Cyanobacterial harmful blooms (CyanoHABs) pose a global ecological problem, and their lipopolysaccharides (LPS) are among the bioactive compounds they release. Previous studies on CyanoHAB-LPS from single cyanobacterial species have shown varying bioactivities in different in vitro cell models. In this study, we isolated LPS from 19 CyanoHAB samples collected at 18 water bodies in the Czech Republic over two consecutive seasons. The proportions of cyanobacteria, Gram-negative bacteria (G-), and other bacteria in the biomass were determined by qPCR, while the cyanobacterial genera were identified using light microscopy. In vitro models of keratinocytes (HaCaT), the intestinal epithelium (co-culture of differentiated Caco-2 cells and peripheral blood mononuclear cells - PBMC), and PBMC alone were treated with isolated LPS at concentrations of 50, 100, and 1 μg/ml, respectively. The endotoxin activities of these concentrations were within the range measured in the aquatic environment. Approximately 85-90% of the samples displayed biological activity. However, the potency of individual LPS effects and response patterns varied across the different in vitro models. Furthermore, the observed activities did not exhibit a clear correlation with the taxonomic composition of the phytoplankton community, the relative share of microbial groups in the biomass, endotoxin activity of the LPS, or LPS migration and staining pattern in SDS-PAGE. These findings suggest that the effects of CyanoHAB-LPS depend on the specific composition and abundance of various LPS structures within the complex environmental sample and their interactions with cellular receptors.
- MeSH
- biomasa MeSH
- Caco-2 buňky MeSH
- leukocyty mononukleární MeSH
- lidé MeSH
- lipopolysacharidy * toxicita MeSH
- sinice * MeSH
- škodlivý vodní květ MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
The occurrence of toxic bloom-forming cyanobacteria, Microcystis aeruginosa, has been frequently reported worldwide. These colony forming toxic cyanobacteria harbour a wide range of heterotrophic bacterial communities. The present study has attempted to understand the bloom dynamics of M. aeruginosa along with isolating their colony-associated culturable heterotrophic bacteria from two freshwater ponds in south India with a persisting cyanobacterial bloom. The monthly monitoring of these study areas revealed the conducive role of warm, stagnant waters with high nutrients in forming M. aeruginosa bloom. The peak values of temperature, nitrate, and phosphate at station 1 reached up to 30.5 °C, 4.48 mg/L, 1.64 mg/L, and at station 2, 31 °C, 3.45 mg/L, and 0.62 mg/L, respectively. Twenty-eight bacterial isolates belonging to Alphaproteobacteria, Betaproteobacteria, Gammaproteobacteria, Actinobacteria, and Firmicutes were obtained during the study. Among these 28 isolates, Firmicutes was dominant with the M. aeruginosa bloom from both the study areas.
- MeSH
- Bacteria klasifikace genetika izolace a purifikace růst a vývoj MeSH
- biodiverzita * MeSH
- DNA bakterií genetika MeSH
- eutrofizace MeSH
- fylogeneze * MeSH
- Microcystis * růst a vývoj klasifikace genetika MeSH
- RNA ribozomální 16S * genetika MeSH
- sladká voda mikrobiologie MeSH
- teplota MeSH
- Publikační typ
- časopisecké články MeSH
- Geografické názvy
- Indie MeSH
Freshwater cyanobacterial harmful blooms (CyanoHABs) produce a variety of toxic and bioactive compounds including lipopolysaccharides (LPSs). The gastrointestinal tract can be exposed to them via contaminated water even during recreational activities. However, there is no evidence of an effect of CyanoHAB LPSs on intestinal cells. We isolated LPSs of four CyanoHABs dominated by different cyanobacterial species and LPSs of four laboratory cultures representing the respective dominant cyanobacterial genera. Two intestinal and one macrophage cell lines were used to detect in vitro pro-inflammatory activity of the LPS. All LPSs isolated from CyanoHABs and laboratory cultures induced cytokines production in at least one in vitro model, except for LPSs from the Microcystis PCC7806 culture. LPSs isolated from cyanobacteria showed unique migration patterns in SDS-PAGE that were qualitatively distinct from those of endotoxins from Gram-negative bacteria. There was no clear relationship between the biological activity of the LPS and the share of genomic DNA of Gram-negative bacteria in the respective biomass. Thus, the total share of Gram-negative bacteria, or the presence of Escherichia coli-like LPSs, did not explain the observed pro-inflammatory activities. The pro-inflammatory properties of environmental mixtures of LPSs from CyanoHABs indicate their human health hazards, and further attention should be given to their assessment and monitoring.
