Assessment of two non-invasive techniques for measuring turbulent benthic fluxes in a shallow lake.

Benthic fluxes refer to the exchange rates of nutrients and other compounds between the water column and the sediment bed in aquatic ecosystems. Their quantification contributes to our understanding of aquatic ecosystem functioning. Near-bed hydrodynamics plays an important role at the sediment-water interface, especially in shallow lakes, but it is poorly considered by traditional measuring techniques of flux quantification, such as sediment incubations. Thus, alternative sampling techniques are needed to characterize key benthic fluxes under in-situ hydrodynamic conditions. This study aimed to evaluate the performance of two promising methods: relaxed eddy accumulation (REA) and mass transfer coefficient (MTC). We applied them in a hyper-eutrophic shallow lake to measure the fluxes of ammonium, phosphate, iron, and manganese ions. For the first time, REA revealed hourly nutrient flux variations, indicating a strong lake biogeochemical dynamics at short time-scales. Daily average fluxes are of similar orders of magnitude for REA and MTC for ammonium (24 and 42 mmol m2 d-1), manganese (1.0 and 0.8), and iron (0.8 and 0.7) ions. They are one order of magnitude higher than fluxes estimated from sediment incubations, due to the difficulty in reproducing in-situ oxygen and hydrodynamic conditions in the laboratory. Although the accuracy of both techniques needs to be improved, the results revealed their potential: REA follows the short-term biogeochemical dynamics of sediments, while MTC could be widely used for lake monitoring because of its simpler implementation.

Bioaccumulation of chemical elements at post-industrial freshwater sites varies predictably between habitats, elements and taxa: A power law approach.

Elevated environmental levels of elements originating from anthropogenic activities threaten natural communities and public health, as these elements can persist and bioaccumulate in the environment. However, their environmental risks and bioaccumulation patterns are often habitat-, species- and element-specific. We studied the bioaccumulation patterns of 11 elements in seven freshwater taxa in post-mining habitats in the Czech Republic, ranging from less polluted mining ponds to highly polluted fly ash lagoons. We found nonlinear, power-law relationships between the environmental and tissue concentrations of the elements, which may explain differences in bioaccumulation factors (BAF) reported in the literature. Tissue concentrations were driven by the environmental concentrations in non-essential elements (Al, As, Co, Cr, Ni, Pb and V), but this dependence was limited in essential elements (Cu, Mn, Se and Zn). Tissue concentrations of most elements were also more closely related to substrate than to water concentrations. Bioaccumulation was habitat specific in eight elements: stronger in mining ponds for Al and Pb, and stronger in fly ash lagoons for As, Cu, Mn, Pb, Se, V and Zn, although the differences were often minor. Bioaccumulation of some elements further increased in mineral-rich localities. Proximity to substrate, rather than trophic level, drove increased bioaccumulation levels across taxa. This highlights the importance of substrate as a pollutant reservoir in standing freshwaters and suggests that benthic taxa, such as molluscs (e.g., Physella) and other macroinvertebrates (e.g., Nepa), constitute good bioindicators. Despite the higher environmental risks in fly ash lagoons than in mining ponds, the observed ability of freshwater biota to sustain pollution supports the conservation potential of post-industrial sites. The power law approach used here to quantify and disentangle the effects of various bioaccumulation drivers may be helpful in additional contexts, increasing our ability to predict the effects of other contaminants and environmental hazards on biota.

Muddying the unexplored post-industrial waters: Biodiversity and conservation potential of freshwater habitats in fly ash sedimentation lagoons.

