Salinity tolerance and desalination properties of a Haematococcus lacustris strain from eastern Hungary.

Nowadays the increasing amount of saline wastewaters has given rise to various biological desalination processes, among which the application possibilities of microalgae represents a priority research area. Next to "real" aquatic species (members of phytoplankton or phytobenthon), species from ephemeral aquatic habitats or aeroterrestrial algae also could be good candidates of research studying salt tolerance or desalination ability, since salinity stress is often referred as "physiological drought" and species from ephemeral habitats can be characterized by high drought tolerance. In this study, the salinity tolerance, salt and nutrient removal ability of a Haematococcus lacustris strain from eastern Hungary were investigated. Vegetative cells showed low salt tolerance, survival was ensured by the formation of cysts up to a sodium-chloride concentration of 2,000 mg l-1. Although relatively moderate (a max. 30%) conductivity reduction and chloride removal were observed, notable (nearly 100%) nitrate and phosphate removal occurred even in the presence of 2,000 mg l-1 NaCl. Carotenoid accumulation was observed earlier and in higher extent in salt treated cultures than in drying out ones, although the amount of astaxanthin-esters was significantly higher in the cultures of drying out experiment than in the corresponding cultures of salt treatment characterized with similar chloride content. Our results suggest that algae isolates from ephemeral aquatic habitats endangered by regular drying out (exposed to special salt stress), could have notable salt tolerance and consequently successful applicability in nutrient removal processes from slightly saline wastewaters. The accumulation of valuable metabolites (such as astaxanthin) as a response to salinity stress, could enhance the economic value of the biomass.

Horseradish Essential Oil as a Promising Anti-Algal Product for Prevention of Phytoplankton Proliferation and Biofouling.

Increased proliferation of algae is a current problem in natural and artificial water bodies. Controlling nutrients is the most sustainable treatment of increased algal proliferation, however in certain cases, it is not sufficiently available, or it does not provide results fast enough. Chemicals derived from natural sources, which could be effective in low concentrations and are biodegradable, may have an advantage over conventional chemical treatments. The main aim of the present study was to investigate the anti-cyanobacterial and anti-algal properties of allyl-isothiocyanate-containing essential oil produced from horseradish roots with a complex approach of the topic: on laboratory strains of cyanobacteria and eukaryotic algae, on microcosms containing natural phytoplankton assemblages, and on semi-natural biofilms. The results show that acute treatment can significantly reduce the viability of all the tested cyanobacteria and eukaryotic algae. Results of microcosm experiments with natural phytoplankton assemblages show that horseradish essential oil from 7.1 × 10-6% (v/v) is applicable to push back phytoplankton proliferation even in natural assemblages. The individual number in the biofilm was dropped down to one-fifth of the original individual number, so 7.1 × 10-6% (v/v) and higher concentration of the essential oil can be considered as a successful treatment against biofouling.

Trait convergence and trait divergence in lake phytoplankton reflect community assembly rules.

Environmental filtering and limiting similarity are those locally acting processes that influence community structure. These mechanisms acting on the traits of species result in trait convergence or divergence within the communities. The role of these processes might change along environmental gradients, and it has been conceptualised in the stress-dominance hypothesis, which predicts that the relative importance of environmental filtering increases and competition decreases with increasing environmental stress. Analysing trait convergence and divergence in lake phytoplankton assemblages, we studied how the concepts of 'limiting similarity' versus 'environmental filtering' can be applied to these microscopic aquatic communities, and how they support or contradict the stress-dominance hypothesis. Using a null model approach, we investigated the divergence and convergence of phytoplankton traits along environmental gradients represented by canonical axes of an RDA. We used Rao's quadratic entropy as a measure of functional diversity and calculated effect size (ES) values for each sample. Negative ES values refer to trait convergence, i.e., to the higher probability of the environmental filtering in community assembly, while positive values indicate trait divergence, stressing the importance of limiting similarity (niche partitioning), that is, the competition between the phytoplankters. Our results revealed that limiting similarity and environmental filtering may operate simultaneously in phytoplankton communities, but these assembly mechanisms influenced the distribution of phytoplankton traits differently, and the effects show considerable changes along with the studied scales. Studying the changes of ES values along with the various scales, our results partly supported the stress-dominance hypothesis, which predicts that the relative importance of environmental filtering increases and competition decreases with increasing environmental stress.

Environmental filtering and limiting similarity as main forces driving diatom community structure in Mediterranean and continental temporary and perennial streams.

