蓝藻毒素的类型及其产毒基因

水体富营养化及蓝藻水华的频繁发生,严重破坏水域生态系统,有毒水华蓝藻产生的藻毒素,污染水体且危害水生动物和人类健康。本文综述了水华蓝藻的次级代谢产物藻毒素及藻毒素合成基因的国内外研究进展,介绍了蓝藻毒素的类型、不同毒素毒性机理、产毒基因以及利用产毒基因进行产毒蓝藻的分子生物学检测等,并对研究中存在的问题和未来的研究方向进行了展望,目的是为建立有毒水华蓝藻的预警机制、科学评价产毒蓝藻及其藻毒素的生态、环境危害及生态环境治理提供理论和技术支撑。     

微囊藻毒素对水环境的影响研究进展

微囊藻毒素是富营养化淡水水体中最常见的藻类毒素,是湖泊蓝藻产生的一类肽类毒素,它的产生受到藻类的遗传和环境因素的共同影响。由于其毒性大,分布广,结构稳定,从而成为水环境中的潜在危害物质。有关微囊藻毒素性质、毒理毒性、在环境中的迁移、转化以及控制预防已成为关注热点。在总结国内外研究的基础上,综述了微囊藻毒素的性质、产生机理以及其与水环境、水生生物(水生植物、鱼类、无脊椎动物)间的相互作用,讨论了微囊藻毒素对水生生物的影响以及水生生物对微囊藻毒素的降解作用,为水体中微囊藻毒素的防治提供科学的依据。     

环境水体藻毒素生物处理技术研究进展

近年来,随着水体富营养化程度加剧,蓝藻水华现象时有发生,蓝藻及其释放的藻毒素对生态环境和人类健康构成严重威胁.本文概述了常见藻毒素的分类及其主要理化性质,总结了生物接触氧化、生物滤池和生物-生态耦合等工艺对藻毒素的去除性能与机制,分析了生物处理工艺的反应条件(温度、pH和水力停留时间)、原水性质及营养限制等影响因素,并对藻毒素去除机理、新工艺研发等方面进行了展望.     

微囊藻毒素生物治理技术研究进展*

近年来,随着全球气候变暖和水体富营养化程度加深,蓝藻水华频繁暴发。微囊藻毒素是有害蓝藻产生及释放的危害最大的一类蓝藻毒素,对生态环境和公众健康造成了严重的威胁。因此,寻求有效的微囊藻毒素降解方法已成为全球科学领域的研究热点。针对微囊藻毒素生物治理技术展开综述,阐述了微囊藻毒素的产生、理化性质及生物毒性,总结了微生物、水生植物、浮游动物等自然生物降解微囊藻毒素的能力。在此基础上概述了生物滤池、人工湿地、生态浮床、膜生物膜反应器等生物治理技术对微囊藻毒素的去除效果,分析了现有微囊藻毒素生物处理方法的优势和局限性,并对今后的研究方向提出展望,为解决水环境中微囊藻毒素的污染问题提供思路。     

微囊藻毒素生物合成基因及其功能研究进展

水体富营养化加剧,导致了蓝藻水华在世界范围内频发。蓝藻产生的微囊藻毒素是最常见的一种藻毒素,对人类和动物造成了很大的危害甚至导致死亡。微囊藻毒素经非核糖体合成途径由多肽合成酶合成。对微囊藻毒素的结构与性质、微囊藻毒素合成基因的功能及其生物合成、微囊藻毒素的分子生物学检测技术进行了评述,对未来的研究方向进行了展望。     