Secondary metabolites of cyanobacteria and algae released during algal blooms often exhibit toxic effects, but only a small number of the metabolites are the subject of routine analytical screenings. Alternatively, ecotoxicological assays offer a better representation of the overall negative effects. The aim of this work was to compare multiple assays in their sensitivity towards cellular algal organic matter (COM) of the toxin-producing cyanobacterium Microcystis aeruginosa. Multiple endpoints were investigated: mortality, growth inhibition, bioluminescence inhibition, genotoxicity, endocrine-disrupting effects, oxidative stress, and the induction of ethoxyresorufin-O-deethylase (EROD). Three rainbow trout (Oncorhynchus mykiss) cell lines as well as representatives of bacteria, yeasts, algae, vascular plants, and crustaceans were employed, and the results were expressed per mg of dissolved organic carbon (DOC) in the COM. M. aeruginosa COM was toxic to the RTgill-W1, RTG-2, and RTL-W1 cell lines (EC50 values ranging from 0.48 ± 0.02 to 1.9 ± 0.1 mgDOC/L), to the crustacean Thamnocephalus platyurus (LC50 = 20 ± 1 mgDOC/L), and to Lepidium sativum (IC50 = 241 ± 13 mgDOC/L). In contrast, no effect was observed for bacteria and yeasts, and the growth of the alga Desmodesmus subspicatus was even stimulated. No genotoxicity, endocrine-disrupting effects or increase in oxidative stress or EROD activity was detected. The content of six microcystins (MC-LR, MC-RR, MC-YR, MC-LY, MC-LW, and MC-LF), anatoxin-a, cylindrospermopsin, and nodularin in the M. aeruginosa COM was determined by liquid chromatography-tandem mass spectrometry. An artificially prepared mixture of the detected cyanotoxins in the corresponding concentrations did not induce response in the O. mykiss cell lines and T. platyurus, suggesting that other cyanobacterial metabolites are responsible for the toxicity of M. aeruginosa.
- MeSH
- eutrofizace MeSH
- Microcystis * MeSH
- mikrocystiny toxicita MeSH
- sinice * MeSH
- Publikační typ
- časopisecké články MeSH
Man-made shallow fishponds in the Czech Republic have been facing high eutrophication since the 1950s. Anthropogenic eutrophication and feeding of fish have strongly affected the physicochemical properties of water and its aquatic community composition, leading to harmful algal bloom formation. In our current study, we characterized the phytoplankton community across three eutrophic ponds to assess the phytoplankton dynamics during the vegetation season. We microscopically identified and quantified 29 cyanobacterial taxa comprising non-toxigenic and toxigenic species. Further, a detailed cyanopeptides (CNPs) profiling was performed using molecular networking analysis of liquid chromatography-tandem mass spectrometry (LC-MS/MS) data coupled with a dereplication strategy. This MS networking approach, coupled with dereplication, on the online global natural product social networking (GNPS) web platform led us to putatively identify forty CNPs: fourteen anabaenopeptins, ten microcystins, five cyanopeptolins, six microginins, two cyanobactins, a dipeptide radiosumin, a cyclooctapeptide planktocyclin, and epidolastatin 12. We applied the binary logistic regression to estimate the CNPs producers by correlating the GNPS data with the species abundance. The usage of the GNPS web platform proved a valuable approach for the rapid and simultaneous detection of a large number of peptides and rapid risk assessments for harmful blooms.
