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This work presents the preliminary result of the multidisciplinary cruise EXPLOSEA2 surveying the northern Mid-Atlantic Ridge and Azores Archipelago from 46◦30′N to 38◦30′N aboard the R/VSarmiento de Gamboaand ROVLusoover 54 days (June 11 to July 27, 2019). In this cruise report, we detail the geophysical, hydrographic, geological, oceanographic, ecological, and microbiological data acquired and a brief of main findings. The cruise addressed the exploration and comprehensive characterization of venting sites, including the water column, the sediments and rocks that host the hydrothermal activity, and the associated mineralizations, biology, and microbiology. Deep hydrothermal chimneys and massive sulfide deposits (up 3,000 m in depth) within the Moytirra hydrothermal active field were identified on slopes that had not been explored previously. Another striking finding made during the EXPLOSEA2 cruise was the field of carbonate chimneys named the “Magallanes-Elcano” field, a potentially relict ultramafic-hosted hydrothermal site sourced by abiotic methane. This field is related to a serpentinite and gabbro rock outcropping on a dome-shaped massif named the “Iberian Massif.” An outstanding finding of the EXPLOSEA2 survey was the identification of the first garden of soft corals growing after active submarine eruptions were reported in the Azores Archipelago composed by a high density of soft corals the suborderAlcyoniinaat the summit and flanks of a recent volcanic cone at 160 m water depth developed during the 1957–1958 eruption of Capelinhos. Several cold-water coral habitats formed by colonial scleractinians (e.g.,Lophelia pertusaandMadrepora oculata), coral gardens composed of mixed assemblages of black corals (Leiopathessp.), and octocorals and dense aggregations of the glass spongePheronema carpenterithat may be classifiedas vulnerable marine ecosystems (VMEs) have been discovered during the EXPLOSEA2 cruise along the northern Mid-Atlantic Ridge. This work reveals the importance of multidisciplinary surveys to the knowledge of deep-sea environments.

In mining areas in which sulfur-containing ores are exposed, the oxidation of sulfides leads to the formation of acidic mine drainage (AMD) waters. Both traditional and molecular microbial studies have shown that chemolithotrophic sulfur- and iron-oxidizing bacteria are responsible for this activity. The Iberian Pyrite Belt (IPB), located in southwestern Spain represents one of the world’s largest accumulations of mine wastes and AMD waters. Mineralogical and textural characteristics of the IPB ores favor the oxidation and dissolution of pyrite and the subsequent formation of AMD waters. Acidic pit lakes in the IPB are a hotspot for this activity. These pit lakes are former open pit mines where ores have been exposed leading the formation of AMD waters. At present, there are more than 25 of these pit lakes between the provinces of Huelva and Seville (SW Spain). This work reports the physical properties, hydrogeochemical characteristics, and microbial diversity of two pit lakes located in the IPB Nuestra Señora del Carmen (NSC), and Concepción (CN). Both pit lakes are acid (pH 2-4) and showed chemical and thermal stratification with well defined chemoclines. One particular characteristic of NSC is that it has developed a chemocline very close to the surface (2 m depth). Microbial community composition of the water column was analyzed by 16S and 18S rRNA gene cloning and sequencing. The microorganisms detected in NSC are characteristic of acid mine drainage (AMD), including iron oxidizing bacteria (Leptospirillum) and facultative iron reducing bacteria and archaea (Acidithiobacillus ferrooxidans, Actinobacteria, Acidimicrobiales, Ferroplasma) detected in the bottom layer. Diversity in CN was higher than in NSC. Microorganisms known from AMD systems (Acidiphilium, Acidobacteria and Ferrovum) and microorganisms never reported from AMD systems were both identified.

