Does microcystin disrupt the induced effect of Daphnia kairomone on colony formation in Scenedesmus?

In natural aquatic system, Scenedesmus and Microcystis species usually coexist. Microcystins are released into water after lysis of Microcystis cells during the collapse of heavy blooms. The released toxins can then come into contact with a wide range of aquatic organisms. In this study, we used filtered Daphnia test water containing kairomone from Daphnia magna to stimulate the inducible colony formation in Scenedesmus obliquus under microcystin-contaminated system, to examine how microcystin affects the induced effect of Daphnia kairomone on colony formation in S. obliquus. The results showed neither microcystin nor Daphnia kairomone affected the growth of S. obliquus. Microcystin neither promoted nor impaired the overall Daphnia-induced colony formation in S. obliquus, except reducing the proportion of eight-celled colonies on day 2, indicating that the effect of microcystin was just short-term and in general did not disrupt grazer-induced colony formation of S. obliquus.     

A review of the global ecology,genomics, and biogeography of the toxic cyanobacterium,Microcystis spp.

This review summarizes the present state of knowledge regarding the toxic, bloom-forming cyanobacterium, Microcystis, with a specific focus on its geographic distribution, toxins, genomics, phylogeny, and ecology. A global analysis found documentation suggesting geographic expansion of Microcystis, with recorded blooms in at least 108 countries, 79 of which have also reported the hepatatoxin microcystin. The production of microcystins (originally “Fast-Death Factor”) by Microcystis and factors that control synthesis of this toxin are reviewed, as well as the putative ecophysiological roles of this metabolite. Molecular biological analyses have provided significant insight into the ecology and physiology of Microcystis, as well as revealed the highly dynamic, and potentially unstable, nature of its genome. A genetic sequence analysis of 27 Microcystis species, including 15 complete/draft genomes are presented. Using the strictest biological definition of what constitutes a bacterial species, these analyses indicate that all Microcystis species warrant placement into the same species complex since the average nucleotide identity values were above 95%, 16S rRNA nucleotide identity scores exceeded 99%, and DNA–DNA hybridization was consistently greater than 70%. The review further provides evidence from around the globe for the key role that both nitrogen and phosphorus play in controlling Microcystis bloom dynamics, and the effect of elevated temperature on bloom intensification. Finally, highlighted is the ability of Microcystis assemblages to minimize their mortality losses by resisting grazing by zooplankton and bivalves, as well as viral lysis, and discuss factors facilitating assemblage resilience.     

Spatial and temporal diversity of microcystins and microcystin-producing genotypes in Oneida Lake, NY

Oneida Lake is a shallow, eutrophic lake with a well-established cyanobacterial population with reported toxic blooms containing hepatotoxic microcystins (MC). Peak bloom events from the summers of 2002 and 2003 were analyzed to determine the principal cyanobacterial genera containing microcystin synthetase (mcy) genes. Sequence analysis of a partial mcyA amplicon targeting Microcystis, Anabaena and Planktothrix sp. indicated that Microcystis sp. was the dominant mcy genotype. This Microcystis clade was split into two distinct sub-clades. Bloom events contained members of both sub-clades with the higher MC concentrations found when both sub-clades were present in near equal proportions. The proportion of Microcystis containing the mcyD gene ranged from 0 to 37% of the total Microcystis population as determined by quantitative PCR (qPCR). The total concentration of Microcystis containing mcyD genes was linearly related to the concentration of MCs (r² = 0.63). The relationship between mcy genotype and physiochemical variables was examined to determine the factor(s) controlling the periodicity in MC production in Oneida Lake. Multivariate statistical analyses, used to correlate the continuous-response variables, revealed a strong relationship between chlorophyll a, MCs and total Microcystis.     

伊乐藻生物碱的GC-MS分析及其对铜绿微囊藻的化感作用

藻类暴发性生长是水体富营养化带来的环境问题之一,利用植物化感作用抑制藻类生长作为一种新型的生物抑藻技术在近年来开始受到研究者的重视,并取得了一定的研究成果。文章采用GC-MS联用技术鉴定出伊乐藻中的9种生物碱成分,还研究了其总生物碱对铜绿微囊藻的化感作用。结果发现添加总生物碱的处理组中铜绿微囊藻生物量均受到了抑制,在总生物碱的浓度为62.0mg/L时,3d后铜绿微囊藻的抑制率为44.0%,表明伊乐藻总生物碱对铜绿微囊藻的生物量增长具有明显的抑制作用。该结论为通过沉水植物恢复富营养化水体提供了重要依据。       

铜锈微囊藻两种表型的生长生理特性及毒素组成比较分析

从滇池蓝藻水华中分离得到的铜锈微囊藻群体在实验室无机营养中解聚成单细胞,结果表明,群体微囊藻的生长速度明显低于单细胞微囊藻;前者具明显可见的胞外酸性多糖胶鞘,而单细胞则几乎没有;按常规方法分析比较两种细胞形态的毒性大小和毒素组成,发现群体微囊藻主要含有三种微囊藻毒素的异构体,而单细胞以MCLR为主;且单细胞微囊藻的毒性约为群体的10倍.二者的LDH和PGM同工酶酶谱也有差异.本研究为解释毒素的合成和调控机理提供了新的证据.       

