IntroductionShallow hydrothermal systems share many characteristics with their deep-sea counterparts, but their accessibility facilitates their study. One of the most studied shallow hydrothermal vent fields lies at Paleochori Bay off the coast of Milos in the Aegean Sea (Greece). It has been studied through extensive mapping and its physical and chemical processes have been characterized over the past decades. However, a thorough description of the microbial communities inhabiting the bay is still missing. MethodsWe present the first in-depth characterization of the prokaryotic communities of Paleochori Bay by sampling eight different seafloor types that are distributed along the entire gradient of hydrothermal influence. We used deep sequencing of the 16S rRNA marker gene and complemented the analysis with qPCR quantification of the 16S rRNA gene and several functional genes to gain insights into the metabolic potential of the communities. ResultsWe found that the microbiome of the bay is strongly influenced by the hydrothermal venting, with a succession of various groups dominating the sediments from the coldest to the warmest zones. Prokaryotic diversity and abundance decrease with increasing temperature, and thermophilic archaea overtake the community. DiscussionRelevant geochemical cycles of the Bay are discussed. This study expands our limited understanding of subsurface microbial communities in acidic shallow-sea hydrothermal systems and the contribution of their microbial activity to biogeochemical cycling.

Submarine hydrothermal activity is responsible for heat and chemical exchanges through the seafloor. Shallow-water hydrothermal systems (SWHS), while identified around the globe, are often studied in a way that is less comprehensive than their deep-ocean counterparts (e.g., along ridges), where systematic optical and acoustic mapping is more prevalent and coupled to in situ observations and sampling. Using aerial drones, an AUV, and temperature measurements at 10-40 cm subseafloor, we investigated in 2019 one of the most extensive SWHS known to date, in Paleochori and nearby Spathi and Agia Kyriaki Bays (south of Milos, Greece). Hydrothermal venting, found from the shore to water depths of almost 500 m, shows emissions of gases and high-temperature fluids, often associated with bacterial mats and/or hydrothermal mineral precipitates. This study provides extensive drone mapping coupled with local AUV surveys for seafloor characterization and ground-truthing from the shore to similar to 20 m water depth. Seafloor photomosaics also provide a detailed context to samples, measurements and observations carried in situ. We interpret the photomosaics to define distinct seafloor types, linked to this hydrothermal activity. White hydrothermal patches (WHPs) often show a clear polygonal organization, together with outflow areas that are both more dispersed and distributed. Polygonal patterns likely result from fluid convection in a sandy porous medium heated from below. These WHPs display elevated subseafloor temperatures, typically >50 degrees C, with maximum values of similar to 75 degrees C. Photomosaics also display textures of biological origin, including seagrass and bioturbation patterns. Widespread bioturbation by burrowing shrimps is often associated with WHPs, bounding them, but also occurs on sandy seafloor away from hydrothermal patterns. Subseafloor temperatures at these bioturbated areas are of similar to 30-40 degrees C, and are thus transitional between hot WHPs and sedimented seafloor unaffected by hydrothermal activity (similar to 24 degrees C). In addition to linking subseafloor temperature data and interpreted seafloor photomosaics, our results provide a comprehensive general overview of this SWHS, of the organization of its hydrothermal outflow through the seafloor, and of the underlying subseafloor fluid circulation. This paper also gives the first perspectives on the heat fluxes of the system, and constitutes a background for other studies on the nature and distribution of microbial communities, controlled by this hydrothermal activity.

Submarine hydrothermal activity is common at the flanks of volcanic islands, and in some cases, occurring at very shallow water (0-100 meter depth). These sites are a key target for systematic seafloor mapping to understand the location, geometry and nature of hydrothermal discharge. These data are also critical for monitoring the temporal variability of these dynamic systems, while providing a context for instrumental measurements, sampling and other observations (e.g., temperature of outflow, chemistry, etc.). Here we present a systematic mapping of the Milos hydrothermal system in the Hellenic volcanic Arc, characterized by submarine gas emissions, high-temperature outflow, bacterial mats, precipitation of hydrothermal minerals, and small hydrothermal constructs and edifices. We have mapped this site at regional scales using satellite imagery (World-View2 images from the DigitalGlobe foundation), complemented with aerial photography acquired with drones, and high-resolution seafloor photomosaics (<1 cm resolution) from underwater imagery acquired by the autonomous underwater vehicle Sparus II (University of Girona). Our drone and AUV mapping ground truths the correlation between patterns in satellite imagery and hydrothermal outflow, associated to mineral precipitates and/or bacterial mats at the seafloor. This mapping also reveals a clear organization of the hydrothermal outflow in sandy areas. In particular, polygonal patterns are common and often associated with inactive or actively bubbling pockmarks. These areas, showing white bacterial mats and hydrothermal precipitates, are rippled, suggesting that the hydrothermal precipitates do not consolidate the sediment. White precipitates display subseafloor temperatures >50°C at depths of 10 to 50 cm. The white areas are bound by bands of seafloor with a hummocky structure due to intense bioturbation, that obliterates the ripples, with widths of up to a few meters. This area shows subseafloor temperatures of 20-40°C, and corresponds to a transition from the high-temperature white zones and the seafloor with ripples and no hydrothermal precipitates. This area exhibits subseafloor temperatures similar to those of seawater, and can be associated with seagrass. These patterns reveal a clear organization of a narrowly focused hydrothermal outflow that controls the biological communities at the seafloor and subseafloor. We will discuss the implications of these observations to quantify hydrothermal fluxes in the study area.

