Deep-sea carbonate mounds can harbor a wide variety of heterotrophic and chemosynthetic microbial communities, providing biodiversity hotspots among the deep-sea benthic ecosystems. This study examined the bacterial and archaeal diversity and community structure in the water column and sediments associated to a recently described giant carbonate mound named Alpha Crucis Carbonate Ridge (ACCR), Southwestern Atlantic. Due to the acoustic evidence of gas chimneys from a previous study, we further evaluated the chemosynthetic primary production through in situ-simulated dark carbon fixation rates. Pelagic microbial communities varied significantly with depth, showing a high abundance of photosynthetic groups in surface waters and taxa related to nitrification in intermediate and deep waters. The benthic communities from the top of the ACCR were very similar along with the sediment depth, while those from the base of ACCR showed a clear stratification pattern, with members in the deep strata mainly related to anoxic and chemosynthetic ecosystems. Dark carbon fixation rates were of the same order of magnitude as those of deep-sea cold seeps and hydrothermal vents. Our study provides the first description of the ACCR microbiome and adds new information to help formulate and implement future conservation and management strategies for vulnerable marine ecosystems.

Ocean and Coastal ResearchPockmarks are geological features that often sustain hydrocarbon-related communities of microorganisms when active, seeping oil or methane. Microbial communities of inactive pockmarks have not been well studied until the present. Plenty of pockmarks in the Southwestern Atlantic Ocean have been discovered. However, no information is available about the organic characteristics in association with the microbial diversity related to methane and nonmethane hydrocarbon consumption. This study examined the identity and potential ecology of the methane-related microbial community in an inactive SW Atlantic Carbonate-enriched Pockmark (SWACP). Undisturbed sediment cores were enriched with CH 4 99.5% at 5°C, with samples harvested at 16h, 120h, 240h, 720h, and 960h, followed by metataxonomics functional prediction, and the correlation of microbial groups with incubation and enrichment types. The SWACP is depleted in organic compounds, and chemosynthetic production is dominant. Incubated mini-cores of sediment were affected by incubation time and enrichment type, which influenced the microbial composition. Although several taxa were shared among all sediment samples, specific groups per enrichment type and incubation time were observed. These communities comprised taxa previously reported in marine bottom waters, carbonate crusts, active cold-seeps, and inactive pockmarks. The methane-enriched taxa were predominantly related to aerobic methanotrophy, methylotrophy, aerobic and anaerobic non-methane hydrocarbon degradation, and fermentation. This study brings the first survey of the key microbial groups in methane fluxes of a Brazilian deep-sea pockmark, providing data for understanding the ecology surrounding the SW Atlantic gas field areas.

Dark carbon fixation (DCF) is a source of new and labile carbon in the deep ocean, while heterotrophic microbial production (HMP) promotes organic matter transfer through the microbial loop. Despite their ecological relevance, there is a scientific gap regarding the estimates of DCF and HMP in the Southwestern Atlantic Ocean. Thus, the aim of this study was to investigate the spatial distribution of DCF and HMP; their relevance to the ocean carbon cycle; their relationship with environmental parameters and amongst themselves on the upper slope of Santos Basin. The samples were collected at three different water depths and sediment layers aboard the R/V Alpha Crucis in November 2019. DCF and HMP rates were measured by 14 C-bicarbonate and 3 H-leucine incorporation, respectively, and incubated in the dark. In the water column, DCF rates varied from 1.51 x 10 1 to 3.24 x 10 2 µg C m -3 h -1 , which were one to two orders of magnitude lower than the HMP rates, from 1.26 x 10 2 to 1.48 x 10 4 µg C m -3 h -1 . In the sediments, the DCF ranged from 1.15 x 10 4 to 1.83 x 10 5 µg C m -3 h -1 , while HMP was one to four orders of magnitude lower, 3.22 x 10 1 to 1.56 x 10 3 µ gC m -3 h -1 . DCF rates were significantly higher in the sediments, due to a higher availability of energy sources than in the oligotrophic water above. The HMP had higher rates in the water column as it is deeply dependent on organic matter derived from photosynthesis. This is the first study to investigate DCF and HMP considering the water column and sediments of the Southwestern Atlantic Ocean, thus contributing to a better understanding of the microbial role in the marine carbon cycle and ecosystem functioning.

The continental margin off the southeastern Brazilian coast is punctuated by a series of geological-geomorphological features, such as subsurface saline diapirs and pockmarks at the seafloor interface, which evidence the abundant presence of oil and gas in the region. In several of these sites, hydrocarbons can be naturally released into the water column, areas are cold seep areas. These are marked by the presence of oil- and gas-dependent ecosystems, where specific organisms are able to fix carbon from hydrocarbon chemosynthesis. In addition, light hydrocarbon fluid flow through the sediment may build up authigenic carbonates that can be further colonized by cold-water corals, generating large carbonate mounds over geological time, normally positioned at the border of these pockmark features. The present work reports on a multidisciplinary oceanographic cruise carried out in the Santos Basin, SW Atlantic, to seek, map, and collect geological, chemical, and biological data from different deep-sea habitats. The cruise occurred in November 2019 on the R/V Alpha Crucis of the Oceanographic Institute of the University of São Paulo (IOUSP). We intended to discover and detail different geomorphological features, characterize free-living and symbiotic microorganisms, determine the chemosynthetic rates in relation to heterotrophic microbial production, and characterize the fauna and study their ecological and evolutionary links within and across ocean basins. All discoveries made during the cruise and their respective results will be presented separately in several papers that comprise this special volume.