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The North Temperate Lakes Long-Term Ecological Research (NTL-LTER) program has been extensively used to improve understanding of how aquatic ecosystems respond to environmental stressors, climate fluctuations, and human activities. Here, we report on the metagenomes of samples collected between 2000 and 2019 from Lake Mendota, a freshwater eutrophic lake within the NTL-LTER site. We utilized the distributed metagenome assembler MetaHipMer to coassemble over 10 terabases (Tbp) of data from 471 individual Illumina-sequenced metagenomes. A total of 95,523,664 contigs were assembled and binned to generate 1,894 non-redundant metagenome-assembled genomes (MAGs) with ≥50% completeness and ≤10% contamination. Phylogenomic analysis revealed that the MAGs were nearly exclusively bacterial, dominated by Pseudomonadota (Proteobacteria, N = 623) and Bacteroidota (N = 321). Nine eukaryotic MAGs were identified by eukCC with six assigned to the phylum Chlorophyta. Additionally, 6,350 high-quality viral sequences were identified by geNomad with the majority classified in the phylum Uroviricota. This expansive coassembled metagenomic dataset provides an unprecedented foundation to advance understanding of microbial communities in freshwater ecosystems and explore temporal ecosystem dynamics.

Although microbes mediate much of the biogeochemical cycling in freshwater, the categories of carbon and nutrients currently used in models of freshwater biogeochemical cycling are too broad to be relevant on a microbial scale. One way to improve these models is to incorporate microbial data. Here, we analyze both genes and genomes from three metagenomic time series and propose specific roles for microbial taxa in freshwater biogeochemical cycles. Our metagenomic time series span multiple years and originate from a eutrophic lake (Lake Mendota) and a humic lake (Trout Bog Lake) with contrasting water chemistry. Our analysis highlights the role of polyamines in the nitrogen cycle, the diversity of diazotrophs between lake types, the balance of assimilatory vs. dissimilatory sulfate reduction in freshwater, the various associations between types of phototrophy and carbon fixation, and the density and diversity of glycoside hydrolases in freshwater microbes. We also investigated aspects of central metabolism such as hydrogen metabolism, oxidative phosphorylation, methylotrophy, and sugar degradation. Finally, by analyzing the dynamics over time in nitrogen fixation genes and Cyanobacteria genomes, we show that the potential for nitrogen fixation is linked to specific populations in Lake Mendota. This work represents an important step towards incorporating microbial data into ecosystem models and provides a better understanding of how microbes may participate in freshwater biogeochemical cycling.

Freshwater ecosystems can serve as model systems that reveal insights into biological invasions. In this article, we summarize nine lessons about aquatic invasive species from the North Temperate Lakes Long-Term Ecological Research program and affiliated projects. The lessons about aquatic invasive species are as follows: Invasive species are more widespread than has been documented; they are usually at low abundance; they can irrupt from low-density populations in response to environmental triggers; they can occasionally have enormous and far-reaching impacts; they can affect microbial communities; reservoirs act as invasive species hotspots; ecosystem vulnerability to invasion can be estimated; invasive species removal can produce long-term benefits; and the impacts of invasive species control may be greater than the impacts of the invasive species. This synthesis highlights how long-term research on a freshwater landscape can advance our understanding of invasions. [ABSTRACT FROM AUTHOR]

Species invasions can disrupt aquatic ecosystems by re‐wiring food webs. A trophic cascade triggered by the invasion of the predatory zooplankter spiny water flea (Bythotrephes cederströmii) resulted in increased phytoplankton due to decreased zooplankton grazing. Here, we show that increased phytoplankton biomass led to an increase in lake anoxia. The temporal and spatial extent of anoxia experienced a step change increase coincident with the invasion, and anoxic factor increased by 11 d. Post‐invasion, anoxia established more quickly following spring stratification, driven by an increase in phytoplankton biomass. A shift in spring phytoplankton phenology encompassed both abundance and community composition. Diatoms (Bacillaryophyta) drove the increase in spring phytoplankton biomass, but not all phytoplankton community members increased, shifting the community composition. We infer that increased phytoplankton biomass increased labile organic matter and drove hypolimnetic oxygen consumption. These results demonstrate how a species invasion can shift lake phenology and biogeochemistry. [ABSTRACT FROM AUTHOR]

Freshwater ecosystems can serve as model systems that reveal insights into biological invasions. In this article, we summarize nine lessons about aquatic invasive species from the North Temperate Lakes Long-Term Ecological Research program and affiliated projects. The lessons about aquatic invasive species are as follows: Invasive species are more widespread than has been documented; they are usually at low abundance; they can irrupt from low-density populations in response to environmental triggers; they can occasionally have enormous and far-reaching impacts; they can affect microbial communities; reservoirs act as invasive species hotspots; ecosystem vulnerability to invasion can be estimated; invasive species removal can produce long-term benefits; and the impacts of invasive species control may be greater than the impacts of the invasive species. This synthesis highlights how long-term research on a freshwater landscape can advance our understanding of invasions., (© The Author(s) 2024. Published by Oxford University Press on behalf of the American Institute of Biological Sciences.)

Ecology and evolution are often viewed as distinct processes, which interact on contemporary time scales in microbiomes. To observe these processes in a natural system, we collected a two-decade, 471-sample freshwater lake time series, creating the longest metagenome dataset to date. Among 2,855 species-representative genomes, diverse species and strains followed cyclical seasonal patterns, and one in five species experienced decadal shifts in strain composition. The most globally abundant freshwater bacterium had constant species-level abundance, but environmental extremes appeared to trigger a shift in strain composition and positive selection of amino acid and nucleic acid metabolism genes. These genes identify organic nitrogen compounds as potential drivers of freshwater responses to global change. Seasonal and long-term strain dynamics could be regarded as ecological processes or equivalently as evolutionary change. Rather than as distinct processes that interact, we propose a conceptualization where ecology and evolution converge along a continuum to better describe change in diverse microbial communities., Competing Interests: Competing interests Authors declare that they have no competing interests.