Biological degradation of algae and microcystins by microbial enrichment on artificial media

The removal efficiency of algae and microcystins (MCs) of a pilot setup based on biological degradation of enrichment microbes by artificial media was studied. The results showed that when chlorophyll-a in Lake Taihu was 3.76–266.1 μg L⁻¹, 62.8% of chlorophyll-a could be removed within a hydraulic retention time of 6–7 days. High-performance liquid chromatography (HPLC) analysis for the detection of MCs was applied. When total microcystins RR and LR (TMC-RR and TMC-LR) and extracellular microcystins RR and LR (EMC-RR and EMC-LR) were 0.23–8.93, 0.14–4.73, 0.12–1.15, 0.02–0.17 μg L⁻¹, respectively, and in source water, the average removal efficiencies of TMC-RR, TMC-LR, EMC-RR, EMC-LR were 67.0%, 40.5%, 40.0% and 66.0%, respectively. The polymerase chain reaction (PCR) electrophoresis chart revealed that there was a large amount of algae-lysing bacteria such as Pseudomonas spp. and Bacillus spp. on the artificial medium. The protozoa number in the assembled medium was higher than in lake water. Enrichment microbes on the artificial medium could effectively degrade algae and microcystins in Lake Taihu.     

Application of Bayesian network including Microcystis morphospecies for microcystin risk assessment in three cyanobacterial bloom-plagued lakes,China

Microcystis spp., which occur as colonies of different sizes under natural conditions, have expanded in temperate and tropical freshwater ecosystems and caused seriously environmental and ecological problems. In the current study, a Bayesian network (BN) framework was developed to access the probability of microcystins (MCs) risk in large shallow eutrophic lakes in China, namely, Taihu Lake, Chaohu Lake, and Dianchi Lake. By means of a knowledge-supported way, physicochemical factors, Microcystis morphospecies, and MCs were integrated into different network structures. The sensitive analysis illustrated that Microcystis aeruginosa biomass was overall the best predictor of MCs risk, and its high biomass relied on the combined condition that water temperature exceeded 24 °C and total phosphorus was above 0.2 mg/L. Simulated scenarios suggested that the probability of hazardous MCs (≥1.0 μg/L) was higher under interactive effect of temperature increase and nutrients (nitrogen and phosphorus) imbalance than that of warming alone. Likewise, data-driven model development using a naïve Bayes classifier and equal frequency discretization resulted in a substantial technical performance (CCI = 0.83, K = 0.60), but the performance significantly decreased when model excluded species-specific biomasses from input variables (CCI = 0.76, K = 0.40). The BN framework provided a useful screening tool to evaluate cyanotoxin in three studied lakes in China, and it can also be used in other lakes suffering from cyanobacterial blooms dominated by Microcystis.     

Isolation and structure determination of two new hydrophobic microcystins from Microcystis sp. (CAWBG11)

Two new hydrophobic microcystins, microcystin-FA (1) and microcystin-WA (2), were isolated from the cyanobacterium Microcystis sp. (CAWBG11). The structures were deduced using one- and two-dimensional nuclear magnetic resonance spectroscopy and tandem mass spectrometry. The absolute stereochemistry of the amino acid residues in 1 and 2 was determined using the Advanced Marfey's method.     

Alteration of intracellular GSH levels and its role in microcystin-LR-induced DNA damage in human hepatoma HepG2 cells

Microcystin-LR (MCLR) is a liver-specific toxin known as a tumour promoter in experimental animals. Its mechanisms of hepatotoxicity have been well documented; however, the mechanisms of other effects, in particular those related to its genotoxicity, are not well understood. In our previous studies, we showed that MCLR-induced DNA strand breaks are transiently present and that the damage is mediated by reactive oxygen species (ROS). In this study, we show that exposure of HepG2 cells to non-cytotoxic doses of MCLR-induced time-dependent alterations in the level of intracellular reduced glutathione (GSH). These comprised a rapid initial decrease followed by a gradual increase, reaching a maximum after 6 h of exposure, before returning to the control level after 8 h. During the first 4 h, expression of glutamate-cysteine ligase (GCL), the rate-limiting enzyme of GSH synthesis, increased, indicating an increased rate of de novo synthesis of GSH. The most important observation of this study, combined with the results of our previous studies is the correlation between the time course of alterations of intracellular GSH content and the formation and disappearance of MCLR-induced DNA damage. When the intracellular GSH level was reduced, MCLR-induced DNA damage was observed to increase. Later, when the level of intracellular GSH was normal or elevated, new DNA damage was not induced and existing damage was repaired. To confirm the role of GSH system in MCLR-induced genotoxicity, the intracellular GSH level was moderated by pre-treatment with buthionine-(S,R)-sulfoximine (BSO), a specific GSH synthesis inhibitor, and with N-acetylcysteine (NAC), a GSH precursor. Pre-treatment with BSO dramatically increased the susceptibility of HepG2 cells to MCLR-induced DNA damage, while pre-treatment with NAC almost completely prevented MCLR-induced DNA damage. Thus, intracellular GSH is shown to play a critical role in the cellular defence against MCLR-induced DNA damage in HepG2 cells.