The Arctic is undergoing rapid changes in climate, altering the status and functioning of high-latitude soils and permafrost. The vast majority of studies on Arctic soils and permafrost are conducted during the summer period due to ease of accessibility, sampling, instrument operation, and making measurements, in comparison to during winter and transition seasons. However, there is increasing evidence that microbial activity continues in Arctic soils outside of the summer period. Moreover, it is becoming clear that understanding the seasonal dynamics of Arctic soils is of critical importance, especially considering that the under-studied winter is the period that is most sensitive to climate warming. Soil biogeochemical models have advanced our understanding of the functioning and fate of soils in the Arctic, however it is vital that seasonality in biotic and abiotic processes is accurately captured in these models. Here we synthesize recent investigations and observations of the year-round functioning of Arctic soils, review soil biogeochemical modelling frameworks, and highlight certain processes and behaviors that are shaped by seasonality and thus warrant particular consideration within these models. More attention to seasonal processes will be critical to improving datasets and soil biogeochemical models that can be used to understand the year-round functioning of soils and the fate of the soil carbon reservoir in the Arctic.

Arctic regions are among the fastest warming areas of the planet. Increasing average temperatures over the last five decades have deepened the thawing of the upper-most layer of permafrost across the Arctic, which contains significant amounts of organic carbon. The progressive deepening of seasonal thawing releases carbon that is used by active microorganisms which also produce greenhouse gases, potentially onsetting a positive feedback on global warming. Despite their importance in controlling organic matter degradation and greenhouse gas fluxes to the atmosphere, there is a lack of data on activity and dynamics of microbial communities in High Arctic soils in response to seasonal thaw. This report describes three specific expeditions performed on the Svalbard archipelago, carried out within the framework of the PRA (Italian Arctic Research Program) project Melting-ICE, performed between February and October 2022, reporting site characteristics and samples collected. The project aims to investigate the diversity and activity of active layer microbial communities across a full season thaw cycle, correlating microbial diversity with gas fluxes and composition. During these expeditions, a total of eight different sites were selected to investigate the microbiology and geochemistry of soils, as well as to estimate the gas fluxes from the soil to the atmosphere. The data collected in the field, combined with the results obtained in the laboratory, will provide a snapshot of the seasonal activity of the microbial communities present in the permafrost’s active layer. The three campaigns will provide data to estimate the impact of permafrost melting on the carbon cycle and the role of microorganisms in the release of greenhouse gases.

Arctic regions are among the fastest warming areas of the planet. Increasing average temperatures over the last five decades have deepened the thawing of the upper-most layer of permafrost across the Arctic, which contains significant amounts of organic carbon. The progressive deepening of seasonal thawing releases carbon that is used by active microorganisms which also produce greenhouse gases, potentially onsetting a positive feedback on global warming. Despite their importance in controlling organic matter degradation and greenhouse gas fluxes to the atmosphere, there is a lack of data on activity and dynamics of microbial communities in High Arctic soils in response to seasonal thaw. This report describes three specific expeditions performed on the Svalbard archipelago, carried out within the framework of the PRA (Italian Arctic Research Program) project Melting-ICE, performed between February and October 2022, reporting site characteristics and samples collected. The project aims to investigate the diversity and activity of active layer microbial communities across a full season thaw cycle, correlating microbial diversity with gas fluxes and composition. During these expeditions, a total of eight different sites were selected to investigate the microbiology and geochemistry of soils, as well as to estimate the gas fluxes from the soil to the atmosphere. The data collected in the field, combined with the results obtained in the laboratory, will provide a snapshot of the seasonal activity of the microbial communities present in the permafrost’s active layer. The three campaigns will provide data to estimate the impact of permafrost melting on the carbon cycle and the role of microorganisms in the release of greenhouse gases.

