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Diatoms, eukaryotic algae with silica cell wall have been proved to be reliable bioindicators and are applied in ecological status assessment of aquatic ecosystems using indices calculated from pollution sensitivity and indicator values of species. Traditional method of identification of diatom species is based on morphological features of frustule that requires in-depth taxonomical knowledge and expertise. Metabarcoding that combines barcodes and high-throughput sequencing offers a promising alternative. In this pilot study we tested the applicability of metabarcoding of benthic diatom assemblages for ecological status assessment of water bodies in Hungary comparing its performance to that of morphology-based identification of species. Samples from 78 rivers and streams arranged along a trophic gradient and 14 soda pans representing unique types of habitats were investigated. Sequences of the barcode region of rbcL gene suggested by Vasselon et al. (2017) were acquired revealing 1135 diatom amplicon sequence variants (ASVs) of which 54% could be assigned at species level using Diat.barcode (Rimet et al. 2019) as reference database. Morphological investigation detected more species and intraspecific taxa in the lotic and lentic samples (413 and 78, respectively) than metabarcoding (190 and 75). Correspondence between taxa detected based on morphology and DNA sequences was relatively low (on average 24% in lotic and 26% in lentic samples) but considerably increased (on average 66% and 56%, respectively) when taking into account only the taxa reaching higher than 5% relative abundance in microscopy counting. The differences mainly derived from that a considerable portion (59% and 32%, respectively) of the morphologically identified taxa were not recorded in the reference database. Community structure of samples from both running and standing waters based on microscopy and DNA sequence analysis showed significant correlation as revealed using Mantel test. For rivers and streams diatom index IPS (Specific Pollution Sensitivity Index, (Coste in Cemagref, 1982) for soda pans indices H (Ziemann et al., 2001) and IBD (Indice Biologique Diatomées, Lenoir and Coste, 1996) were calculated from the results of the two aspects. Strong correlation was found between morphology- and sequence-based indices and in about half of the samples, the ecological status class obtained with the two methods coincided. Our results suggest that metabarcoding inheres a great opportunity and could be successfully applied in benthic diatom-based ecological status assessment of Hungarian waters after the reference database is supplemented with taxa frequently occurring in these habitats. References Cemagref, 1982. Etude des méthodes biologiques quantitativesd’appréciation de la qualité des eaux, Rapport Division Qualité des Eaux Lyon. Agence financière de Bassin Rhône-Méditerranée-Corse, Pierre-Bénite, France. Lenoir, A., Coste, M., 1996. Development of a practical diatom index of overall water quality applicable to the French National Water Board Network. Presented at the International symposium, Volksbildungsheim Grilhof Vill, AUT, 17-19 September 1995, Universität Innsbruck, pp. 29–43. Rimet, F., Gusev, E., Kahlert, M., Kelly, M.G., Kulikovskiy, M., Maltsev, Y., Mann, D.G., Pfannkuchen, M., Trobajo, R., Vasselon, V., Zimmermann, J., Bouchez, A., 2019. Diat.barcode, an open-access curated barcode library for diatoms. Sci. Rep. 9, 15116. Vasselon, V., Rimet, F., Tapolczai, K., Bouchez, A., 2017. Assessing ecological status with diatoms DNA metabarcoding: Scaling-up on a WFD monitoring network (Mayotte island, France). Ecol. Indic. 82, 1–12. Ziemann, H., Kies, L., Schulz, C.-J., 2001. Desalinization of running waters: III. Changes in the structure of diatom assemblages caused by a decreasing salt load and changing ion spectra in the river Wipper (Thuringia, Germany). Limnologica 31, 257–280.

Although metacommunity dynamics of lentic phytoplankton are relatively well-documented, studies on the role of environmental and spatial processes in shaping phytoplankton communities of large rivers are still scarce. Here, we examined six phytoplankton data sets, which were collected in 1978-2017 from large river-scale segments (mean spatial extent 1117 km) in the Danube River. Our aim was to elucidate role of climatic, spatial and temporal predictors in variation of phytoplankton beta diversity using variance partitioning for compositions of species and functional groups sensu Reynolds. We hypothesised that phytoplankton beta diversity (measured as average distance to group centroid) would be positively related to both climatic heterogeneity and spatial extent used as a proxy for dispersal limitation. Additionally, we tested alternative dispersal models to evaluate different spatial processes structuring phytoplankton community. Our results revealed that spatial variables were more important than climatic factors in controlling both species and functional group composition. Climatic heterogeneity showed significant positive relationship with beta diversity. In contrast, there was no significant relationship between beta diversity and spatial extent, suggesting that spatial effect on beta-diversity was attenuated by anthropogenic disturbance. The better performance of non-directional model compared to model of water directionality suggested that spatial dynamics of phytoplankton metacommunity was in large part regulated by differences in the regional species pools. Spatial and temporal variables outperformed environmental (including climatic) factors in explaining phytoplankton metacommunity structure, indicating that phytoplankton exhibited strong biogeographical patterns. Thus, dispersal limitation interfered with species-sorting processes in determining phytoplankton community structure. In conclusion, our findings revealed that the development of a more reliable bioassessment program of the Danube River should be based on separation into basin regions.

