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.

• Klíčová slova
• Blooms, Ecological stoichiometry, Ecological succession, Nutrients, Phytoplankton, Wind,
• 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
• Názvy látek
• dusík MeSH
• fosfor MeSH
• křemík MeSH

Stagnant freshwaters can be affected by anthropogenic pollution and eutrophication that leads to massive growth of cyanobacteria and microalgae forming complex water blooms. These can produce various types of bioactive compounds, some of which may cause embryotoxicity, teratogenicity, endocrine disruption and impair animal or human health. This study focused on potential co-occurrence of estrogenic and retinoid-like activities in diverse stagnant freshwaters affected by phytoplankton blooms with varying taxonomic composition. Samples of phytoplankton bloom biomass and its surrounding water were collected from 17 independent stagnant water bodies in the Czech Republic and Hungary. Total estrogenic equivalents (EEQ) of the most potent samples reached up to 4.9 ng·g-1 dry mass (dm) of biomass extract and 2.99 ng·L-1 in surrounding water. Retinoic acid equivalent (REQ) measured by in vitro assay reached up to 3043 ng·g-1 dm in phytoplankton biomass and 1202 ng·L-1in surrounding water. Retinoid-like and estrogenic activities at some sites exceeded their PNEC and effect-based trigger values, respectively. The observed effects were not associated with any particular species of cyanobacteria or algae dominating the water blooms nor related to phytoplankton density. We found that taxonomically diverse phytoplankton communities can produce and release retinoid-like compounds to surrounding water, while estrogenic potency is likely related to estrogens of anthropogenic origin adsorbed to phytoplankton biomass. Retinoids occurring in water blooms are ubiquitous signalling molecules, which can affect development and neurogenesis. Selected water bloom samples (both water and biomass extracts) with retinoid-like activity caused effects on neurodifferentiation in vitro corresponding to those of equivalent all-trans-retinoic acid concentrations. Co-occurrence of estrogenic and retinoid-like activities in stagnant water bodies as well as the potential of compounds produced by water blooms to interfere with neural differentiation should be considered in the assessment of risks associated with water blooms, which can comprise complex mixtures of natural and anthropogenic bioactive compounds.

• Klíčová slova
• Endocrine disruption, Estrogenic activity, Neural differentiation, Phytoplankton water bloom, Retinoid-like activity,
• MeSH
• estrogeny analýza   MeSH
• estron MeSH
• eutrofizace MeSH
• fytoplankton MeSH
• komplexní směsi MeSH
• lidé MeSH
• retinoidy * MeSH
• rostlinné extrakty MeSH
• sinice * MeSH
• tretinoin MeSH
• voda MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• estrogeny MeSH
• estron MeSH
• komplexní směsi MeSH
• retinoidy * MeSH
• rostlinné extrakty MeSH
• tretinoin MeSH
• voda MeSH

Stagnant water bodies have generally received little attention regarding the presence of endocrine disruptive compounds, although they can integrate diverse pollutants from multiple different sources. Many compounds of anthropogenic as well as natural origin can contribute to the overall estrogenicity of surface waters and some of them can exhibit adverse effects on aquatic biota even in very low concentrations. This study focused on freshwater ponds and reservoirs affected by water blooms and determined the estrogenic activity of water by in vitro bioassay as well as concentrations of several important groups of estrogenic compounds (estrogenic hormones, alkylphenols, and phytoestrogens) by LC-MS/MS analyses. Estrogenic hormones were found at concentrations up to 7.1 ng.L-1, similarly to flavonoids, whose concentrations did not exceed 12.5 ng.L-1. Among alkylphenols, only bisphenol A and 4-tert-octylphenol were detected in levels reaching 100 ng.L-1 at maximum. Estrogenic activity of water samples varied from below the quantification limit to 1.95 ng.L-1. There does not seem to be any general causal link of the massive phytoplankton occurrence with the estrogenicity of water or concentration of phytoestrogens, since they showed no direct relationship with the phytoplankton abundance or composition across sites. The contribution of the analysed compounds to the estrogenic activity was calculated in three scenarios. In minimum scenario, just the compounds above quantification limit (LOQ) were taken into account and for most samples, only minor part (<6%) of the biological activity could be explained. In the mean and maximum scenarios, we included also compounds below LOQ into the calculations at the level of LOQ/2 and LOQ, respectively. In these cases, a considerable part of the estrogenic activity could be attributed to the possible presence of steroid estrogens below LOQ. However, for the samples with estrogenic activity greater than 1 ng.L-1, more than 50% of the estrogenic activity remained unexplained even in the maximum scenario. Probably other compounds or possible interactions between individual substances cause the estrogenic activity in these types of water bodies and in this case, the results of LC-MS/MS analyses cannot sufficiently predict the biological effects. A complex approach including bioassays is needed when assessing the estrogenicity of these types of surface waters.