This report describes the GRL22 expedition to Disko Island (West Greenland) to sample the thermal springs generated by residual heat associated with formation of the Icelandic hotspot. Despite the large number of springs present on Disko Island, our knowledge of the microbiology of these springs and how they relate to cycling of volatile elements and controls on their budgets is limited. The aim of the project is to understand the diversity of microbial communities brought to Earth’s surface by spring fluids and to determine their role in biogeochemical cycling. The long-term goal of the project is to combine these data with hot spring data from East Greenland and Iceland. This approach will create a transect in “space and time” across the Icelandic hotspot track, that will allow us to evaluate the potential influences of the plume on the subsurface microbial communities. This leg of the expedition took place between July 25 and August 2, 2022, and nine distinct sites were sampled for microbiology and geochemistry.

Margaret Cramm explains how quantitative stable isotope probing is used to understand microbial activities in the environment.

The deep biosphere is the largest microbial habitat on Earth and features abundant bacterial endospores. Whereas dormancy and survival at theoretical energy minima are hallmarks of microbial physiology in the subsurface, ecological processes such as dispersal and selection in the deep biosphere remain poorly understood. We investigated the biogeography of dispersing bacteria in the deep sea where upward hydrocarbon seepage was confirmed by acoustic imagery and geochemistry. Thermophilic endospores in the permanently cold seabed correlated with underlying seep conduits reveal geofluid-facilitated cell migration pathways originating in deep petroleum-bearing sediments. Endospore genomes highlight adaptations to life in anoxic petroleum systems and bear close resemblance to oil reservoir microbiomes globally. Upon transport out of the subsurface, viable thermophilic endospores reenter the geosphere by sediment burial, enabling germination and environmental selection at depth where new petroleum systems establish. This microbial dispersal loop circulates living biomass in and out of the deep biosphere.

Summary paragraph The deep biosphere is the largest microbial habitat on Earth and features abundant bacterial endospores 1,2 . Whereas dormancy and survival at theoretical energy minima are hallmarks of subsurface microbial populations 3 , the roles of fundamental ecological processes like dispersal and selection in these environments are poorly understood 4 . Here we combine geophysics, geochemistry, microbiology and genomics to investigate biogeography in the subsurface, focusing on bacterial endospores in a deep-sea setting characterized by thermogenic hydrocarbon seepage. Thermophilic endospores in permanently cold seabed sediments above petroleum seep conduits were correlated with the presence of hydrocarbons, revealing geofluid-facilitated cell migration pathways originating in deep oil reservoirs. Genomes of thermophilic bacteria highlight adaptations to life in anoxic petroleum systems and reveal that these dormant populations are closely related to oil reservoir microbiomes from around the world. After transport out of the subsurface and into the deep-sea, thermophilic endospores re-enter the geosphere by sedimentation. Viable thermophilic endospores spanning the top several metres of the seabed correspond with total endospore counts that are similar to or exceed the global average. Burial of dormant cells enables their environmental selection in sedimentary formations where new petroleum systems establish, completing a geological microbial loop that circulates living biomass in and out of the deep biosphere.

Increases in transportation of diesel and crude oil in the Labrador Sea will pose a significant threat to remote benthic and shoreline environments, where coastal communities and wildlife are particularly vulnerable to oil spill contaminants. Whereas marine microbiology has not been incorporated into environmental assessments in the Labrador Sea, there is a growing demand for microbial biodiversity evaluations given the pronounced impact of climate change in this region.

