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The increasing frequency of cyanobacteria blooms in waterbodies caused by ecosystem eutrophication could endanger human health. This risk can be mitigated by effective monitoring incorporating molecular methods. To date, most molecular studies on toxigenic cyanobacteria have been limited to microcystins (MCs), disregarding other cyanotoxins, to freshwater planktic habitats while ignoring benthic habitats, and to limited geographic areas (usually one or a few specific waterbodies). In this study, we used PCR-based methods including PCR product sequencing and chemical-analytical methods (LC-MS/MS) to screen many plankton (n = 123) and biofilm samples (n = 113) originating from 29 Alpine lakes and 18 rivers for their cyanotoxin production potential. Both mcyE (indicating MC synthesis) and anaC (indicating anatoxin (ATX) synthesis) gene fragments were able to qualitatively predict MC or ATX occurrence. The abundance of mcyE gene fragments was significantly related to MC concentrations in plankton samples (R 2 = 0.61). mcyE gene fragments indicative of MC synthesis were most abundant in planktic samples (65 %) and were assigned to the genera Planktothrix and Microcystis. However, mcyE rarely occurred in biofilms of lakes and rivers, i.e., 4 % and 5 %, respectively, and were assigned to Microcystis, Planktothrix, and Nostoc. In contrast, anaC gene fragments occurred frequently in planktic samples (14 % assigned to Tychonema, Phormidium (Microcoleus), and Oscillatoria), but also in biofilms of lakes (49 %) and rivers (18 %) and were assigned to the genera Phormidium, Oscillatoria, and Nostocales. The cyrJ gene fragment indicating cylindrospermopsin synthesis occurred only once in plankton (assigned to Dolichospermum), while saxitoxin synthesis potential was not detected. For plankton samples, monomictic and less eutrophic conditions were positively related to mcyE/MC occurrence frequency, while oligomictic conditions were related to anaC/ATX frequency. The anaC/ATX frequency in biofilm was related to the lake habitats generally showing higher biodiversity as revealed from metabarcoding in a parallel study., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

Plastic debris is thought to be widespread in freshwater ecosystems globally 1 . However, a lack of comprehensive and comparable data makes rigorous assessment of its distribution challenging 2,3 . Here we present a standardized cross-national survey that assesses the abundance and type of plastic debris (>250 μm) in freshwater ecosystems. We sample surface waters of 38 lakes and reservoirs, distributed across gradients of geographical position and limnological attributes, with the aim to identify factors associated with an increased observation of plastics. We find plastic debris in all studied lakes and reservoirs, suggesting that these ecosystems play a key role in the plastic-pollution cycle. Our results indicate that two types of lakes are particularly vulnerable to plastic contamination: lakes and reservoirs in densely populated and urbanized areas and large lakes and reservoirs with elevated deposition areas, long water-retention times and high levels of anthropogenic influence. Plastic concentrations vary widely among lakes; in the most polluted, concentrations reach or even exceed those reported in the subtropical oceanic gyres, marine areas collecting large amounts of debris 4 . Our findings highlight the importance of including lakes and reservoirs when addressing plastic pollution, in the context of pollution management and for the continued provision of lake ecosystem services., (© 2023. The Author(s), under exclusive licence to Springer Nature Limited.)

In many regions across the globe, extreme weather events such as storms have increased in frequency, intensity, and duration due to climate change. Ecological theory predicts that such extreme events should have large impacts on ecosystem structure and function. High winds and precipitation associated with storms can affect lakes via short-term runoff events from watersheds and physical mixing of the water column. In addition, lakes connected to rivers and streams will also experience flushing due to high flow rates. Although we have a well-developed understanding of how wind and precipitation events can alter lake physical processes and some aspects of biogeochemical cycling, our mechanistic understanding of the emergent responses of phytoplankton communities is poor. Here we provide a comprehensive synthesis that identifies how storms interact with lake and watershed attributes and their antecedent conditions to generate changes in lake physical and chemical environments. Such changes can restructure phytoplankton communities and their dynamics, as well as result in altered ecological function (e.g., carbon, nutrient and energy cycling) in the short- and long-term. We summarize the current understanding of storm-induced phytoplankton dynamics, identify knowledge gaps with a systematic review of the literature, and suggest future research directions across a gradient of lake types and environmental conditions., (© 2020 The Authors. Global Change Biology published by John Wiley & Sons Ltd.)

