004) were less favorable for the cyanobacterial water blooms. Detectable MCs (>0.125 μg/L) were found in 145 waters (70% of all samples) with a median concentration of 0.67 μg/L. The total median, including samples with no detectable MCs, was 0.26 μg/L. The seasonal trend showed an increase from July to September at most localities (maximum concentrations of about 8 μg/L and extremes up to 37 μg/L). However, a peak at the beginning of the season or continuously elevated MC concentrations were also observed at some localities. Positive correlations between increasing concentrations of MCs and the dominancy of Microcystis spp. have been observed. The survey provides the first large-scale study of the occurrence and seasonal variability of extracellular microcystins that are of particular interest for drinking water management.     

关于从中国滇池微囊藻中分离出微晶体的初报

Isolates of Microcystis blooms collected from Dianchi Lake of South-western China in different seasons were tested by mouse bioassays. The LD₅₀ of the vcdcrocystis isolates corre-sponding M. wesenbergii, M.viridis and M. aeruginosa were 221 mg · kg ^-1, 188 mg · kg ^-1, and 198 mg · kg ^-1 respectively. The cells of the three samples contained hepatotoxins or microcystins. The HPLC (high performance liquid chromatography) analysis showed that all the three Microcystis species contained microcystin LR. In addition, M. wesenbergii contained microcystin RR, M. viridis contained microcystinYR, M. aeruginosa contained both microcystin RR and YR. The results suggested that in different seasons the toxins which were produced by the dominating Microcystis species were fluctuent with regard to the composition and content of the toxins and consen-quently were obvious factors that aroused the water quality to decrease and change.     

Occurrence of the First Toxic Microcystis Bloom in a Recent Moroccan Reservoir

In order to understand initial conditions that promote the first occurrence of a Microcystis aeruginosa bloom and to evaluate its toxic potential, we studied spatio-temporal dynamics of phytoplankton, including Microcystis, microcystins concentrations, the relative proportion of potentially microcystin producing cells and physicochemical parameters in the recently setup Moroccan reservoir “Yaacoub Al Mansour.” High summer temperatures, thermal stability, hydrological stability of the water column and the decrease of nutrient contributions seem to be the major environmental factors responsible of this first bloom occurrence. Despite the low cellular abundance, high microcystins concentrations, associated with low percentage of toxic strains, were measured. Thus, microcystins analysis and associated management of the reservoir are necessary from the beginning of the setup and can help managers to find the better environmental strategy against early cyanobacteria proliferations.     

Contamination of some reservoirs and lakes in Republic of Bulgaria by microcystins

This paper presents the results from analyses of water samples from 15 Bulgarian reservoirs and lakes for detection of microcystins presence by high‐performance liquid chromatography (HPLC) coupled to a ultraviolet (UV) and photo diode array (PDA) detector. In 3 of the studied drinking water reservoirs cyanobacterial blooms were not detected. Cyanobacterial blooms were observed in 6 of 12 investigated water bodies used for fishing and water sports. Microcystis was the dominant cyanobacterial genus. The concentration of total microcystins in the biomasses ranged from 8 to 1070 μg g^–1 (d.w.). Microcystin‐LR and microcystin‐RR were the predominant microcystins, followed by microcystin‐YR. Presence of dissolved microcystins was obtained in one water sample with concentration of total microcystins 1.64 μg L^–1. Presence of microcystin‐LF and ‐LW or unknown peaks possessing characteristic microcystin‐like UV‐spectra were not detected. Those first results for occurrence of microcystins in some Bulgarian reservoirs and lakes are a contribution to the knowledge on their distribution.     

Cyanobacterial blooms in estuarine ecosystems: characteristics and effects on Laeonereis acuta (Polychaeta, Nereididae)

In January of 2003, a cyanobacterial bloom in the Patos' Lagoon (Southern Brazil) (32 degrees 05'S-52 degrees 12'W) was observed. Water samples were taken to identify the composition and abundance of the bloom, as well as the occurrence of toxins. The effects of this occurrence on the estuarine worm Laeonereis acuta (Polychaeta, Nereididae) was also evaluated. Predominance of cyanobacteria, particularly Anabaena trichomes ( approximately 2.5.10(6) individuals per liter) was observed, and low concentrations of microcystins and anticholinesterasic toxins were detected. Augmented levels of lipid hydroperoxides (LPO) and glutathione-S-transferase activity, and lowering of total protein content were also observed in organisms collected during the bloom event. Although non-toxic, the cyanobacterial bloom could augment the cycle of hyper-oxygenation and hypoxia in the water. During hyperoxia, L. acuta, an oxyconformer, should consume more oxygen, thus augmenting the rate of reactive oxygen species generation. A repeated cycle of hyper-oxygenation and hypoxia would finally induce oxidative stress, as evidenced by the high levels of LPO and glutathione-S-transferase activity.