• Klíčová slova
• Alkylphenols, Estrogenicity, Estrogens, Flavonoids, Surface water,
• MeSH
• chemické látky znečišťující vodu analýza   metabolismus   MeSH
• chromatografie kapalinová MeSH
• endokrinní disruptory analýza   metabolismus   MeSH
• estrogeny analýza   metabolismus   MeSH
• fytoplankton chemie   metabolismus   MeSH
• sladká voda chemie   MeSH
• tandemová hmotnostní spektrometrie MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• chemické látky znečišťující vodu MeSH
• endokrinní disruptory MeSH
• estrogeny MeSH

Some freshwater phytoplankton species have been suggested to produce estrogenic compounds in concentrations which could cause adverse effects to aquatic biota, while other studies showed no estrogenic effects after exposure to phytoplankton extracts or pointed out possible sources of the overestimation of the estrogenic activity. This study aimed to clarify these research inconsistencies by investigating estrogenicity of biomass extracts from both environmental freshwater blooms and laboratory cyanobacterial and algae cultures by in vitro reporter bioassay. Biomasses of 8 cyanobacterial and 3 algal species from 7 taxonomic orders were extracted and tested. Next to this, samples of environmental water blooms collected from 8 independent water bodies dominated by phytoplankton species previously assessed as laboratory cultures were tested. The results showed undetectable or low estrogenicity of both freshwater blooms and laboratory cultures with E2 equivalent concentration (EEQ) in a range from LOQ up to 4.5 ng EEQ/g of dry mass. Moreover, the co-exposure of biomass extracts with environmentally relevant concentration of model estrogen (steroid hormone 17β-estradiol; E2), commonly occurring in surface waters, showed simple additive interaction. However, some of the biomass extracts elicited partially anti-estrogenic effects in co-exposure with higher E2 concentration. In conclusion, our study documents undetectable or relatively low estrogenic potential of biomass extracts from both environmental freshwater blooms and studied laboratory cultured cyanobacterial and algae species. Nevertheless, in case of very high-density water blooms, even this low estrogenicity (detected for two cyanobacterial species) could lead to EEQ content in biomass reaching effect-based trigger values indicating potential risk, if recalculated per water volume at field sites. However, these levels would not occur in water under realistic environmental scenarios and the potential estrogenic effects would be most probably minor compared to other toxic effects caused by massive freshwater blooms of such high densities.

• Klíčová slova
• Anti-Estrogenicity, Aquatic ecosystem, Cyanobacteria, Estrogen, Freshwater bloom, Phytoplankton,
• MeSH
• estradiol toxicita   MeSH
• estrogeny * toxicita   analýza   MeSH
• estron MeSH
• fytoplankton MeSH
• sinice * MeSH
• voda MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• estradiol MeSH
• estrogeny * MeSH
• estron MeSH
• voda MeSH

The release of phosphorus (P) from bed sediments to the overlying water can delay the recovery of lakes for decades following reductions in catchment contributions, preventing water quality targets being met within timeframes set out by environmental legislation (e.g. EU Water Framework Directive: WFD). Therefore supplementary solutions for restoring lakes have been explored, including the capping of sediment P sources using a lanthanum (La)-modified bentonite clay to reduce internal P loading and enhance the recovery process. Here we present results from Loch Flemington where the first long-term field trial documenting responses of phytoplankton community structure and abundance, and the UK WFD phytoplankton metric to a La-bentonite application was performed. A Before-After-Control-Impact (BACI) analysis was used to distinguish natural variability from treatment effect and confirmed significant reductions in the magnitude of summer cyanobacterial blooms in Loch Flemington, relative to the control site, following La-bentonite application. However this initial cyanobacterial response was not sustained beyond two years after application, which implied that the reduction in internal P loading was short-lived; several possible explanations for this are discussed. One reason is that this ecological quality indicator is sensitive to inter-annual variability in weather patterns, particularly summer rainfall and water temperature. Over the monitoring period, the phytoplankton community structure of Loch Flemington became less dominated by cyanobacteria and more functionally diverse. This resulted in continual improvements in the phytoplankton compositional and abundance metrics, which were not observed at the control site, and may suggest an ecological response to the sustained reduction in filterable reactive phosphorus (FRP) concentration following La-bentonite application. Overall, phytoplankton classification indicated that the lake moved from poor to moderate ecological status but did not reach the proxy water quality target (i.e. WFD Good Ecological Status) within four years of the application. As for many other shallow lakes, the effective control of internal P loading in Loch Flemington will require further implementation of both in-lake and catchment-based measures. Our work emphasizes the need for appropriate experimental design and long-term monitoring programmes, to ascertain the efficacy of intervention measures in delivering environmental improvements at the field scale.

