Physical structure of the environment contributes to the development of diversity of microalgal assemblages.

Aquatic macrophytes form a three dimensional complex structure in the littoral zones of lakes, with many physical, chemical and biological gradients and interactions. This special habitat harbours a unique microalgal assemblage called metaphyton, that differs both from the phytoplankton of the pelagial and from the benthic assemblages whose elements are tightly attached to the substrates. Since metaphytic assemblages significantly contribute to the diversity of lakes' phytoplankton, it is crucial to understand and disentangle those mechanisms that ensure their development. Therefore, we focused on the question of how a single solid physical structure contribute to maintaining metaphytic assemblages. Using a laboratory experiment we studied the floristic and functional differences of microalgal assemblages in microcosms that simulated the conditions that an open water, a complex natural macrophyte stand (Utricularia vulgaris L.), or an artificial substrate (cotton wool) provide for them. We inoculated the systems with a species rich (> 326 species) microalgal assemblage collected from a eutrophic oxbow lake, and studied the diversity, trait and functional group composition of the assemblages in a 24 day long experimental period. We found that both natural and artificial substrates ensured higher species richness than the open water environment. Functional richness in the open water environment was lower than in the aquaria containing natural macrophyte stand but higher than in which cotton wool was placed. This means that the artificial physical structure enhanced functional redundancy of the resident functional groups. Elongation measures of microalgal assemblages showed the highest variation in the microcosms that simulated the open water environment. Our results suggest that assembly of metaphytic algal communities is not a random process, instead a deterministic one driven by the niche characteristics of the complex three dimensional structure created by the stands of aquatic macrophytes.

Uncertainties of cell number estimation in cyanobacterial colonies and the potential use of sphere packing.

Cyanobacteria are notorious bloom formers causing various water quality concerns, such as toxin production, extreme diurnal variation of oxygen, pH, etc., therefore, their monitoring is essential to protect the ecological status of aquatic systems. Cyanobacterial cell counts and biovolumes are currently being used in water management and water quality alert systems. In this study, we investigated the accuracy of traditional colonial biovolume and cell count estimation approaches used in everyday practice. Using shape realistic 3D images of cyanobacterial colonies, we demonstrated that their shape cannot be approximated by ellipsoids. We also showed that despite the significant relationship between overall colony volume and cell biovolumes, because of the considerable scatter of cell count data the regressions give biased estimates for cyanobacterial cell counts. We proposed a novel approach to estimate cell counts in colonies that was based on the random close sphere packing method. This method provided good results only in those cases when overall colony volumes could be accurately measured. The visual investigation of colonies done by skilled experts has given precise but lower estimates for cell counts. The estimation results of several experts were surprisingly good, which suggests that this capability can be improved and estimation bias can be reduced to the level acceptable for water quality estimations.

Estimating nutrient thresholds for eutrophication management: Novel insights from understudied lake types.

Nutrient targets based on pressure-response models are essential for defining ambitions and managing eutrophication. However, the scale of biogeographical variation in these pressure-response relationships is poorly understood, which may hinder eutrophication management in regions where lake ecology is less intensively studied. In this study, we derive ecology-based nutrient targets for five major ecoregions of Europe: Northern, Central-Baltic, Alpine, Mediterranean and Eastern Continental. As a first step, we developed regressions between nutrient concentrations and ecological quality ratios (EQR) based on phytoplankton and macrophyte communities. Significant relationships were established for 13 major lake types; in most cases, these relationships were stronger for phosphorus than for nitrogen, and stronger for phytoplankton than for macrophytes. Using these regressions, we estimated the total phosphorus (TP) and total nitrogen (TN) concentrations at which lakes of different types are likely to achieve good ecological status. However, in the very shallow lakes of the Eastern Continental region, relations between nutrient and biological communities were weak or non-significant. This can be attributed to high nutrient concentrations (in the asymptotic zone of phosphorus-phytoplankton models) suggesting other factors (light, grazing) limit primary production. However, we also show that fish stocking is a major pressure on Eastern Continental lakes, negatively affecting ecological status: lakes with low fish stocking show low chlorophyll-a concentrations and good ecological status despite high nutrient levels, while the lakes with high fish stocking show high chlorophyll-a and low ecological status. This study highlights the need to better understand lakes in biogeographic regions that have been, for historical reasons, less studied. This, in turn, helps reveal factors that challenge the dominant paradigms of lake assessment and management.

