Description: © The Author(s), 2021. This article is distributed under the terms of the Creative Commons Attribution License. The definitive version was published in Castro, S. P., Borton, M. A., Regan, K., de Angelis, I. H., Wrighton, K. C., Teske, A. P., Strous, M., & Ruff, S. E. Degradation of biological macromolecules supports uncultured microbial populations in Guaymas Basin hydrothermal sediments. Isme Journal. (2021), https://doi.org/10.1038/s41396-021-01026-5.

Description: Hydrothermal sediments contain large numbers of uncultured heterotrophic microbial lineages. Here, we amended Guaymas Basin sediments with proteins, polysaccharides, nucleic acids or lipids under different redox conditions and cultivated heterotrophic thermophiles with the genomic potential for macromolecule degradation. We reconstructed 20 metagenome-assembled genomes (MAGs) of uncultured lineages affiliating with known archaeal and bacterial phyla, including endospore-forming Bacilli and candidate phylum Marinisomatota. One Marinisomatota MAG had 35 different glycoside hydrolases often in multiple copies, seven extracellular CAZymes, six polysaccharide lyases, and multiple sugar transporters. This population has the potential to degrade a broad spectrum of polysaccharides including chitin, cellulose, pectin, alginate, chondroitin, and carrageenan. We also describe thermophiles affiliating with the genera Thermosyntropha, Thermovirga, and Kosmotoga with the capability to make a living on nucleic acids, lipids, or multiple macromolecule classes, respectively. Several populations seemed to lack extracellular enzyme machinery and thus likely scavenged oligo- or monomers (e.g., MAGs affiliating with Archaeoglobus) or metabolic products like hydrogen (e.g., MAGs affiliating with Thermodesulfobacterium or Desulforudaceae). The growth of methanogens or the production of methane was not observed in any condition, indicating that the tested macromolecules are not degraded into substrates for methanogenesis in hydrothermal sediments. We provide new insights into the niches, and genomes of microorganisms that actively degrade abundant necromass macromolecules under oxic, sulfate-reducing, and fermentative thermophilic conditions. These findings improve our understanding of the carbon flow across trophic levels and indicate how primary produced biomass sustains complex and productive ecosystems.

Description: We are grateful to the captain and crew of the R/V Atlantis AT37-06 as well as the crew of the human occupied vehicle Alvin for their tireless support. Sampling at Guaymas Basin was supported by NSF (OCE-1357238).

Description: © The Author(s), 2020. This article is distributed under the terms of the Creative Commons Attribution License. The definitive version was published in Kuloyo, O., Ruff, S. E., Cahill, A., Connors, L., Zorz, J. K., de Angelis, I. H., Nightingale, M., Mayer, B., & Strous, M. Methane oxidation and methylotroph population dynamics in groundwater mesocosms. Environmental Microbiology. (2020), doi:10.1111/1462-2920.14929.

Description: Extraction of natural gas from unconventional hydrocarbon reservoirs by hydraulic fracturing raises concerns about methane migration into groundwater. Microbial methane oxidation can be a significant methane sink. Here, we inoculated replicated, sand‐packed, continuous mesocosms with groundwater from a field methane release experiment. The mesocosms experienced thirty‐five weeks of dynamic methane, oxygen and nitrate concentrations. We determined concentrations and stable isotope signatures of methane, carbon dioxide and nitrate and monitored microbial community composition of suspended and attached biomass. Methane oxidation was strictly dependent on oxygen availability and led to enrichment of 13C in residual methane. Nitrate did not enhance methane oxidation under oxygen limitation. Methylotrophs persisted for weeks in the absence of methane, making them a powerful marker for active as well as past methane leaks. Thirty‐nine distinct populations of methylotrophic bacteria were observed. Methylotrophs mainly occurred attached to sediment particles. Abundances of methanotrophs and other methylotrophs were roughly similar across all samples, pointing at transfer of metabolites from the former to the latter. Two populations of Gracilibacteria (Candidate Phyla Radiation) displayed successive blooms, potentially triggered by a period of methane famine. This study will guide interpretation of future field studies and provides increased understanding of methylotroph ecophysiology.

Description: The authors acknowledge funding from the Alberta Innovates Technology Futures (AITF), and University of Calgary Eyes High Doctoral Scholarships (O.O.K., J.K.Z.) and AITF/Eyes High Postdoctoral Fellowships (S.E.R.), as well as the PROMOS Internship Abroad Scholarship by the German Academic Exchange Service (I.H.d.A.). Additional support was provided by the Natural Sciences and Engineering Research Council of Canada (NSERC), Strategic Project Grant no. 463045‐14, the Campus Alberta Innovation Chair Program (M.S.), Alberta Innovates, The Canadian Foundation for Innovation (M.S.), the Alberta Small Equipment Grant Program (M.S.) and an NSERC Discovery Grant (M.S. and B.M.).

