Description: Selected multicorer cores of R/V Sonne cruise SO268-2 were scanned by a Toshiba Aquilion 64 computer tomograph (CT) at the hospital Klinikum Bremen-Mitte with an x-ray source voltage of 120 kV and a current of 600 mA. The CT image stacks have a resolution of 0.35 mm in x/y-direction and 0.5 mm resolution in z-direction (0.3 mm reconstruction unit). Images were reconstructed using Toshiba's patented helical cone beam reconstruction technique (TCOT) and are provided in DICOM-format. The data were processed with the ZIB edition of the Amira software (version 2021.03; Stalling et al., 2005; http://amira.zib.de). Within Amira, the core liners, including about 2 mm of the core rims, were deleted from the data set. Mangan nodules with diameters 〉~1 mm, open bioturbation traces, an unidentified organism (only present in core SO268-2-200-1 MC18), the natural bioturbated and surficial homogenised sediment were segmented by a marker-based watershed segmentation and visualized in 3D. Subsequently, the sediment components were quantified (MaterialStatistics module; volume per slice) and the mean and standard deviation of the sediment x-ray attenuation (MaterialStatistics module; statistics per slice per label; to avoid marginal averaging effects the sediment label was reduced by three voxels) were determined.

Description: Underway temperature and salinity data was collected along the cruise track with two autonomous measurement systems, called self-cleaning monitoring boxes (SMBs). Usually, the SMBs are changed after ~24 hours. While temperature is taken at the water inlet in about 4 m depth, salinity is estimated within the SMB from conductivity and interior temperature. No temperature and salinity calibration were performed. For details to all processing steps see Data Processing Report.

Description: Deep-sea mining for polymetallic nodules is expected to have severe environmental impacts because not only nodules but also benthic fauna and the upper reactive sediment layer are removed through the mining operation and blanketed by resettling material from the suspended sediment plume. This study aims to provide a holistic assessment of the biogeochemical recovery after a disturbance event by applying prognostic simulations based on an updated diagenetic background model and validated against novel data on microbiological processes. It was found that the recovery strongly depends on the impact type; complete removal of the reactive surface sediment reduces benthic release of nutrients over centuries, while geochemical processes after resuspension and mixing of the surface sediment are near the pre-impact state 1 year after the disturbance. Furthermore, the geochemical impact in the DISturbance and reCOLonization (DISCOL) experiment area would be mitigated to some degree by a clay-bound Fe(II)-reaction layer, impeding the downward diffusion of oxygen, thus stabilizing the redox zonation of the sediment during transient post-impact recovery. The interdisciplinary (geochemical, numerical and biological) approach highlights the closely linked nature of benthic ecosystem functions, e.g. through bioturbation, microbial biomass and nutrient fluxes, which is also of great importance for the system recovery. It is, however, important to note that the nodule ecosystem may never recover to the pre-impact state without the essential hard substrate and will instead be dominated by different faunal communities, functions and services.

Description: Deep-seabed polymetallic nodule mining can have multiple adverse effects on benthic communities, such as permanent loss of habitat by removal of nodules and habitat modification of sediments. One tool to manage biodiversity risks is the mitigation hierarchy, including avoidance, minimization of impacts, rehabilitation and/or restoration, and offset. We initiated long-term restoration experiments at sites in polymetallic nodule exploration contract areas in the Clarion-Clipperton Zone that were (i) cleared of nodules by a preprototype mining vehicle, (ii) disturbed by dredge or sledge, (iii) undisturbed, and (iv) naturally devoid of nodules. To accommodate for habitat loss, we deployed 〉2000 artificial ceramic nodules to study the possible effect of substrate provision on the recovery of biota and its impact on sediment biogeochemistry. Seventy-five nodules were recovered after eight weeks and had not been colonized by any sessile epifauna. All other nodules will remain on the seafloor for several years before recovery. Furthermore, to account for habitat modification of the top sediment layer, sediment in an epibenthic sledge track was loosened by a metal rake to test the feasibility of sediment decompaction to facilitate soft-sediment recovery. Analyses of granulometry and nutrients one month after sediment decompaction revealed that sand fractions are proportionally lower within the decompacted samples, whereas total organic carbon values are higher. Considering the slow natural recovery rates of deep-sea communities, these experiments represent the beginning of a ~30-year study during which we expect to gain insights into the nature and timing of the development of hard-substrate communities and the influence of nodules on the recovery of disturbed sediment communities. Results will help us understand adverse long-term effects of nodule removal, providing an evidence base for setting criteria for the definition of “serious harm” to the environment. Furthermore, accompanying research is needed to define a robust ecosystem baseline in order to effectively identify restoration success.