Here we report the results of our decades-long study on epiphytic communities from two tributary systems of the Szigetk&ouml, z section of the Danube River. The main goal of the investigation was to detect changes in the epiphytic communities at structural (core species, changes in the relative abundance of common species) and functional (trait changes) levels as a result of the most important anthropogenic effects on Szigetk&ouml, z, i.e., hydro-morphological modifications. We also examined the impact of rehabilitation on the tributary systems in terms of ecological potential. We discovered that mainly motile diatom species characterized the epiphyton due to reduced water volume were introduced into the tributary system after the diversion of the Danube. The ecosystem stabilized in the rehabilitated section, while the non-rehabilitated section showed a worsening tendency, mainly in the parapotamic branches. Our long-term data sets may provide a good basis for comparisons of different aquatic ecosystems, to define changes in the abundance of core species and in the structure of community in response to different anthropogenic pressures. It is fundamental to determine adaptive traits in assessing the impact of global warming stressors on biodiversity.

While phytoplankton studies on large potamal rivers have increased in number in recent years, upper river sections have received considerably less attention. However, in order to better understand processes that govern the development of dominance of euplanktonic elements in the lower river sections, detailed studies of the upstream areas are necessary. We studied the composition, diversity and recruitment of the planktonic algal communities in the River Sajo and in its main tributary River Hernad, two characteristic rhithral river systems in Central Europe. Results revealed that diatoms are dominant elements of the phytoplankton in the upper river segments both in terms of taxa richness, and relative abundance. We found that composition of the phytoplankton showed the closest resemblance to that of benthic communities of soft substrates, highlighting the key role of riverbed characteristics in river phytoplankton recruitment. The occurrence of euplanktonic elements was not restricted to human impacted river segments, these taxa also occurred in pristine headwaters. Accordingly, planktonic algae potentially colonise small headwater streams naturally, although their dominance is expected to occur only at downstream reaches following impounded river segments. Diatom metrics used in ecological state assessment were calculated for the rithroplankton. These metrics fell in the range of values calculated for diatom flora on the hard substrates. However, the potential use of rithroplankton for quality assessment might be limited, because the high variability of index values at site level would result in misclassification of the ecological status.

Dissipation of the herbicide active ingredient glyphosate was investigated in natural waters. To assess combined effects, glyphosate was applied in its pure form (glyphosate isopropylammonium salt) and in preparation Roundup Classic® formulated with polyethoxylated tallowamines (POEA). Standing and running surface water samples originated from Lake Balaton and River Danube between early May and mid-June of 2015. The kinetics of dissipation of glyphosate, measured by high-performance liquid chromatography combined with UV-VIS absorbance detection or tandem mass spectrometry, was investigated under laboratory conditions in aquaria with or without the presence of biofilms. The quantity and the biofilm structure of algal biomass were determined by in vivo fluorimetry and scanning electron microscopy. The presence of POEA affected the dissipation of glyphosate, and dissipation profiles differed in the investigated natural waters. Significantly higher initial concentrations of glyphosate were measured i...

Specimens of a very small diatom were found in a dead oxbow of Tisza River (Hungary), in Eula River (Germany) and Parroy Pond (France). After SEM studies we identified all of them as Eolimna archibaldii J.C. Taylor et Lange-Bert., which is currently regarded as endemic species to South Africa. Furthermore, the results of Principal Components Analysis of morphometric characters clearly demonstrated an overlap between the type material and European populations. Therefore this diatom cannot be considered endemic to South Africa. Moreover, E. archibaldii and a very similar species, E. becaresii are transferred to the genus Sellaphora. Our results confirm the benefit of SEM for verifying the identity of small-sized diatoms, especially for routine monitoring, and allow more precise ecological assessment.

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.