• Klíčová slova
• Cyanobacteria, Eutrophication, Lake restoration, Lanthanum-modified bentonite, Recovery,
• MeSH
• bentonit chemie   MeSH
• eutrofizace MeSH
• fosfor MeSH
• fytoplankton * MeSH
• jezera chemie   MeSH
• lanthan chemie   MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• bentonit MeSH
• fosfor MeSH
• lanthan MeSH

Biological processes tend to dominate the oxygen regime of productive waters. However, in shallow aquatic ecosystems, it is unclear whether the oxygen regime is driven by oxygen production and consumption in the water column or by sediment oxygen demand (SOD). In managed eutrophic ecosystems, this question is especially important in the context of extreme daily oscillations of dissolved oxygen (DO) that could breach physiological limits of heterotrophic aerobic organisms. High-frequency measurement of DO, temperature, global radiation (Gl.Rad.), and pH in a 0.6 m deep, 22 ha eutrophic fishpond Rod (Czech Republic) shows that the oxygen regime depended on the ecosystem state. Over the clearwater period in the early season, the DO level reflected ecosystem heterotrophy with relatively low daily DO oscillations. However, during the summer phytoplankton bloom, the fishpond was primarily autotrophic with extreme DO fluctuation. During late summer, a collapse of the phytoplankton bloom and an associated shift towards heterotrophy and DO deficit frequently occur. In-situ mesocosm experiments in Rod fishpond were conducted throughout 2018 and 2019 growing seasons, to address the importance of SOD to the oxygen regime. We enclosed the water column in transparent and opaque/dark plastic cylinders open or closed to the sediment. The results show that the proportional contribution of SOD to total respiration decreased from 70 to 90% at low phytoplankton biomass (expressed as Chlorophyll-a (Chl-a) concentration) to approximately 10% at phytoplankton bloom. At night, the difference between the oxygen consumption in the cylinders with or without sediment was statistically significant, when the concentration of Chl-a was <100 μg·L-1. On the contrary, the difference was not significant when the concentration of Chl-a was >100 μg·L-1. This revealed that the impact of SOD is negligible at high phytoplankton biomass.

• Klíčová slova
• Dissolved oxygen, Eutrophic water, Fishpond, Oxygen deficit, Sediment oxygen demand, Shallow aquatic ecosystem,
• MeSH
• biomasa MeSH
• dýchání MeSH
• ekosystém * MeSH
• eutrofizace MeSH
• fytoplankton MeSH
• kyslík * analýza   MeSH
• roční období MeSH
• voda MeSH
• Publikační typ
• časopisecké články MeSH
• Geografické názvy
• Česká republika MeSH
• Názvy látek
• kyslík * MeSH
• voda MeSH

Standard ISO method for chlorophyll a quantification (extraction into ethanol, spectrophotometrical quantification at 665 and 750 nm), spectrofluorometry (reader for 96 wells, excitation 410 nm, emission 670 nm), and a submersible fluorescence probe for in situ phytoplankton quantification (excitation 410, 525, 570, 590, and 610 nm, emission 685 nm) were compared in different freshwater environments-reservoirs and rivers. The ISO method is accepted as a standard method but requires sample handling and transport to the laboratory. Spectrofluorometry is a sensitive method, even for natural phytoplankton populations. Nevertheless, it cannot be recommended for the quantification of cyanobacterial water blooms because colonial and filamentous species such as Microcystis, Anabaena, or Aphanizomenon display unacceptable variability (18-33%). The submersible probe featured high correlation with a standard ISO method (r=0.97, P<0.05). This probe can provide the selective measurement of technologically important phytoplankton groups like cyanobacteria, diatoms, green algae, and cryptophytes in lake vertical profiles of up to 100 m. The limitation of this instrument is the possible reabsorption of the light signal, e.g. in the presence of humic substances, or dense algal blooms. The use of submersible probes for in situ phytoplankton quantification can be recommended as a sensitive tool for water management, especially in the case of drinking water resources.

