Search

Previous studies have reported the role of some species of acidophilic bacteria in accelerating the dissolution of goethite under aerobic and anaerobic conditions. This has relevance for environments impacted by acid mine drainage and for the potential bioleaching of limonitic laterite ores. In this study, natural wellcharacterized goethite mineral samples and synthetic goethite were used in aerobic and anaerobic laboratory batch culture incubation experiments with ferric iron-reducing, acidophilic bacteria, including the lithoautotrophic species Acidithiobacillus (At.) thiooxidans, At. ferrooxidans, and At. caldus, as well as two strains of the organoheterotrophic species Acidiphilium cryptum. All bacteria remained alive throughout the experiments and efficiently reduced soluble ferric iron in solution in positive control assays. However, goethite dissolution was low to negligible in all experimental assays with natural goethite, while some dissolution occurred with synthetic goethite in agreement with previous publications. The results indicate that ferric iron-reducing microbial activity at low pH is less relevant for goethite dissolution than the oxidation of elemental sulfur to sulfuric acid. Microbial ferric iron reduction enhances but does not initiate goethite dissolution in very acidic liquors. [ABSTRACT FROM AUTHOR]

The extraction of nickel, cobalt, and other metals from laterite ores via bioleaching with sulfur-oxidizing and ferric iron-reducing, autotrophic, acidophilic bacteria (e.g. Acidithiobacillus species) has been demonstrated under anaerobic as well as aerobic conditions in experiments in different laboratories. This study demonstrated the bioleaching of laterites from Brazil with the addition of elemental sulfur in 2-L stirred-tank bioreactors with pure and mixed cultures of Acidithiobacillus and Sulfobacillus species under aerobic conditions. In particular, a potential disturbance of mineral dissolution under aerobic conditions by ferrous iron-oxidizing acidophiles likely introduced as contaminants in an applied bioleaching process was investigated with Leptospirillum ferrooxidans at 30°C and Leptospirillum ferriphilum at 40°C, at maintained pH 1.5 or without maintained pH leading to an increase in acidity (with pH values <1.0) due to the biological production of sulfuric acid. Despite the proportion of ferrous iron to the total amount of extracted iron in the solution being drastically reduced in the presence of Leptospirillum species, there was a negligible effect on the extraction efficiency of nickel and cobalt, which is positive news for laterite bioleaching under aerobic conditions. [ABSTRACT FROM AUTHOR]

Dibenzothiophene (DBT) is a major form of organo-sulfur compound chiefly found in coal. Acidithiobacillus ferrooxidans cells played the role of catalyst in degrading DBT to 2-hydroxy biphenyl (2HBP) through the formation of dibenzothiophene-sulfone and releasing the sulfur present in it. Experiments on the biodegradation of DBT by A. ferrooxidans bacterium using 9K medium with DBT at 3 g/L in a 500 mL shake flask scale achieved degradation of DBT up to 95% in 12 days. The A. ferrooxidans cells degrade DBT to 2-hydroxy biphenyl and release the sulfur atom in DBT through the formation of the intermediate dibenzothiophene-sulfone. The formation of 2HBP from microbial degradation of DBT followed the chemical control model of shrinking core reactions model in solid-liquid interactions. The kinetics of the DBT degradation by A. ferrooxidans followed first-order for a low initial concentration of DBT (3 g/L), and the kinetics were shifted to second-order with an increase in DBT concentration to 5 g/L. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

Previous studies have reported the role of some species of acidophilic bacteria in accelerating the dissolution of goethite under aerobic and anaerobic conditions. This has relevance for environments impacted by acid mine drainage and for the potential bioleaching of limonitic laterite ores. In this study, natural well-characterized goethite mineral samples and synthetic goethite were used in aerobic and anaerobic laboratory batch culture incubation experiments with ferric iron-reducing, acidophilic bacteria, including the lithoautotrophic species Acidithiobacillus (At.) thiooxidans, At. ferrooxidans, and At. caldus, as well as two strains of the organoheterotrophic species Acidiphilium cryptum. All bacteria remained alive throughout the experiments and efficiently reduced soluble ferric iron in solution in positive control assays. However, goethite dissolution was low to negligible in all experimental assays with natural goethite, while some dissolution occurred with synthetic goethite in agreement with previous publications. The results indicate that ferric iron-reducing microbial activity at low pH is less relevant for goethite dissolution than the oxidation of elemental sulfur to sulfuric acid. Microbial ferric iron reduction enhances but does not initiate goethite dissolution in very acidic liquors.