Biovolume and surface area calculations for microalgae, using realistic 3D models.

Morphology and spatial dimensions of microalgal units (cells or colonies) are among the most relevant traits of planktic algae, which have a pronounced impact on their basic functional properties, like access to nutrients or light, the velocity of sinking or tolerance to grazing. Although the shape of algae can be approximated by geometric forms and thus, their volume and surface area can be calculated, this approach cannot be validated and might have uncertainties especially in the case of complicated forms. In this study, we report on a novel approach that uses real-like 3D mesh objects to visualize microalgae and calculates their volume and surface area. Knowing these dimensions and their intraspecific variabilities, we calculated specific shape and surface area constants for more than 300 forms, covering more than two thousand taxa. Using these constants, the accurate volume and surface area can be quickly computed for each taxon and having these values, morphology-related metrics like surface area/volume ratio, the diameter of spherical equivalent can also be given quickly and accurately. Besides their practical importance, the volume and surface area constants can be considered as size-independent morphological traits that are characteristic for the microalgal shapes, and provide new possibilities of data analyses in the field of phytoplankton ecology.

Freshwater phytoplankton diversity: models, drivers and implications for ecosystem properties.

Our understanding on phytoplankton diversity has largely been progressing since the publication of Hutchinson on the paradox of the plankton. In this paper, we summarise some major steps in phytoplankton ecology in the context of mechanisms underlying phytoplankton diversity. Here, we provide a framework for phytoplankton community assembly and an overview of measures on taxonomic and functional diversity. We show how ecological theories on species competition together with modelling approaches and laboratory experiments helped understand species coexistence and maintenance of diversity in phytoplankton. The non-equilibrium nature of phytoplankton and the role of disturbances in shaping diversity are also discussed. Furthermore, we discuss the role of water body size, productivity of habitats and temperature on phytoplankton species richness, and how diversity may affect the functioning of lake ecosystems. At last, we give an insight into molecular tools that have emerged in the last decades and argue how it has broadened our perspective on microbial diversity. Besides historical backgrounds, some critical comments have also been made.

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.

Cell Size Decrease and Altered Size Structure of Phytoplankton Constrain Ecosystem Functioning in the Middle Danube River Over Multiple Decades.

Reduced body size is among the universal ecological responses to global warming. Our knowledge on how altered body size affects ecosystem functioning in ectothermic aquatic organisms is still limited. We analysed trends in the cell size structure of phytoplankton in the middle Danube River over a 34-year period at multiple levels: (1) average cell size of assemblages (ACS), (2) within the centric diatom community and (3) in the dominant centric diatom taxon: Stephanodiscus. We asked whether global warming and human impacts affected the average cell size of phytoplankton. Also, whether the altered size structure affected how chlorophyll-a, as an ecosystem functioning measure, relates to the ACS of phytoplankton. The cell size of phytoplankton decreased significantly at all organisation levels, and the assemblages became more dispersed in cell size over time. Environmental variables related to global warming and human impacts affected the ACS of phytoplankton significantly. The relationship between chlorophyll-a and the ACS of phytoplankton shifted from negative linear to broad and then narrow hump shape over time. Longer water residence time, warming and decline in nutrients and suspended solids decrease the ACS of phytoplankton in the middle Danube and expectedly in other large rivers. Our results suggest that cell size decrease in phytoplankton, especially of centric diatoms, constrains planktic algal biomass production in large rivers, independently of algal density. Such cell size decrease may also affect higher trophic levels and enhance the more frequent occurrence of "clear-water" plankton in large, human-impacted rivers under global change.

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.

Trait-based community assembly of epiphytic diatoms in saline astatic ponds: a test of the stress-dominance hypothesis.