应用环境微生物治理淡水湖泊微囊藻毒素污染的研究进展

近年来,随着水体富营养化程度的加剧,蓝藻水华现象时有发生,蓝藻及其释放的藻毒素对生态环境和人类健康构成严重威胁。在各种藻毒素中,以微囊藻毒素(Microcystins,MCs)毒性最强,对人类危害也最大,微囊藻毒素的化学性质相对稳定且难以通过常规水处理方法消除,因此如何有效去除环境中的MCs是国内外普遍关注的难题。研究发现自然界中的微生物能够有效降解和消除MCs污染,由此产生的环保技术极具应用价值。本文主要概述了微囊藻毒素的产生机理、化学结构以及毒性危害,总结了微囊藻毒素的自然分解过程以及微生物群落对微囊藻毒素的响应机制,重点分析了微生物群落在微囊藻毒素污染控制技术中的潜在应用,并对应用微生物技术治理微囊藻毒素污染的技术瓶颈提出了建议,以期加速微囊藻毒素微生物降解技术的完善和应用。     

环境水体微囊藻毒素微生物降解技术研究进展

湖库水体富营养化及其产生的藻毒素污染已对生态环境和人类健康构成极大威胁,而目前常规水污染控制技术存在一定的局限性,因此水环境中藻毒素处理新工艺亟待研发.鉴于环境水体中的微囊藻毒素可被微生物降解为无毒或低毒的中间产物,本文综述了微囊藻毒素的降解菌株、生物降解过程影响因素与机理、降解产物及其结构特性等,总结了目前微囊藻毒素降解菌株在水环境修复中的应用,并对今后微生物降解微囊藻毒素的研究方向进行了展望,以期为解决我国日益严峻的湖库水体藻毒素污染和饮用水安全问题提供技术思路.     

国内简讯

中国科学院青岛生物能源与过程研究所蓝藻甲烷化与微囊藻毒素降解研究取得新进展水体富营养化及其产生的蓝藻是沿湖地区所面临的一个主要环境问题,厌氧消化被认为是现阶段快速处置大量打捞蓝藻的可行性技术之一。中国科学院青岛生物能源与过程研究所生物制氢与沼气团队研究     

淡水湖泊中微囊藻水华的成因分析

我国藻型富营养化湖泊中一个重要的生物类群是微囊藻,结合国内近几年来对微囊藻形成水华的机制的研究,从微囊藻的结构和生理特点以及环境中温度、光照、pH和营养盐等生态因子综合作用上分析了微囊藻水华的成因。     

微囊藻毒素-LR的离体鳃灌流研究

随着水体富营养化进程的加剧, 有毒蓝藻的频繁暴发已成为一个世界性的难题。自1878年Francis1报道了世界上首例由于接触有毒蓝藻而引发的牲畜中毒事件以来, 已经有许多关于鱼类、爬行动物、鸟类和哺乳动物接触蓝藻而中毒甚至死亡的案例25。在为数众多的藻毒素中, 微囊藻毒素(Microcystins, MCs)因具有很强的肝毒性6, 7和促癌作用8, 而被认为是最具危害的种类之一。       

全细胞多重PCR检测蓝藻、微囊藻及产毒微囊藻方法初探

选取三对分别针对微囊藻、蓝藻16S rDNA及微囊藻毒素合成酶基因mcyB的保守序列的特异性引物209F/409R、27F1/409R、MTR/MTF,其中409R为一条共用引物。设计并优化了一种可以同时检测蓝藻和微囊藻的两重全细胞PCR方法和一种可以同时检测蓝藻、微囊藻和可产毒微囊藻的三重全细胞PCR方法,并且测试了这两种PCR反应的灵敏度区间,分别为10⁵～10³cell·mL^-1、10⁵～10²cell·mL^-1。对采集水库水样检测结果表明双重全细胞PCR方法可以直接应用于对天然水样的检测,三重全细胞PCR方法可用于实验室培养藻细胞的筛查。全细胞多重PCR方法具有快速、简便、准确等特点,在水体微囊藻毒素检测预警方面具有应用价值。     

Profiles of toxic and non-toxic oligopeptides of Radiocystis fernandoii (Cyanobacteria) exposed to three different light intensities