Description: Groundwater ecosystems are globally widespread yet still poorly understood. We investigated the microbiology of 〉100 groundwater samples from 90 monitoring wells (〈250m depth) located in 14 aquifers in the Canadian Prairie. This dataset contains the nucleotide sequences and taxonomy of archaeal 16S rRNA gene amplicons retrieved from groundwater samples of aquifers in the Canadian Prairie, collected between 2015 and 2020. The study area comprised 14 major aquifers and a geographic area of ~210.000 km2. The goal of the study was to understand the links between the biogeochemistry and microbial ecology of groundwater ecosystems in diverse geological settings on a broad spatial scale. Details concerning DNA extraction, sequencing and sequence analyses can be found in the associated publication by Ruff et al. 2023.

Description: Groundwater ecosystems are globally widespread yet still poorly understood. We investigated the microbiology of 〉100 groundwater samples from 90 monitoring wells (〈250m depth) located in 14 aquifers in the Canadian Prairie. This dataset contains the nucleotide sequences and taxonomy of bacterial 16S rRNA gene amplicons retrieved from groundwater samples of aquifers in the Canadian Prairie, collected between 2015 and 2020. The study area comprised 14 major aquifers and a geographic area of ~210.000 km2. The goal of the study was to understand the links between the biogeochemistry and microbial ecology of groundwater ecosystems in diverse geological settings on a broad spatial scale. Details concerning DNA extraction, sequencing and sequence analyses can be found in the associated publication by Ruff et al. 2023.

Description: Groundwater ecosystems are globally widespread yet still poorly understood. We investigated the microbiology of 〉100 groundwater samples from 90 monitoring wells (〈250m depth) located in 14 aquifers in the Canadian Prairie. This dataset contains relative abundances of archaeal lineages based on 16S rRNA gene amplicons retrieved from 52 groundwater samples of aquifers in the Canadian Prairie, collected between 2015 and 2020. The study area comprised 14 major aquifers and a geographic area of ~210.000 km2. The goal of the study was to understand the links between the biogeochemistry and microbial ecology of groundwater ecosystems in diverse geological settings on a broad spatial scale. Details concerning DNA extraction, sequencing and sequence analyses can be found in the associated publication by Ruff et al. 2023.

Description: Groundwater ecosystems are globally widespread yet still poorly understood. We therefore investigated the total microbial cell counts of 78 groundwater samples from aquifers in the Canadian Prairie, collected between 2015 and 2020. The study area comprised 14 major aquifers and a geographic area of ~210.000 km2. The goal of the study was to understand the links between the biogeochemistry and microbial ecology of groundwater ecosystems in diverse geological settings on a broad spatial scale. Details concerning microscopy and cell enumeration can be found in the associated publication by Ruff et al. 2023.

Description: Groundwater ecosystems are globally widespread yet still poorly understood. We investigated the microbiology of groundwater samples from various monitoring wells (〈250m depth) located in 14 aquifers in the Canadian Prairie. This dataset contains archaeal community diversity indices based on 16S rRNA gene amplicons retrieved from 52 groundwater samples of aquifers in the Canadian Prairie, collected between 2015 and 2020. The study area comprised 14 major aquifers and a geographic area of ~210.000 km2. The goal of the study was to understand the links between the biogeochemistry and microbial ecology of groundwater ecosystems in diverse geological settings on a broad spatial scale. Details concerning DNA extraction, sequencing and sequence analyses can be found in the associated publication by Ruff et al. 2023.

Description: Groundwater ecosystems are globally widespread yet still poorly understood. This dataset contains values and calculations to estimate microbial oxygen consumption in groundwater samples from aquifers in the Canadian Prairie, collected between 2015 and 2020. The study area comprised 14 major aquifers and a geographic area of ~210.000 km2. The goal of the study was to understand the links between the biogeochemistry and microbial ecology of groundwater ecosystems in diverse geological settings on a broad spatial scale. Details concerning microbial cell counts and oxygen consumption can be found in the associated publication by Ruff et al. 2023.

Description: Groundwater ecosystems are globally widespread yet still poorly understood. We investigated the microbiology of groundwater samples from various monitoring wells (〈250m depth) located in 14 aquifers in the Canadian Prairie. This dataset contains bacterial community diversity indices based on 16S rRNA gene amplicons retrieved from 119 groundwater samples of aquifers in the Canadian Prairie, collected between 2015 and 2020. The study area comprised 14 major aquifers and a geographic area of ~210.000 km2. The goal of the study was to understand the links between the biogeochemistry and microbial ecology of groundwater ecosystems in diverse geological settings on a broad spatial scale. Details concerning DNA extraction, sequencing and sequence analyses can be found in the associated publication by Ruff et al. 2023.

Description: Groundwater ecosystems are globally widespread yet still poorly understood. We investigated the microbiology of 〉100 groundwater samples from 90 monitoring wells (〈250m depth) located in 14 aquifers in the Canadian Prairie. This dataset contains relative abundances of bacterial lineages based on 16S rRNA gene amplicons retrieved from 119 groundwater samples of aquifers in the Canadian Prairie, collected between 2015 and 2020. The study area comprised 14 major aquifers and a geographic area of ~210.000 km2. The goal of the study was to understand the links between the biogeochemistry and microbial ecology of groundwater ecosystems in diverse geological settings on a broad spatial scale. Details concerning DNA extraction, sequencing and sequence analyses can be found in the associated publication by Ruff et al. 2023.