This work describes recent research carried out in an extremely acidic (pH 0.61–0.82) and hypersaline (e.g., 134 g/L SO4 2-, 74 g/L Fe, 7.5 g/L Al, 3 g/L Mg, 2 g/L Cu, 1 g/L Zn) leachate which seeps from a pyrite pile in San Telmo mine (Huelva, SW Spain) and forms evaporative pools of ultra-concentrated water in which attractive crystals of Zn-rich melanterite (FeIISO4 7H2O) are formed. Geochemical modeling with the Pitzer method indicates that the acidic brine was near saturation with respect to melanterite (SIMel = 0 ± 0.2). The microbiological investigation has revealed a surprisingly high biomass (1.4 × 106 cells mL−1) and an exotic ecosystem composed of acidophilic, Fe-oxidizing archaea (mainly Ferroplasma spp., representing 52% of the microbial population), and minor numbers of acidophilic bacteria (including Leptospirillum spp. (3.2%), Acidithiobacillus spp. (1.6%), and Alphaproteobacteria (2.8%)). The microbial production of FeIII allows the oxidative dissolution of pyrite and other sulphides, which results in additional inputs of FeII, SO4 2- and acidity to the system. The surfaces of the pyrite crystals show a typical etch-pitted texture, as well as blobs of elemental sulphur, which are both compatible with this indirect, microbially mediated oxidation mechanism. The composition of the acidic leachate seems to result from the combination of several processes which include: (1) formation of melanterite within the pile during relatively dry seasons, (2) subsequent dissolution of melanterite during rainy episodes, (3) microbial oxidation of FeII, (4) sulphide oxidation mediated by FeIII, (5) dissolution of chlorite and other aluminosilicates present in the pile, and (6) cooling and/or evaporation of seepage from the pile and consequent melanterite precipitation.

The exposure of fresh sulfide-rich lithologies by the retracement of the Nevado Pastoruri glacier (Central Andes, Peru) is increasing the presence of heavy metals in the water as well as decreasing the pH, producing an acid rock drainage (ARD) process in the area. We describe the microbial communities of an extreme ARD site in Huascaran National Park as well as their correlation with the water physicochemistry. Microbial biodiversity was analyzed by FLX 454 sequencing of the 16S rRNA gene. The suggested geomicrobiological model of the area distinguishes three different zones. The proglacial zone is located in the upper part of the valley, where the ARD process is not evident yet. Most of the OTUs detected in this area were related to sequences associated with cold environments (i.e., psychrotolerant species of Cyanobacteria or Bacteroidetes). After the proglacial area, an ARD-influenced zone appeared, characterized by the presence of phylotypes related to acidophiles (Acidiphilium) as well as other species related to acidic and cold environments (i.e., acidophilic species of Chloroflexi, Clostridium and Verrumicrobia). Sulfur- and iron-oxidizing acidophilic bacteria (Acidithiobacillus) were also identified. The post-ARD area was characterized by the presence of OTUs related to microorganisms detected in soils, permafrost, high mountain environments, and deglaciation areas (Sphingomonadales, Caulobacter or Comamonadaceae).

The Iberian Pyrite Belt (IPB) hosts one of the world's largest accumulations of acidic mine wastes and pit lakes. The mineralogical and textural characteristics of the IPB ores have favored the oxidation and dissolution of metallic sulfides, mainly pyrite, and the subsequent formation of acidic mining drainages. This work reports the physical properties, hydrogeochemical characteristics, and microbial diversity of two pit lakes located in the IPB. Both pit lakes are acidic and showed high concentrations of sulfate and dissolved metals. Concentrations of sulfate and heavy metals were higher in the Nuestra Señora del Carmen lake (NSC) by one order of magnitude than in the Concepción (CN) lake. The hydrochemical characteristics of NSC were typical of acid mine waters and can be compared with other acidic environments. When compared to other IPB acidic pit lakes, the superficial water of CN is more diluted than that of any of the others due, probably, to the strong influence of runoff water. Both pit lakes showed chemical and thermal stratification with well defined chemoclines. One particular characteristic of NSC is that it has developed a chemocline very close to the surface (2 m depth). Microbial community composition of the water column was analyzed by 16S and 18S rRNA gene cloning and sequencing. The microorganisms detected in NSC were characteristic of acid mine drainage (AMD), including iron oxidizing bacteria (Leptospirillum, Acidithiobacillus ferrooxidans) and facultative iron reducing bacteria and archaea (Acidithiobacillus ferrooxidans, Acidiphilium, Actinobacteria, Acidimicrobiales, Ferroplasma) detected in the bottom layer. Diversity in CN was higher than in NSC. Microorganisms known from AMD systems (Acidiphilium, Acidobacteria and Ferrovum) and microorganisms never reported from AMD systems were identified. Taking into consideration the hydrochemical characteristics of these pit lakes and the spatial distribution of the identified microorganisms, a model explaining their geomicrobiology is advanced., This work has been supported by grants from the MINECO-Spain (www.mineco.gob.es) (CGL2011-22540 and CGL2009-11059), IGME 478 Grant (www.igme.es) and an ERC project ( erc.europa.eu) (250-350-IPBSL).