(1) Urbanization significantly influences the ecosystems of rivers in various ways, including the so-called loading effect of wastewater production. Benthic diatoms are used in ecological status assessments of waters. Beside species composition, traits can be used as indicators. We aimed to evaluate how the loading of the large city of Budapest manifests in the physico-chemical variables of the River Danube and what species composition and trait response this loading results in for the benthic diatom communities. (2) Weekly samplings were performed at points upstream and downstream of Budapest on both riverbanks. Samples were compared, based on general physical-chemical variables and the concentration of thirty-four elements, as well as species composition and seven traits of species of diatom communities. Ecological status was assessed using the Specific Pollution Sensitivity Index (IPS). (3) Only a few measured environmental variables showed differences between the sampling points, suggesting that the nutrient loading has significantly decreased due to the installation of several efficiently working wastewater treatment plants since the introduction of the European Union Water Framework Directive. In contrast, the species composition and traits of species showed the effect of land use. Benthic diatoms indicate the environmental changes caused by land use in the longer-term, while chemical measurements reflect instantaneous status.

The spatial response of epiphytic diatom communities to environmental stress was studied in a moderately saline wetland area located in the plain of Danube-Tisza Interfluve, Hungary. The area is characterised by World War II bomb crater ponds and can be regarded as an excellent ecological model system where the dispersion of species is slightly limited by distance. To study the effect of environmental variables on the communities, canonical correspondence analysis was applied. Salinity, pH, total suspended solids, total phosphorous and depth proved to be significant environmental drivers in this analysis. The ecological status of the ponds was assessed with Ziemann’s halobity index, as the trophity-depending metric cannot be applied to these habitats (due to the naturally high phosphorus content). Ponds in “good” ecological status significantly differed from those appertaining to water quality category of “not-good” ecological status considering characteristic of natural astatic soda pans (e.g. salinity, pH, ammonium, total phosphorous concentration, nitrogen:phosphorous ratio and turbidity). The differences between epiphytic diatom communities inhabiting the ponds were detected using non-parametric multidimensional scaling. The samples formed three groups according to the types of ponds (“transparent”, “transitional” and “turbid”) based on the width of the macrophyte belt around them. Indicator species related to the ecological status of the ponds and diatom communities contributing to the separation of groups of ponds were identified. One of the indicator species differed from species already described. Light and scanning electron microscopy features and phylogenetic analyses based on three genes (18S and 28S rRNA genes, rbcL) proved that it was a new species of Nitzschia genus, closely related to Nitzschia frustulum and Nitzschia inconspicua. Therefore, description of a new species, Nitzschia reskoi Ács, Duleba, C.E.Wetzel & Ector is proposed. We concluded that the increasing abundance of Nitzschia reskoi was a signal of the degradation of the intermittent saline wetlands.

A detailed scanning electron microscopic investigation was carried out to clarify the taxonomic status of a small sigmoid Nitzschia species, a potential indicator of Central European soda waters. We found this taxon to be one of the dominant epiphytic diatoms collected from sodic bomb crater ponds at Apaj (Hungary). The large population allowed for a morphometric comparison based on frustule ultrastructure with the type material of the most similar species, Nitzschia austriaca Hustedt that was originally described from a soda pan in the region. The results clearly demonstrated an overlap between the Apaj population and the type material of N. austriaca (based on NMDS analysis), therefore we argue that they represent the same taxon. An emended diagnosis of N. austriaca is given. Total suspended solids and total phosphorous proved to be the most important factors predicting the occurrence of the species, with possible interactive effects of conductivity and pH. We then expanded the distribution of the species by revisiting data originating from previous large-scale surveys targeting sodic habitats in Hungary. On the basis of our results, N. austriaca is a characteristic species for Central European soda waters, including the protected astatic soda pans, indicating their typical chemical and physical characteristics.

An unknown small naviculoid diatom was found in the epiphyton samples from Hungary (in Kanyari Holt-Tisza oxbow) and in the epilithon samples from France (Canal de Berry at Épineuil-le-Fleuriel). Detailed observations using light and scanning electron microscopy were used to describe Brevilinea kevei Ács & Ector sp. nov. It can be separated from Brevilinea pocosinensis Siver et al. (which was the only described species in the genus) by its size (smaller than B. pocosinensis), shape (elliptic valve, never capitate) and ecological requirements. Brevilinea pocosinensis lives in acidic conditions while B. kevei in neutral ones. Differential diagnostic criteria with respect to similar taxa, especially Fallacia indifferens (Hustedt) D.G. Mann, together with ecological implications of this finding are briefly discussed.