Abyssivirga alkaniphila strain L81T, recently isolated from a black smoker biofilm at the Loki’s Castle hydrothermal vent field, was previously described as a mesophilic, obligately anaerobic heterotroph able to ferment carbohydrates, peptides, and aliphatic hydrocarbons. The strain was classified as a new genus within the family Lachnospiraceae. Herein, its genome is analyzed and A. alkaniphila is reassigned to the genus Vallitalea as a new strain of V. guaymasensis, designated V. guaymasensis strain L81. The 6.4 Mbp genome contained 5651 protein encoding genes, whereof 4043 were given a functional prediction. Pathways for fermentation of mono-saccharides, di-saccharides, peptides, and amino acids were identified whereas a complete pathway for the fermentation of n-alkanes was not found. Growth on carbohydrates and proteinous compounds supported methane production in co-cultures with Methanoplanus limicola. Multiple confurcating hydrogen-producing hydrogenases, a putative bifurcating electron-transferring flavoprotein—butyryl-CoA dehydrogenase complex, and a Rnf-complex form a basis for the observed hydrogen-production and a putative reverse electron-transport in V. guaymasensis strain L81. Combined with the observation that n-alkanes did not support growth in co-cultures with M. limicola, it seemed more plausible that the previously observed degradation patterns of crude-oil in strain L81 are explained by unspecific activation and may represent a detoxification mechanism, representing an interesting ecological function. Genes encoding a capacity for polyketide synthesis, prophages, and resistance to antibiotics shows interactions with the co-occurring microorganisms. This study enlightens the function of the fermentative microorganisms from hydrothermal vents systems and adds valuable information on the bioprospecting potential emerging in deep-sea hydrothermal systems.

A strictly anaerobic, mesophilic, syntrophic, alkane-degrading strain, L81T, was isolated from a biofilm sampled from a black smoker chimney at the Loki's Castle vent field. Cells were straight, rod-shaped, Gram-positive-staining and motile. Growth was observed at pH 6.2-9.5, 14-42 °C and 0.5-6 % (w/w) NaCl, with optima at pH 7.0-8.2, 37 °C and 3% (w/w) NaCl. Proteinaceous substrates, sugars, organic acids and hydrocarbons were utilized for growth. Thiosulfate was used as an external electron acceptor during growth on crude oil. Strain L81T was capable of syntrophic hydrocarbon degradation when co-cultured with a methanogenic archaeon, designated strain LG6, isolated from the same enrichment. Phylogenetic analysis based on the 16S rRNA gene sequence indicated that strain L81T is affiliated with the family Lachnospiraceae, and is most closely related to the type strains of Natranaerovirga pectinivora (92 % sequence similarity) and Natranaerovirga hydrolytica (90%). The major cellular fatty acids of strain L81T were C15 : 0, anteiso-C15 : 0 and C16 : 0, and the profile was distinct from those of the species of the genus Natranaerovirga. The polar lipids were phosphatidylglycerol, diphosphatidylglycerol, three unidentified phospholipids, four unidentified glycolipids and two unidentified phosphoglycolipids. The G+C content of genomic DNA was determined to be 31.7 mol%. Based on our phenotypic, phylogenetic and chemotaxonomic results, strain L81T is considered to represent a novel species of a new genus of the family Lachnospiraceae, for which we propose the name Abyssivirga alkaniphila gen. nov., sp. nov. The type strain of Abyssivirga alkaniphila is L81T (=DSM 29592T=JCM 30920T). We also provide emended descriptions of Natranaerovirga pectinivora and Natranaerovirga hydrolytica.