Effluent from La Zarza-Perrunal, a mine on the Iberian Pyrite Belt, was chosen to be geomicrobiologically characterized along a 1,200-m stream length. The pH at the origin was 3.1, which decreased to 1.9 at the final downstream sampling site. The total iron concentration showed variations along the effluent, resulting from (i) significant hydrolysis and precipitation of Fe(III) (especially along the first reach of the stream) and (ii) concentration induced by evaporation (mostly in the last reach). A dramatic increase in iron oxidation was observed along the course of the effluent [from Fe(III)/Fetotal = 0.11 in the origin to Fe(III)/Fetotal = 0.99 at the last sampling station]. A change in the O2 content along the effluent, from nearly anoxic at the origin to saturation with oxygen at the last sampling site, was also observed. Prokaryotic and eukaryotic diversity throughout the effluent was determined by microscopy and 16S rRNA gene cloning and sequencing. Sulfate-reducing bacteria (Desulfosporosinus and Syntrophobacter) were detected only near the origin. Some iron-reducing bacteria (Acidiphilium, Acidobacterium, and Acidosphaera) were found throughout the river. Iron-oxidizing microorganisms (Leptospirillum spp., Acidithiobacillus ferrooxidans, and Thermoplasmata) were increasingly detected downstream. Changes in eukaryotic diversity were also remarkable. Algae, especially Chlorella, were present at the origin, forming continuous, green, macroscopic biofilms, subsequently replaced further downstream by sporadic Zygnematales filaments. Taking into consideration the characteristics of this acidic extreme environment and the physiological properties and spatial distribution of the identified microorganisms, a geomicrobiological model of this ecosystem is advanced., A.A. and V.S.-E. were supported by a Ramón y Cajal contract from the Ministerio de Educación y Ciencia. E.G.-T. is supported by Instituto Nacional de Tecnica Aeroespacial (INTA). This work has been supported by grants from the MICINN-Spain (CGL2008-02298/BOS) and the Centro de Astrobiología (CSIC-INTA).

The Arcas and River Moscas lake complexes, protected natural areas in Cuenca province, are located in interstratified karst developed in Cretaceous-Tertiary boundary gypsum. These sites are in the central part of the province, near the town of Cuenca, each covering an area of some 3 km2 and distributed between the municipalities of Arcas del Villar, Cuenca, Fuentes, Valdetortola and Villar de Olalla. This paper presents certain geomorphological aspects established in the framework of a broader study on the hydrogeology of the two complexes, fed chiefly by groundwater discharge from this gypsiferous aquifer, whose hydrodynamic behaviour is determined by its fractures and faults.

Iron terraces that form in acidic mine drainage settings are unique and extreme geomicrobiological systems that can provide highly relevant information about the interaction between microbes and their surrounding aqueous environments. These singular systems can represent, additionally, potential models for the study of ancient geological formations (e.g., banded iron formations, stromatolites) and/or for the cycling of iron on Mars. This work describes geochemical, mineralogical, morphological, and microbiological evidence obtained in the highly acidic and Fe-rich Tintillo River (Riotinto mines, Huelva, SW Spain), which can be used to speculate about the origin and nature of the terraced iron formations (TIFs) that are being currently formed in acid mine drainage environments. The size (up to 36 m long and 1 m thick) and continuity (strong development over 3.5 km) of the iron terraces offer a unique opportunity to study the different organic (mainly microbial) and inorganic processes involved in the construction of these characteristic, travertine-like, sedimentary structures. Evidence presented in this study suggests that both types of processes appear to be controlling factors in the formation and internal arrangement of the TIFs, although no defi nitive evidence has been found to support the prevalence of any of these mechanisms with respect to another. The photosynthetic production of dissolved oxygen by eukaryotic microorganisms (green algae, euglenophytes, and diatoms) and the Fe-oxidizing metabolism of acidophilic prokaryotes are critical factors for the formation of TIFs, whereas abiotic parameters, such as water composition, fl ow rate and velocity, or stream channel geometry, also appear to be essential variables.