Seabed hydrocarbon seeps present natural laboratories for investigating responses of marine ecosystems to petroleum input. A hydrocarbon seep near Scott Inlet, Baffin Bay, was visited for in situ observations and sampling in the summer of 2018. Video evidence of an active hydrocarbon seep was confirmed by methane and hydrocarbon analysis of the overlying water column, which is 260 m at this site. Elevated methane concentrations in bottom water above and down current from the seep decreased to background seawater levels in the mid-water column >150 m above the seafloor. Seafloor microbial mats morphologically resembling sulfide-oxidizing bacteria surrounded areas of bubble ebullition. Calcareous tube worms, brittle stars, shrimp, sponges, sea stars, sea anemones, sea urchins, small fish and soft corals were observed near the seep, with soft corals showing evidence for hydrocarbon incorporation. Sediment microbial communities included putative methane-oxidizing Methyloprofundus, sulfate-reducing Desulfobulbaceae and sulfide-oxidizing Sulfurovum. A metabolic gene diagnostic for aerobic methanotrophs (pmoA) was detected in the sediment and bottom water above the seep epicentre and up to 5 km away. Both 16S rRNA gene and pmoA amplicon sequencing revealed that pelagic microbial communities oriented along the geologic basement rise associated with methane seepage (running SW to NE) differed from communities in off-axis water up to 5 km away. Relative abundances of aerobic methanotrophs and putative hydrocarbon-degrading bacteria were elevated in the bottom water down current from the seep. Detection of bacterial clades typically associated with hydrocarbon and methane oxidation highlights the importance of Arctic marine microbial communities in mitigating hydrocarbon emissions from natural geologic sources.

Microorganisms in marine subsurface sediments substantially contribute to global biomass. Sediments warmer than 40°C account for roughly half the marine sediment volume, but the processes mediated by microbial populations in these hard-to-access environments are poorly understood. We investigated microbial life in up to 1.2-kilometer-deep and up to 120°C hot sediments in the Nankai Trough subduction zone. Above 45°C, concentrations of vegetative cells drop two orders of magnitude and endospores become more than 6000 times more abundant than vegetative cells. Methane is biologically produced and oxidized until sediments reach 80° to 85°C. In 100° to 120°C sediments, isotopic evidence and increased cell concentrations demonstrate the activity of acetate-degrading hyperthermophiles. Above 45°C, populated zones alternate with zones up to 192 meters thick where microbes were undetectable.

Dormant endospores of anaerobic, thermophilic bacteria found in cold marine sediments offer a useful model for studying microbial biogeography, dispersal, and survival. The dormant endospore phenotype confers resistance to unfavorable environmental conditions, allowing dispersal to be isolated and studied independently of other factors such as environmental selection. To study the resilience of thermospores to conditions relevant for survival in extreme cold conditions, their viability following different freezing treatments was tested. Marine sediment was frozen at either −80°C or −20°C for 10 days prior to pasteurization and incubation at +50°C for 21 days to assess thermospore viability. Sulfate reduction commenced at +50°C following both freezing pretreatments indicating persistence of thermophilic endospores of sulfate-reducing bacteria. The onset of sulfate reduction at +50°C was delayed in −80°C pretreated microcosms, which exhibited more variability between triplicates, compared to −20°C pretreated microcosms and parallel controls that were not frozen in advance. Microbial communities were evaluated by 16S rRNA gene amplicon sequencing, revealing an increase in the relative sequence abundance of thermophilic endospore-forming Firmicutes in all microcosms. Different freezing pretreatments (−80°C and −20°C) did not appreciably influence the shift in overall bacterial community composition that occurred during the +50°C incubations. Communities that had been frozen prior to +50°C incubation showed an increase in the relative sequence abundance of operational taxonomic units (OTUs) affiliated with the class Bacilli, relative to unfrozen controls. These results show that freezing impacts but does not obliterate thermospore populations and their ability to germinate and grow under appropriate conditions. Indeed the majority of the thermospore OTUs detected in this study (21 of 22) could be observed following one or both freezing treatments. These results are important for assessing thermospore viability in frozen samples and following cold exposure such as the very low temperatures that would be encountered during panspermia.

