In August 2019, three dogs died after bathing in or drinking from Mandichosee, a mesotrophic reservoir of the River Lech (Germany). The dogs showed symptoms of neurotoxic poisoning and intoxication with cyanotoxins was considered. Surface blooms were not visible at the time of the incidents. Benthic Tychonema sp., a potential anatoxin-a (ATX)-producing cyanobacterium, was detected in mats growing on the banks, as biofilm on macrophytes and later as aggregations floating on the lake surface. The dogs’ pathological examinations showed lung and liver lesions. ATX and dihydroanatoxin-a (dhATX) were detected by LC-MS/MS in the stomachs of two dogs and reached concentrations of 563 and 1207 µg/L, respectively. Anatoxins (sum of ATX and dhATX, ATXs) concentrations in field samples from Mandichosee ranged from 0.1 µg/L in the open water to 68,000 µg/L in samples containing a large amount of mat material. Other (neuro)toxic substances were not found. A molecular approach was used to detect toxin genes by PCR and to reveal the cyanobacterial community composition by sequencing. Upstream of Mandichosee, random samples were taken from other Lech reservoirs, uncovering Tychonema and ATXs at several sampling sites. Similar recent findings emphasize the importance of focusing on the investigation of benthic toxic cyanobacteria and applying appropriate monitoring strategies in the future.

Bacterioplankton plays an essential role in aquatic ecosystems, and cyanobacteria are an influential part of the microbiome in many water bodies. In freshwaters used for recreational activities or drinking water, toxic cyanobacteria cause concerns due to the risk of intoxication with cyanotoxins, such as microcystins. In this study, we aimed to unmask relationships between toxicity, cyanobacterial community composition, and environmental factors. At the same time, we assessed the correlation of a genetic marker with microcystin concentration and aimed to identify the main microcystin producer. We used Illumina MiSeq sequencing to study the bacterioplankton in two recreational lakes in South Germany. We quantified a microcystin biosynthesis gene (mcyB) using qPCR and linked this information with microcystin concentration to assess toxicity. Microcystin biosynthesis gene (mcyE)-clone libraries were used to determine the origin of microcystin biosynthesis genes. Bloom toxicity did not alter the bacterial community composition, which was highly dynamic at the lowest taxonomic level for some phyla such as Cyanobacteria. At the OTU level, we found distinctly different degrees of temporal variation between major bacteria phyla. Cyanobacteria and Bacteroidetes showed drastic temporal changes in their community compositions, while the composition of Actinobacteria remained rather stable in both lakes. The bacterial community composition of Alpha- and Beta-proteobacteria remained stable over time in Lake Klostersee, but it showed temporal variations in Lake Bergknappweiher. The presence of potential microcystin degraders and potential algicidal bacteria amongst prevalent Bacteroidetes and Alphaproteobacteria implied a role of those co-occurring heterotrophic bacteria in cyanobacterial bloom dynamics. Comparison of both lakes studied revealed a large shared microbiome, which was shaped toward the lake specific community composition by environmental factors. Microcystin variants detected were microcystin-LR, -RR, and -YR. The maximum microcystin concentrations measured was 6.7 μg/L, a value still acceptable for recreational waters but not drinking water. Microcystin concentration correlated positively with total phosphorus and mcyB copy number. We identified low abundant Microcystis sp. as the only microcystin producer in both lakes. Therefore, risk assessment efforts need to take into account the fact that non-dominant species may cause toxicity of the blooms observed.

The Osterseen Lake District in Bavaria consists of 19 small interconnected lakes that exhibit a pronounced trophic gradient from eutrophic to oligotrophic. It therefore presents a unique model system to address ecological questions regarding niche adaptation and Baas Becking's long standing hypothesis of “everything is everywhere, but the environment selects.” Here, we present the first assessment of the microbial diversity in these lakes. We sampled the lakes in August and December and used 454 pyrosequencing of 16S rRNA amplicons to analyze the microbial diversity. The diversity patterns between lakes and seasons were compared and the bacterial community composition was correlated with key chemical and physical parameters. Distinct patterns of bacterial diversity only emerged at the level of individual OTUs (operational taxonomic units), but not at the level of the major bacterial phyla. This emphasizes the high functional and physiological diversity among bacterial species within a phylum and calls for analysis of biodiversity at the level of OTUs in order to understand fine-scale biogeography. We were able to identify a number of cosmopolitan OTUs as well as specialist OTUs that were restricted to certain lakes or seasons, suggesting adaptation to specific ecological niches.

We analyzed the depth distributions of benthic diatoms in two adjacent, but hydrologically distinct subalpine lakes (Lakes Soiernseen, S-Germany). Lake Unterer Soiernsee is affected by marked water-level fluctuations and is light-penetrated to the bottom most of the year, while Lake Oberer Soiernsee provides more stable conditions and an extended aphotic zone. Mixed samples of epiphytic, epilithic, epipsammic and epipelic periphyton were taken in one-meter depth steps by scuba divers. Most of the common benthic diatoms occurred in distinct depth-areas. RDA analyses showed that depth was strongly correlated with species distribution in both lakes. Depth-constrained cluster analyses indicated three distinct diatom community zones in each lake. A shallow littoral zone hosting mainly epiphytic and epilithic species and a deeper littoral zone with mainly epipsammic and epipelic taxa existed in both lakes. Additionally, a highly disturbed near-shore littoral zone with diatoms adapted to unstable conditions (aerophilic taxa, pioneer species) was found in Lake Unterer Soiernsee, and a deep-water pelagic zone with mainly planktonic taxa in Lake Oberer Soiernsee. Light availability, substrate, physical stressors and nutrient concentrations were linked closely with water depth. While light availability affected the ratio of benthic and planktonic diatoms, substrate type influenced benthic diatom assemblage structures. Diatoms occurring in surficial sediments of the aphotic zone represent an ideal cross-section of the recent diatom assemblage of the lake, including benthic and planktonic species. However, sediment samples taken in light-flooded depths are inappropriate for studies based on shifts between benthic and planktonic taxa, because in situ benthic species dominate the surface-sediment assemblages, while settled tychoplanktonic and planktonic species occur less frequently. A diatom-inferred depth model was created for each lake to prove the usability for down-core studies using weighted-averaging approaches. For both lakes these models are highly appropriate to reconstruct past fluctuations in water-transparency or lake-level. With regard to the development of diatom-based TP-transfer-functions for Bavarian mountain lakes, we found it is highly important to consider lake depth and transparency. Based on the findings of this study we recommend the creation of two different training-sets, one for deep or low-transparency lakes with an aphotic zone including both benthic and planktonic diatoms, and another one for shallow, clear water lakes solely using benthic diatoms.

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