Phthalocyanines (Pcs) are promising photosensitizers for use in various branches of science and industry. In the presence of visible light and diatomic oxygen, phthalocyanines can react to produce singlet oxygen, a member of reactive oxygen species able to damage different molecules and tissues. The aim of this study was to investigate the ability of phthalocyanines to degrade natural toxins in the presence of visible light. As the representative of hardly degradable toxins, a group of cyanobacterial peptide toxins--microcystin-LR--was chosen for this study. According to our results, phthalocyanines are able to degrade 61.5% of microcystins within a 48-hour incubation (38% of microcystins was degraded after 24 h and 24% after 12 h of incubation). Although other oxidants like hydrogen peroxide or ozone are able to degrade microcystins within several hours, we assume that by optimizing the spectrum emitted by light source and by changing the absorption characteristics of Pcs, microcystins degradation by phthalocyanines could be more effective in the near future.
- MeSH
- Water Pollutants, Chemical chemistry MeSH
- Water Pollution, Chemical prevention & control MeSH
- Enzyme-Linked Immunosorbent Assay MeSH
- Indoles chemistry MeSH
- Enzyme Inhibitors chemistry MeSH
- Microcystins chemistry MeSH
- Oxidants chemistry MeSH
- Singlet Oxygen MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
Benthic cyanobacteria recognized as producers of natural products, including cyanotoxins, have been neglected for systematic toxicological studies. Thus, we have performed a broad study investigating cyanotoxin potential of 311 non-planktic nostocacean representatives combining molecular and chemical analyses. Out of these, a single strain Nostoc sp. Treb K1/5, was identified as a new microcystin producer. Microcystins [Asp3]MC-YR, [Asp3]MC-FR, [Asp3]MC-HtyR and Ala-Leu/Ile-Asp-Arg-Adda-Glu-Mdha are reported for the first time from the genus Nostoc. All the studied strains were also analyzed for the occurrence of nodularins, cylindrospermopsin and (homo)anatoxin-a, yet no novel producer has been discovered. Our findings indicate rare occurrence of the common cyanotoxins in non-planktic nostocaceae which is in contrast with frequent reports of cyanotoxin producers among phylogenetically closely related planktic cyanobacteria.
- MeSH
- Phylogeny MeSH
- Microcystins chemistry metabolism MeSH
- Nostoc genetics metabolism MeSH
- Publication type
- Journal Article MeSH
Microcystins (MCs) are primarily hepatotoxins produced by cyanobacteria and are responsible for intoxication in humans and animals. There are many incidents of chronic exposure to MCs, which have been attributed to the inappropriate treatment of water supplies or contaminated food. Using RAW 264.7 macrophages, we showed the potency of microcystin-LR (MC-LR) to stimulate production of pro-inflammatory cytokines (tumor necrosis factor α and interleukin-6) as a consequence of fast nuclear factor κB and nitrogen-activated protein kinase activation. In contrast to other studies, the observed effects were not attributed to the intracellular inhibition of protein phosphatases 1/2A due to lack of specific transmembrane transporters for MCs. However, the MC-LR-induced activation of macrophages was effectively inhibited by a specific peptide that blocks signaling of receptors, which play a pivotal role in the innate immune responses. Taken together, we showed for the first time that MC-LR could interfere with macrophage receptors that are responsible for triggering the above-mentioned signaling pathways. These findings provide an interesting mechanistic explanation of some adverse health outcomes associated with toxic cyanobacteria and MCs.