The extraction of nickel, cobalt, and other metals from laterite ores via bioleaching with sulfur-oxidizing and ferric iron-reducing, autotrophic, acidophilic bacteria (e.g. Acidithiobacillus species) has been demonstrated under anaerobic as well as aerobic conditions in experiments in different laboratories. This study demonstrated the bioleaching of laterites from Brazil with the addition of elemental sulfur in 2-L stirred-tank bioreactors with pure and mixed cultures of Acidithiobacillus and Sulfobacillus species under aerobic conditions. In particular, a potential disturbance of mineral dissolution under aerobic conditions by ferrous iron-oxidizing acidophiles likely introduced as contaminants in an applied bioleaching process was investigated with Leptospirillum ferrooxidans at 30°C and Leptospirillum ferriphilum at 40°C, at maintained pH 1.5 or without maintained pH leading to an increase in acidity (with pH values

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.

During the last decades, technologies for agricultural drainage water remediation had been widely explored to oppose the arising intimidation of their pollutants on health, biotas and farming sustainability. The present study targeted to augment the remediation competence of inorganic pollutants and faecal coliforms from agricultural drainage water using different types of clay minerals. Kaolinite, Bentonite, Ball clay and Aswan clay were used either solely or in amalgamation with some biological premeditative amendments (cultures of Phosphate dissolving Bacteria (PDB) and/or Acidithiobacillus sp.] for drainage water remediation of samples delivered from Belbies and Rahawy agricultural drains. Nineteen trails and four groups were implemented for each drain together with a control sample. Water samples were analysed before and after clay minerals' trails application for detection of Cd(II), Cu(II), Ni(II), Zn(II) and As(V) by Atomic Spectroscopy. Total faecal bacteria presence was also quantified through direct culturing on MacConkey agar plates. The obtained results confirmed a significant decrease in the concentrations of Cd, Cu, Ni, Zn and As pollutants as well as a decrease in the faecal coliform intensities under all trailed treatments compared to control samples. Worthy to mention that 4th group and especially T18 (Bentonite + Acidithiobacillus+ phosphate dissolving bacteria) was the most efficient in minimising the hazards of PTEs and the existence of faecal microbes. [ABSTRACT FROM AUTHOR]

Conventional sewage treatment plants aim to minimize environmental impacts, but generate by-products, such as sludge, which also must be properly treated and dewatered. From the resource recovery perspective, a promising alternative is a reuse as fertilizer in agriculture, due to the nutrients in its composition, but the feasibility of this reuse depends on sludge treatment technologies that, in addition to promoting dewatering, treat toxic pollutants, pathogens and metals. Many techniques have been employed in an attempt to promote effective dewatering and remove pollutants from sludge, but the high cost of physicochemical treatments and risk of causing secondary pollution, make it difficult to apply on a larger scale, as well as reuse the sludge. Therefore, biological treatments, such as bioleaching, have become an increasingly popular choice, as they have economic and environmental benefits, which with improvements can enable the use of sludge as a resource. The objective of the present study was to investigate the scientific production of the last twelve years, regarding the microbiology of extracellular polymeric substances influence on sewage sludge dewatering during the bioleaching process. The specific objectives were to elucidate the influence of extracellular polymeric substances fractions on sludge dewatering, to identify the main microorganisms involved in extracellular polymeric substances generation and identify the main microorganisms involved in its removal during bioleaching. This literature review will help to understand the microbiological aspects of dewatering during the bioleaching process and will provide useful information to improve future research on the application of bioleaching in sewage sludge treatment. [ABSTRACT FROM AUTHOR]

