Daniel Jančula

Multimodal Action and Selective Toxicity of Zerovalent Iron Nanoparticles against Cyanobacteria

Cyanobacteria pose a serious threat to water resources around the world. This is compounded by the fact that they are extremely resilient, having evolved numerous protective mechanisms to ensure their dominant position in their ecosystem. We show that treatment with nanoparticles of zerovalent iron (nZVI) is an effective and environmentally benign method for destroying and preventing the formation of cyanobacterial water blooms. The nanoparticles have multiple modes of action, including the removal of bioavailable phosphorus, the destruction of cyanobacterial cells, and the immobilization of microcystins, preventing their release into the water column. Ecotoxicological experiments showed that nZVI is a highly selective agent, having an EC(50) of 50 mg/L against cyanobacteria; this is 20-100 times lower than its EC(50) for algae, daphnids, water plants, and fishes. The primary product of nZVI treatment is nontoxic and highly aggregated Fe(OH)(3), which promotes flocculation and gradual settling of the decomposed cyanobacterial biomass.

Metabolic activity and membrane integrity changes in Microcystis aeruginosa – new findings on hydrogen peroxide toxicity in cyanobacteria

The aim of our study was to investigate the intracellular toxicity mechanisms of the photoactive, potentially anti-cyanobacterial agent hydrogen peroxide (H2O2) in Microcystis aeruginosa, which represents one of the most significant toxin-producing cyanobacterial species in European water bodies. Metabolic activity and cell membrane integrity were evaluated by flow cytometry in cyanobacteria exposed to H2O2 in the dark or light; the relationships between exposure effects and the kinetics of hydrogen peroxide decomposition were studied. In the light (irradiance 140 µmol m−2 s−1), cyanobacteria were exposed to initial H2O2 concentrations of 0.00 (control), 0.75, 2.00, and 4.00 mg l−1 respectively, while in the dark concentrations were ten times higher. Flow cytometry and chlorophyll a fluorescence measurements suggested that hydrogen peroxide exposure elicits an immediate decline of metabolic (esterase) activity, measured as a decrease in fluorescein fluorescence after hydrolysis of fluorescein diacetate (FDA), and immediate changes of chlorophyll a fluorescence parameters, followed later by an increase in the percentage of membrane-compromised (SYTOX Green positive) cells. When the concentration of H2O2 used was lethal (in the two highly exposed treatments in the light), a significant drop in total cell counts was detected, whereas in other treatments no drop was observed during the entire experimental period (72 h). Our study also confirmed that light is one of the critical factors affecting H2O2 decomposition and thus greatly influences its toxicity. Whereas in the light, M. aeruginosa exposed to 0.75 mg l−1 H2O2 recovered after all the H2O2 had decomposed, in the dark H2O2 decomposed relatively slowly and its toxic effects on the cyanobacteria were observed over the whole 72-h period, though without cell lysis in any experimental concentration.

Algicidal activity of phthalocyanines—Screening of 31 compounds

Phthalocyanines and their analogues show great potential as photodynamic agents producing reactive oxygen species (ROS), especially in medicine. However, their biocidal effects may also be employed to inhibit various undesirable organisms. This study explores their potential algicidal effects. The laboratory tests concern the effects of various phthalocyanine derivatives on the green alga Pseudokirchneriella subcapitata and cyanobacterium Synechococcus nidulans. Their effects on one example of the sensitive nontarget aquatic organism—crustacean Daphnia magna were also screened. Among 31 tested compounds, the cationic phthalocyanines substituted with heterocycle exhibited the strongest effects on phytoplankton species, some of them even below the level of 1 mg/L, while effects on crustaceans ranged from 3.6 to more than 50 mg/L. These results show that some phthalocyanine derivatives can act as potent algicides.

Controlling internal phosphorus loading in lakes by physical methods to reduce cyanobacterial blooms: a review

The aim of this paper was to summarize the current knowledge on how physical methods can reduce or control internal P release from sediments in lakes and reservoirs. Particular emphasis is given to the role of internal phosphorus load in fueling cyanobacterial blooms which are predicted to increase in frequency and intensity in response to climate change and eutrophication. We present selective case studies (both successful and unsuccessful) to assess the applicability and efficiency of major physical approaches used for decades to reduce internal loading in different systems of various morphology. In particular, we concentrate on where and when (1) hypolimnetic aeration/oxygenation, (2) hypolimnetic withdrawal and (3) sediment dredging are likely to reduce cyanobacterial blooms and whether these methods have an adverse impact on other organisms. We conclude that each method has its strength and weakness depending on the system considered. Sufficient knowledge of all lake nutrient sources and their dynamics together with detailed lake and sediment characteristics is an essential prerequisite for choosing an appropriate control method. We also report that many experiences demonstrated that a combination of restoration methods is often more successful than a single method.