- MeSH
- Cell Line drug effects MeSH
- Water Pollutants, Chemical toxicity MeSH
- Immunologic Factors toxicity MeSH
- Interleukin-6 metabolism MeSH
- Macrophages drug effects metabolism pathology MeSH
- Microcystins toxicity MeSH
- Mice MeSH
- NF-kappa B metabolism MeSH
- Immunity, Innate drug effects MeSH
- Protein Phosphatase 2 metabolism MeSH
- Cyanobacteria pathogenicity MeSH
- Toxicity Tests, Chronic methods MeSH
- Tumor Necrosis Factor-alpha metabolism MeSH
- Inflammation chemically induced immunology metabolism MeSH
- Water Supply MeSH
- Animals MeSH
- Check Tag
- Mice MeSH
- Animals MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
OBJECTIVES: Apart from infections and habitat loss, environmental pollution is another major factor of global decline of amphibians. Using the model of Xenopus laevis embryos, we test the hypothesis that combined exposure of amphibians to natural toxins and anthropogenic pollutants induces more pronounced adverse effects than single exposures. METHODS: Experimental procedures adhered to Frog Embryo Teratogenesis Assay - Xenopus standards (FETAX). Exposure groups included controls, solvent (dimethyl sulfoxide) controls, and embryos exposed for 96 h to single, double and triple action of paraoxon (P), bromadiolone (B), and microcystin-LR (M), added to the FETAX medium at a dose of 300, 350, and 500 μg.L(-1), respectively. Studied responses of X. laevis embryos included mortality and malformations, head-to-tail length, total antioxidant capacity, lipid peroxidation, and caspase-3 activity. RESULTS: The triple combination induced the highest mortality. Malformations in embryos significantly prevailed only in B-, and B+P-exposure groups. Apart from the single exposure to B, the tested substances and their combinations inhibited the embryonic growth. Triple exposure had the most pronounced effect both on the growth inhibition and total antioxidant capacity. Lipid peroxidation was increased after B+M exposure, while single and combined exposures to B and P had an opposite effect. CONCLUSIONS: This study helps to understand adverse effects of environmental pollution by natural toxins and agrochemicals in amphibians. The results allow for risk assessment of environmental pollution and findings of low concentrations of contaminants in aquatic environments. Further research to address issues such as mixture toxicity to metamorphosing and adult amphibians is necessary.
- MeSH
- 4-Hydroxycoumarins toxicity MeSH
- Abnormalities, Drug-Induced * MeSH
- Anticoagulants toxicity MeSH
- Cholinesterase Inhibitors toxicity MeSH
- Embryo, Nonmammalian drug effects embryology MeSH
- Enzyme Inhibitors toxicity MeSH
- Caspase 3 drug effects MeSH
- Microcystins toxicity MeSH
- Paraoxon toxicity MeSH
- Lipid Peroxidation drug effects MeSH
- Toxicity Tests MeSH
- Body Size drug effects MeSH
- Xenopus laevis embryology MeSH
- Animals MeSH
- Check Tag
- Animals MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
Microcystins are cyclic peptide toxins with hepatotoxic and tumor-promoting properties, which are produced in significant quantities (up to tens of μg/L) in freshwater cyanobacterial water blooms. Several studies reported microcystin accumulation in fish with possible food transfer to humans. These compounds are further metabolized to cysteine and glutathione conjugates which can be present in tissues in significant concentrations. In this study, we focused on the development and evaluation of robust and highly sensitive SPE-LC-MS/MS method for the analysis of microcystin conjugates in fish tissue samples. For the first time, we demonstrate the use of isotopically labeled internal standards which are essential for accurate and precise determination of analytes in complex biotic matrices. LLOQs of respective microcystin conjugates (signal-to-noise ratio; S/N > 10, peak-to-peak method) ranged from 3.3 to 5.0 ng/g of tissue fresh weight (FW). The calibration was linear within a range of concentrations from 1 to 70 ng/mL for all analyzed conjugates. The precision and repeatability of the method were very good with recoveries in the range of 88.5-107.6% and relative standard deviations between 8.8 and 13.2% for all analytes. In the follow-up study, fully validated method was used for the determination of microcystin conjugate levels in common carp exposed to microcystin-containing cyanobacterial biomass under controlled conditions. Significant amounts of microcystin conjugates (up to 55 ng/g) were found in the tissues of fish after 7 weeks of exposure. Our method was shown to be robust, sensitive, selective, and suitable for the determination of trace levels of microcystin conjugates in fish tissues.