Simple Summary: The application of extremophilic, acidophilic microbial populations for the bio-oxidation of gold-bearing sulfide concentrates in industry is a source of great interest in the research of these populations. Understanding the effects of different factors on the activity of such acidophilic populations may allow for the performance of the bio-oxidation of sulfide concentrates to be regulated. In the present work, we studied the effects of different temperatures as well as carbon sources (carbon dioxide and molasses) on the rate of the bio-oxidation of a gold-bearing pyrite–arsenopyrite concentrate as well as on the composition of the microbial population performing this process. It was shown that an increase in the temperature from 40 to 50 °C led to a decrease in the intensity of bio-oxidation, while the application of additional carbon dioxide as a carbon source made it possible to prevent the inhibition of bio-oxidation due to a temperature increase. An analysis of the populations formed under different experimental conditions revealed that both temperature and carbon dioxide affected the composition of the microbial population which, in turn, may explain the effect on the bio-oxidation performance. Thus, the use of additional carbon dioxide may be proposed as the method to increase the efficiency of bio-oxidation and to prevent the negative effect of a temperature increase on the activity of bio-oxidation. In the present study, the effect of additional carbon sources (carbon dioxide and molasses) on the bio-oxidation of a pyrite–arsenopyrite concentrate at temperatures of 40–50 °C was studied, and novel data regarding the patterns of the bio-oxidation of gold-bearing sulfide concentrates and the composition of the microbial populations performing these processes were obtained. At 40 °C, additional carbon sources did not affect the bio-oxidation efficiency. At the same time, the application of additional carbon dioxide improved the bio-oxidation performance at temperatures of 45 and 50 °C and made it possible to avoid the inhibition of bio-oxidation due to an increase in the temperature. Therefore, the use of additional carbon dioxide may be proposed to prevent the negative effect of an increase in temperature on the bio-oxidation of sulfide concentrates. 16S rRNA gene profiling revealed archaea of the family Thermoplasmataceae (Acidiplasma, Ferroplasma, Cuniculiplasma, and A-plasma group) and bacteria of the genera Leptospirillum, with Sulfobacillus and Acidithiobacillus among the dominant groups in the community. Temperature influenced the composition of the communities to a greater extent than the additional sources of carbon and the mode of operation of the bioreactor. Elevating the temperature from 40 °C to 50 °C resulted in increases in the shares of Acidiplasma and Sulfobacillus and decreases in the relative abundances of Ferroplasma, Leptospirillum, and Acidithiobacillus, while Cuniculiplasma and A-plasma were more abundant at 45 °C. A metagenomic analysis of the studied population made it possible to characterize novel archaea belonging to an uncultivated, poorly-studied group of Thermoplasmatales which potentially plays an important role in the bio-oxidation process. Based on an analysis of the complete genome, we propose describing the novel species and novel genus as "Candidatus Carboxiplasma ferriphilum" gen. nov., spec. nov. [ABSTRACT FROM AUTHOR]