- MeSH
- bakteriální toxiny analýza toxicita MeSH
- hmotnostní spektrometrie s elektrosprejovou ionizací * MeSH
- hodnocení rizik MeSH
- metabolomika MeSH
- mikrobiologie vody MeSH
- mikrobiota MeSH
- monitorování životního prostředí * MeSH
- mořské toxiny analýza toxicita MeSH
- online sociální sítě * MeSH
- populační dynamika MeSH
- roční období MeSH
- rybníky mikrobiologie MeSH
- sinice klasifikace růst a vývoj metabolismus MeSH
- škodlivý vodní květ * MeSH
- tandemová hmotnostní spektrometrie * MeSH
- vysokoúčinná kapalinová chromatografie * MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Geografické názvy
- Česká republika MeSH
Anthropogenic eutrophication of freshwater bodies increases the occurrence of toxic cyanobacterial blooms. The cyanobacterial toxin cylindrospermopsin (CYN) is detected in the environment with increasing frequency, driving the scientific effort to assess emerging health risks from CYN-producing blooms. Oral exposure to CYN results primarily in hepatotoxicity. Nevertheless, extrahepatic manifestations of CYN toxicity have been reported. Furthermore, cyanotoxins have been detected in aerosols and dust particles, suggesting potential toxic effects in the respiratory tract. To assess the susceptibility of airway epithelia towards cyanotoxins, monolayers of immortalized human bronchial epithelial cells HBE1 and 16HBE14o- were exposed to a concentration range of 0.1-10 μM CYN. Cytotoxic endpoints were assessed as morphologic alterations, resazurin reduction capacity, esterase activity, neutral red uptake, and by impedimetric real-time cell analysis. Depending on the endpoint assessed, EC50 values ranged between 0.7 and 1.8 μM (HBE1) and 1.6-4.8 μM (16HBE14o-). To evaluate alterations of other cellular events by subcytotoxic concentration of CYN (1 μM), phosphorylation of mitogen-activated protein kinases ERK and p38 was determined. Only a slight increase in p38 phosphorylation was induced by CYN in HBE1 cell line after 48 h, while activities of both ERK1/2 and p38 gradually and significantly increased in 16HBE14o- cells during 8-48 h exposure. This study suggests possible hazards of inhalation CYN exposures, which may severely impact the integrity of airway epithelia and epithelial cell signaling. Further research of CYN-induced toxicity and underlying mechanisms is needed, as well as more data on environmental concentrations of cyanotoxins in aerosols for exposure assessment.
- MeSH
- bakteriální toxiny farmakologie MeSH
- buněčné linie MeSH
- dýchací soustava cytologie MeSH
- epitelové buňky účinky léků MeSH
- eutrofizace * MeSH
- lidé MeSH
- mikrocystiny farmakologie MeSH
- mořské toxiny farmakologie MeSH
- uracil analogy a deriváty farmakologie MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
In order to understand of the processes controlling phytoplankton successions in the NE Black Sea, long-term data series are needed. We compiled 15 years (2002-2017) of measurements from which the existence emerges of a tight link between phytoplankton species dominance and nutrients concentrations. The latter is strongly influenced by wind direction. The link between algal dominance and nutrients is mediated by the growth strategy adopted by algal species. In spring, when nutrients are abundant, small diatoms such as Pseudo-nitzschia pseudodelicatissima, with a "rapid growth strategy", prevail. In late spring and early summer, when N is low and P and Si are high, coccolithophorids such as Emiliania huxhleyi dominate, thanks to an "affinity growth strategy". Large diatoms, especially Pseudosolenia calcar-avis, dominate in summer and autumn, when their "storage growth strategy" allows the exploitation of discontinuous upwelling of nutrients. These seasonal changes of dominant species influence the structure of the food web.
- MeSH
- dusík analýza MeSH
- eutrofizace * MeSH
- fosfor analýza MeSH
- fytoplankton růst a vývoj MeSH
- křemík analýza MeSH
- monitorování životního prostředí MeSH
- mořská voda chemie MeSH
- pohyb vody MeSH
- roční období MeSH
- rozsivky růst a vývoj MeSH
- vítr MeSH
- živiny MeSH
- Publikační typ
- časopisecké články MeSH
- Geografické názvy
- Černé moře MeSH
Massive toxic blooms of cyanobacteria represent a major threat to water supplies worldwide. Here, the biological activities of lipopolysaccharide (LPS) isolated from Microcystis aeruginosa, the most prominent cyanobacteria in water bloom, were studied. LPS was isolated from complex environmental water bloom samples dominated by M. aeruginosa, and from laboratory cultures of non-axenic as well as axenic M. aeruginosa strains PCC7806 and HAMBI/UHCC130. Employing human blood-based in vitro tests, the LPS isolated from complex water bloom revealed the priming of both major blood phagocyte population monocytes and polymorphonuclear leukocytes documented by the increased surface expression of CD11b and CD66b. This was accompanied by a water bloom LPS-mediated dose-dependent induction of tumor necrosis factor α, interleukin-1β, and interleukin-6 production. In accordance with its priming effects, water bloom LPS induced significant activation of p38 and ERK1/2 kinases, as well as NF-κB phosphorylation, in isolated polymorphonuclear leukocytes. Interestingly, the pro-inflammatory potential of LPS from the axenic strain of M. aeruginosa was not lower compared to that of LPS isolated from non-axenic strains. In contrast to the biological activity, water bloom LPS revealed almost twice higher pyrogenicity levels compared to Escherichia coli LPS, as analyzed by the PyroGene test. Moreover, LPS from the non-axenic culture exhibited higher endotoxin activity in comparison to LPS from axenic strains. Taking the above findings together, M. aeruginosa LPS can contribute to the health risks associated with contamination by complex water bloom mass.