- MeSH
- Biomass MeSH
- Chromatography, Liquid methods MeSH
- Cysteine analysis MeSH
- Glutathione analysis MeSH
- Limit of Detection MeSH
- Microcystins analysis chemistry MeSH
- Radioisotope Dilution Technique MeSH
- Reproducibility of Results MeSH
- Cyanobacteria chemistry MeSH
- Tandem Mass Spectrometry methods MeSH
- Publication type
- Journal Article MeSH
In mid-July and August 2003 and 2004, 18 reservoirs in the Czech Republic were sampled for phytoplankton species composition and concentration of intracellular microcystins (MCs). As a consequence of high nutrient loading, most of the reservoirs experienced cyanobacterial blooms of various intensities, with the prevalence of cyanobacteria increasing markedly in August, along with a conspicuous shift in species composition toward dominance of Microcystis spp. Microcystins were detected in 90% of the samples, and their amount also increased considerably in August, reflecting the cyanobacterial biomass. In Microcystis-dominated samples, a significantly higher amount of MCs (p < 0.001) occurred than in samples in which other taxa prevailed. Microcystins were positively correlated with chlorophyll a and cyanobacterial biovolume (p < 0.05, R2 = 0.61 and 0.66, respectively), with the strongest correlation found for Microcystis spp. biovolume (p < 0.001, R2 = 0.87). This taxon was the most important producer of MCs in Czech reservoirs. The main structural variants of MCs were MC-LR, MC-RR, and MC-YR. This study's data also indicate that the relative share of MC variants (MC-LR and MC-RR) varies considerably with time, most likely as a consequence of different species and strain compositions during the summer. This study clearly demonstrates a high prevalence of MC-producing cyanobacteria in Czech reservoirs. Therefore, regular monitoring of these reservoirs is highly desirable in an effort to minimize potential health risks to the human population. Copyright 2006 Wiley Periodicals, Inc.
- MeSH
- Biomass MeSH
- Peptides, Cyclic metabolism MeSH
- Eutrophication MeSH
- Water Microbiology MeSH
- Microcystins MeSH
- Environmental Monitoring MeSH
- Population Dynamics MeSH
- Seasons MeSH
- Cyanobacteria growth & development MeSH
- Fresh Water chemistry microbiology MeSH
- Water Supply MeSH
- Publication type
- Research Support, Non-U.S. Gov't MeSH
As human co-exposure to natural toxins through food and water is inevitable, risk assessments to safeguard health are necessary. Aflatoxin B1 and fumonisin B1, frequent co-contaminants of maize and microcystin-LR, produced in freshwater by cyanobacteria are all naturally occurring potent toxins that threaten human health. Populations in the poorest regions of the world may suffer repeated simultaneous exposure to these contaminants. Using High Content Analysis, multiple cytotoxicity endpoints were measured for the individual toxins and mixtures in various cell lines. Results highlighted that significant cytotoxic effects were observed for aflatoxin B1 in all cell lines while no cytotoxic effects were observed for fumonisin B1 or microcystin-LR. Aflatoxin B1/microcystin-LR was cytotoxic in the order HepG2 > Caco-2 > MDBK. Fumonisin B1/microcystin-LR affected MDBK cells. The ternary mixture was cytotoxic to all cell lines. Most combinations were additive, however antagonism was observed for binary and ternary mixtures in HepG2 and MDBK cell lines at low and high concentrations. Synergy was observed in all cell lines, including at low concentrations. The combination of these natural toxins may pose a significant risk to populations in less developed countries. Furthermore, the study highlights the complexity around trying to regulate for human exposure to multiple contaminants.