High-throughput sequencing (HTS) was used to analyze the seasonal variations in the bacterioplankton community composition (BCC) in the euphotic layer of a large and deep lake south of the Alps (Lake Garda). The BCC was analyzed throughout two annual cycles by monthly samplings using the amplification and sequencing of the V3-V4 hypervariable region of the 16S rRNA gene by the MiSeq Illumina platform. The dominant and most diverse bacterioplankton phyla were among the more frequently reported in freshwater ecosystems, including the Proteobacteria, Cyanobacteria, Bacteroidetes, Verrucomicrobia, Actinobacteria, and Planctomycetes. As a distinctive feature, the development of the BCC showed a cyclical temporal pattern in the two analyzed years and throughout the euphotic layer. The recurring temporal development was controlled by the strong seasonality in water temperature and thermal stratification, and by cyclical temporal changes in nutrients and, possibly, by the remarkable annual cyclical development of cyanobacteria and eukaryotic phytoplankton hosting bacterioplankton that characterizes Lake Garda. Further downstream analyses of operational taxonomic units associated to cyanobacteria allowed confirming the presence of the most abundant taxa previously identified by microscopy and/or phylogenetic analyses, as well as the presence of other small Synechococcales/Chroococcales and rare Nostocales never identified so far in the deep lakes south of the Alps. The implications of the high diversity and strong seasonality are relevant, opening perspectives for the definition of common and discriminating patterns characterizing the temporal and spatial distribution in the BCC, and for the application of the new sequencing technologies in the monitoring of water quality in large and deep lakes.

Insight into how environmental change determines the production and distribution of cyanobacterial toxins is necessary for risk assessment. Management guidelines currently focus on hepatotoxins (microcystins). Increasing attention is given to other classes, such as neurotoxins (e.g., anatoxin-a) and cytotoxins (e.g., cylindrospermopsin) due to their potency. Most studies examine the relationship between individual toxin variants and environmental factors, such as nutrients, temperature and light. In summer 2015, we collected samples across Europe to investigate the effect of nutrient and temperature gradients on the variability of toxin production at a continental scale. Direct and indirect effects of temperature were the main drivers of the spatial distribution in the toxins produced by the cyanobacterial community, the toxin concentrations and toxin quota. Generalized linear models showed that a Toxin Diversity Index (TDI) increased with latitude, while it decreased with water stability. Increases in TDI were explained through a significant increase in toxin variants such as MC-YR, anatoxin and cylindrospermopsin, accompanied by a decreasing presence of MC-LR. While global warming continues, the direct and indirect effects of increased lake temperatures will drive changes in the distribution of cyanobacterial toxins in Europe, potentially promoting selection of a few highly toxic species or strains., Competing Interests: The authors declare no conflict of interest.

Lake water level fluctuations (WLF) are an important factor driving the selection and seasonal dynamics of phytoplankton and potentially toxigenic cyanobacteria. Nevertheless, the relative importance of environmental drivers connected to WLF may be completely different, depending on the typology and use of waterbodies, latitude and climatic regimes. In this study, we investigated the impact of WLF in a large subtropical reservoir in south-eastern China (Hongfeng Reservoir, Guizhou Province). The study was based on monthly samplings carried out in 2014 in six stations. The strong increase in the water level observed in early summer caused a radical shift in the phytoplankton community. While in the pre-flooding period phytoplankton was composed of large diatoms, chrysophytes and Oscillatoriales (mostly Limnothrix sp.), the post-flooding period showed an increase in smaller and more competitive chlorophytes, smaller diatoms and cryptophytes better adapted to a fast colonisation of new and nutrient-rich environments. The environmental drivers that drove the change were dilution, flushing and interference with the seasonal water stratification processes. We concluded that, because WLF represents a complex variable integrating different physical effects in one explanatory descriptor, its value as a predictor of phytoplankton and cyanobacteria dynamics in lake ecosystems is difficult to generalise and needs to be investigated on a case-by-case basis. For this reason, considering the year-to-year hydrological variability that potentially characterise reservoirs, definite indications for management should be outlined considering more than 1-year study.