X-ray computed tomography (XCT) can be used to identify lithologies and deformation structures within geological core, with the potential for the identification processes to be applied automatically. However, because of drilling disturbance and other artifacts, the use of large XCT-datasets in automated processes requires methods of quality control that can be applied systematically. We propose a new systematic method for quality control of XCT data that applies numerical measures to CT slices, and from this obtains data reflective of core quality. Because the measures are numerical they can be applied quickly and consistently between different sections and cores. This quality control processing protocol produces downhole radiodensity profiles from mean CT-values that can be used for geological interpretation. The application of this quality control protocols was applied to XCT data from International Ocean Discovery Program (IODP) Expedition 370 Site C0023 located at the toe of the Nankai accretionary complex. The evaluation of core quality based on this protocol was found to be a good fit to standard-evaluations based on the visual description of core, and could be used to select samples free from drilling disturbance or contamination. The quality-controlled downhole mean CT-value profile has features that can be used to identify lithologies within a formation, the presence and type of deformation structures and to distinguish formations.

SummaryThermophilic endospores are widespread in cold marine sediments where the temperature is too low to support growth and activity of thermophiles in situ. These endospores are likely expelled from warm subsurface environments and subsequently dispersed by ocean currents. The endospore upper temperature limit for survival is 140°C, which can be tolerated in repeated short exposures, potentially enabling transit through hot crustal fluids. Longer‐term thermal tolerance of endospores, and how long they could persist in an environment hotter than their maximum growth temperature, is less understood. To test whether thermophilic endospores can survive prolonged exposure to high temperatures, sediments were incubated at 80–90°C for 6, 12 or 463 days. Sediments were then cooled by 10–40°C, mimicking the cooling in subsurface oil reservoirs subjected to seawater injection. Cooling the sediments induced sulfate reduction, coinciding with an enrichment of endospore‐forming Clostridia. Different Desulfofundulus, Desulfohalotomaculum, Desulfallas, Desulfotomaculum and Desulfofarcimen demonstrated different thermal tolerances, with some Desulfofundulus strains surviving for >1 year at 80°C. In an oil reservoir context, heat‐resistant endospore‐forming sulfate‐reducing bacteria have a survival advantage if they are introduced to, or are resident in, an oil reservoir normally too hot for germination and growth, explaining observations of reservoir souring following cold seawater injection.

The study investigates the in-situ strength of sediments across a plate boundary décollement using drilling parameters recorded when a 1180-m-deep borehole was established during International Ocean Discovery Program (IODP) Expedition 370, Temperature - Limit of the Deep Biosphere off Muroto ( T - Limit ). Information of the in-situ strength of the shallow portion in/around a plate boundary fault zone is critical for understanding the development of accretionary prisms and of the décollement itself. Studies using seismic reflection surveys and scientific ocean drillings have recently revealed the existence of high pore pressure zones around frontal accretionary prisms, which may reduce the effective strength of the sediments. A direct measurement of in-situ strength by experiments, however, has not been executed due to the difficulty in estimating in-situ stress conditions. In this study, we derived a depth profile for the in-situ strength of a frontal accretionary prism across a décollement from drilling parameters using the recently established equivalent strength (EST) method. At site C0023, the toe of the accretionary prism area off Cape Muroto, Japan, the EST gradually increases with depth but undergoes a sudden change at ~ 800 mbsf, corresponding to the top of the subducting sediment. At this depth, directly below the décollement zone, the EST decreases from ~ 10 to 2 MPa, with a change in the baseline. This mechanically weak zone in the subducting sediments extends over 250 m (~ 800–1050 mbsf), corresponding to the zone where the fluid influx was discovered, and high-fluid pressure was suggested by previous seismic imaging observations. Although the origin of the fluids or absolute values of the strength remain unclear, our investigations support previous studies suggesting that elevated pore pressure beneath the décollement weakens the subducting sediments.

