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Toxins ◽  
2022 ◽  
Vol 14 (1) ◽  
pp. 61
Author(s):  
Wannes Hugo R. Van Hassel ◽  
Mirjana Andjelkovic ◽  
Benoit Durieu ◽  
Viviana Almanza Marroquin ◽  
Julien Masquelier ◽  
...  

In the context of increasing occurrences of toxic cyanobacterial blooms worldwide, their monitoring in Belgium is currently performed by regional environmental agencies (in two of three regions) using different protocols and is restricted to some selected recreational ponds and lakes. Therefore, a global assessment based on the comparison of existing datasets is not possible. For this study, 79 water samples from a monitoring of five lakes in Wallonia and occasional blooms in Flanders and Brussels, including a canal, were analyzed. A Liquid Chromatography with tandem mass spectrometry (LC-MS/MS) method allowed to detect and quantify eight microcystin congeners. The mcyE gene was detected using PCR, while dominant cyanobacterial species were identified using 16S RNA amplification and direct sequencing. The cyanobacterial diversity for two water samples was characterized with amplicon sequencing. Microcystins were detected above limit of quantification (LOQ) in 68 water samples, and the World Health Organization (WHO) recommended guideline value for microcystins in recreational water (24 µg L−1) was surpassed in 18 samples. The microcystin concentrations ranged from 0.11 µg L−1 to 2798.81 µg L−1 total microcystin. For 45 samples, the dominance of the genera Microcystis sp., Dolichospermum sp., Aphanizomenon sp., Cyanobium/Synechococcus sp., Planktothrix sp., Romeria sp., Cyanodictyon sp., and Phormidium sp. was shown. Moreover, the mcyE gene was detected in 75.71% of all the water samples.


Toxins ◽  
2021 ◽  
Vol 13 (12) ◽  
pp. 838
Author(s):  
Donata Overlingė ◽  
Anna Toruńska-Sitarz ◽  
Marija Kataržytė ◽  
Renata Pilkaitytė ◽  
Greta Gyraitė ◽  
...  

Microcystins (MCs) are the most widely distributed and structurally diverse cyanotoxins that can have significant health impacts on living organisms, including humans. The identification of MC variants and their quantification is very important for toxicological assessment. Within this study, we explored the diversity of MCs and their potential producers from the Curonian Lagoon. MC profiles were analyzed by liquid chromatography-tandem mass spectrometry (LC-MS/MS) method, while the potential producers were detected based on the presence of genus-specific mcyE gene sequences. Among the numerous MCs detected, one new potential MC variant with m/z 1057 was partially characterized. Moreover, two other MCs with m/z 1075 and m/z 1068 might belong to new variants with serine (Ser), rarely detected in position one of the peptides. They might also represent MC-Y(OMe)R and MC-WR, respectively. However, the application of a low-resolution MS/MS system made the unambiguous identification of the MCs impossible. Based on this example, the problems of peptide structure identification are discussed in the work. Genetic analysis revealed that potential MCs producers include Dolichospermum/Anabaena, Microcystis spp., and Planktothrix agardhii. The diversity and temporal variations in MC profiles may indicate the presence of several chemotypes of cyanobacteria in the Curonian Lagoon.


Toxins ◽  
2021 ◽  
Vol 13 (2) ◽  
pp. 133
Author(s):  
Maša Zupančič ◽  
Polona Kogovšek ◽  
Tadeja Šter ◽  
Špela Remec Rekar ◽  
Leonardo Cerasino ◽  
...  

Due to increased frequency of cyanobacterial blooms and emerging evidence of cyanotoxicity in biofilm, reliable methods for early cyanotoxin threat detection are of major importance for protection of human, animal and environmental health. To complement the current methods of risk assessment, this study aimed to evaluate selected qPCR assays for detection of potentially toxic cyanobacteria in environmental samples. In the course of one year, 25 plankton and 23 biofilm samples were collected from 15 water bodies in Slovenia. Three different analyses were performed and compared to each other; qPCR targeting mcyE, cyrJ and sxtA genes involved in cyanotoxin production, LC-MS/MS quantifying microcystin, cylindrospermopsin and saxitoxin concentration, and microscopic analyses identifying potentially toxic cyanobacterial taxa. qPCR analyses detected potentially toxic Microcystis in 10 lake plankton samples, and potentially toxic Planktothrix cells in 12 lake plankton and one lake biofilm sample. A positive correlation was observed between numbers of mcyE gene copies and microcystin concentrations. Potential cylindrospermopsin- and saxitoxin-producers were detected in three and seven lake biofilm samples, respectively. The study demonstrated a potential for cyanotoxin production that was left undetected by traditional methods in both plankton and biofilm samples. Thus, the qPCR method could be useful in regular monitoring of water bodies to improve risk assessment and enable timely measures.


Botanica ◽  
2021 ◽  
pp. 77-94
Author(s):  
Blagoy Uzunov ◽  
Katerina Stefanova ◽  
Mariana Radkova ◽  
Jean-Pierre Descy ◽  
Georg Gärtner ◽  
...  

