Dataset Information

Investigation of the Occurrence of Cyanotoxins in Lake Karaoun (Lebanon) by Mass Spectrometry, Bioassays and Molecular Methods.

ABSTRACT: Lake Karaoun is the largest artificial lake in Lebanon and serves multiple purposes. Recently, intensive cyanobacterial blooms have been reported in the lake, raising safety and aesthetic concerns related to the presence of cyanotoxins and cyanobacterial taste and odor (T&O) compounds, respectively. Here, we communicate for the first time results from a recent investigation by LC-MS/MS covering multiple cyanotoxins (microcystins (MCs), anatoxin-a, cylindrospermopsin, nodularin) in water and fish collected between 2019 and 2020. Eleven MCs were identified reaching concentrations of 211 and 199 μg/L for MC-LR and MC-YR, respectively. Cylindrospermopsin, anatoxin-a and nodularin were not detected. The determination of the total MCs was also carried out by ELISA and Protein Phosphatase Inhibition Assay yielding comparable results. Molecular detection of cyanobacteria (16S rRNA) and biosynthetic genes of toxins were carried out by qPCR. Untargeted screening analysis by GC-MS showed the presence of T&O compounds, such as β-cyclocitral, β-ionone, nonanal and dimethylsulfides that contribute to unpleasant odors in water. The determination of volatile organic compounds (VOCs) showed the presence of anthropogenic pollutants, mostly dichloromethane and toluene. The findings are important to develop future monitoring schemes in order to assess the risks from cyanobacterial blooms with regard to the lake's ecosystem and its uses.

SUBMITTER: Hammoud NA

PROVIDER: S-EPMC8540339 | biostudies-literature | 2021 Oct

REPOSITORIES: biostudies-literature

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Investigation of the Occurrence of Cyanotoxins in Lake Karaoun (Lebanon) by Mass Spectrometry, Bioassays and Molecular Methods.

Hammoud Noura Alice NA Zervou Sevasti-Kiriaki SK Kaloudis Triantafyllos T Christophoridis Christophoros C Paraskevopoulou Aikaterina A Triantis Theodoros M TM Slim Kamal K Szpunar Joanna J Fadel Ali A Lobinski Ryszard R Hiskia Anastasia A

Toxins 20211010 10

Lake Karaoun is the largest artificial lake in Lebanon and serves multiple purposes. Recently, intensive cyanobacterial blooms have been reported in the lake, raising safety and aesthetic concerns related to the presence of cyanotoxins and cyanobacterial taste and odor (T&O) compounds, respectively. Here, we communicate for the first time results from a recent investigation by LC-MS/MS covering multiple cyanotoxins (microcystins (MCs), anatoxin-a, cylindrospermopsin, nodularin) in water and fish ...[more]

PMID: 34679009

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Project description:The proliferation of cyanobacteria can result in algal blooms, which may cause environmental and biological harm due to the production and release of secondary metabolites, or cyanotoxins, into the affected waterway. Cyanobacteria can produce multiple classes of cyanotoxins; therefore, to understand the full toxic load of algal blooms, it is necessary to perform analyses that quantify each class. These classes are generally monitored individually due to the challenges associated with the differing physicochemical properties of the cyanotoxins. Hydrophilic interaction liquid chromatography (HILIC) is a form of chromatography capable of retaining multiple classes of cyanotoxins that differ in physicochemical properties. Here an HILIC-MS/MS method was developed and validated to detect 3 microcystins, 11 saxitoxins, and 2 anatoxins. The chromatographic conditions were optimized to allow for the separation of multiple pairs of saxitoxin epimers, and in-source fragmentation in the MS interface was used to develop unique MRMs between the pairs. The method was validated and had low limits of detection (LODs, between 0.00770 and 9.75 µg L-1) and limits of quantification (LOQs, between 0.0257 and 32.5 µg L-1) for all compounds. All analytes exhibited good linearity (R2 values ≥ 0.991) and low percentage relative standard deviations for retention time (0%-1.74%) and peak area (4.54%-27.6%), with spiked recoveries ranging from 75.6% to 117% for all compounds. A multiclass sample preparation method to extract the three classes of analytes from cyanobacterial samples was developed and validated, with 80:20 acetonitrile:water and 0.1% formic acid as the optimal extraction solvent. The newly developed sample preparation and analysis methods were applied to cultured cyanobacteria and field samples, with microcystins and saxitoxins detected. The multiclass sample preparation and analysis methods developed here improve on individual methods as they reduce the complexity and time of sample preparation and analysis and will assist ecotoxicologists in assessing the full toxic risk of cyanobacterial blooms.

Project description:Endocrine disrupting chemicals (EDCs) are man-made compounds interfering with hormone signaling and thereby adversely affecting human health. Recent reports provide evidence for the presence of EDCs in commercially available bottled water, including steroid receptor agonists and antagonists. However, since these findings are based on biological data the causative chemicals remain unidentified and, therefore, inaccessible for toxicological evaluation. Thus, the aim of this study is to assess the antiestrogenic and antiandrogenic activity of bottled water and to identify the causative steroid receptor antagonists. We evaluated the antiestrogenic and antiandrogenic activity of 18 bottled water products in reporter gene assays for human estrogen receptor alpha and androgen receptor. Using nontarget high-resolution mass spectrometry (LTQ-Orbitrap Velos), we acquired corresponding analytical data. We combined the biological and chemical information to determine the exact mass of the tentative steroid receptor antagonist. Further MS(n) experiments elucidated the molecule's structure and enabled its identification. We detected significant antiestrogenicity in 13 of 18 products. 16 samples were antiandrogenic inhibiting the androgen receptor by up to 90%. Nontarget chemical analysis revealed that out of 24520 candidates present in bottled water one was consistently correlated with the antagonistic activity. By combining experimental and in silico MS(n) data we identified this compound as di(2-ethylhexyl) fumarate (DEHF). We confirmed the identity and biological activity of DEHF and additional isomers of dioctyl fumarate and maleate using authentic standards. Since DEHF is antiestrogenic but not antiandrogenic we conclude that additional, yet unidentified EDCs must contribute to the antagonistic effect of bottled water. Applying a novel approach to combine biological and chemical analysis this is the first study to identify so far unknown EDCs in bottled water. Notably, dioctyl fumarates and maleates have been overlooked by science and regulation to date. This illustrates the need to identify novel toxicologically relevant compounds to establish a more holistic picture of the human exposome.

Dataset Information

Investigation of the Occurrence of Cyanotoxins in Lake Karaoun (Lebanon) by Mass Spectrometry, Bioassays and Molecular Methods.

Publications

Investigation of the Occurrence of Cyanotoxins in Lake Karaoun (Lebanon) by Mass Spectrometry, Bioassays and Molecular Methods.

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