The stress dominance hypothesis (SDH) postulates that strong environmental gradients drive trait convergence in communities over limiting similarity. Previous studies, conducted mostly with terrestrial plant communities, found controversial evidence for this prediction. We provide here the first test for SDH for epiphytic diatoms. We studied community assembly in diatom communities of astatic ponds. These water bodies serve as a good model system for testing SDH because they exhibit stress gradients of various environmental factors. Functional diversity of diatom communities was assessed based on four traits: (1) combined trait reflecting the trade-off between stress tolerance and competitive dominance, (2) cell size, (3) oxygen requirement and (4) N-uptake strategy. According to our results, salinity, pH and the width of the macrophyte belt appeared as significant predictors of the trait convergence/divergence patterns presumably acting through influencing the availability of carbon dioxide and turbidity. Lower trait diversity was found in turbid, more saline and more alkaline ponds and functional diversity was higher in transparent, less saline and less alkaline ponds. Overall, our results supported the stress dominance hypothesis. In habitats representing increased environmental stress, environmental filtering was the most important community assembly rule, while limiting similarity became dominant under more favourable conditions.

Microcystis Chemotype Diversity in the Alimentary Tract of Bigheaded Carp.

Most cyanobacterial organisms included in the genus Microcystis can produce a wide repertoire of secondary metabolites. In the mid-2010s, summer cyanobacterial blooms of Microcystis sp. occurred regularly in Lake Balaton. During this period, we investigated how the alimentary tract of filter-feeding bigheaded carps could deliver different chemotypes of viable cyanobacteria with specific peptide patterns. Twenty-five Microcystis strains were isolated from pelagic plankton samples (14 samples) and the hindguts of bigheaded carp (11 samples), and three bloom samples were collected from the scums of cyanobacterial blooms. An LC-MS/MS-based untargeted approach was used to analyze peptide patterns, which identified 36 anabaenopeptin, 17 microginin, and 13 microcystin variants. Heat map clustering visualization was used to compare the identified chemotypes. A lack of separation was observed in peptide patterns of Microcystis that originated from hindguts, water samples, and bloom-samples. Except for 13 peptides, all other congeners were detected from the viable and cultivated chemotypes of bigheaded carp. This finding suggests that the alimentary tract of bigheaded carps is not simply an extreme habitat, but may also supply the cyanobacterial strains that represent the pelagic chemotypes.

Groups of small lakes maintain larger microalgal diversity than large ones.

The question of whether one large, continuous area or many smaller habitats maintain more species is one of the most relevant questions in conservation ecology, and it is referred to as the SLOSS (Single Large Or Several Small) dilemma in the literature. This question has not yet been raised in the case of microscopic organisms, therefore we investigated whether or not the SLOSS dilemma could apply to phytoplankton and benthic diatom metacommunities. Benthic diatom and phytoplankton diversity in pools and ponds of different sizes (ranging between 10-2-107 m2) was studied. Species richness of water bodies belonging to neighbouring size categories was compared step by step across the whole size gradient. With the exception of the 104-105 m2 and 105-106 m2 size categories, where phytoplankton and benthic diatom richness values of the SL water bodies were higher than that of the SS ones, findings showed that the diversity of several smaller (SS) sized waters was higher than that in single large water bodies (SL) throughout the whole studied size range. The proportion of the various functional groups of algae, including both the benthic diatoms and phytoplankton, showed remarkable changes from the smaller water bodies to large sized ones.

Protecting and restoring Europe's waters: An analysis of the future development needs of the Water Framework Directive.

The Water Framework Directive (WFD) is a pioneering piece of legislation that aims to protect and enhance aquatic ecosystems and promote sustainable water use across Europe. There is growing concern that the objective of good status, or higher, in all EU waters by 2027 is a long way from being achieved in many countries. Through questionnaire analysis of almost 100 experts, we provide recommendations to enhance WFD monitoring and assessment systems, improve programmes of measures and further integrate with other sectoral policies. Our analysis highlights that there is great potential to enhance assessment schemes through strategic design of monitoring networks and innovation, such as earth observation. New diagnostic tools that use existing WFD monitoring data, but incorporate novel statistical and trait-based approaches could be used more widely to diagnose the cause of deterioration under conditions of multiple pressures and deliver a hierarchy of solutions for more evidence-driven decisions in river basin management. There is also a growing recognition that measures undertaken in river basin management should deliver multiple benefits across sectors, such as reduced flood risk, and there needs to be robust demonstration studies that evaluate these. Continued efforts in 'mainstreaming' water policy into other policy sectors is clearly needed to deliver wider success with WFD goals, particularly with agricultural policy. Other key policy areas where a need for stronger integration with water policy was recognised included urban planning (waste water treatment), flooding, climate and energy (hydropower). Having a deadline for attaining the policy objective of good status is important, but even more essential is to have a permanent framework for river basin management that addresses the delays in implementation of measures. This requires a long-term perspective, far beyond the current deadline of 2027.

