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 classification genetics isolation & purification growth & development MeSH
- Biodiversity * MeSH
- DNA, Bacterial genetics MeSH
- Eutrophication MeSH
- Phylogeny * MeSH
- Microcystis * growth & development classification genetics MeSH
- RNA, Ribosomal, 16S * genetics MeSH
- Fresh Water microbiology MeSH
- Temperature MeSH
- Publication type
- Journal Article MeSH
- Geographicals
- India MeSH
Our previous studies have shown that CyanoHAB LPS (lipopolysaccharides) and LPS from cyanobacterial cultures induce pro-inflammatory effects on intestinal epithelial and immune cells in vitro. To expand our understanding, we investigated their impact on human keratinocytes, which are targeted during water recreational activities. LPS samples were isolated from CyanoHAB biomasses dominated by Microcystis, Aphanizomenon, Planktothrix, and Dolichospermum, or from axenic cultures of these genera. We identified two CyanoHAB biomasses containing a high proportion of Gram-negative bacteria, including potentially pathogenic genera. These biomasses showed the highest induction of interleukin (IL) 8, IL-6, C-C motif chemokine ligand (CCL) 2 (also known as MCP-1), and CCL20 production by HaCaT cells. Interestingly, all CyanoHAB-derived LPS and LPS from axenic cultures (except for Microcystis) accelerated cell proliferation and migration. Our findings highlight the role of G- bacteria composition and LPS structural disparities in influencing these effects, with implications for skin health during recreational activities.
- MeSH
- Lakes MeSH
- Keratinocytes MeSH
- Skin MeSH
- Humans MeSH
- Lipopolysaccharides toxicity MeSH
- Microcystis * MeSH
- Cyanobacteria * chemistry MeSH
- Check Tag
- Humans MeSH
- Publication type
- Journal Article 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.
Retinoids are newly detected compounds in aquatic ecosystems associated with cyanobacterial water blooms. Their potential health risks are only scarcely described despite numerous detections of all-trans retinoic acid (ATRA) and its derivatives in the environment. Besides the known teratogen ATRA there is only little or no information about their potency and namely their effects in vivo. We characterize ATRA and 8 other retinoids reported to occur in the environment for their bioactivity and teratogenicity using four in vitro reporter gene assays and zebrafish (Danio rerio) embryotoxicity assay. Our results document the ability of these compounds to interfere with retinoid signalling and cause teratogenicity at environmentally relevant levels with EC50 values at nM (hundreds of ng/L) levels and teratogenic indexes ranging from 2.8 (9cis retinoic acid) to 15.8 (retinal). The relative potency of individual compounds for teratogenicity ranged from 0.059 (retinal) to 0.96 (5,6-epoxy ATRA) when compared to ATRA. An environmentally relevant mixture of retinoids was tested showing good predictability of teratogenicity from the in vitro activities and additive toxicity of the mixture. The high teratogenicity of the newly described compounds associated with cyanobacteria presents a concern for developmental stages due to high conservation of the retinoid signalling across vertebrates.
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
- Eutrophication MeSH
- Microcystis * MeSH
- Microcystins toxicity MeSH
- Cyanobacteria * MeSH
- Publication type
- Journal Article MeSH
Some phytoplankton species were shown to produce teratogenic retinoids. This study assessed the variability in the extracellular production of compounds with retinoid-like activity for 50 independent cultivations of wide spectra of species including 12 cyanobacteria (15 strains) and 4 algae of different orders. Extracellular retinoid-like activity was detected for repeated cultivations of six cyanobacteria. The results were consistent for some species including Microcystis aeruginosa and Aphanizomenon gracile. The detected retinoid-like activities ranged from below the limit of quantification of 16 ng/L to over 6 µg all-trans retinoic acid (ATRA) equivalent/L. Nontargeted virtual fractionation together with suspect screening approach enabled to identify some retinoid-like compounds in exudates, including ATRA, 9/13-cis retinoic acid, all-trans 5,6-epoxy retinoic acid, 4keto-ATRA, 4keto-retinal, 4hydroxy-ATRA, and retinal. Most of them were for the first time repeatedly detected in exudates of all studied algae (at ng/L levels) and cyanobacteria. Their relative potencies ranged from 0.018 (retinal) to 1 compared to ATRA. They accounted for less than 0.1-50% of total detected retinoid-like activity. The high detected activities and concentrations of retinoids in some samples and their direct accessibility from exudates document potential risk of developmental toxicity for organisms in proximity of massive water blooms.
- MeSH
- Aphanizomenon * MeSH
- Phytoplankton MeSH
- Microcystis * MeSH
- Retinoids MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
Cyanobacteria routinely release potentially harmful bioactive compounds into the aquatic environment. Several recent studies suggested a potential link between the teratogenicity of effects caused by cyanobacteria and production of retinoids. To investigate this relationship, we analysed the teratogenicity of field-collected cyanobacterial bloom samples by means of an in vivo zebrafish embryo test, an in vitro reporter gene bioassay and by the chemical analysis of retinoids. Extracts of biomass from cyanobacterial blooms with the dominance of Microcystis aeruginosa and Aphanizomenon klebahnii were collected from water bodies in the Czech Republic and showed significant retinoid-like activity in vitro, as well as high degrees of teratogenicity in vivo. Chemical analysis was then used to identify a set of retinoids in ng per gram of dry weight concentration range. Subsequent fractionation and bioassay-based characterization identified two fractions with significant in vitro retinoid-like activity. Moreover, in most of the retinoids eluted from these fractions, teratogenicity with malformations typical for retinoid signalling disruption was observed in zebrafish embryos after exposure to the total extracts and these in vitro effective fractions. The zebrafish embryo test proved to be a sensitive toxicity indicator of the biomass extracts, as the teratogenic effects occurred at even lower concentrations than those expected from the activity detected in vitro. In fact, teratogenicity with retinoid-like activity was detected at concentrations that are commonly found in biomasses and even in bulk water surrounding cyanobacterial blooms. Overall, these results provide evidence of a link between retinoid-like activity, teratogenicity and the retinoids produced by cyanobacterial water blooms in the surrounding environment.
