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Ratios of glycerol dialkyl glycerol tetraethers (GDGT), which are membrane lipids of bacteria and archaea, are at the base of several paleoenvironmental proxies. They are frequently applied to soils as well as lake- and marine sediments to generate records of past temperature and soil pH. To derive meaningful environmental information from these reconstructions, high analytical reproducibility is required. Based on submitted results by 39 laboratories from across the world, which employ a diverse range of analytical and quantification methods, we explored the reproducibility of brGDGT-based proxies (MBT′5ME, IR, and #ringstetra) measured on four soil samples and four soil lipid extracts. Correct identification and integration of 5- and 6-methyl brGDGTs is a prerequisite for the robust calculation of proxy values, but this can be challenging as indicated by the large inter-interlaboratory variation. The exclusion of statistical outliers improves the reproducibility, where the remaining uncertainty translates into a temperature offset from median proxy values of 0.3–0.9°C and a pH offset of 0.05–0.3. There is no apparent systematic impact of the extraction method and sample preparation steps on the brGDGT ratios. Although reported GDGT concentrations are generally consistent within laboratories, they vary greatly between laboratories. This large variability in brGDGT quantification may relate to variations in ionization efficiency or specific mass spectrometer settings possibly impacting the response of brGDGTs masses relative to that of the internal standard used. While ratio values of GDGT are generally comparable, quantities can currently not be compared between laboratories.

Volcanic ash significantly contributes to marine sediments, especially in regions with active onshore volcanoes. Alteration of volcanic ash releases bicarbonate and cations, which drive precipitation of authigenic carbonate and clay minerals. Furthermore, volcanic ashes are commonly enriched in reactive iron (Fe[III]), suggesting that ash alteration as a source of reactants plays an important role in (bio-)geochemical processes in marine sediments. Volcanic ash layers are ubiquitous in sediments of Site C0023, which was established down to 1180 m below seafloor (mbsf) in the Nankai Trough off Japan during International Ocean Discovery Program Expedition 370. Shipboard measurements show a release of dissolved Fe between 200 and 600 mbsf, coinciding with a high abundance of ash layers [1]. The release of Fe can be related to microbial reduction of structural Fe(III) in smectite promoting the smectite-to-illite transition, as recently proposed [2]. By combining shipboard pore-water data with sequential extractions of reactive Fe pools on ash layers and surrounding mud rock and stable Fe isotope (δ56Fe) analyses, we elucidate the role of ash alteration on (bio-)geochemical cycling at Site C0023. Our results demonstrate that reactive Fe(III) is unexpectedly lower in ash layers compared to the surrounding mud rock (0.6 and 1.2 wt%, respectively). This indicates that (1) Fe(III) originally deposited with tephra has either been used or (2) Fe(III) in tephra is generally lower due to a different chemical composition in the volcanic source material. The δ56Fe signature of hydroxylamine-extracted Fe, which represents easily reducible Fe-oxides and Fe bound in phyllosilicates, is isotopically light (-0.08 to -0.42‰) compared to terrestrial background values (~0.09‰; [3]). This suggests that this pool is diagenetically overprinted by the precipitation of authigenic smectite formed as a result of ash alteration and/or secondary Fe-oxides. Pore-water Fe is extremely negative with

The deep subseafloor biosphere represents one of the Earth’s largest, but also least understood ecosystems with diverse species and mostly uncharacterized microbial communities. International Ocean Discovery Program (IODP) Expedition 370 (Temperature Limit of the Deep Biosphere off Muroto) established Site C0023 down to 1180 mbsf in the Nankai Trough off Japan to explore the upper temperature limit of microbial life in the deep sedimentary biosphere [1]. Site C0023 is characterized by a complex lithostratigraphic and depositional history with strongly changing sedimentation rates. Volcanic ash layers are ubiquitous in all lithological units. However, the highest abundance of ash layers could be observed between 400 and 700 mbsf. Previous studies have shown that volcanic ashes represent hotspots for microbial life [2] and are commonly characterized by high Fe(III) and Mn(IV) contents [3]. Onboard measurements show a release of dissolved Fe in the depth interval associated with the highest abundance of ash layers [1]. Therefore, we hypothesized that the release is related to microbial Fe reduction fueled by the mineralogy of the volcanic ash. In order to identify the source and reaction pathway of the liberated Fe, we applied sequential extractions of differently reactive Fe oxide pools on mud rock and ash layer samples as well as stable iron isotope (δ56Fe) analyses on pore-water and solid-phase samples. Microbial Fe reduction leads to Fe isotope fractionation with an enrichment of light isotopes in the released Fe and a respective enrichment of heavy isotopes in the residual ferric substrate. Therefore, the δ56Fe signals of different reactive Fe pools and the pore water are used to identify the pools actually involved in microbial respiration processes. Our results show that the total Fe content in mud rock of Site C0023 is relatively constant at ~4.2 wt%. Reactive Fe oxides represent 25% of the total Fe. The bulk Fe content in the ash layers varies between 1.4 and