Mobile genetic elements (MGEs) are relevant agents in bacterial adaptation and evolutionary diversification. Stable appropriation of these DNA elements depends on host factors, among which are the nucleoid-associated proteins (NAPs). NAPs are highly abundant proteins that bind and bend DNA, altering its topology and folding, thus affecting all known cellular DNA processes from replication to expression. Even though NAP coding genes are found in most prokaryotic genomes, their functions in host chromosome biology and xenogeneic silencing are only known for a few NAP families. Less is known about the occurrence, abundance, and roles of MGE-encoded NAPs in foreign elements establishment and mobility. In this study, we used a combination of comparative genomics and phylogenetic strategies to gain insights into the diversity, distribution, and functional roles of NAPs within the class Acidithiobacillia with a special focus on their role in MGE biology. Acidithiobacillia class members are aerobic, chemolithoautotrophic, acidophilic sulfur-oxidizers, encompassing substantial genotypic diversity attributable to MGEs. Our search for NAP protein families (PFs) in more than 90 genomes of the different species that conform the class, revealed the presence of 1,197 proteins pertaining to 12 different NAP families, with differential occurrence and conservation across species. Pangenome-level analysis revealed 6 core NAP PFs that were highly conserved across the class, some of which also existed as variant forms of scattered occurrence, in addition to NAPs of taxa-restricted distribution. Core NAPs identified are reckoned as essential based on the conservation of genomic context and phylogenetic signals. In turn, various highly diversified NAPs pertaining to the flexible gene complement of the class, were found to be encoded in known plasmids or, larger integrated MGEs or, present in genomic loci associated with MGE-hallmark genes, pointing to their role in the stabilization/maintenance of these elements in strains and species with larger genomes. Both core and flexible NAPs identified proved valuable as markers, the former accurately recapitulating the phylogeny of the class, and the later, as seed in the bioinformatic identification of novel episomal and integrated mobile elements. [ABSTRACT FROM AUTHOR]

Abstract Background The genus Acidithiobacillus has been widely concerned due to its superior survival and oxidation ability in acid mine drainage (AMD). However, the contribution of insertion sequence (IS) to their biological evolution and environmental adaptation is very limited. ISs are the simplest kinds of mobile genetic elements (MGEs), capable of interrupting genes, operons, or regulating the expression of genes through transposition activity. ISs could be classified into different families with their own members, possessing different copies. Results In this study, the distribution and evolution of ISs, as well as the functions of the genes around ISs in 36 Acidithiobacillus genomes, were analyzed. The results showed that 248 members belonging to 23 IS families with a total of 10,652 copies were identified within the target genomes. The IS families and copy numbers among each species were significantly different, indicating that the IS distribution of Acidithiobacillus were not even. A. ferrooxidans had 166 IS members, which may develop more gene transposition strategies compared with other Acidithiobacillus spp. What’s more, A. thiooxidans harbored the most IS copies, suggesting that their ISs were the most active and more likely to transpose. The ISs clustered in the phylogenetic tree approximately according to the family, which were mostly different from the evolutionary trends of their host genomes. Thus, it was suggested that the recent activity of ISs of Acidithiobacillus was not only determined by their genetic characteristics, but related with the environmental pressure. In addition, many ISs especially Tn3 and IS110 families were inserted around the regions whose functions were As/Hg/Cu/Co/Zn/Cd translocation and sulfur oxidation, implying that ISs could improve the adaptive capacities of Acidithiobacillus to the extremely acidic environment by enhancing their resistance to heavy metals and utilization of sulfur. Conclusions This study provided the genomic evidence for the contribution of IS to evolution and adaptation of Acidithiobacillus, opening novel sights into the genome plasticity of those acidophiles.

Background: The genus Acidithiobacillus has been widely concerned due to its superior survival and oxidation ability in acid mine drainage (AMD). However, the contribution of insertion sequence (IS) to their biological evolution and environmental adaptation is very limited. ISs are the simplest kinds of mobile genetic elements (MGEs), capable of interrupting genes, operons, or regulating the expression of genes through transposition activity. ISs could be classified into different families with their own members, possessing different copies. Results: In this study, the distribution and evolution of ISs, as well as the functions of the genes around ISs in 36 Acidithiobacillus genomes, were analyzed. The results showed that 248 members belonging to 23 IS families with a total of 10,652 copies were identified within the target genomes. The IS families and copy numbers among each species were significantly different, indicating that the IS distribution of Acidithiobacillus were not even. A. ferrooxidans had 166 IS members, which may develop more gene transposition strategies compared with other Acidithiobacillus spp. What's more, A. thiooxidans harbored the most IS copies, suggesting that their ISs were the most active and more likely to transpose. The ISs clustered in the phylogenetic tree approximately according to the family, which were mostly different from the evolutionary trends of their host genomes. Thus, it was suggested that the recent activity of ISs of Acidithiobacillus was not only determined by their genetic characteristics, but related with the environmental pressure. In addition, many ISs especially Tn3 and IS110 families were inserted around the regions whose functions were As/Hg/Cu/Co/Zn/Cd translocation and sulfur oxidation, implying that ISs could improve the adaptive capacities of Acidithiobacillus to the extremely acidic environment by enhancing their resistance to heavy metals and utilization of sulfur. Conclusions: This study provided the genomic evidence for the contribution of IS to evolution and adaptation of Acidithiobacillus, opening novel sights into the genome plasticity of those acidophiles. [ABSTRACT FROM AUTHOR]