The toxicity of phthalocyanines to the aquatic plant Lemna minor (duckweed) – Testing of 31 compounds

Phthalocyanines are prospective chemicals that have applications in industry, medicine and biology due especially to their architectural flexibility and production of reactive oxygen species. Although they are used in so many areas of human activities nowadays, there is still little knowledge of their ecotoxicity. Here we present the first observation of their toxic effects on representatives of the aquatic plants Lemna minor. The tested phthalocyanines possess a wide spectrum of phytotoxicity ranging from seldom (>50 mg L(-1)) to highly toxic 0.11 mg L(-1). Moreover, the potential of phthalocyanines to be used as selective cyanocides or herbicides is discussed as well.

Existing and emerging cyanocidal compounds : New perspectives for cyanobacterial bloom mitigation

To help ban the use of general toxic algicides, research efforts are now directed towards the discovery of compounds that are specifically acting as cyanocides. Here, we review the past and look forward into the future, where the less desirable general algicides like copper sulphate, diuron or endothall may become replaced by compounds that show better specificity for cyanobacteria and are biodegradable or transform into non-toxic products after application. For a range of products, we review the activity, the mode of action, effectiveness, durability, toxicity towards non-target species, plus costs involved, and discuss the experience with and prospects for small water volume interventions up to the mitigation of entire lakes; we arrive at recommendations for a series of natural products and extracted organic compounds or derived synthetic homologues with promising cyanocidal properties, and briefly mention emerging nanoparticle applications. Finally, we detail on the recently introduced application of hydrogen peroxide for the selective killing of cyanobacteria in freshwater lakes.

Evaluation of antibacterial properties of novel phthalocyanines against Escherichia coli--comparison of analytical methods.

We analyzed antibacterial effects of several novel phthalocyanines against Escherichia coli and evaluated the suitability of flow cytometry for the detection of antibacterial effects of phthalocyanines in comparison with routinely used cultivation. After 3h of exposure under cool white light eight cationic phthalocyanines showed very high antibacterial activity in the concentration of 2.00 mg L(-1) and four of them were even efficient in the concentration of 0.20 mg L(-1). Antibacterial activity of neutral and anionic compounds was considerably lower or even negligible. No antibacterial effect was detected when bacteria were exposed without illumination. Binding affinity to bacterial cells was found to represent an important parameter influencing phthalocyanine antibacterial activity that can be modified by total charge of peripheral substituents and by the presence of suitable functional groups inside them. Agglomeration of cells observed in suspensions treated with a higher concentration of certain cationic phthalocyanines (the strongest binders to bacterial membrane) affected cytometric measurements of total cell counts, thus without appropriate pretreatment of the sample before analysis this parameter seems not to be fully valid in the evaluation of phthalocyanine antibacterial activity. Cytometric measurement of cell membrane integrity appears to be a suitable and even more sensitive parameter than cultivation.

In search of the main properties of phthalocyanines participating in toxicity against cyanobacteria

Phthalocyanines are promising photosensitizers for use in various branches of science including nanotechnology. In the presence of visible light and diatomic oxygen, phthalocyanines can react to produce singlet oxygen (1O2*), which has known inhibitory effects on cellular growth and metabolic activity, although other mechanisms may be involved. The present work focuses on the properties of phthalocyanines (atom charge densities, singlet oxygen production, inhibition effects at various irradiances) contributing to toxicity against the cyanobacteria, Synechococcus nidulans. Our results indicate that positive charge densities at peripheral parts of substituents exhibit greater inhibitory effects against S. nidulans than the amount of singlet oxygen produced, potentially by binding to negatively charged membranes on the cell surface. The weak effect of 1O2* was further demonstrated by a 10% increase in phthalocyanine toxicity (the maximal inhibition detected) when the irradiance increased 3-fold from 1200 to 4000 lux.

Degradation of natural toxins by phthalocyanines–example of cyanobacterial toxin, microcystin

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.

Seven years from the first application of polyaluminium chloride in the Czech Republic – effects on phytoplankton communities in three water bodies

This study determines the efficiency of polyaluminium chloride application on phytoplankton species as a consequence of five reservoir restorations in the Czech Republic during the years 2005 and 2008, including the first ever large-scale application. Although polyaluminium chloride has been used in water treatment plants across the world, information about its application toward cyanobacterial blooms in nature is poor. Although the application of polyaluminium chloride did not cause any fundamental long-term changes in the composition of phytoplankton species or phosphorus load, instead causing fast and acute removal of the phytoplankton community, it may act as an algicidal compound with fast removal efficiency. All treated water bodies described in our study remained unaffected by cyanobacterial blooms and the hygienic limit for the purposes of recreation was not exceeded in any particular season. This article should serve as notice of the advantages and disadvantages of polyaluminium chloride application, and also warn against the uniform usage of this chemical as a method of reducing phytoplankton species in all types of water bodies where cyanobacteria are present. Moreover, data about the effects on non-target (invertebrates) species and microcystin release from cyanobacterial cells are also mentioned.