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Active cold seeps in the Okinawa Trough (OT) have been widely identified, but the sediment microbial communities associated with these sites are still poorly understood. Here, we investigated the distribution and biomass of the microbial communities, particularly those associated with the anaerobic oxidation of methane (AOM), in sediments from an active cold seep in the mid-Okinawa Trough. Methane-oxidizing archaea, including ANME-1a, ANME-1b, ANME-2a/b, ANME-2c, and ANME-3, were detected in the OT cold seep sediments. Vertical stratification of anaerobic methanotrophic archaea (ANME) communities was observed in the following order: ANME-3, ANME-1a, and ANME-1b. In addition, the abundance of methyl coenzyme M reductase A (mcrA) genes corresponded to high levels of dissolved iron, suggesting that methane-metabolizing archaea might participate in iron reduction coupled to methane oxidation (Fe-AOM) in the OT cold seep. Furthermore, the relative abundance of ANME-1a was strongly related to the concentration of dissolved iron, indicating that ANME-1a is a key microbial player for Fe-AOM in the OT cold seep sediments. Co-occurrence analysis revealed that methane-metabolizing microbial communities were mainly associated with heterotrophic microorganisms, such as JS1, Bathy-1, and Bathy-15. [ABSTRACT FROM AUTHOR]

• The diatom assemblages are closely correlated with the environmental variables. • Three assemblages are distinguished with different hydrological characteristics. • Important for palaeoceanographic studies in the northern and middle Okinawa Trough. Determining the relationship between diatoms in surface sediments and the local sedimentary environment is important for paleoenvironmental reconstruction, including in marine settings. In this study, we used canonical correspondence analysis of 41 surface sediment samples from the northern and middle Okinawa Trough to analyze the relationship between diatom species and environmental variables. The principal results are as follows. (i) The diatom assemblages are closely correlated with the environmental variables, specifically, July sea surface salinity (SSS-Jul), February sea surface temperature (SST-Feb), and June sea surface temperature (SST-Jun). (ii) Three diatom assemblages are distinguished, representing environments with different hydrological characteristics. The high-temperature diatom assemblage may indicate a warm-water mass (the Kuroshio Current), and the polyhaline diatom assemblage can be used as an indicator of a high salinity water mass. The mixed diatom assemblage is associated with the warm-water masses of the Kuroshio Current, and shelf water masses with a low salinity and low temperature. (iii) The high-temperature and mixed diatom assemblages occur on the boundary of the continental shelf and Okinawa Trough, and are temperature sensitive; the high-temperature assemblage is located further upstream of the Kuroshio Current, indicating higher temperatures. Changes in the distribution of the polyhaline diatom assemblage within sediment cores can be regarded as an indicator of changes in water masses, and of possible salinity variations, in this area. These three assemblages are potentially important for future palaeoceanographic studies. [ABSTRACT FROM AUTHOR]

• Cold seeps were linked to fault scarps and dome-structures in the Okinawa Trough. • Numerical models were used to quantify CH 4 fluxes and biogeochemical processes. • Cold seeps are C sources to the seawater, and Seep-wide budgets were constrained. The methane-enriched fluids in cold seeps are likely to crystallize as gas hydrates and serve as crucial sources of carbon to seawater. In this research, we analyzed the pore-water composition in terms of CH 4 , dissolved inorganic carbon (DIC), Cl−, Br−, SO 4 2−, Na+, Mg2+, Ca2+, Sr2+, and NH 4 +, and the δ13C DIC , δ13C CH4 , and δD CH4 values of two gravity cores and six remotely operated vehicle (ROV) video-guided push cores retrieved from fault scarps and dome-like structures (DSs) on the western slope of the mid-Okinawa Trough. In addition, a reaction–transport model was applied to quantify the methane fluxes and related biogeochemical processes. Active seepage of biogenic (δ13C CH4 ~ –70‰ V-PDB) and thermogenic (δ13C CH4 = −40‰ to −56‰ V-PDB) methane was identified on fault scarps and dome structures, respectively. Methane seepage was controlled by the transport and dissolution of the ascending gas rather than by clay dehydration or gas hydrate dissociation-induced fluid advection. The high methane concentrations and shallow sulfate–methane transition zones (SMTZs; between 0.1 and 0.4 mbsf) at sites R3-C2, R4-C4, and R6-C1 suggest strong methane seepage at three of the four studied DS (the highest gas dissolution rates are 6450, 1475, and 515 mmol m−2 yr−1). Site GC08, located along a fault scarp, exhibits a moderate methane seepage; the SMTZ is located at ~2.5 mbsf, and the rate of anaerobic oxidation of methane (AOM) is 130 mmol m−2 yr−1. The methane migrating from depth is mainly consumed by AOM. However, ~12%–66% of the methane released from the two most intensive seep sites escapes to the water column. The precipitation of high–Mg calcite (at all sites) and aragonite (only at site R3-C2) has fixed 27%–50% (average = 39%) of the DIC. Therefore, the carbon outputs to the water column have been reduced. In the study area, the area-weighted seafloor CH 4 and DIC fluxes are ~30 and 20 mmol m−2 yr−1, respectively. Together, they correspond to ~7%–14% of the organic carbon burial rate, indicating that sediments could not be simply regarded as a stable carbon sink because they provide methane and a certain amount of DIC to the water column. Our findings contribute to the results of the ongoing efforts in understanding carbon cycling in submarine cold seep systems. [ABSTRACT FROM AUTHOR]