Mobile genetic elements (MGEs) are relevant agents in bacterial adaptation and evolutionary diversification. Stable appropriation of these DNA elements depends on host factors, among which are the nucleoid-associated proteins (NAPs). NAPs are highly abundant proteins that bind and bend DNA, altering its topology and folding, thus affecting all known cellular DNA processes from replication to expression. Even though NAP coding genes are found in most prokaryotic genomes, their functions in host chromosome biology and xenogeneic silencing are only known for a few NAP families. Less is known about the occurrence, abundance, and roles of MGE-encoded NAPs in foreign elements establishment and mobility. In this study, we used a combination of comparative genomics and phylogenetic strategies to gain insights into the diversity, distribution, and functional roles of NAPs within the class Acidithiobacillia with a special focus on their role in MGE biology. Acidithiobacillia class members are aerobic, chemolithoautotrophic, acidophilic sulfur-oxidizers, encompassing substantial genotypic diversity attributable to MGEs. Our search for NAP protein families (PFs) in more than 90 genomes of the different species that conform the class, revealed the presence of 1,197 proteins pertaining to 12 different NAP families, with differential occurrence and conservation across species. Pangenome-level analysis revealed 6 core NAP PFs that were highly conserved across the class, some of which also existed as variant forms of scattered occurrence, in addition to NAPs of taxa-restricted distribution. Core NAPs identified are reckoned as essential based on the conservation of genomic context and phylogenetic signals. In turn, various highly diversified NAPs pertaining to the flexible gene complement of the class, were found to be encoded in known plasmids or, larger integrated MGEs or, present in genomic loci associated with MGE-hallmark genes, pointing to their role in the stabilization/maintenance of these elements in strains and species with larger genomes. Both core and flexible NAPs identified proved valuable as markers, the former accurately recapitulating the phylogeny of the class, and the later, as seed in the bioinformatic identification of novel episomal and integrated mobile elements.

Aiming at recovery of uranium (U) from low-grade uranium ore with a high fluorine content, column bioleaching of uranium extraction with an indigenous mesophilic consortium was studied. The performance of the bacterial community and the effect of chemical parameters on column bioleaching of uranium were investigated. The results showed that U extraction reached a high value of 95.69%, including acid preleaching (65.53%) and bioleaching (30.16%), in a 124-day period of the leaching process. The composition and diversity of the bacterial community were found to be significantly changing in the acid preleaching and bioleaching stages; nevertheless, they existed in residue and pregnant leaching solution samples for the same stage and showed little difference. Additionally, dominant genera in the solid samples, including Stenotrophomonas, Pseudomonas, Brevundimonas Clostridium_sensu_stricto, Bacillus, Clostridium XI, and Acidithiobacillus, and the Acidithiobacillus genus were ubiquitous throughout the leaching column, which was in favour of U extraction. [ABSTRACT FROM AUTHOR]

Banana crops require large amounts of fertilizers to maintain nutrients levels and achieve satisfactory yields. The use of biological fertilizers combined with organic waste is an efficient and sustainable option for agricultural production. Therefore, this study evaluated the use of a biological fertilizer combined with sewage sludge to make nutrients available in banana crops. We conducted a field experiment in a randomized block design, with four blocks and five repetitions per block in a subdivided plot scheme. The experiment lasted two years. We performed the soil and plant chemical analysis and we also assessed the plant biometric data. The lowest dose of biological fertilizer combined with sewage sludge increased P content in the soil in the first and second collections, but decreased in the third collection. The highest nutrient levels in the soil and plant were attained when biological fertilizer was applied with sewage sludge; however, high Ca content caused an imbalance of exchangeable bases in the soil solution. [ABSTRACT FROM AUTHOR]