Cyanobacteria produce a high variety of bioactive oligopeptides, which function, ecological, physiological roles and responses to environmental changes are still unclear. The influence of light intensity on the cell quota and the diversity of oligopeptides of two strains of the cyanobacterium Radiocystis fernandoii were experimentally tested. The peptides were quantified by HPLC and identified by a MALDI-TOF-TOF. Microcystins (MC) were generally more abundant in the treatment with low light. A compensatory mechanism was observed for the different variants of microcystin, whereby MC-RR responses were contrary to those observed for the other three variants and showed higher concentration in the treatment with intermediate light. Two microviridins were also produced at higher amounts at intermediate irradiance. For cyanopeptolins and a third microviridin no significant difference among treatments was found. The absence of a similar response for all peptides suggests that these compounds may have unique cellular functions, which better understanding could help explaining changes in toxicity. Finally, we observed that each chemical profile reflected in physiological differences between strains, strengthening the idea that chemotypes may act as distinct ecotypes in nature.     

Amino acid availability determines the ratio of microcystin variants in the cyanobacterium Planktothrix agardhii

Cyanobacteria are capable of producing multiple microcystin variants simultaneously. The mechanisms that determine the composition of microcystin variants in cyanobacteria are still debated. [Asp³]microcystin-RR contains arginine at the position where the more toxic [Asp³]microcystin-LR incorporates leucine. We cultured the filamentous cyanobacterium Planktothrix agardhii strain 126/3 with and without external addition of leucine and arginine. Addition of leucine to the growth medium resulted in a strong increase of the [Asp³]microcystin LR/RR ratio, while addition of arginine resulted in a decrease. This demonstrates that amino acid availability plays a role in the synthesis of different microcystin variants. Environmental changes affecting cell metabolism may cause differences in the intracellular availability of leucine and arginine, which can thus affect the production of microcystin variants. Because leucine contains one nitrogen atom while arginine contains four nitrogen atoms, we hypothesized that low nitrogen availability might shift the amino acid composition in favor of leucine, which might explain seasonal increases in the [Asp³]microcystin LR/RR ratio in natural populations. However, when a continuous culture of P. agardhii was shifted from nitrogen-saturated to a nitrogen-limited mineral medium, leucine and arginine concentrations decreased, but the leucine/arginine ratio did not change. Accordingly, while the total microcystin concentration of the cells decreased, we did not observe changes in the [Asp³]microcystin LR/RR ratio in response to nitrogen limitation.     

Potent toxins in Arctic environments – Presence of saxitoxins and an unusual microcystin variant in Arctic freshwater ecosystems

Cyanobacteria are the predominant phototrophs in freshwater ecosystems of the polar regions where they commonly form extensive benthic mats. Despite their major biological role in these ecosystems, little attention has been paid to their physiology and biochemistry. An important feature of cyanobacteria from the temperate and tropical regions is the production of a large variety of toxic secondary metabolites. In Antarctica, and more recently in the Arctic, the cyanobacterial toxins microcystin and nodularin (Antarctic only) have been detected in freshwater microbial mats. To date other cyanobacterial toxins have not been reported from these locations. Five Arctic cyanobacterial communities were screened for saxitoxin, another common cyanobacterial toxin, and microcystins using immunological, spectroscopic and molecular methods. Saxitoxin was detected for the first time in cyanobacteria from the Arctic. In addition, an unusual microcystin variant was identified using liquid chromatography–mass spectrometry. Gene expression analyses confirmed the analytical findings, whereby parts of the sxt and mcy operon involved in saxitoxin and microcystin synthesis, were detected and sequenced in one and five of the Arctic cyanobacterial samples, respectively. The detection of these compounds in the cryosphere improves the understanding of the biogeography and distribution of toxic cyanobacteria globally. The sequences of sxt and mcy genes provided from this habitat for the first time may help to clarify the evolutionary origin of toxin production in cyanobacteria.     

Evidence for positive selection acting on microcystin synthetase adenylation domains in three cyanobacterial genera

Background  

Cyanobacteria produce a wealth of secondary metabolites, including the group of small cyclic heptapeptide hepatotoxins that constitutes the microcystin family. The enzyme complex that directs the biosynthesis of microcystin is encoded in a single large gene cluster (mcy). mcy genes have a widespread distribution among cyanobacteria and are likely to have an ancient origin. The notable diversity within some of the Mcy modules is generated through various recombination events including horizontal gene transfer.     

