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.     

两座抽水型水库蓝藻种群与微囊藻毒素的比较分析

2008年每隔两月调查1次广东省两座抽水型水库:凤凰山水库和南屏水库的蓝藻组成与微囊藻毒素分布特征.结果共检测到蓝藻有14属,主要为丝状体和群体种类,常见的有假鱼腥藻属(Pseudanabaena)、蓝纤维藻属(Dactylococcopsis)、湖丝藻属(Limnothrix)、拟柱孢藻属(Cylindrospermopsis)和色球藻属(Chroococcus).两座水库的蓝藻细胞密度为119～137373 cells mL-1,微囊藻毒素浓度为0～2.26μg L-1.凤凰山水库的全年微囊藻毒素浓度均低于国家安全标准(1 μg L-1),最高浓度出现在3月份;南屏水库在丰水期微囊藻毒素浓度高于凤凰山水库,最高浓度出现在7月(2.26μgL-1).长水力滞留时间的凤凰山水库的蓝藻细胞密度比短水力滞留时间的南屏水库高出1个数量级.两座水库蓝藻细胞密度的峰值都出现在枯水期的3月,这主要与温度和调水量变化引起的营养盐季节性变化有关.假鱼腥藻、湖丝藻和拟柱孢藻是凤凰山水库的优势蓝藻种类,大多数情况下相对丰度超过50%.在南屏水库,枯水期蓝藻以丝状体种类为优势,丰水期则以群体种类为优势.温度和调水量的季节性变化引起的营养盐季节性变化可能是引起南屏水库不同生活类型优势蓝藻种类季节演替的主要因素.     

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。       

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.     

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.       

Interspecific allelopathy in cyanobacteria: Cylindrospermopsin and Cylindrospermopsis raciborskii effect on the growth and metabolism of Microcystis aeruginosa

The biological role of cyanobacteria secondary metabolites is relatively unknown although several possible hypotheses have been discussed. In the following study the effect of cylindrospermopsin (CYN) and metabolites of non-CYN producing Cylindrospermopsis raciborskii strain on growth, alkaline phosphatase (ALP) activity and microcystin-LR (MC-LR) production in Microcystis aeruginosa was evaluated. Higher concentrations of CYN (10 and 50 μg L⁻¹) induced toxicity effects demonstrated by significant growth inhibition and M. aeruginosa cell necrosis. Lower concentrations of CYN (1 and 5 μg L⁻¹) slightly decreased growth rates but significantly up-regulated ALP activity. Moreover, under all studied CYN concentrations MC-LR production strongly decreased. Spent C. raciborskii medium mimicked the CYN action by inducing strong inhibition of M. aeruginosa growth and MC-LR production and through up-regulation of ALP activity. On the other hand, spent M. aeruginosa medium did not affect C. raciborskii growth and no alterations in ALP activity were observed. Co-culturing of these two species resulted in an increase of C. raciborskii contribution at the expense of M. aeruginosa. From the results we conclude that CYN can be involved in interspecific competition in cyanobacteria and that non-CYN producing C. raciborskii strains may produce a hitherto unknown bioactive compound(s) which can mimic CYN action.     

Detection of persistent microcystin toxins at the land–sea interface in Monterey Bay,California

Blooms of toxin-producing Microcystis aeruginosa occur regularly in freshwater systems throughout California, but until recently potential impacts in the coastal ocean have been largely ignored. Twenty-one sites in and around Monterey Bay were surveyed for evidence of microcystin toxin (2010–2011) at the land–sea interface. Following this initial survey four major watersheds in the Monterey Bay area were surveyed (2011–2013) for microcystin concentration, nutrients, alkalinity and water temperature to identify potential environmental factors correlated with the abundance of microcystin at the land–sea interface. During the first year microcystin was detected in 15 of 21 sites. Data from years two and three were analyzed by principal components analysis and mixed effects model. Results indicated that coastal nutrient loading (nitrate, phosphate silicate, ammonium, urea), were statistically significant predictors of the microcystin concentrations in the watersheds with clear evidence for seasonality at some sites. Microcystin was frequently at highest concentration in the autumn; however, at some locations high levels of toxin were measured during spring. Because this toxin has the ability to biomagnify and persist within food webs, elevated levels within the watershed may decrease potential for health and survival of wildlife and humans exposed to freshwater and marine waters. The widespread occurrence of microcystin at low to moderate levels throughout the year and throughout the sampled watersheds demonstrates the potential difficulty for management.     

Cyanotoxin production and phylogeny of benthic cyanobacterial strains isolated from the northeast of Brazil

Most of the knowledge about cyanobacteria toxin production is traditionally associated with planktonic cyanobacterial blooms. However, some studies have been showing that benthic cyanobacteria can produce cyanotoxins. According to this, we aimed to evaluate the production of microcystins and saxitoxins in benthic cyanobacteria isolated from aquatic ecosystems in the Northeast of Brazil and to use a polyphasic approach for their identification. Forty-five clonal strains were isolated from rivers and water supply reservoirs, and identified using morphological and molecular phylogenetic characteristics. In order to evaluate the toxins production, the strains were screened for genes involved in the biosynthesis of microcystins and saxitoxins, positive results were confirmed and cyanotoxins quantified using HPLC. Eight species were identified belonging to the Phormidiaceae, Pseudanabaenaceae and Nostocaceae families. This is the first study in Brazil that shows that strains from the Geitlerinema genus correspond to at least three phylogenetic lineages, which possibly correspond to three distinct species to be subsequently reclassified. The strains that showed one of the genes involved in the cyanotoxins production were analyzed by HPLC and Geitlerinema amphibium, Geitlerinema lemmermannii, Cylindrospermum stagnale and Phormidium uncinatum were identified as producing one or more saxitoxins variants. Thus, this is the first report of saxitoxins production for those first three species and the first report in Brazil for P. uncinatum.     

iTRAQ-based proteomic study of the effects of microcystin-LR on medaka fish liver

Microcystins are cyanotoxins that occur in ground water and thus pose a potential health risk. Microcystin-LR (microcystin-leucine-arginine) is a potent hepatotoxin, and is suspected of being a tumour promoter. Poisoning with this toxin causes several dysfunctions in hepatocytes by inhibiting protein phosphatases 1 and 2A, and notably produces oxidative stress, disrupts the cytoskeleton, and deregulates mitogen-activated protein kinase pathway. Medaka fish (Oryzias latipes) was chosen as a model for studying the effects of this cyanotoxin on liver proteins using a gel-free approach, iTRAQ. Fish were gavaged with microcystin-LR. Two hours later, 325 proteins could be identified by Scaffold Q+ and 32 proteins revealed statistically significant variations above a ∣0.2∣ threshold of log(2) ratio by comparison with control. These proteins are mostly involved in the translation and maturation of proteins, lipid metabolism and detoxification. Notably, apolipoproteins are deregulated which indicates a possible alteration of chylomicron-mediated transport.