At deep-sea hydrothermal vents, magmatically driven rock-water reactions in the crust generate gases and other reduced compounds that subseafloor microorganisms use for chemolithoautotrophy. In this study, microbial autotrophs from three diffuse flow hydrothermal vents at Axial Seamount in 2013 and 2014 were isotopically labeled using RNA stable isotope probing, targeting subseafloor autotrophic mesophiles (30°C), thermophiles (55°C), and hyperthermophiles (80°C). We constructed taxonomic and functional profiles of active chemolithoautotrophs, examined population distributions across sites, and linked primary producers to their specific metabolic strategies within the subseafloor community. Dominant autotrophs exhibited hydrogen-dependent dissimilatory metabolisms, such as sulfur and nitrate reduction and methanogenesis, as well as microaerophilic sulfide oxidation even at 80°C, consistent with fluid chemistries at each site. While hydrogenotrophic methanogenic archaea (Methanothermococcus) were restricted in their distribution and activity, hydrogenotrophic sulfur and nitrate reducers from the Aquificota (Thermovibrio) and Campylobacterota (Nautiliaceae, Hydrogenimonas, and Desulfurobacteriaceae) were consistently active and present at all sites and years at both the population and community levels. Hydrogenase transcripts were significantly differentially expressed, and diverse hydrogenases were found in metagenome-assembled genomes of Aquificota members, highlighting the importance and versatility of their hydrogen utilization strategies, which likely contribute to their cosmopolitan distribution across geochemically disparate subseafloor sites. Together, this study provides new insights into the functional dynamics and distribution of key subseafloor autotrophic microbial communities in young oceanic crust at deep-sea hydrothermal vents. IMPORTANCE:Deep-sea hydrothermal vents are hotspots for life in the dark ocean, where rich animal ecosystems are supported by microbial primary producers utilizing the abundant chemical energy supplied by high-temperature water-rock reactions. Despite increasing knowledge about the geochemistry and microbiology of deep-sea hydrothermal vents, there is still a gap in our understanding of the key microbial players who fix much of the carbon at these sites, especially in the productive subseafloor. In this study, stable isotope probing was used to label active microbial autotrophs in diffuse flow venting fluids from three sites over 2 years and was combined with metatranscriptomic sequencing to identify their specific metabolic strategies. This research highlights the microbial community composition, function, gene regulation, and population dynamics that enable hydrothermal ecosystems to persist.

Depressurization and sample processing delays may impact the outcome of shipboard microbial incubations of samples collected from the deep sea. To address this knowledge gap, we developed a remotely operated vehicle (ROV)-powered incubator instrument to carry out and compare results from in situ and shipboard RNA stable isotope probing (RNA-SIP) experiments to identify the key chemolithoautotrophic microbes and metabolisms in diffuse, low-temperature venting fluids from Axial Seamount. All the incubations showed microbial uptake of labeled bicarbonate primarily by thermophilic autotrophic Epsilonbacteraeota that oxidized hydrogen coupled with nitrate reduction. However, the in situ seafloor incubations showed higher abundances of transcripts annotated for aerobic processes, suggesting that oxygen was lost from the hydrothermal fluid samples prior to shipboard analysis. Furthermore, transcripts for thermal stress proteins such as heat shock chaperones and proteases were significantly more abundant in the shipboard incubations, suggesting that depressurization induced thermal stress in the metabolically active microbes in these incubations. Together, the results indicate that while the autotrophic microbial communities in the shipboard and seafloor experiments behaved similarly, there were distinct differences that provide new insight into the activities of natural microbial assemblages under nearly native conditions in the ocean.IMPORTANCE Diverse microbial communities drive biogeochemical cycles in Earth's ocean, yet studying these organisms and processes is often limited by technological capabilities, especially in the deep ocean. In this study, we used a novel marine microbial incubator instrument capable of in situ experimentation to investigate microbial primary producers at deep-sea hydrothermal vents. We carried out identical stable isotope probing experiments coupled to RNA sequencing both on the seafloor and on the ship to examine thermophilic, microbial autotrophs in venting fluids from an active submarine volcano. Our results indicate that microbial communities were significantly impacted by the effects of depressurization and sample processing delays, with shipboard microbial communities being more stressed than seafloor incubations. Differences in metabolism were also apparent and are likely linked to the chemistry of the fluid at the beginning of the experiment. Microbial experimentation in the natural habitat provides new insights into understanding microbial activities in the ocean.