Lake Erie Microcystis: Relationship between microcystin production, dynamics of genotypes and environmental parameters in a large lake

Cyanobacteria of genus Microcystis sp. have been commonly found in Lake Erie waters during recent summer seasons. In an effort to elucidate relationships between microcystin production, genotypic composition of Microcystis community and environmental parameters in a large lake ecosystem, we collected DNA samples and environmental data during a three-year (2003–2005) survey within Lake Erie and used the data to perform a series of correlation analyses. Cyanobacteria and Microcystis genotypes were quantified using quantitative real-time PCR (qPCR). Our data show that Microcystis in Lake Erie forms up to 42% of all cyanobacteria, and that Microcystis exists as a mixed population of potentially toxic and (primarily) non-toxic genotypes. In the entire lake, the total abundance of Microcystis as well as the abundance of microcystin-producing Microcystis is strongly correlated with the abundance of cyanobacteria suggesting that Microcystis is a significant component of the cyanobacterial community in Lake Erie during summer seasons. The proportion of total Microcystis of all cyanobacteria was strongly linked to the microcystin concentrations, while the percentage of microcystin-producing genotypes within Microcystis population showed no correlation with microcystin concentrations. Correlation analysis indicated that increasing total phosphorus concentrations correlate strongly with increasing microcystin concentrations as well as with the total abundance of Microcystis and microcystin-producing Microcystis.     

Evaluating putative ecological drivers of microcystin spatiotemporal dynamics using metabarcoding and environmental data

Microcystin is a cyanobacterial hepatotoxin of global concern. Understanding the environmental factors that cause high concentrations of microcystin is crucial to the development of lake management strategies that minimize harmful exposures. While the literature is replete with studies linking cyanobacterial production of microcystin to changes in various nutrients, abiotic stressors, grazers, and competitors, no single biotic or abiotic factor has been shown to be reliably predictive of microcystin concentrations in complex ecosystems. We performed random forest regression analyses with 16S and 18S rRNA gene sequencing data and environmental data to determine which putative ecological drivers best explained spatiotemporal variation in total microcystin and several individual congeners in a eutrophic freshwater reservoir. Model performance was best for predicting concentrations of the congener MC-LR, with ca. 88% of spatiotemporal variance explained. Most of the variance was associated with changes in the relative abundance of the cyanobacterial genus Microcystis. Follow-up RF regression analyses revealed that factors that were the most important in predicting MC-LR were also the most important in predicting Microcystis population dynamics. We discuss how these results relate to prevailing ecological hypotheses regarding the function of microcystin.     

Deciphering the genetic basis of microcystin tolerance

Background

Cyanobacteria constitute a serious threat to freshwater ecosystems by producing toxic secondary metabolites, e.g. microcystins. These microcystins have been shown to harm livestock, pets and humans and to affect ecosystem service and functioning. Cyanobacterial blooms are increasing worldwide in intensity and frequency due to eutrophication and global warming. However, Daphnia, the main grazer of planktonic algae and cyanobacteria, has been shown to be able to suppress bloom-forming cyanobacteria and to adapt to cyanobacteria that produce microcystins. Since Daphnia’s genome was published only recently, it is now possible to elucidate the underlying molecular mechanisms of microcystin tolerance of Daphnia.

Results

Daphnia magna was fed with either a cyanobacterial strain that produces microcystins or its genetically engineered microcystin knock-out mutant. Thus, it was possible to distinguish between effects due to the ingestion of cyanobacteria and effects caused specifically by microcystins. By using RNAseq the differentially expressed genes between the different treatments were analyzed and affected KOG-categories were calculated. Here we show that the expression of transporter genes in Daphnia was regulated as a specific response to microcystins. Subsequent qPCR and dietary supplementation with pure microcystin confirmed that the regulation of transporter gene expression was correlated with the tolerance of several Daphnia clones.