- MeSH
- Aflatoxin B1 administration & dosage chemistry toxicity MeSH
- Biomarkers urine MeSH
- Toxins, Biological MeSH
- Cell Line MeSH
- Fumonisins administration & dosage chemistry toxicity MeSH
- Food Contamination MeSH
- Humans MeSH
- Microcystins administration & dosage chemistry toxicity MeSH
- Cattle MeSH
- Dose-Response Relationship, Drug MeSH
- Animals MeSH
- Check Tag
- Humans MeSH
- Cattle MeSH
- Animals MeSH
- Publication type
- Journal Article MeSH
Nile tilapia (Oreochromis niloticus) were fed by diets supplemented with cyanobacteria containing in part the cyanotoxin microcystin-LR (MC-LR) to determine the potential impacts on detoxification. Four different diets were prepared based on a commercial diet: (1) control, (2) MC-5% (containing 5% dried Microcystis sp. biomass with 4.92 μg MC-LR g(-1) diet), (3) MC-20% (containing 20% dried Microcystis sp. biomass with 19.54 μg MC-LR g(-1) diet), and (4) Arthr-20% (containing 20% dried Arthrospira sp. biomass without MC-LR). Blood and liver samples were taken after one, 7, and 28 days and protein has been determined in plasma and liver. In the liver, impacts on detoxification were measured by glutathione-S-transferase (GST) activities and gene expression of multi drug resistance protein (MDRP). Plasma protein did not change between all four diets at any sampling time whereas liver protein was significantly elevated already after one day in Arthr-20% and after 28 days in both, MC-20% and Arthr-20%. Biochemical measurements of GST activities revealed no significant impact at any sampling time. In order to characterize the potential effect of MC-LR on MDRP, RT-qPCR method was established. However, as for GST activities no significant changes in MDRP gene expression have been observed. Thus, in summary, oral exposure of MC-LR containing cyanobacteria to Nile tilapia via feed ingestion did not impact significantly detoxification in liver concerning GST activities and MDRP expression despite biochemical composition concerning liver protein was significantly elevated by the diets containing 20% cyanobacteria biomass, regardless whether they contained MC-LR or not.
- MeSH
- Biomass MeSH
- Cichlids metabolism MeSH
- Diet MeSH
- Gene Expression drug effects MeSH
- Liver chemistry enzymology metabolism MeSH
- Inactivation, Metabolic MeSH
- Microcystins blood metabolism MeSH
- Tissue Distribution MeSH
- Animals MeSH
- Check Tag
- Animals MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
In this study, a simple and low-cost method to synthesize iron(III) oxide nanopowders in large quantity was successfully developed for the photocatalytic degradation of microcystin-LR (MC-LR). Two visible light-active iron(III) oxide samples (MG-9 calcined at 200 °C for 5 h and MG-11 calcined at 180 °C for 16 h) with a particle size of 5-20 nm were prepared via thermal decomposition of ferrous oxalate dihydrate in air without any other modifications such as doping. The synthesized samples were characterized by X-ray powder diffraction, 57Fe Mössbauer spectroscopy, transmission electron microscopy, Brunauer-Emmett-Teller (BET) specific surface area analysis, and UV-visible diffuse reflectance spectroscopy. The samples exhibited similar phase composition (a mixture of α-Fe2O3 and γ-Fe2O3), particle size distribution (5-20 nm), particle morphology, and degree of agglomeration, but different specific surface areas (234 m2 g-1 for MG-9 and 207 m2 g-1 for MG-11). The results confirmed higher photocatalytic activity of the catalyst with higher specific surface area. The highest photocatalytic activity of the sample to decompose MC-LR was observed at solution pH of 3.0 and catalyst loading of 0.5 g L-1 due to large amount of MC-LR adsorption, but a little iron dissolution of 0.0065 wt% was observed. However, no iron leaching was observed at pH 5.8 even though the overall MC-LR removal was slightly lower than at pH 3.0. Thus, the pH 5.8 could be an appropriate operating condition for the catalyst to avoid problems of iron contamination by the catalyst. Moreover, magnetic behavior of γ-Fe2O3 gives a possibility for an easy separation of the catalyst particles after their use.