• MeSH
• chlorofyl a MeSH
• chlorofyl analýza   MeSH
• eutrofizace * MeSH
• fluorescence MeSH
• fytoplankton chemie   MeSH
• monitorování životního prostředí přístrojové vybavení   metody   MeSH
• populační dynamika MeSH
• řeky MeSH
• senzitivita a specificita MeSH
• zásobování vodou MeSH
• Publikační typ
• časopisecké články MeSH
• hodnotící studie MeSH
• srovnávací studie MeSH
• Názvy látek
• chlorofyl a MeSH
• chlorofyl MeSH

BACKGROUND: The phytoplankton spring bloom in freshwater habitats is a complex, recurring, and dynamic ecological spectacle that unfolds at multiple biological scales. Although enormous taxonomic shifts in microbial assemblages during and after the bloom have been reported, genomic information on the microbial community of the spring bloom remains scarce. RESULTS: We performed a high-resolution spatio-temporal sampling of the spring bloom in a freshwater reservoir and describe a multitude of previously unknown taxa using metagenome-assembled genomes of eukaryotes, prokaryotes, and viruses in combination with a broad array of methodologies. The recovered genomes reveal multiple distributional dynamics for several bacterial groups with progressively increasing stratification. Analyses of abundances of metagenome-assembled genomes in concert with CARD-FISH revealed remarkably similar in situ doubling time estimates for dominant genome-streamlined microbial lineages. Discordance between quantitations of cryptophytes arising from sequence data and microscopic identification suggested the presence of hidden, yet extremely abundant aplastidic cryptophytes that were confirmed by CARD-FISH analyses. Aplastidic cryptophytes are prevalent throughout the water column but have never been considered in prior models of plankton dynamics. We also recovered the first metagenomic-assembled genomes of freshwater protists (a diatom and a haptophyte) along with thousands of giant viral genomic contigs, some of which appeared similar to viruses infecting haptophytes but owing to lack of known representatives, most remained without any indication of their hosts. The contrasting distribution of giant viruses that are present in the entire water column to that of parasitic perkinsids residing largely in deeper waters allows us to propose giant viruses as the biological agents of top-down control and bloom collapse, likely in combination with bottom-up factors like a nutrient limitation. CONCLUSION: We reconstructed thousands of genomes of microbes and viruses from a freshwater spring bloom and show that such large-scale genome recovery allows tracking of planktonic succession in great detail. However, integration of metagenomic information with other methodologies (e.g., microscopy, CARD-FISH) remains critical to reveal diverse phenomena (e.g., distributional patterns, in situ doubling times) and novel participants (e.g., aplastidic cryptophytes) and to further refine existing ecological models (e.g., factors affecting bloom collapse). This work provides a genomic foundation for future approaches towards a fine-scale characterization of the organisms in relation to the rapidly changing environment during the course of the freshwater spring bloom. Video Abstract.

• MeSH
• Bacteria MeSH
• Eukaryota genetika   MeSH
• metagenom * MeSH
• plankton MeSH
• sladká voda MeSH
• viry * genetika   MeSH
• voda MeSH
• Publikační typ
• audiovizuální média MeSH
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• voda 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.

• MeSH
• biomasa MeSH
• cyklické peptidy metabolismus   MeSH
• eutrofizace * MeSH
• mikrobiologie vody * MeSH
• mikrocystiny MeSH
• monitorování životního prostředí * MeSH
• populační dynamika MeSH
• roční období MeSH
• sinice růst a vývoj   MeSH
• sladká voda chemie   mikrobiologie   MeSH
• zásobování vodou MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Geografické názvy
• Česká republika MeSH
• Názvy látek
• cyklické peptidy MeSH
• microcystin MeSH Prohlížeč
• mikrocystiny MeSH

In vivo fluorescence methods have been accepted as a quick, simple, and useful tool for quantification of phytoplankton organisms. In this paper, we present a case study in which fluorescence methods were employed for the selective detection of potentially toxic cyanobacteria in raw water at the drinking water treatment plant. The occurrence of cyanobacteria in the drinking water reservoir and in raw water was monitored by phycocyanin fluorescence measurements and by standard methods for phytoplankton quantification (cell counts, chlorophyll a). A special attention was paid to the most critical parts of the season -- spring recruitment of cyanobacteria from sediment to water column and autumn bloom collapse. All methods showed similar patterns within the season. Phycocyanin fluorescence was found to be a simple and sensitive indicator of cyanobacteria in water and can serve as a tool that can provide an early warning about the presence of potentially toxic cyanobacterial metabolites in water.

• MeSH
• fluorescence * MeSH
• fykokyanin chemie   MeSH
• mikrobiologie vody * MeSH
• sinice izolace a purifikace   MeSH
• voda MeSH
• zásobování vodou analýza   MeSH
• Publikační typ
• časopisecké články MeSH
• hodnotící studie MeSH
• práce podpořená grantem MeSH
• Názvy látek
• fykokyanin MeSH
• voda MeSH