Buoyant microplastic marine debris (MPMD) is a pollutant in the ocean surface boundary layer (OSBL) that is submerged by wave-driven turbulent transport processes. This study analyzes observed MPMD surface concentrations in the Atlantic and Pacific Oceans to reveal a significant increase in concentrations during surface heating and a decrease during surface cooling. Turbulence-resolving large-eddy simulations of the OSBL for an idealized diurnal heating cycle suggest that turbulent downward fluxes of buoyant tracers are enhanced at night, facilitating deep submergence of plastics, and suppressed in heating conditions, resulting in surface-trapped MPMD. Simulations agree better with observations if enhanced mixing due to wave-driven Langmuir turbulence (LT) is included. The simulated time-dependent OSBL response results in hysteresis effects so that surface concentrations depend also on the phase of the diurnal heating cycle. The results demonstrate the controlling influence of surface heat fluxes and LT on turbulent transport in the OSBL and on vertical distributions of buoyant marine particles.

An in-situ chemical sensor system was developed for long term monitoring of hydrothermal vent fluids as part of the US NSF Ocean Observatory Initiative (OOI) Regional Scale Nodes (RSN) in the Northeast Pacific Ocean. The sensor wand design is based on Yittria-stabilized zirconia (YSZ) ceramic membrane electrode in conjunction with Ag|AgCl reference electrode, Au or Pt redox electrode and Ag|Ag2S (H2S) electrode. The YSZ ceramic electrode was constructed with a titanium tube for the long term deployment application. To achieve high measurement accuracy, the high impedance signal of the YSZ ceramic electrode was converted to the low impedance signal by a pre-amplifier. The instrument utilizes the power supply and bandwidth of the fiber optic ocean cable to conduct continuous monitoring of the hot vent fluids. The instrument was deployed on Regional Scale Nodes (RSN) Axial Seamount site via MJ03C mid-power junction box. A modified version of the instrument was also deployed multi times by DSV Alvin during AT26-17 cruise on Axial Seamount and Main Endeavour Field (MEF) vent field, Juan De Fuca Ridge. This integrated electrochemical system has successfully measured pH, dissolved H2 and H2S of hydrothermal vent fluids at 300 °C and 220 bars.

This paper is the second of a two-part series that investigates passive buoyant tracers in the ocean surface boundary layer (OSBL). The first part examines the influence of equilibrium wind-waves on vertical tracer distributions, based on large eddy simulations (LESs) of the wave-averaged Navier-Stokes equation. Motivated by observations of buoyant microplastic marine debris (MPMD), this study applies the LES model and the parametric one-dimensional column model from part one to examine the vertical distributions of MPMD. MPMD is widely distributed in vast regions of the subtropical gyres and has emerged as a major open ocean pollutant whose distribution is subject to upper ocean turbulence. The models capture shear-driven turbulence, Langmuir turbulence (LT), and enhanced turbulent kinetic energy input due to breaking waves (BWs). Model results are only consistent with observations of MPMD profiles and the relationship between surface concentrations and wind speed if LT effects are included. Neither BW nor shear-driven turbulence is capable of deeply submerging MPMD, suggesting that the observed vertical MPMD distributions are a characteristic signature of wave-driven LT. Thus, this study demonstrates that LT substantially increases turbulent transport in the OSBL, resulting in deep submergence of buoyant tracers. The parametric model is applied to 11 years of observations in the North Atlantic and North Pacific subtropical gyres to show that surface measurements substantially underestimate MPMD concentrations by a factor of 3-13.

Microplastics (particles less than 5 mm) numerically dominate marine debris and occur from coastal waters to mid‐ocean gyres, where surface circulation concentrates them. Given the prevalence of plastic marine debris (PMD) and the rise in plastic production, the impacts of plastic on marine ecosystems will likely increase. Microscopic life (the “Plastisphere”) thrives on these tiny floating “islands” of debris and can be transported long distances. Using next‐generation DNA sequencing, we characterized bacterial communities from water and plastic samples from the North Pacific and North Atlantic subtropical gyres to determine whether the composition of different Plastisphere communities reflects their biogeographic origins. We found that these communities differed between ocean basins – and to a lesser extent between polymer types – and displayed latitudinal gradients in species richness. Our research reveals some of the impacts of microplastics on marine biodiversity, demonstrates that the effects and fate of PMD may vary considerably in different parts of the global ocean, and suggests that PMD mitigation will require regional management efforts.

We present an extensive survey of floating plastic debris in the eastern North and South Pacific Oceans from more than 2500 plankton net tows conducted between 2001 and 2012. From these data we defined an accumulation zone (25 to 41 °N, 130 to 180 °W) in the North Pacific subtropical gyre that closely corresponds to centers of accumulation resulting from the convergence of ocean surface currents predicted by several oceanographic numerical models. Maximum plastic concentrations from individual surface net tows exceeded 10(6) pieces km(-2), with concentrations decreasing with increasing distance from the predicted center of accumulation. Outside the North Pacific subtropical gyre the median plastic concentration was 0 pieces km(-2). We were unable to detect a robust temporal trend in the data set, perhaps because of confounded spatial and temporal variability. Large spatiotemporal variability in plastic concentration causes order of magnitude differences in summary statistics calculated over short time periods or in limited geographic areas. Utilizing all available plankton net data collected in the eastern Pacific Ocean (17.4 °S to 61.0 °N; 85.0 to 180.0 °W) since 1999, we estimated a minimum of 21,290 t of floating microplastic.