Deposits of fly ash and other coal combustion wastes are common remnants of the energy industry. Despite their environmental risks from heavy metals and trace elements, they have been revealed as refuges for threatened terrestrial biodiversity. Surprisingly, freshwater biodiversity of fly ash sedimentation lagoons remains unknown despite such lack of knowledge strongly limits the efficient restoration of fly ash deposits. We bring the first comprehensive survey of freshwater biodiversity, including nekton, benthos, zooplankton, phytoplankton, and macrophytes, in fly ash lagoons across industrial regions of the Czech Republic. To assess their conservation potential, we compared their biodiversity with abandoned post-mining ponds, the known strongholds of endangered aquatic species in the region with a shortage of natural ponds. Of 28 recorded threatened species, 15 occurred in the studied fly ash lagoons, some of which were less abundant or even absent in the post-mining ponds. These are often species of nutrient-poor, fishless waters with rich vegetation, although some are specialised extremophiles. Species richness and conservation value of most groups in the fly ash lagoons did not significantly differ from the post-mining ponds, except for species richness of benthos, zooplankton, and macrophytes, which were slightly lower in the fly ash lagoons. Although the concentrations of some heavy metals (mainly Se, V, and As) were significantly higher in the fly ash lagoons, they did not significantly affect species richness or conservation value of the local communities. The differences in species composition therefore does not seem to be caused by water chemistry. Altogether, we have shown that fly ash lagoons are refuges for threatened aquatic species, and we thus suggest maintaining water bodies during site restoration after the cessation of fly ash deposition. Based on our analyses of environmental variables, we discuss suitable restoration practices that efficiently combine biodiversity protection and environmental risk reduction.

Contrasting catchment soil pH and Fe concentrations influence DOM distribution and nutrient dynamics in freshwater systems.

Organic matter (OM) quantity, quality, and nutrient dynamics within twelve shallow lakes in the Czech Republic were assessed in the context of catchment soil pH and iron (Fe) concentration. The catchments of the lakes were classified into two categories: (i) slightly acidic (soil pH = 5.1-6.3) with Fe-rich soils (H_Fe; Fe = 315-344 mg kg-1 in Mehlich 3 extract); and (ii) neutral (soil pH = 6.8-7.6) with Fe-poor soils (L_Fe; Fe = 126-259 mg kg-1 in Mehlich 3 extract). The quality of OM in the two lake types was characterized using a combination of spectroscopic techniques (UV-Vis, fluorescence, and Fourier Transform Infrared spectroscopy). We show that dissolved nutrient and dissolved organic carbon (DOC) concentrations, as well as the amount of aromatic and protein-like compounds in the water column and sediment porewater were significantly (p < 0.01) lower in the H_Fe lakes compared to the waterbodies located within L_Fe catchments. The FTIR analyses of the H_Fe sediments contained higher relative concentrations of aromatic compounds with hydroxyl-containing functional groups and carbohydrates, while more aliphatic and oxidised OM was found in the L_Fe lake sediments. These results suggest that the pH value of catchment soils and, particularly, their Fe content have profound geochemical effects on the mobility of OM and nutrients in the sediments of recipient waters. Because the OM-Fe association stabilises OM in sediments, waterbodies within L_Fe catchments are likely more vulnerable to increasing eutrophication and oxygen depletion compared to those in H_Fe catchments and this has important implications for water quality management, risk assessment, and predictions of aquatic ecosystem vulnerability under conditions of accelerating climate change.

Correction to: Hunters or farmers? Microbiome characteristics help elucidate the diet composition in an aquatic carnivorous plant.

AbstractFollowing publication of the original article [1], the author reported an error in Fig. 3.

Hunters or farmers? Microbiome characteristics help elucidate the diet composition in an aquatic carnivorous plant.

BACKGROUND: Utricularia are rootless aquatic carnivorous plants which have recently attracted the attention of researchers due to the peculiarities of their miniaturized genomes. Here, we focus on a novel aspect of Utricularia ecophysiology-the interactions with and within the complex communities of microorganisms colonizing their traps and external surfaces. RESULTS: Bacteria, fungi, algae, and protozoa inhabit the miniature ecosystem of the Utricularia trap lumen and are involved in the regeneration of nutrients from complex organic matter. By combining molecular methods, microscopy, and other approaches to assess the trap-associated microbial community structure, diversity, function, as well as the nutrient turn-over potential of bacterivory, we gained insight into the nutrient acquisition strategies of the Utricularia hosts. CONCLUSIONS: We conclude that Utricularia traps can, in terms of their ecophysiological function, be compared to microbial cultivators or farms, which center around complex microbial consortia acting synergistically to convert complex organic matter, often of algal origin, into a source of utilizable nutrients for the plants.