Description: The Deep Ocean Observing Strategy (DOOS) is an international, community-driven initiative that facilitates collaboration across disciplines and fields, elevates a diverse cohort of early career researchers into future leaders, and connects scientific advancements to societal needs. DOOS represents a global network of deep-ocean observing, mapping, and modeling experts, focusing community efforts in the support of strong science, policy, and planning for sustainable oceans. Its initiatives work to propose deep-sea Essential Ocean Variables; assess technology development; develop shared best practices, standards, and cross-calibration procedures; and transfer knowledge to policy makers and deep-ocean stakeholders. Several of these efforts align with the vision of the UN Ocean Decade to generate the science we need to create the deep ocean we want. DOOS works toward (1) a healthy and resilient deep ocean by informing science-based conservation actions, including optimizing data delivery, creating habitat and ecological maps of critical areas, and developing regional demonstration projects; (2) a predicted deep ocean by strengthening collaborations within the modeling community, determining needs for interdisciplinary modeling and observing system assessment in the deep ocean; (3) an accessible deep ocean by enhancing open access to innovative low-cost sensors and open-source plans, making deep-ocean data Findable, Accessible, Interoperable, and Reusable, and focusing on capacity development in developing countries; and finally (4) an inspiring and engaging deep ocean by translating science to stakeholders/end users and informing policy and management decisions, including in international waters.

Description: Automated sampling technologies can enhance the temporal and spatial resolution of marine microbial observations, particularly in remote and inaccessible areas. A critical aspect of automated microbiome sampling is the preservation of nucleic acids over long-term autosampler deployments. Understanding the impact of preservation method on microbial metabarcoding is essential for implementing genomic observatories into existing infrastructure, and for establishing best practices for the regional and global synthesis of data. The present study evaluates the effect of two preservatives commonly used in autosampler deployments (mercuric chloride and formalin) and two extraction kits (PowerWater and NucleoSpin) on amplicon sequencing of 16S and 18S rRNA gene over 50 weeks of sample storage. Our results suggest the combination of mercuric chloride preservation and PowerWater extraction as most adequate for 16S and 18S rRNA gene amplicon-sequencing from the same seawater sample. This approach provides consistent information on species richness, diversity and community composition in comparison to control samples (nonfixed, filtered and frozen) when stored up to 50 weeks at in situ temperature. Preservation affects the recovery of certain taxa, with specific OTUs becoming overrepresented (SAR11 and diatoms) or underrepresented (Colwellia and pico-eukaryotes) after preservation. In case eukaryotic sequence information is the sole target, formalin preservation and NucleoSpin extraction performed best. Our study contributes to the design of long-term autonomous microbial observations in remote ocean areas, allowing cross-comparison of microbiome dynamics across sampling devices (e.g., water and particle samplers) and marine realms.

Description: The trace metal iron (Fe) is an essential micronutrient that controls phytoplankton productivity, which subsequently affects organic matter cycling with feedback on the cycling of macronutrients. Along the continental margin of the US West Coast, high benthic Fe release has been documented, in particular from deep anoxic basins in the Southern California Borderland. However, the influence of this Fe release on surface primary production remains poorly understood. In the present study from the Santa Barbara Basin, in situ benthic Fe fluxes were determined along a transect from shallow to deep sites in the basin. Fluxes ranged between 0.23 and 4.9mmolm-2d-1, representing some of the highest benthic Fe fluxes reported to date. To investigate the influence of benthic Fe release from the oxygen-deficient deep basin on surface phytoplankton production, we combined benthic flux measurements with numerical simulations using the Regional Ocean Modeling System coupled to the Biogeochemical Elemental Cycling (ROMS-BEC) model. For this purpose, we updated the model Fe flux parameterization to include the new benthic flux measurements from the Santa Barbara Basin. Our simulations suggest that benthic Fe fluxes enhance surface primary production, supporting a positive feedback on benthic Fe release by decreasing oxygen in bottom waters. However, a reduction in phytoplankton Fe limitation by enhanced benthic fluxes near the coast may be partially compensated for by increased nitrogen limitation further offshore, limiting the efficacy of this positive feedback.