Climatic extreme events such as droughts (unpredictable), dry periods (predictable) or even flush floods, threaten freshwater ecosystems worldwide. The filtering mechanisms of these events and their strength on communities, however, can be different among regions. While time-for-adaptation theory defines whether or not water scarcity can be considered as disturbance, the stress-dominance theory predicts an increase in importance of environmental filtering and a decrease in the role of biotic interactions in communities with increasing environmental stress. Here, we tested whether environmental filtering (leading to trait convergence) or limiting similarity (leading to trait divergence) is the main assembly rule shaping the structure and trait composition of benthic diatom assemblages in Mediterranean (Portuguese) and continental (Hungarian) temporary and perennial streams. We assumed that the trait composition of diatom assemblages in the two stream types would be less different in the Mediterranean than in the continental region (addressed to time-for-adaptation theory). We also hypothesized that trait composition would be shaped by environmental filtering in the Hungarian streams while by biotic interactions in Portuguese streams (addressed to stress-dominance theory). Our results supported our first hypothesis since traits, which associated primarily to temporary streams were found only in the continental region. Our findings, however, only partially proved the stress-dominance hypothesis. In the continental region, where drying up of streams were induced by unpredictable droughts, biotic interactions were the main assembly rules shaping community structure. In contrast, environmental filtering was nearly as important as limiting similarity in structuring trait composition in the Mediterranean region during the predictable dry phase with no superficial flow. These analyses also highlighted that drought events (both predictable and unpredictable ones) have a complex and strong influence on benthic diatom assemblages resulting even in irreversible changes in trait composition and thereby in ecosystem functioning.

Effects of non-steroidal anti-inflammatory drugs on cyanobacteria and algae in laboratory strains and in natural algal assemblages.

In recent years measurable concentrations of non-steroidal anti-inflammatory drugs (NSAIDs) have been shown in the aquatic environment as a result of increasing human consumption. Effects of five frequently used non-steroidal anti-inflammatory drugs (diclofenac, diflunisal, ibuprofen, mefenamic acid and piroxicam in 0.1 mg ml(-1) concentration) in batch cultures of cyanobacteria (Synechococcus elongatus, Microcystis aeruginosa, Cylindrospermopsis raciborskii), and eukaryotic algae (Desmodesmus communis, Haematococcus pluvialis, Cryptomonas ovata) were studied. Furthermore, the effects of the same concentrations of NSAIDs were investigated in natural algal assemblages in microcosms. According to the changes of chlorophyll-a content, unicellular cyanobacteria seemed to be more tolerant to NSAIDs than eukaryotic algae in laboratory experiments. Growth of eukaryotic algae was reduced by all drugs, the cryptomonad C. ovata was the most sensitive to NSAIDs, while the flagellated green alga H. pluvialis was more sensitive than the non-motile green alga D. communis. NSAID treatments had weaker impact in the natural assemblages dominated by cyanobacteria than in the ones dominated by eukaryotic algae, confirming the results of laboratory experiments. Diversity and number of functional groups did not change notably in cyanobacteria dominated assemblages, while they decreased significantly in eukaryotic algae dominated ones compared to controls. The results highlight that cyanobacteria (especially unicellular ones) are less sensitive to the studied, mostly hardly degradable NSAIDs, which suggest that their accumulation in water bodies may contribute to the expansion of cyanobacterial mass productions in appropriate environmental circumstances by pushing back eukaryotic algae. Thus, these contaminants require special attention during wastewater treatment and monitoring of surface waters.

Effects of Cylindrospermopsin Producing Cyanobacterium and Its Crude Extracts on a Benthic Green Alga-Competition or Allelopathy?

Cylindrospermopsin (CYN) is a toxic secondary metabolite produced by filamentous cyanobacteria which could work as an allelopathic substance, although its ecological role in cyanobacterial-algal assemblages is mostly unclear. The competition between the CYN-producing cyanobacterium Chrysosporum (Aphanizomenon) ovalisporum, and the benthic green alga Chlorococcum sp. was investigated in mixed cultures, and the effects of CYN-containing cyanobacterial crude extract on Chlorococcum sp. were tested by treatments with crude extracts containing total cell debris, and with cell debris free crude extracts, modelling the collapse of a cyanobacterial water bloom. The growth inhibition of Chlorococcum sp. increased with the increasing ratio of the cyanobacterium in mixed cultures (inhibition ranged from 26% to 87% compared to control). Interestingly, inhibition of the cyanobacterium growth also occurred in mixed cultures, and it was more pronounced than it was expected. The inhibitory effects of cyanobacterial crude extracts on Chlorococcum cultures were concentration-dependent. The presence of C. ovalisporum in mixed cultures did not cause significant differences in nutrient content compared to Chlorococcum control culture, so the growth inhibition of the green alga could be linked to the presence of CYN and/or other bioactive compounds.