Iron and nitrogen cycling, bacterioplankton community composition and mineral transformations involving phosphorus stabilisation in the ferruginous hypolimnion of a post-mining lake.

Lake Medard is an oligotrophic post-mining lake characterised by ferruginous bottom waters, with marked redox gradients resulting from iron (Fe) and nitrogen (N) speciation and accompanying depth-dependent variations in the abundance of volatile fatty acids (VFAs), pH and alkalinity. The lacustrine system is meromictic, featuring a dysoxic hypolimnion and an anoxic monimolimnion with relatively high concentrations of sulfate (SO42-, 19 ± 2 mM) and Fe(ii) (127 ± 17 µM). An increase in dissolved manganese is also observed with increasing depth, together with a general lack of sulfide, which can only be detected at the sediment-water interface at concentrations of ∼0.30 µM. In the hypolimnion, nitrate (NO3-) becomes progressively depleted and ammonium (NH4+) dominates the dissolved N inventory (up to 185 ± 13 µM). Here we describe the biogeochemical disequilibrium conditions governing critical mineralogical transformations involving Fe and phosphorus (P) co-precipitation in the dysoxic-to-anoxic bottom water column. A combination of mineral equilibrium modelling and synchrotron-based diffraction and spectroscopic techniques was applied to investigate the minerals comprising the upper anoxic sediments. The combined dataset indicates that elemental recycling on and below the hypolimnion promote the precipitation of FeOOH polymorphs that accumulate as heterogeneous mineral clusters. Changes in the relative abundance of bacterioplankton taxa with increasing water depth point to a link between the activity of certain members of Proteobacteria and the co-recycling of carbon (C), N, and Fe stocks. Such a redox recycling process seems to lead to P stabilisation into organic-rich Fe-(oxyhydr)oxides near and above the anoxic sediment-water interface (SWI).

Approach for predicting P sorption/desorption behaviour of potentially eroded topsoil in watercourses.

This study focuses on the quantification of the impact of potentially eroded topsoil particles on the available watercourse P concentration. We used 56 topsoil samples for determining the relation existing between the molar ratio of sorption-active Fe- and Al-(hydr)oxides to plant available P, as determined by the commonly used oxalate and Mehlich 3 extractions (PM3). Our sample set covers the most common combinations of non-calcareous soil types, land and agricultural uses. By using Freundlich isotherms, we found that the concentration of adsorbed/desorbed P (Q) by soil particles in water with specific soluble reactive phosphorus (SRP) concentrations are significantly correlated to PM3 (Q=a∗PM3+b) and also to the degree of P saturation as measured by oxalate extraction (DPSox; Q=c∗DPSox+d). The observed relations varied in slope and intercept parameters for discrete equilibrium P concentrations in water. However, in the environmentally important range of PO4-P concentrations, i.e., from 20 to 220µgL-1, a strong non-linear correlation was found between the individual parameters and equilibrium P concentration. Accordingly, we derived equations useful for quantitatively predict P exchange between topsoil particles when potentially eroded into watercourse and the recipient water of either known or targeted P concentrations. This approach might refine erosion models, and thus facilitates the quantification of the impact of soil erosion on the in-stream bioavailable P concentrations.

Tuning Surface Chemistry of Polyetheretherketone by Gold Coating and Plasma Treatment.