The summer phytoplankton structure of ten Bulgarian waterbodies was studied by HPLC analysis of marker pigments, light microscopy (LM) and PCR amplification of mcyB and mcyE gene sequences. The aim was to detect biodiversity and spread of toxigenic strains of potential microcystin producers and the important bloom-forming genus Microcystis in particular. The screening was done in three waterbodies, where Microcystis had already been found (Lakes Ezerets and Durankulak and Reservoir Koprinka), three waterbodies from which it had not been reported (Reservoirs Shilkovtsi, Zhrebchevo, Suedinenie) and four reservoirs that were sampled for the first time (Malka Smolnitsa, Plachidol 2, Preselka, Duvanli). LM and HPLC data similarly showed that cyanoprokaryotes contributed significantly to the total phytoplankton composition (29%) and biomass (15–87%) in nine sampled waterbodies. Microcystis aeruginosa, M. natans, M. smithii, M. wesenbergii, Microcystis spp., M. cf. comperei and M. pseudofilamentosa, were identified using LM (the last two tropical species were found for the first time in the country). Despite the low contribution of Microcystis to the phytoplankton diversity (1–4 taxa per sample) and to the total phytoplankton biomass (< 0.01–0.5%), 57 toxigenic strains of this genus were revealed by PCR, most of which demonstrated high similarity with NCBI M. aeruginosa and M. wesenbergii strains.


2020 ◽  
Vol 18 (6) ◽  
pp. 983-994
Author(s):  
Maria Fernanda Falcone-Dias ◽  
Marianna Vaz Rodrigues ◽  
Jeppe Lund Nielsen ◽  
Nadieh de Jonge ◽  
Niels O. G. Jørgensen ◽  
...  

Abstract Fish farming can have a negative impact on water quality and aquatic organisms due to emerging blooms of Cyanobacteria and the production of cyanotoxins. In this study, the effect of aquaculture in hydroelectric reservoirs in Brazil was evaluated in six fish farms and in upstream and downstream water through analysis of the microbiome, Cyanobacteria and microcystin concentrations. Synechococcus and Microcystis were observed at all six locations, while Limnothrix was also observed abundantly at two locations. An increase in the relative abundance of Cyanobacteria inside the fish farms was observed at two locations, while an increase of Cyanobacteria was observed in downstream at five of the six locations. Microcystins were detected in significant and high values in all locations, with concentrations up to 1.59 μg/L. The trend in microcystin concentrations was mirrored in copy numbers of the mcyE gene (encodes microcystin synthetase) and presence of Microcystis, but not in any of the other observed cyanobacterial groups. In summary, the study shows that aquaculture production influenced the water microbiome inside and downstream the fish farms, and a direct correlation was found between mcyE gene copies, microcystin production and abundance of Microcystis, but not for the total abundance of Cyanobacteria.


Toxins ◽  
2020 ◽  
Vol 12 (6) ◽  
pp. 382 ◽  
Author(s):  
Alexandra Galetović ◽  
Joana Azevedo ◽  
Raquel Castelo-Branco ◽  
Flavio Oliveira ◽  
Benito Gómez-Silva ◽  
...  

Edible Llayta are cyanobacterial colonies consumed in the Andes highlands. Llayta and four isolated cyanobacteria strains were tested for cyanotoxins (microcystin, nodularin, cylindrospermopsin, saxitoxin and β-N-methylamino-L-alanine—BMAA) using molecular and chemical methods. All isolates were free of target genes involved in toxin biosynthesis. Only DNA from Llayta amplified the mcyE gene. Presence of microcystin-LR and BMAA in Llayta extracts was discarded by LC/MS analyses. The analysed Llayta colonies have an incomplete microcystin biosynthetic pathway and are a safe food ingredient.


Toxins ◽  
2020 ◽  
Vol 12 (4) ◽  
pp. 211 ◽  
Author(s):  
Kristel Panksep ◽  
Marju Tamm ◽  
Evanthia Mantzouki ◽  
Anne Rantala-Ylinen ◽  
Reet Laugaste ◽  
...  

Global warming, paired with eutrophication processes, is shifting phytoplankton communities towards the dominance of bloom-forming and potentially toxic cyanobacteria. The ecosystems of shallow lakes are especially vulnerable to these changes. Traditional monitoring via microscopy is not able to quantify the dynamics of toxin-producing cyanobacteria on a proper spatio-temporal scale. Molecular tools are highly sensitive and can be useful as an early warning tool for lake managers. We quantified the potential microcystin (MC) producers in Lake Peipsi using microscopy and quantitative polymerase chain reaction (qPCR) and analysed the relationship between the abundance of the mcyE genes, MC concentration, MC variants and toxin quota per mcyE gene. We also linked environmental factors to the cyanobacteria community composition. In Lake Peipsi, we found rather moderate MC concentrations, but microcystins and microcystin-producing cyanobacteria were widespread across the lake. Nitrate (NO3−) was a main driver behind the cyanobacterial community at the beginning of the growing season, while in late summer it was primarily associated with the soluble reactive phosphorus (SRP) concentration. A positive relationship was found between the MC quota per mcyE gene and water temperature. The most abundant variant—MC-RR—was associated with MC quota per mcyE gene, while other MC variants did not show any significant impact.