Alterations of phytoplankton assemblages treated with chlorinated hydrocarbons: effects of dominant species sensitivity and initial diversity.

Changes in composition of phytoplankton assemblages due to short-chained chlorinated hydrocarbons (tetrachloroethane, tetrachloroethylene and trichloroethylene) were studied in microcosm experiments with different initial diversities. Diversity decreased further during treatments in the less diverse 2011 summer assemblages, dominated by the euglenid Trachelomonas volvocinopsis (its relative abundance was nearly 70 %). Diversity did not change significantly during treatments in the more diverse 2012 summer assemblages, dominated by cryptomonads (their relative abundance was 40 %). The dominant Trachelomonas volvocinopsis in 2011, due to its insensitivity to the treatment and presumably high competition skills, filled released habitats occurring when sensitive species were not detectable any more. In contrast, cryptomonads were extremely sensitive to the treatments, their abundance decreased under detection limit in the treated assemblages, regardless of diversity conditions. Our results showed that population dynamics of dominant species determine the response to the contamination of the entire community, if these species display high resistance or resilience. If the dominant species was highly sensitive and recovered slowly, compensatory growth of rare species maintained high levels of ecosystem performance.

Zinc tolerance and zinc removal ability of living and dried biomass of Desmodesmus communis.

Effects of zinc on growth, cell morphology, oxidative stress, and zinc removal ability of the common phytoplankton species Desmodesmus communis were investigated at a concentration range of 0.25-160 mg L(-1) zinc. Cell densities and chlorophyll content decreased in treated cultures, changes in coenobia morphology and elevated lipid peroxidation levels appeared above 2.5 mg L(-1) zinc. The most effective zinc removal was observed at 5 mg L(-1) zinc concentration, while maximal amount of removed zinc appeared in 15 mg L(-1) zinc treated culture. Removed zinc is mainly bound on the cell surface. Dead biomass adsorbed more zinc than living biomass relative to unit of dry mass, but living biomass was more effective, relative to initial zinc content. This study comprehensively examines the zinc tolerance and removal ability of D. communis and demonstrates, in comparison with published literature, that these characteristics of different isolates of the same species can vary within a wide range.

Appearance of Planktothrix rubescens bloom with [D-Asp3, Mdha7]MC-RR in gravel pit pond of a shallow lake-dominated area.

Blooms of toxic cyanobacteria are well-known phenomena in many regions of the world. Microcystin (MC), the most frequent cyanobacterial toxin, is produced by entirely different cyanobacteria, including unicellular, multicellular filamentous, heterocytic, and non-heterocytic bloom-forming species. Planktothrix is one of the most important MC-producing genera in temperate lakes. The reddish color of cyanobacterial blooms viewed in a gravel pit pond with the appearance of a dense 3 cm thick layer (biovolume: 28.4 mm(3) L(-1)) was an unexpected observation in the shallow lake-dominated alluvial region of the Carpathian Basin. [D-Asp(3), Mdha(7)]MC-RR was identified from the blooms sample by MALDI-TOF and NMR. Concentrations of [D-Asp(3), Mdha(7)]MC-RR were measured by capillary electrophoresis to compare the microcystin content of the field samples and the isolated, laboratory-maintained P. rubescens strain. In analyzing the MC gene cluster of the isolated P. rubescens strain, a deletion in the spacer region between mcyE and mcyG and an insertion were located in the spacer region between mcyT and mcyD. The insertion elements were sequenced and partly identified. Although some invasive tropical cyanobacterial species have been given a great deal of attention in many recent studies, our results draw attention to the spread of the alpine organism P. rubescens as a MC-producing, bloom-forming species.

Observation of sward destruction caused by irrigation with toxic Microcystis morphospecies containing water in Southern Hungary.