Polyetheretherketone (PEEK) has good chemical and biomechanical properties that are excellent for biomedical applications. However, PEEK exhibits hydrophobic and other surface characteristics which cause limited cell adhesion. We have investigated the potential of Ar plasma treatment for the formation of a nanostructured PEEK surface in order to enhance cell adhesion. The specific aim of this study was to reveal the effect of the interface of plasma-treated and gold-coated PEEK matrices on adhesion and spreading of mouse embryonic fibroblasts. The surface characteristics (polarity, surface chemistry, and structure) before and after treatment were evaluated by various experimental techniques (gravimetry, goniometry, X-ray photoelectron spectroscopy (XPS), and electrokinetic analysis). Further, atomic force microscopy (AFM) was employed to examine PEEK surface morphology and roughness. The biological response of cells towards nanostructured PEEK was evaluated in terms of cell adhesion, spreading, and proliferation. Detailed cell morphology was evaluated by scanning electron microscopy (SEM). Compared to plasma treatment, gold coating improved PEEK wettability. The XPS method showed a decrease in the carbon concentration with increasing time of plasma treatment. Cell adhesion determined on the interface between plasma-treated and gold-coated PEEK matrices was directly proportional to the thickness of a gold layer on a sample. Our results suggest that plasma treatment in a combination with gold coating could be used in biomedical applications requiring enhanced cell adhesion.

Vegetation dynamics at the upper elevational limit of vascular plants in Himalaya.

A rapid warming in Himalayas is predicted to increase plant upper distributional limits, vegetation cover and abundance of species adapted to warmer climate. We explored these predictions in NW Himalayas, by revisiting uppermost plant populations after ten years (2003-2013), detailed monitoring of vegetation changes in permanent plots (2009-2012), and age analysis of plants growing from 5500 to 6150 m. Plant traits and microclimate variables were recorded to explain observed vegetation changes. The elevation limits of several species shifted up to 6150 m, about 150 vertical meters above the limit of continuous plant distribution. The plant age analysis corroborated the hypothesis of warming-driven uphill migration. However, the impact of warming interacts with increasing precipitation and physical disturbance. The extreme summer snowfall event in 2010 is likely responsible for substantial decrease in plant cover in both alpine and subnival vegetation and compositional shift towards species preferring wetter habitats. Simultaneous increase in summer temperature and precipitation caused rapid snow melt and, coupled with frequent night frosts, generated multiple freeze-thaw cycles detrimental to subnival plants. Our results suggest that plant species responses to ongoing climate change will not be unidirectional upward range shifts but rather multi-dimensional, species-specific and spatially variable.

Cadmium toxicity investigated at the physiological and biophysical levels under environmentally relevant conditions using the aquatic model plant Ceratophyllum demersum.

Cadmium (Cd) is an important environmental pollutant and is poisonous to most organisms. We aimed to unravel the mechanisms of Cd toxicity in the model water plant Ceratophyllum demersum exposed to low (nM) concentrations of Cd as are present in nature. Experiments were conducted under environmentally relevant conditions, including nature-like light and temperature cycles, and a low biomass to water ratio. We measured chlorophyll (Chl) fluorescence kinetics, oxygen exchange, the concentrations of reactive oxygen species and pigments, metal binding to proteins, and the accumulation of starch and metals. The inhibition threshold concentration for most parameters was 20 nM. Below this concentration, hardly any stress symptoms were observed. The first site of inhibition was photosynthetic light reactions (the maximal quantum yield of photosystem II (PSII) reaction centre measured as Fv /Fm , light-acclimated PSII activity ΦPSII , and total Chl). Trimers of the PSII light-harvesting complexes (LHCIIs) decreased more than LHC monomers and detection of Cd in the monomers suggested replacement of magnesium (Mg) by Cd in the Chl molecules. As a consequence of dysfunctional photosynthesis and energy dissipation, reactive oxygen species (superoxide and hydrogen peroxide) appeared. Cadmium had negative effects on macrophytes at much lower concentrations than reported previously, emphasizing the importance of studies applying environmentally relevant conditions. A chain of inhibition events could be established.

