Browsing by Author "Vera, Mario"

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    Characterization of Extracellular Polymeric Substances Produced by an Acidianus Species and Their Relevance to Bioleaching
    (2023) Castro, Camila; Donati, Edgardo R.; Vera, Mario
    Extracellular polymeric substances (EPS) produced by microorganisms play a crucial role in various bioprocesses, including bioleaching. The microbial leaching of metal sulfides requires an initial cell attachment, which is facilitated by EPS. These substances are mixtures of polysaccharides, proteins, lipids, and other compounds, and their composition and properties can vary depending on the species, growth conditions, and environmental factors. Despite the significance of iron/sulfur oxidizing species in biomining processes, the knowledge of the interfacial processes between thermoacidophilic archaeal species and mineral surfaces is limited. This study examines the cell surface characteristics and EPS produced by an Acidianus strain. The research was conducted using microscopic techniques, Zeta-potential measurements, spectrophotometric methods, Fourier transform infrared spectroscopy, and fluorescence lectin-binding analysis. The results suggest that non-soluble substrates, such as sulfur or pyrite, induce changes in cell surface structures, including the presence of cell appendages, wider cell envelopes, higher hydrophobicities, and increased EPS production, compared to cells grown with soluble substrates such as tetrathionate or ferrous iron. The EPS mainly consist of proteins and carbohydrates, including glucose, manose, N-acetylgalactosamine, and N-acetylglucosamine residues. This study contributes to a better understanding of the relationship between thermophilic archaea and mineral surfaces in biomining processes.
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    Development of an implantable three-dimensional model of a functional pathogenic multispecies biofilm to study infected wounds
    (2022) Cárdenas Calderón, Camila Valentina; Veloso Giménez, Valentina del Carmen; González, Tamara; Wozniak Banchero, Aniela; García, Patricia; San Martín, Sebastián; Varas, Juan F.; Carrasco-Wong, Ivo; Vera, Mario; Egaña, José T.
    Chronic wounds cannot heal due to impairment of regeneration, mainly caused by the persistent infection of multispecies biofilms. Still, the effects of biofilm wound infection and its interaction with the host are not fully described. We aimed to study functional biofilms in physiological conditions in vitro, and their potential effects in health and regeneration in vivo. Therefore, Pseudomonas aeruginosa, Staphylococcus aureus and Enterococcus faecalis were seeded in collagen-based scaffolds for dermal regeneration. After 24 h, scaffolds had bacterial loads depending on the initial inoculum, containing viable biofilms with antibiotic tolerance. Afterwards, scaffolds were implanted onto full skin wounds in mice, together with daily supervision and antibiotic treatment. Although all mice survived their health was affected, displaying fever and weight loss. After ten days, histomorphology of scaffolds showed high heterogeneity in samples and within groups. Wounds were strongly, mildly, or not infected according to colony forming units, and P. aeruginosa had higher identification frequency. Biofilm infection induced leucocyte infiltration and elevated interferon-γ and interleukin-10 in scaffolds, increase of size and weight of spleen and high systemic pro-calcitonin concentrations. This functional and implantable 3D biofilm model allows to study host response during infection, providing a useful tool for infected wounds therapy development.
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    Diffusible signal factor signaling controls bioleaching activity and niche protection in the acidophilic, mineral-oxidizing leptospirilli
    (2021) Bellenberg, Soren; Salas, Beatriz; Ganji, Suresh; Jorquera-Roman, Cristian; Valenzuela, Maria Luisa; Buetti-Dinh, Antoine; Unelius, C. Rikard; Dopson, Mark; Vera, Mario
    Bioleaching of metal sulfide ores involves acidophilic microbes that catalyze the chemical dissolution of the metal sulfide bond that is enhanced by attached and planktonic cell mediated oxidation of iron(II)-ions and inorganic sulfur compounds. Leptospirillum spp. often predominate in sulfide mineral-containing environments, including bioheaps for copper recovery from chalcopyrite, as they are effective primary mineral colonizers and oxidize iron(II)-ions efficiently. In this study, we demonstrated a functional diffusible signal factor interspecies quorum sensing signaling mechanism in Leptospirillum ferriphilum and Leptospirillum ferrooxidans that produces (Z)-11-methyl-2-dodecenoic acid when grown with pyrite as energy source. In addition, pure diffusible signal factor and extracts from supernatants of pyrite grown Leptospirillum spp. inhibited biological iron oxidation in various species, and that pyrite grown Leptospirillum cells were less affected than iron grown cells to self inhibition. Finally, transcriptional analyses for the inhibition of iron-grown L. ferriphilum cells due to diffusible signal factor was compared with the response to exposure of cells to N- acyl-homoserine-lactone type quorum sensing signal compounds. The data suggested that Leptospirillum spp. diffusible signal factor production is a strategy for niche protection and defense against other microbes and it is proposed that this may be exploited to inhibit unwanted acidophile species.