Conclusions

Here, we were able to identify new candidate genes that specifically respond to microcystins by separating cyanobacterial effects from microcystin effects. The involvement of these candidate genes in tolerance to microcystins was validated by correlating the difference in transporter gene expression with clonal tolerance. Thus, the prevention of microcystin uptake most probably constitutes a key mechanism in the development of tolerance and adaptation of Daphnia. With the availability of clear candidate genes, future investigations examining the process of local adaptation of Daphnia populations to microcystins are now possible.

Electronic supplementary material

The online version of this article (doi:10.1186/1471-2164-15-776) contains supplementary material, which is available to authorized users.     

Cyanotoxins in inland lakes of the United States: Occurrence and potential recreational health risks in the EPA National Lakes Assessment 2007

A large nation-wide survey of cyanotoxins (1161 lakes) in the United States (U.S.) was conducted during the EPA National Lakes Assessment 2007. Cyanotoxin data were compared with cyanobacteria abundance- and chlorophyll-based World Health Organization (WHO) thresholds and mouse toxicity data to evaluate potential recreational risks. Cylindrospermopsins, microcystins, and saxitoxins were detected (ELISA) in 4.0, 32, and 7.7% of samples with mean concentrations of 0.56, 3.0, and 0.061 μg/L, respectively (detections only). Co-occurrence of the three cyanotoxin classes was rare (0.32%) when at least one toxin was detected. Cyanobacteria were present and dominant in 98 and 76% of samples, respectively. Potential anatoxin-, cylindrospermopsin-, microcystin-, and saxitoxin-producing cyanobacteria occurred in 81, 67, 95, and 79% of samples, respectively. Anatoxin-a and nodularin-R were detected (LC/MS/MS) in 15 and 3.7% samples (n = 27). The WHO moderate and high risk thresholds for microcystins, cyanobacteria abundance, and total chlorophyll were exceeded in 1.1, 27, and 44% of samples, respectively. Complete agreement by all three WHO microcystin metrics occurred in 27% of samples. This suggests that WHO microcystin metrics based on total chlorophyll and cyanobacterial abundance can overestimate microcystin risk when compared to WHO microcystin thresholds. The lack of parity among the WHO thresholds was expected since chlorophyll is common amongst all phytoplankton and not all cyanobacteria produce microcystins.     

Sublethal Microcystin Exposure and Biochemical Outcomes among Hemodialysis Patients

Cyanobacteria are commonly-occurring contaminants of surface waters worldwide. Microcystins, potent hepatotoxins, are among the best characterized cyanotoxins. During November, 2001, a group of 44 hemodialysis patients were exposed to microcystins via contaminated dialysate. Serum microcystin concentrations were quantified with enzyme-linked immunosorbent assay which measures free serum microcystin LR equivalents (ME). We describe serum ME concentrations and biochemical outcomes among a subset of patients during 8 weeks following exposure. Thirteen patients were included; 6 were males, patients’ median age was 45 years (range 16–80), one was seropositive for hepatitis B surface antigen. The median serum ME concentration was 0.33 ng/mL (range: <0.16–0.96). One hundred thirty nine blood samples were collected following exposure. Patients’ biochemical outcomes varied, but overall indicated a mixed liver injury. Linear regression evaluated each patient’s weekly mean biochemical outcome with their maximum serum ME concentration; a measure of the extrinsic pathway of clotting function, prothrombin time, was negatively and significantly associated with serum ME concentrations. This group of exposed patients’ biochemical outcomes display evidence of a mixed liver injury temporally associated with microcystin exposure. Interpretation of biochemical outcomes are complicated by the study population’s underlying chronic disease status. It is clear that dialysis patients are a distinct ‘at risk’ group for cyanotoxin exposures due to direct intravenous exposure to dialysate prepared from surface drinking water supplies. Careful monitoring and treatment of water supplies used to prepare dialysate is required to prevent future cyanotoxin exposure events.