Dinitrogen fixation associated with shoots of aquatic carnivorous plants: is it ecologically important?

BACKGROUND AND AIMS: Rootless carnivorous plants of the genus Utricularia are important components of many standing waters worldwide, as well as suitable model organisms for studying plant-microbe interactions. In this study, an investigation was made of the importance of microbial dinitrogen (N2) fixation in the N acquisition of four aquatic Utricularia species and another aquatic carnivorous plant, Aldrovanda vesiculosa. METHODS: 16S rRNA amplicon sequencing was used to assess the presence of micro-organisms with known ability to fix N2. Next-generation sequencing provided information on the expression of N2 fixation-associated genes. N2 fixation rates were measured following (15)N2-labelling and were used to calculate the plant assimilation rate of microbially fixed N2. KEY RESULTS: Utricularia traps were confirmed as primary sites of N2 fixation, with up to 16 % of the plant-associated microbial community consisting of bacteria capable of fixing N2. Of these, rhizobia were the most abundant group. Nitrogen fixation rates increased with increasing shoot age, but never exceeded 1·3 µmol N g(-1) d. mass d(-1). Plant assimilation rates of fixed N2 were detectable and significant, but this fraction formed less than 1 % of daily plant N gain. Although trap fluid provides conditions favourable for microbial N2 fixation, levels of nif gene transcription comprised <0·01 % of the total prokaryotic transcripts. CONCLUSIONS: It is hypothesized that the reason for limited N2 fixation in aquatic Utricularia, despite the large potential capacity, is the high concentration of NH4-N (2·0-4·3 mg L(-1)) in the trap fluid. Resulting from fast turnover of organic detritus, it probably inhibits N2 fixation in most of the microorganisms present. Nitrogen fixation is not expected to contribute significantly to N nutrition of aquatic carnivorous plants under their typical growth conditions; however, on an annual basis the plant-microbe system can supply nitrogen in the order of hundreds of mg m(-2) into the nutrient-limited littoral zone, where it may thus represent an important N source.

What do results of common sequential fractionation and single-step extractions tell us about P binding with Fe and Al compounds in non-calcareous sediments?

Correct identification of P forms together with their main Fe and Al binding partners in non-calcareous sediments is of crucial importance for evaluation of P cycling in water bodies. In this paper, we assess extraction methods frequently used for this purpose, i.e., a sequential five-step fractionation (water, bicarbonate buffered dithionite solution (BD), NaOH, HCl, nitric-perchloric acid), ascorbate extraction (pH ~7.5), and oxalate extraction (pH ~3), directly on a range of laboratory prepared Fe and Al minerals enriched with adsorbed P. Extraction selectivity and efficiency for particular P, Fe and Al forms were also verified by specific combinations of these extraction methods applied on freshwater sediment samples. In the sequential fractionation, BD was highly effective in dissolving both amorphous and crystalline Fe (hydr)oxides and the associated P, while neither FeS nor Al (hydr)oxides were dissolved. The following NaOH extraction effectively dissolved both amorphous and crystalline Al (hydr)oxides. The high solubilizing power of BD and NaOH to dissolve crystalline Fe and Al oxides that have only a small P-sorption ability prevents the use of resulting Fe/P and Al/P ratios as simple predictors of total P sorption capacity of sediments and soils. Ascorbate non-selectively extracted small proportions of FeS and amorphous Fe and Al (hydr)oxides, but significant amounts of adsorbed P, which hinders its use for the characterization of P forms in non-calcareous sediments. Similar nonselective characteristics were found for oxalate extractions. As oxalate extracts most of the adsorbed phosphate, it is not possible to use it unambiguously to determine specific Fe/P and Al/P ratios of active complexes. However, this method is convenient (and more selective than NaOH step in the sequential fractionation) for the determination of amorphous Al (hydr)oxides.

Ecological implications of organic carbon dynamics in the traps of aquatic carnivorous Utricularia plants.