- MeSH
- antigeny CD11b metabolismus MeSH
- CD antigeny metabolismus MeSH
- cytokiny krev MeSH
- eutrofizace MeSH
- GPI-vázané proteiny metabolismus MeSH
- kultivované buňky MeSH
- laboratoře MeSH
- leukocyty mononukleární účinky léků metabolismus MeSH
- lidé MeSH
- lipopolysacharidy toxicita MeSH
- Microcystis * MeSH
- molekuly buněčné adheze metabolismus MeSH
- přirozená imunita účinky léků MeSH
- pyrogeny toxicita MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
The loss of biodiversity is thought to have adverse effects on multiple ecosystem functions, including the decline of community stability. Decreased diversity reduces the strength of the portfolio effect, a mechanism stabilizing community temporal fluctuations. Community stability is also expected to decrease with greater variability in individual species populations and with synchrony of their fluctuations. In semi-natural meadows, eutrophication is one of the most important drivers of diversity decline; it is expected to increase species fluctuations and synchrony among them, all effects leading to lower community stability. With a 16-year time series of biomass data from a temperate species-rich meadow with fertilization and removal of the dominant species, we assessed population biomass temporal (co)variation under different management types and competition intensity, and in relation to species functional traits and to species diversity. Whereas the effect of dominant removal was relatively small (with a tendency toward lower stability), fertilization markedly decreased community stability (i.e., increased coefficient of variation in the total biomass) and species diversity. On average, the fluctuations of individual populations were mutually independent, with a slight tendency toward synchrony in unfertilized plots, and a tendency toward compensatory dynamics in fertilized plots and no effects of removal. The marked decrease of synchrony with fertilization, contrary to the majority of the results reported previously, follows the predictions of increased compensatory dynamics with increased asymmetric competition for light in a more productive environment. Synchrony increased also with species functional similarity stressing the importance of shared ecological strategies in driving similar species responses to weather fluctuations. As expected, the decrease of temporal stability of total biomass was mainly related to the decrease of species richness, with its effect remaining significant also after accounting for fertilization. The weakening of the portfolio effect with species richness decline is a crucial driver of community destabilization. However, the positive effect of species richness on temporal stability of total biomass was not due to increased compensatory dynamics, since synchrony increased with species richness. This shows that the negative effect of eutrophication on community stability does not operate through increasing synchrony, but through the reduction of diversity.
- MeSH
- biodiverzita * MeSH
- biomasa MeSH
- ekologie MeSH
- ekosystém * MeSH
- eutrofizace MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Cyanobacterial species produce wide range of bioactive compounds. This study characterized production of retinoid-like compounds with embryotoxic and teratogenic potential by commonly occurring cyanobacterial species with tendency to form massive water blooms. The major goal was to simultaneously assess the intracellular and extracellular retinoid-like activity from several independent cultivations of one coccal (Microcystis aeruginosa) and four filamentous cyanobacteria (Aphanizomenon gracile, Cylindrospermopsis raciborskii, Limnothrix redekeii, and Planktothrix agardhii) and characterize the variability in its production among cultivations. The retinoid-like activity was evaluated by in vitro assay along with chemical analyses of nine retinoids: all-trans retinoic acid (ATRA), 9-cis retinoic acid (9cis-RA), 13cis-RA, 13cis-RA methyl ester, 5,6 epoxy-RA, 4keto-ATRA, 4keto-retinal, 4hydoxy-retinoic acid (4OH-ATRA), retinal and retinol. The production of retinoid-like compounds was recalculated per volume, per biomass dry weight and per cell to provide relevant data for risk assessment in relation to occurrence of massive water blooms in the environment. Total produced retinoid-like activity of five selected species ranged from 170 to 25,600ng ATRA-equivalents (REQ)/g dm corresponding to 0.001-0.392ng REQ/106 cyanobacterial cells. Results from chemical analyses showed that all tested extracts contained 4keto-ATRA and retinal. All-trans retinoic acid, 9/13cis-retinoic acid and 5,6 epoxy-retinoic acid were detected in most exudate and extract samples. The reported results of recalculated total retinoid-like activity enable potential predictions of its production by the studied species in water blooms of known cell densities relevant for risk assessment.