In the summer of 2006 bloom-like phenomenon occurred in a garden pond in Szeged, Southern Hungary. After regular watering of a sward with pond water containing the algal mass, destruction of garden grass occurred. Microcystis aeruginosa, Microcystis viridis, Microcystis ichthyoblabe, and Microcystis wesenbergii were identified by light microscopy in the water sample; microcystin-FR, -LR, -RR and -YR were determined by matrix-assisted laser desorption/ionization--time-of-flight analysis. There was an 80% decrease in the green mass (87% in chlorophyll-content) of the grass in a 1 m² area of the garden irrigated with pond water.

Application of external micro-spectrophotometric detection to improve sensitivity on microchips.

The goal of this work was to increase the sensitivity of a UV-Vis spectrophotometer by decreasing the background noise and lengthening the optical path. A microphotometer has been modified to precisely select very small parts of a microfluidic channel pattern of a chip and to measure light absorbance on a magnified area of the selected part of the channel. The viability of combining a projection microscope and a spectrophotometer for external absorbance measurements on disposable PDMS chips was studied. Besides the external direct detection above a microfluidic channel, the optical pathlength was lengthened by detecting in the region of the perpendicular exit port. Increasing the cross-sectional area of the zone of irradiation improved the signal-to-noise ratio and the limits of detection (LOD).

Cylindrospermopsin induces alterations of root histology and microtubule organization in common reed (Phragmites australis) plantlets cultured in vitro.

We aimed to study the histological and cytological alterations induced by cylindrospermopsin (CYN), a protein synthesis inhibitory cyanotoxin in roots of common reed (Phragmites australis). Reed is an ecologically important emergent aquatic macrophyte, a model for studying cyanotoxin effects. We analyzed the histology and cytology of reed roots originated from tissue cultures and treated with 0.5-40 microg ml(-1) (1.2-96.4 microM) CYN. The cyanotoxin decreased root elongation at significantly lower concentrations than the elongation of shoots. As general stress responses of plants to phytotoxins, CYN increased root number and induced the formation of a callus-like tissue and necrosis in root cortex. Callus-like root cortex consisted of radially swollen cells that correlated with the reorientation of microtubules (MTs) and the decrease of MT density in the elongation zone. Concomitantly, the cyanotoxin did not decrease, rather it increased the amount of beta-tubulin in reed plantlets. CYN caused the formation of double preprophase bands; the disruption of mitotic spindles led to incomplete sister chromatid separation and disrupted phragmoplasts in root tip meristems. This work shows that CYN alters reed growth and anatomy through the alteration of MT organization.

Microcystin-LR, a cyanobacterial toxin, induces growth inhibition and histological alterations in common reed (Phragmites australis) plants regenerated from embryogenic calli.

The aim of this study was to establish the histological effects of exposure to microcystin-LR (MC-LR), a cyanotoxin, on axenic Phragmites australis plantlets. Plantlets were regenerated from embryogenic reed calli by tissue culture methods. Microcystin-LR inhibited the growth and development of embryogenic calli and the growth of reed plantlets. The 50% plantlet growth inhibitory concentration value (IC50) of MC-LR was 12 microg ml(-1) (12.07 microM) on mineral medium and 36 microg ml(-1) (36.22 microM) on Murashige-Skoog medium. In the case of roots, the IC50 value was 4.1 microg ml(-1) (4.12 microM) on both media. Microcystin-LR induced aerenchyma obturation, altered lignification of cell walls in the axial organs, root necrosis and the capture of lateral or adventitious roots in the tissues of axial organs of reed plantlets. Cyanotoxin induced the premature development of lateral roots, root coalescence and early aerenchyma formation. Our data suggest that microcystin-LR, a cyanotoxin, induced developmental and histological alterations leading to growth inhibition of reed, and the induced harms have an impact on understanding reed decay in eutrophic fresh waters.

Alteration of cylindrospermopsin production in sulfate- or phosphate-starved cyanobacterium Aphanizomenon ovalisporum.

The effect of sulfate and phosphate deprivation on cell growth and cylindrospermopsin level was studied in Aphanizomenon ovalisporum ILC-164. Sulfate starvation induced a characteristic reduction of cylindrospermopsin pool size on the basis of cell number and unit of dry mass of culture. Phosphorous starvation of A. ovalisporum cultures induced a lesser reduction of cylindrospermopsin pool size. This divergence in the pool size of cylindrospermopsin may be the consequence of different growth rate. To show the metabolic changes concomitant with reduction of cylindrospermopsin pool size were obtained by measurement of ATP sulfurylase and alkaline phosphatase activity. The present study is the first concerning the cylindrospermopsin content under sulfate starvation and discusses it in relation to phosphorous starvation.