Description: The recently discovered cryptic methane cycle in the sulfate-reducing zone of marine and wetland sediment couples methylotrophic methanogenesis to anaerobic oxidation of methane (AOM). Here we present evidence of cryptic methane cycling activity within the upper regions of the sulfate-reducing zone, along a depth transect within the Santa Barbara Basin, off the coast of California, USA. The top 0-20 cm of sediment from each station was subjected to geochemical analyses and radiotracer incubations using 35S-SO42-14C-mono-methylamine, and 14C-CH4 to find evidence of cryptic methane cycling. Methane concentrations were consistently low (3 to 16 μM) across the depth transect, despite AOM rates increasing with decreasing water depth (from max 0.05 nmol cm-3 d-1 at the deepest station to max 1.8 nmol cm-3 d-1 at the shallowest station). Porewater sulfate concentrations remained high (23 to 29 mM), despite the detection of sulfate reduction activity from 35S-SO42- incubations with rates up to 134 nmol cm-3 d-1. Metabolomic analysis showed that substrates for methanogenesis (i.e., acetate, methanol and methylamines) were mostly below the detection limit in the porewater, but some samples from the 1-2 cm depth section showed non-quantifiable evidence of these substrates, indicating their rapid turnover. Estimated methanogenesis from mono-methylamine ranged from 0.2 to 0.5 nmol cm-3 d-1. Discrepancies between the rate constants (k) of methanogenesis (from 14C-mono-methylamine) and AOM (from either 14C-mono-methylamine-derived 14C-CH4 or from directly injected 14C-CH4) suggest the activity of a separate, concurrent metabolic process directly metabolizing mono-methylamine to inorganic carbon. We conclude that the results presented in this work show strong evidence of cryptic methane cycling occurring within the top 20 cm of sediment in the Santa Barbara Basin. The rapid cycling of carbon between methanogenesis and methanotropy likely prevents major build-up of methane in the sulfate-reducing zone. Furthermore, our data suggest that methylamine is utilized by both methanogenic archaea capable of methylotrophic methanogenesis and non-methanogenic microbial groups. We hypothesize that sulfate reduction is responsible for the additional methylamine turnover, but further investigation is needed to elucidate this metabolic activity.

Description: The Santa Barbara Basin naturally experiences transient deoxygenation due to its unique geological setting in the southern California Borderland and seasonal changes in ocean currents. Long-term measurements of the basin showed that anoxic events and subsequent nitrate exhaustion in the bottom waters have been occurring more frequently and lasting longer over the past decade. One characteristic of the Santa Barbara Basin is the seasonal development of extensive mats of benthic nitrate-reducing sulfur-oxidizing bacteria, which are found at the sediment-water interface when the basin's bottom waters reach anoxia but still provide some nitrate. To assess the mat's impact on the benthic and pelagic redox environment, we collected biogeochemical sediment and benthic flux data in November 2019, after anoxia developed in the deepest waters of the basin and dissolved nitrate was depleted (down to 9.9μM). We found that the development of mats was associated with a shift from denitrification to dissimilatory nitrate reduction to ammonium. The zone of sulfate reduction appeared near the sediment-water interface in sediment hosting these ephemeral white mats. We found that an exhaustion of iron oxides in the surface sediment was an additional prerequisite for mat proliferation. Our research further suggests that cycles of deoxygenation and reoxygenation of the benthic environment result in extremely high benthic fluxes of dissolved iron from the basin's sediment. This work expands our understanding of nitrate-reducing sulfur-oxidizing mats and their role in sustaining and potentially expanding marine anoxia.