The Ecophysiological Response of Two Invasive Submerged Plants to Light and Nitrogen.

Two submerged Elodea species have small differences in their ecophysiological responses when exposed to individual environmental factors. However, field observations showed that under eutrophic conditions with low light availability, Elodea canadensis could be displaced by Elodea nuttallii. Here we investigated the combined effect of environmental factors on the ecophysiological response of the two species in order to explain the differences in their invasion successes. We cultivated the plants in aquaria containing five different nitrogen (N) concentrations and incubated at five different light intensities. For both species increasing nitrogen concentrations resulted in increased relative growth rate, chlorophyll concentration, and actual photochemical efficiency of photosystem II (ΦPSII), however, they produced less roots. Lowering light intensity resulted in a lower relative growth rate, root production, and nutrient removal. In contrast, chlorophyll concentration in the leaves, and ΦPSII increased. The main difference between the two Elodea species was that the light compensation point (I c) and weight loss in the dark were significantly higher and photochemical efficiency and chlorophyll concentration were significantly lower for E. canadensis than for E. nuttallii, indicating that the latter can survive under much more shady and hypertrophic conditions. The change in nitrogen concentration of the media and in tissue concentration of the plants indicated that E. nuttallii has a higher nitrogen removal capacity. The ecophysiological differences between the two species can be an explanation for invasion success of E. nuttallii over E. canadensis and thus may explain why the latter is replaced by the first.

Resisting annihilation: relationships between functional trait dissimilarity, assemblage competitive power and allelopathy.

Allelopathic species can alter biodiversity. Using simulated assemblages that are characterised by neutrality, lumpy coexistence and intransitivity, we explore relationships between within-assemblage competitive dissimilarities and resistance to allelopathic species. An emergent behaviour from our models is that assemblages are more resistant to allelopathy when members strongly compete exploitatively (high competitive power). We found that neutral assemblages were the most vulnerable to allelopathic species, followed by lumpy and then by intransitive assemblages. We find support for our modeling in real-world time-series data from eight lakes of varied morphometry and trophic state. Our analysis of this data shows that a lake's history of allelopathic phytoplankton species biovolume density and dominance is related to the number of species clusters occurring in the plankton assemblages of those lakes, an emergent trend similar to that of our modeling. We suggest that an assemblage's competitive power determines its allelopathy resistance.

Functional redundancy modifies species-area relationship for freshwater phytoplankton.

Although species-area relationship (SAR) is among the most extensively studied patterns in ecology, studies on aquatic and/or microbial systems are seriously underrepresented in the literature. We tested the algal SAR in lakes, pools and ponds of various sizes (10-2-108 m2) and similar hydromorphological and trophic characteristics using species-specific data and functional groups. Besides the expectation that species richness increases monotonously with area, we found a right-skewed hump-shaped relationship between the area and phytoplankton species richness. Functional richness however did not show such distortion. Differences between the area dependence of species and functional richness indicate that functional redundancy is responsible for the unusual hump-backed SAR. We demonstrated that the Small Island Effect, which is a characteristic for macroscopic SARs can also be observed for the phytoplankton. Our results imply a so-called large lake effect, which means that in case of large lakes, wind-induced mixing acts strongly against the habitat diversity and development of phytoplankton patchiness and finally results in lower phytoplankton species richness in the pelagial. High functional redundancy of the groups that prefer small-scale heterogeneity of the habitats is responsible for the unusual humpback relationship. The results lead us to conclude that although the mechanisms that regulate the richness of both microbial communities and communities of macroscopic organisms are similar, their importance can be different in micro- and macroscales.

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.