Microorganisms in marine subsurface sediments substantially contribute to global biomass.Sediments warmer than 40°C account for roughly half the marine sediment volume, but theprocesses mediated by microbial populations in these hard-to-access environments are poorlyunderstood. We investigated microbial life in up to 1.2-kilometer-deep and up to 120°C hotsediments in the Nankai Trough subduction zone. Above 45°C, concentrations of vegetativecells drop two orders of magnitude and endospores become more than 6000 times more abundantthan 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 concentrationsdemonstrate the activity of acetate-degrading hyperthermophiles. Above 45°C, populated zonesalternate with zones up to 192 meters thick where microbes were undetectable

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.

IODP Expedition 370 (Temperature Limit of the Deep Biosphere off Muroto) established Site C0023 down to 1180 mbsf in the Nankai Trough off Shikoku Island, Japan, to explore the upper temperature limit of microbial life in deep subseafloor sediments. Part of the scientific program is to investigate the availability of nutrients and energy substrates and to identify unique geochemical and microbial signatures that differentiate the biotic and abiotic realms and/or their transitions (Heuer et al., 2017). Iron (Fe) reduction is considered one of the most ancient forms of microbial respiration (Vargas et al., 1998). In addition, Fe reducers can grow under high temperature and pressure conditions (Kashefi and Lovley, 2003), suggesting that microbes that use Fe oxides as energy substrates are potential candidates to survive close to the temperature limit of the deep biosphere. In this study, we aim at assessing the role of Fe oxides for microbial respiration and the related diagenetic alterations in deep sediments of Site C0023 by applying sequential extractions of Fe oxide and sulfide minerals. Volcanic ash layers, which are ubiquitous in sediments of Site C0023, are of particular interest as they have been identified earlier as hotspots for microbial life (e.g., Inagaki et al., 2003). Torres et al. (2015) further showed that ash layers at a different site in the Nankai Trough are typically rich in Fe and Mn oxides. Their results support the findings of Treude et al. (2014) who postulate a coupling of microbial processes to mineralogy. In addition, on-board measurements show a release of dissolved Fe into the pore water in the depth interval associated with volcanic ash layers (Heuer et al., 2017), suggesting that the observed liberation of dissolved Fe is related to an alteration of Fe phases in these ash layers. Our results show that the total Fe content in sediments of Site C0023 is relatively constant at ~4.2 wt%. The reactive Fe oxide content represents 25% of the to

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. Keywords: Plate boundary décollement, Nankai Trough, Excess fluid pressure, Equivalent strength, Drilling pa, Japan Society for the Promotion of Science (16H04064), Japan Society for the Promotion of Science (15H05717)

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 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 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 sed

We thank the authors for their comment and welcome the opportunity to clarify our interpretations in view of the concerns raised. First, Pujalte and Schmitz dismiss our dataset from Tendrui by claiming that there is no Claret Conglomerate (CC) at this location. The Tendrui section of Domingo et al. (2009) and Manners et al. (2013) can be located using the coordinates recorded in Manners et al. (2013; N 042° 10′07.2″, E 000° 51′25.3″). When sampling at this location the CC was indeed exposed, as presented in the logs of Domingo et al. (2009) and Manners et al. (2013). It may be possible that the Tendrui section without the CC, referred to by Pujalte and Schmitz (2014), is exposed at a different location in the vicinity of Tendrui. Unfortunately their locality cannot be identified as its precise location was not provided by Schmitz and Pujalte (2007) or Pujalte et al. (2014). Second, Pujalte and Schmitz (2014) claim that vegetation covers the critical interval below the CC at the Claret road section (Fig. 1 of their comment), making the sampling through this “covered” interval, as presented in Manners et al. (2013), impossible. There is vegetation in this area, but when sampled in March 2010, this was not so dense as to preclude trenching back into in-situ material (Fig. 1). This is how our samples were taken, with the express aim of providing a more complete record of the carbon isotope excursion (CIE) onset below the CC.