- MeSH
- Aphanizomenon pathogenicity MeSH
- Zebrafish embryology genetics MeSH
- Embryo, Nonmammalian drug effects MeSH
- Microcystis pathogenicity MeSH
- Genes, Reporter MeSH
- Retinoids biosynthesis toxicity MeSH
- Cyanobacteria chemistry pathogenicity MeSH
- Teratogens toxicity MeSH
- Animals MeSH
- Check Tag
- Animals MeSH
- Publication type
- Journal Article MeSH
- Geographicals
- Czech Republic MeSH
Ozonation pretreatment is typically implemented to improve algal cell coagulation. However, knowledge on the effect of ozonation on the characteristics and coagulation of associated algal organic matter, particularly cellular organic matter (COM), which is extensively released during algal bloom decay, is limited. Hence, this study aimed to elucidate the impact of ozonation applied before the coagulation of dissolved COM from the cyanobacteria Microcystis aeruginosa. Additionally, the degradation of microcystins (MCs) naturally present in the COM matrix was investigated. A range of ozone doses (0.1-1.0 mg O3/mg of dissolved organic carbon - DOC) and ozonation pH values (pH 5, 7 and 9) were tested, while aluminium and ferric sulphate coagulants were used for subsequent coagulation. Despite negligible COM removal, ozonation itself eliminated MCs, and a lower ozone dose was required when performing ozonation at acidic or neutral pH (0.4 mg O3/mg DOC at pH 5 and 7 compared to 0.8 mg O3/mg DOC at pH 9). Enhanced MC degradation and a similar pattern of pH dependence were observed after preozonation-coagulation, whereas coagulation alone did not sufficiently remove MCs. In contrast to the benefits of MC depletion, preozonation using ≥ 0.4 mg O3/mg DOC decreased the coagulation efficiency (from 42%/48% to 28%-38%/41%-44% using Al/Fe-based coagulants), which was more severe with increasing ozone dosage. Coagulation was also influenced by the preozonation pH, where pH 9 caused the lowest reduction in COM removal. The results indicate that ozonation efficiently removes MCs, but its employment before COM coagulation is disputable due to the deterioration of coagulation.
- MeSH
- Water Purification * MeSH
- Microcystis * MeSH
- Ozone * MeSH
- Publication type
- Journal Article MeSH
Coagulation followed by floc separation is a key process for the removal of algal organic matter (AOM) in water treatment. Besides optimizing coagulation parameters, knowledge of the properties of AOM-flocs is essential to maximizing AOM removal. However, the impact of AOM on the floc properties remains unclear. This study investigated how peptides/proteins derived from the cellular organic matter (COM) of the cyanobacterium Microcystis aeruginosa influenced the size, structure, and shape of flocs formed at different shear rates (G). Flocs formed by kaolinite, COM-peptides/proteins and a mixture of the same were studied, and the effect of intermolecular interactions between floc components on floc properties was assessed. The coagulation experiments were performed in a Taylor-Couette reactor, with aluminum (Al) or ferric sulphate (Fe) utilized as coagulants. Image analysis was performed to gauge floc size and obtain data on fractal dimension. It was found that floc properties were affected by the presence of the COM-peptides/proteins and the coagulant used. COM-peptides/proteins increased floc size and porosity and widened floc size distributions. The Fe coagulant produced larger and less compact flocs than Al coagulant. Moreover, the decrease in floc size that occurred in parallel with increase in shear rate was not smooth in progress. A rapid change for the kaolinite-coagulant suspension and two rapid changes for the suspensions containing COM were observed. These were attributed to various intermolecular interactions between floc components participating in coagulation at different G. Based on the results obtained, shear rates suitable for efficient separation of flocs containing COM were suggested.
- MeSH
- Bacterial Proteins MeSH
- Water Purification methods MeSH
- Flocculation MeSH
- Microcystis * MeSH
- Water Microbiology * MeSH
- Peptides MeSH
- Publication type
- Journal Article MeSH
Physical floc properties were systematically investigated by analysing the structure of algal and cyanobacterial flocs produced by five species (green algae (Chlorella vulgaris) and cyanobacteria (Microcystis aeruginosa (strain CS-564), Microcystis aeruginosa (strain CS-555/01), Dolichospermum circinale and Cylindrospermopsis raciborskii) using aluminium sulphate (alum) at different doses and pH values. The properties of spherical, compact flocs were determined using a laser diffraction instrument and a new in situ image analysis technique was validated to analyse the structure of more complex flocs. The incorporation of algal-derived organic matter (AOM) into the flocs was inferred by evaluating the dissolved organic matter concentration character before and after flocculation using liquid chromatography with organic carbon detection (LC-OCD). D. circinale, C. raciborskii, and M. aeruginosa (CS-564) produced large flocs (2-9 mm), while M. aeruginosa (CS-555) and C. vulgaris produced smaller flocs (<2 mm). While differences in physical floc properties were observed to result from changes in coagulation mechanism, the cell morphology and the AOM composition were the most influential factors. Examination of floc properties can give a rapid insight at the plant for trouble shooting, particularly through the use of the in situ techniques and provide a mechanism by which floc properties can be tailored to downstream processes.
- MeSH
- Chlorella vulgaris * MeSH
- Water Purification * MeSH
- Flocculation MeSH
- Microcystis * MeSH
- Cyanobacteria * MeSH
- Publication type
- Journal Article MeSH