2019 ◽  
Vol 16 (4(Suppl.)) ◽  
pp. 0978
Author(s):  
Abed Et al.

Several toxigenic cyanobacteria produce the cyanotoxin (microcystin). Being a health and environmental hazard, screening of water sources for the presence of microcystin is increasingly becoming a recommended environmental procedure in many countries of the world. This study was conducted to assess the ability of freshwater cyanobacterial species Westiellopsis prolifica to produce microcystins in Iraqi freshwaters via using molecular and immunological tools. The toxigenicity of W. prolifica was compared via laboratory experiments with other dominant bloom-forming cyanobacteria isolated from the Tigris River: Microcystis aeruginosa, Chroococcus turigidus, Nostoc carneum, and Lyngbya sp. significant expression of mcyE gene and microcystin production was most evident in W. prolifica. Contrary to the prevailing concept that M. aeruginosa is a main microcystin producer in freshwaters around the world, no significant microcystin production was observed with this species throughout the time points studied in our laboratory methods. As for C. turigidus, N. carneum and Lyngbya sp., neither mcyE expression nor microcystin production was significant. Data from mcyE expression by RT-qPCR were generally in agreement with those obtained from microcystin quantification by ELISA. Interestingly, W. prolifica, which showed clear microcystin-producing ability in this study and which was not reported before in the literature to produce microcystin, can be added as a new microcystin producer to the list of toxigenic cyanobacteria.


Author(s):  
Latife Köker ◽  
Reyhan Akçaalan ◽  
Meriç Albay ◽  
Brett A. Neilan

<p>Blooms of cyanobacteria are an increasingly frequent phenomenon in freshwater ecosystems worldwide as a result of eutrophication. Many species can produce hepatotoxins that cause severe health hazards to humans. The aim of this study was to identify the bloom forming cyanobacteria species by molecular methods and to amplify genes responsible for hepatotoxin biosynthesis from the environmental samples and isolated strains of cyanobacteria from Küçükçekmece Lagoon, Sapanca, İznik, Manyas and Taşkısı Lakes. A total of 10 bloom samples and 11 isolated strains were examined and <em>Microcystis</em> spp., <em>Planktothrix</em> spp., <em>Nodularia</em> <em>spumigena</em>, <em>Anabaenopsis</em> <em>elenkinii</em>, <em>Sphaerospermopsis</em> <em>aphanizomenoides</em>, <em>Cylindrospermopsis</em> <em>raciborskii</em> were identified. Hepatotoxin genes were detected in 60% of the bloom samples and 45% of the strains. Two Microcystis strains were obtained from Küçükçekmece Lagoon. While the strain assigned to <em>Microcystis</em> <em>flosaquae</em> was non-toxic, <em>Microcystis</em> <em>aeruginosa</em> strain produced microcystin. According to PCR results, the <em>M. aeruginosa</em> and <em>Planktothrix</em> <em>agardhii</em> bloom samples of Küçükçekmece Lagoon contained the microcystin synthetase gene E (mcyE) indicative of microcystin production, however, no microcystin was detected by HPLC. The mcyE gene was also found in <em>Microcystis</em> <em>wesenbergii</em> isolated from Taşkısı Lake, and in all <em>Planktothrix</em> <em>rubescens</em> bloom samples from Sapanca Lake. To our knowledge, this is the first detailed study for identifiying different toxic cyanobacteria species and their hepatotoxin production from several waterbodies in Turkey using molecular methods.</p>


2016 ◽  
Vol 11 (3) ◽  
pp. 690-699
Author(s):  
Rohaslinda Mohamad ◽  
Mohd Rafatullah ◽  
Tengku Yusof ◽  
Yi Sim ◽  
Norli Ismail ◽  
...  

Toxic cyanobacteria blooms became a worldwide problems as many countries encounter the presence of the blooms in most of water bodies. As part to develop monitoring of cyanobacterial toxins in Malaysia, samples taken in twelve points in five different lakes in Miri, Sarawak. Polymerase chain reaction (PCR) amplification of cyanobacterial 16S rRNA were carried out to detect the presence of cyanobacteria in the water samples. Cyanobacterial 16S rRNA were detected in all the samples collected. While molecular analysis for detection of cyanobacterial toxin encoding gene were done using specific primers. PCR amplification of cyanobacterial toxin-encoding gene were carried using the combination of forward primer; mcyE-F2 and reverse primer; mcyE-R4 to amplify generic microcystin (mcyE) gene in the samples. Out of twelve samples collected, microcystin (mcyE) producing gene was detected in one of the samples tested. Presence of microcystin encoding gene indicates the risk of cyanobacterial toxins in Miri, Sarawak.


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