Microbial processes within the subseafloor can be examined during the ephemeral and uncommonly observed phenomena known as snowblower venting. Snowblowers are characterized by the large quantity of white floc that is expelled from the seafloor following mid-ocean ridge eruptions. During these eruptions, rapidly cooling lava entrains seawater and hydrothermal fluids enriched in geochemical reactants, creating a natural bioreactor that supports a subseafloor microbial "bloom." Previous studies hypothesized that the eruption-associated floc was made by sulfide-oxidizing bacteria; however, the microbes involved were never identified. Here we present the first molecular analysis combined with microscopy of microbial communities in snowblower vents from samples collected shortly after the 2011 eruption at Axial Seamount, an active volcano on the Juan de Fuca Ridge. We obtained fluid samples and white flocculent material from active snowblower vents as well as orange flocculent material found on top of newly formed lava flows. Both flocculent types revealed diverse cell types and particulates when examined by phase contrast and scanning electron microscopy (SEM). Distinct archaeal and bacterial communities were detected in each sample type through Illumina tag sequencing of 16S rRNA genes and through sequencing of the sulfide oxidation gene, soxB. In fluids and white floc, the dominant bacteria were sulfur-oxidizing Epsilonproteobacteria and the dominant archaea were thermophilic Methanococcales. In contrast, the dominant organisms in the orange floc were Gammaproteobacteria and Thaumarchaeota Marine Group I. In all samples, bacteria greatly outnumbered archaea. The presence of anaerobic methanogens and microaerobic Epsilonproteobacteria in snowblower communities provides evidence that these blooms are seeded by subseafloor microbes, rather than from microbes in bottom seawater. These eruptive events thus provide a unique opportunity to observe subseafloor microbial communities.

Bacteria and archaea in the dark ocean (>200 m) comprise 0.3–1.3 billion tons of actively cycled marine carbon. Many of these microorganisms have the genetic potential to fix inorganic carbon (autotrophs) or assimilate single-carbon compounds (methylotrophs). We identified the functions of autotrophic and methylotrophic microorganisms in a vent plume at Axial Seamount, where hydrothermal activity provides a biogeochemical hot spot for carbon fixation in the dark ocean. Free-living members of the SUP05/Arctic96BD-19 clade of marine gamma-proteobacterial sulfur oxidizers (GSOs) are distributed throughout the northeastern Pacific Ocean and dominated hydrothermal plume waters at Axial Seamount. Marine GSOs expressed proteins for sulfur oxidation (adenosine phosphosulfate reductase, sox (sulfur oxidizing system), dissimilatory sulfite reductase and ATP sulfurylase), carbon fixation (ribulose-1,5-bisphosphate carboxylase oxygenase (RuBisCO)), aerobic respiration (cytochrome c oxidase) and nitrogen regulation (PII). Methylotrophs and iron oxidizers were also active in plume waters and expressed key proteins for methane oxidation and inorganic carbon fixation (particulate methane monooxygenase/methanol dehydrogenase and RuBisCO, respectively). Proteomic data suggest that free-living sulfur oxidizers and methylotrophs are among the dominant primary producers in vent plume waters in the northeastern Pacific Ocean.

Mantle rocks exposed on the seafloor constitute a highly reactive chemical and thermal system, in which interaction with seawater to produce serpentinite has major consequences for lithospheric cooling, global geochemical cycles, and microbial activity. Serpentinite-hosted hydrothermal activity is exemplified by the Lost City Hydrothermal Field (30°N, Mid-Atlantic Ridge) where fluid–rock reactions in the underlying ultramafic rocks result in high concentrations of abiotic hydrogen, methane, C2+ alkanes, and formate. Such systems have been proposed as possible analogs to the Early Earth environments that gave rise to the first biochemical pathways. Thus, characterizing the local microbial communities and their potential link with abiogenic compounds is of particular significance. Here we demonstrate that in active carbonate chimneys where microbial sulfate reduction is important, up to 50% of the microbial biomass is synthesized from mantle carbon. Conversely, mantle carbon contributes only ∼10% of the biomass in areas with minimal sulfate reduction. We attribute this difference to greater incorporation of formate or methane by the dominant microbial species, the Lost City Methanosarcinales, in locations where sulfate reducers are able to facilitate this assimilation. The ability of autotrophic communities at Lost City to capitalize on the steady stream of chemical products resulting from serpentinization reactions and to utilize abiogenic mantle carbon lend credence to the hypothesis that early biosynthetic pathways could have developed in similar environments.