This work allowed assessing a widespread occurrence of Tychonema bourrellyi in the largest lakes south of the Alps (Garda, Iseo, Como and Maggiore). The taxonomy of the species was confirmed adopting a polyphasic approach, which included microscopic examinations, molecular (16S rRNA and rbcLX sequences) and (Lake Garda) ecological characterisations. Over 70% of the 36 isolates of Tychonema sampled from the four lakes tested positive for the presence of genes implicated in the biosynthesis of anatoxins (anaF and/or anaC) and for the production of anatoxin-a (ATX) and homoanatoxin-a (HTX). A detailed analysis carried out in Lake Garda showed strong ongoing changes in the cyanobacterial community, with populations of Tychonema developing with higher biovolumes compared to the microcystins (MCs) producer Planktothrix rubescens Moreover, the time × depth distribution of Tychonema was paralleled by a comparable distribution of ATX and HTX. The increasing importance of Tychonema in Lake Garda was also suggested by the opposite trends of ATX and MCs observed since 2009. These results suggest that radical changes are occurring in the largest lakes south of the Alps. Their verification and implications will require to be assessed by extending a complete experimental work to the other large perialpine lakes., (© FEMS 2016. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

A growing concern exists about the effects of chloride (Cl) on freshwater systems. Increasing Cl concentrations have been observed in the last few decades in several rivers and lakes, mainly in northern countries. In Italy, present levels and temporal changes of sodium (Na) and Cl in water bodies have rarely been assessed. Based on long-term data for the lakes of the subalpine district in Italy (Maggiore, Lugano, Como, Iseo, Garda), we analyzed trends affecting Cl and Na concentrations during the last 25 years, with the aim of identifying temporal changes and assessing possible causes. An in-depth analysis is presented for Lake Maggiore. Positive temporal Na and Cl trends were evident in all studied lakes, with the trends increasing since early 2000s. Data for Lake Maggiore tributaries showed a clear seasonality (higher values in winter and early spring). The NaCl used as road de-icing agent, together with Cl discharge from wastewater treatment plants, were identified as the main causes for the observed trends. Chloride concentrations in the lakes are below the threshold limit for reduced water quality and below concentrations known to harm aquatic biota. However, considering the relevance of deep subalpine lakes, representing almost 80% of the total freshwater volume in Italy, these trends indicate an important chemical change, which warrants further analysis.

In order to identify the cyanobacterial species responsible of anatoxin-a (ATX) production in Lake Garda (Northern Italy), an intensive isolation and culturing of filamentous cyanobacteria were established since 2014 from environmental samples. In this work, we report a detailed account of the strategy adopted, which led to the discovery of a new unexpected producer of ATX, Tychonema bourrellyi. So far, this species is the first documented example of cultured Oscillatoriales able to produce ATX isolated from pelagic freshwater ecosystems. The isolated filaments were identified adopting a polyphasic approach, which included microscopic species identification, genetic characterisation and phylogenetic analyses based on 16S rRNA genes. The taxonomic identification was further confirmed by the high (>99%) rbcLX sequence similarities of the T. bourrellyi strains of Lake Garda with those deposited in DNA sequence databases. More than half of the isolates were shown to produce a significant amount of ATX, with cell quota ranging between 0.1 and 2.6 μg mm(-3), and 0.01 and 0.35 pg cell(-1). The toxic isolates were tested positive for anaC of the anatoxin-a synthetase (ana) gene cluster. These findings were confirmed with the discovery of one ATX producing T. bourrellyi strain isolated in Norway. This strain and a further non-ATX producing Norwegian Tychonema bornetii strain tested positive for the presence of the anaF gene of the ana gene cluster. Conversely, none of the Italian and Norwegian Tychonema strains were positive for microcystins (MCs), which was also confirmed by the absence of mcyE PCR products in all the samples analysed. This work suggests that the only reliable strategy to identify cyanotoxins producers should be based on the isolation of strains and their identification with a polyphasic approach associated to a concurrent metabolomic profiling., (Copyright © 2014 Elsevier Ltd. All rights reserved.)