International Ocean Discovery Program (IODP) Expedition 370 aimed to explore the limits of life in the deep subseafloor biosphere at a location where temperature increases with depth at an intermediate rate and exceeds the known temperature maximum of microbial life (~120°C) at the sediment/basement interface ~1.2 km below the seafloor. Drilling Site C0023 is located in the vicinity of Ocean Drilling Program (ODP) Sites 808 and 1174 at the protothrust zone in the Nankai Trough off Cape Muroto at a water depth of 4776 m. ODP Leg 190 in 2000, revealed the presence of microbial cells at Site 1174 to a depth of ~600 meters below seafloor (mbsf), which corresponds to an estimated temperature of ~70°C, and reliably identified a single zone of higher cell concentrations just above the decollement at around 800 mbsf, where temperature presumably reached 90°C; no cell count data was reported for other sediment layers in the 70°–120°C range, because the limit of manual cell count for low-biomass samples was not high enough. With the establishment of Site C0023, we aimed to detect and investigate the presence or absence of life and biological processes at the biotic–abiotic transition with unprecedented analytical sensitivity and precision. Expedition 370 was the first expedition dedicated to subseafloor microbiology that achieved time-critical processing and analyses of deep biosphere samples by simultaneous shipboard and shore-based investigations. Our primary objectives during Expedition 370 were to study the relationship between the deep subseafloor biosphere and temperature. We aimed to comprehensively study the factors that control biomass, activity, and diversity of microbial communities in a subseafloor environment where temperatures increase from ~2°C at the seafloor to ~120°C at the sediment/basement interface and thus likely encompasses the biotic–abiotic transition zone. We also aimed to determine geochemical, geophysical, and hydrogeological characteristics in sediment and the underlying basaltic basement and elucidate if the supply of fluids containing thermogenic and/or geogenic nutrient and energy substrates may support subseafloor microbial communities in the Nankai accretionary complex. To address these primary scientific objectives and questions, we penetrated 1180 m and recovered 112 cores across the sediment/basalt interface. More than 13,000 samples were collected, and selected samples were transferred to the Kochi Core Center by helicopter for simultaneous microbiological sampling and analysis in laboratories with a super-clean environment. Following the coring operations, a temperature observatory with 13 thermistor sensors was installed in the borehole to 863 mbsf.

International Ocean Discovery Program (IODP) Expedition 370 explored the limits of the biosphere in the deep subseafloor where temperature exceeds the known temperature maximum of microbial life (~120°C) at the sediment/basement interface ~1.2 km below the seafloor. Site C0023 is located in the protothrust zone in the Nankai Trough off Cape Muroto at a water depth of 4776 m, in the vicinity of Ocean Drilling Program (ODP) Sites 808 and 1174. In 2000, ODP Leg 190 revealed the presence of microbial cells at Site 1174 to a depth of ~600 meters below seafloor (mbsf ), which corresponds to an estimated temperature of ~70°C, and reliably identified a single zone of elevated cell concentrations just above the décollement at around 800 mbsf, where temperature presumably reached 90°C; no cell count data was reported for other sediment layers in the 70°–120°C range because the detection limit of manual cell counting for low-biomass samples was not low enough. With the establishment of Site C0023, we aimed to detect and investigate the presence or absence of life and biological processes at the biotic–abiotic transition utilizing unprecedented analytical sensitivity and precision. Expedition 370 was the first expedition dedicated to subseafloor microbiology that achieved time-critical processing and analyses of deep biosphere samples, conducting simultaneous shipboard and shore-based investigations. Our primary objective during Expedition 370 was to study the relationship between the deep subseafloor biosphere and temperature. We comprehensively studied the controls on biomass, activity, and diversity of microbial communities in a subseafloor environment where temperatures increase from ~2°C at the seafloor to ~120°C and thus likely encompasses the biotic–abiotic transition zone. This included investigating whether the supply of fluids containing thermogenic and/or geogenic nutrient and energy substrates may support subseafloor microbial communities in the Nankai accretionary complex. To address these primary scientific objectives and questions, we penetrated 1180 m and recovered 112 cores of sediment and across the sediment/basalt interface. More than 13,000 samples were collected, and selected samples were transferred to the Kochi Core Center by helicopter for simultaneous microbiological sampling and analysis in laboratories with a superclean environment. In addition to microbiological measurements, we determined the geochemical, geophysical, and hydrogeological characteristics of the sediment and the underlying basement and installed a 13-thermistor sensor borehole temperature observatory into the borehole at Site C0023. V.B. Heuer et al. Expedition 370 summary