Rootless aquatic carnivorous Utricularia exude up to 25% of their photosynthates into the trap lumen, which also harbours a complex microbial community thought to play a role in enhancing Utricularia nutrient acquisition. We investigated the composition of organic carbon in the trap fluid, its availability for microbial uptake, the influence of plant nutrient status and trap age on its biodegradability, and the composition of prokaryotic assemblages within the traps of three aquatic Utricularia species. Using ion chromatography and basal respiration rate measurements we confirmed that up to 30% of total dissolved organic carbon in Utricularia trap fluid in oligotrophic conditions was easily biodegradable compounds commonly found in plant root exudates (mainly glucose, fructose and lactate). The proportion of these compounds and their microbial utilisation decreased with increasing mineral nutrient supply and trap age. Fluorescence in situ hybridisation analyses showed that microbial trap assemblages are dominated by alpha and beta Proteobacteria, and that the assemblage composition is affected by changes in the ambient mineral nutrient supply. We suggest that organic carbon dynamics within the traps, involving both the plant and associated microbial assemblages, underlies the acquisition of key nutrients by Utricularia and may help explain the evolutionary success of the genus.

Utricularia carnivory revisited: plants supply photosynthetic carbon to traps.

The rootless, aquatic Utricularia species belong to the largest and most cosmopolitan carnivorous plant genus. Populations of Utricularia plants are an important component of many standing, nutrient-poor, and humic waters. Carbon (C) allocation is an aspect of Utricularia's ecophysiology that has not been studied previously and there is considerable uncertainty about the functional and ecological benefit of the trap-associated microbial community and the potential role played by C exudation in enhancing plant-microbe interactions. A 13C-labelling experiment was conducted in greenhouse conditions to determine the C allocation between plant tissues of increasing age and trap fluid in two Utricularia species. Both species allocated a majority of the newly fixed C into the fast growing shoot apex (46.1+/-8.6% in U. vulgaris and 56.1% in U. australis). Carbon allocation rapidly decreased with increasing age of the shoot, constituting only 8.0+/-4.0% and 6.7% of the total newly fixed C in the oldest analysed segments in U. vulgaris and U. australis, respectively. In the trap-bearing shoot segments, the ratio of C exuded into the trap fluid to that in plant tissues increased markedly with age--in the oldest analysed segments twice as much newly fixed C was allocated into the trap fluid than the plant tissue. Overall, a significant amount of the newly fixed C, approximately 25% (U. vulgaris) and 20% (U. australis), was allocated to the trap fluid. The importance of C exudation for the development of the microbial community associated with the traps as well as for the growth and ecology of aquatic Utricularia is discussed.

Aluminum control of phosphorus sorption by lake sediments.

Release of reactive (phosphate-like) phosphorus (P) from freshwater sediments represents a significant internal P source for many lakes. Hypolimnetic P release occurs under reducing conditions that cause reductive dissolution of ferric hydroxide [Fe(OH)3]. This hypolimnetic P release may be naturally low or artificially reduced by sediment with naturally high or artificially elevated concentrations of aluminum hydroxide [Al(OH)3]. We presentfield and laboratory data for a common extraction analysis of sediments from 43 lakes differing in trophic status, pH regime, climate, and P loading. The results indicate that a simple sequential extraction of sediment may be a useful predictor of sediment's ability to release P. Sequential extractions of sediment P, Al, and Fe by water (H2O), bicarbonate-dithionite (BD), and NaOH (at 25 degrees C) showed that negligible amounts of P would be released from lake sediments during hypolimnetic anoxia if either (1) the molar Al(NaOH-25):Fe(BD) ratio is > 3 or (2) the molar Al(NaOH-25):P(H2O+BD) ratio is > 25. These ratios can be used as operational targets for estimation of sediment P release potential and Al dosing of P-rich sediment to prevent hypolimnetic P release under anoxic conditions.