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    Membrane vesicles in Acidithiobacillia class extreme acidophiles: influence on collective behaviors of 'Fervidacidithiobacillus caldus'
    (2024) Rossoni, Stefano; Beard, Simon; Segura-Bidermann, Maria Ignacia; Duarte-Ramirez, Juan; Osorio, Francisco Kirhman; Varas-Godoy, Manuel; Martinez-Bellange, Patricio; Vera, Mario; Quatrini, Raquel; Castro, Matias
    Membrane vesicles (MVs) are envelope-derived extracellular sacs that perform a broad diversity of physiological functions in bacteria. While considerably studied in pathogenic microorganisms, the roles, relevance, and biotechnological potential of MVs from environmental bacteria are less well established. Acidithiobacillaceae family bacteria are active players in the sulfur and iron biogeochemical cycles in extremely acidic environments and drivers of the leaching of mineral ores contributing to acid rock/mine drainage (ARD/AMD) and industrial bioleaching. One key aspect of such a role is the ability of these bacteria to tightly interact with the mineral surfaces and extract electrons and nutrients to support their chemolithotrophic metabolism. Despite recent advances in the characterization of acidithiobacilli biofilms and extracellular matrix (ECM) components, our understanding of its architectural and mechanistic aspects remains scant. Using different microscopy techniques and nano-tracking analysis we show that vesiculation is a common phenomenon in distant members of the Acidithiobacillaceae family, and further explore the role of MVs in multicellular colonization behaviors using 'Fervidacidithiobacillus caldus' as a bacterial model. Production of MVs in 'F. caldus' occurred in both planktonic cultures and biofilms formed on sulfur surfaces, where MVs appeared individually or in chains resembling tube-shaped membranous structures (TSMSs) important for microbial communication. Liquid chromatography-mass spectrometry data and bioinformatic analysis of the MV-associated proteome revealed that 'F. caldus' MVs were enriched in proteins involved in cell-cell and cell-surface processes and largely typified the MVs as outer MVs (OMVs). Finally, microbiological assays showed that amendment of 'F. caldus' MVs to cells and/or biofilms affects collective colonizing behaviors relevant to the ecophysiology and applications of these acidophiles, providing grounds for their exploitation in biomining.
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    Novel Indigenous Strains and Communities with Copper Bioleaching Potential from the Amolanas Mine, Chile
    (2024) Casas-Vargas, Julian C.; Martinez-Bussenius, Cristobal; Videla, Alvaro; Vera, Mario
    Bioleaching, a process catalyzed by acidophilic microorganisms, offers a sustainable approach to metal extraction from sulfide minerals. Chalcopyrite, the world's most abundant copper sulfide, presents challenges due to surface passivation limiting its bioleaching efficiency. Also, indigenous species and microbial communities may present high copper extraction rates and offer new possibilities for application in bioleaching processes. This study examines the bioleaching potential of microbial isolates and communities obtained from Amolanas Mine in Chile. Samples were collected, cultivated, and identified by Sanger sequencing. The bioleaching potential and biofilm formation of isolates and enrichments were evaluated on pyrite and chalcopyrite. The results show the isolation of nine Leptospirillum and two Acidithiobacillus strains. The bioleaching experiments demonstrated good copper bioleaching potentials of the Leptospirillum I2CS27 strain and EICA consortium (composed mainly of Leptospirillum ferriphilum, Acidiphilium sp., and Sulfobacillus thermosulfidooxidans), with 11% and 25% copper recovery rates, respectively. Microbial attachment to the surface mineral was not mandatory for increasing the bioleaching rates. Our findings underscore the importance of indigenous microbial communities in enhancing copper bioleaching efficiency.
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    Quorum Sensing Signaling Molecules Positively Regulate c-di-GMP Effector PelD Encoding Gene and PEL Exopolysaccharide Biosynthesis in Extremophile Bacterium Acidithiobacillus thiooxidans
    (2021) Diaz, Mauricio; San Martin, Diego; Castro, Matias; Vera, Mario; Guiliani, Nicolas
    Acidithiobacillus species are fundamental players in biofilm formation by acidophile bioleaching communities. It has been previously reported that Acidithiobacillus ferrooxidans possesses a functional quorum sensing mediated by acyl-homoserine lactones (AHL), involved in biofilm formation, and AHLs naturally produced by Acidithiobacillus species also induce biofilm formation in Acidithiobacillus thiooxidans. A c-di-GMP pathway has been characterized in Acidithiobacillus species but it has been pointed out that the c-di-GMP effector PelD and pel-like operon are only present in the sulfur oxidizers such as A. thiooxidans. PEL exopolysaccharide has been recently involved in biofilm formation in this Acidithiobacillus species. Here, by comparing wild type and Delta pelD strains through mechanical analysis of biofilm-cells detachment, fluorescence microscopy and qPCR experiments, the structural role of PEL exopolysaccharide and the molecular network involved for its biosynthesis by A. thiooxidans were tackled. Besides, the effect of AHLs on PEL exopolysaccharide production was assessed. Mechanical resistance experiments indicated that the loss of PEL exopolysaccharide produces fragile A. thiooxidans biofilms. qRT-PCR analysis established that AHLs induce the transcription of pelA and pelD genes while epifluorescence microscopy studies revealed that PEL exopolysaccharide was required for the development of AHL-induced biofilms. Altogether these results reveal for the first time that AHLs positively regulate pel genes and participate in the molecular network for PEL exopolysaccharide biosynthesis by A. thiooxidans.