Acidophilic bacteria and archaea inhabit extreme geochemical "islands" that can tell us when and how geographic barriers affect the biogeography of microorganisms. Here, we describe microbial communities from extremely acidic (pH 0 to 1) biofilms, known as snottites, from hydrogen sulfide-rich caves. Given the extreme acidity and subsurface location of these biofilms, and in light of earlier work showing strong geographic patterns among snottite Acidithiobacillus populations, we investigated their structure and diversity in order to understand how geography might impact community assembly. We used 16S rRNA gene cloning and fluorescence in situ hybridization (FISH) to investigate 26 snottite samples from four sulfidic caves in Italy and Mexico. All samples had very low biodiversity and were dominated by sulfur-oxidizing bacteria in the genus Acidithiobacillus. Ferroplasma and other archaea in the Thermoplasmatales ranged from 0 to 50% of total cells, and relatives of the bacterial genera Acidimicrobium and Ferrimicrobium were up to 15% of total cells. Rare phylotypes included Sulfobacillus spp. and members of the phyla "Candidatus Dependentiae" and "Candidatus Saccharibacteria" (formerly TM6 and TM7). Although the same genera of acidophiles occurred in snottites on separate continents, most members of those genera represent substantially divergent populations, with 16S rRNA genes that are only 95 to 98% similar. Our findings are consistent with a model of community assembly where sulfidic caves are stochastically colonized by microorganisms from local sources, which are strongly filtered through environmental selection for extreme acid tolerance, and these different colonization histories are maintained by dispersal restrictions within and among caves. [ABSTRACT FROM AUTHOR]

In the present study, the effect of additional carbon sources (carbon dioxide and molasses) on the bio-oxidation of a pyrite–arsenopyrite concentrate at temperatures of 40–50 °C was studied, and novel data regarding the patterns of the bio-oxidation of gold-bearing sulfide concentrates and the composition of the microbial populations performing these processes were obtained. At 40 °C, additional carbon sources did not affect the bio-oxidation efficiency. At the same time, the application of additional carbon dioxide improved the bio-oxidation performance at temperatures of 45 and 50 °C and made it possible to avoid the inhibition of bio-oxidation due to an increase in the temperature. Therefore, the use of additional carbon dioxide may be proposed to prevent the negative effect of an increase in temperature on the bio-oxidation of sulfide concentrates. 16S rRNA gene profiling revealed archaea of the family Thermoplasmataceae (Acidiplasma, Ferroplasma, Cuniculiplasma, and A-plasma group) and bacteria of the genera Leptospirillum, with Sulfobacillus and Acidithiobacillus among the dominant groups in the community. Temperature influenced the composition of the communities to a greater extent than the additional sources of carbon and the mode of operation of the bioreactor. Elevating the temperature from 40 °C to 50 °C resulted in increases in the shares of Acidiplasma and Sulfobacillus and decreases in the relative abundances of Ferroplasma, Leptospirillum, and Acidithiobacillus, while Cuniculiplasma and A-plasma were more abundant at 45 °C. A metagenomic analysis of the studied population made it possible to characterize novel archaea belonging to an uncultivated, poorly-studied group of Thermoplasmatales which potentially plays an important role in the bio-oxidation process. Based on an analysis of the complete genome, we propose describing the novel species and novel genus as “Candidatus Carboxiplasma ferriphilum” gen. nov., spec. nov.

La trehalosa es un tipo de carbohidrato, que en procariotas protege contra diferentes tipos de estrés y también se utiliza como fuente de almacenamiento de carbono. Hay cuatro formas diferentes de sintetizar trehalosa en Acidithiobacillus ferrivorans y dos en Acidithiobacillus ferrooxidans, pero su propósito sigue siendo desconocido. Este estudio tuvo como objetivo medir la producción de trehalosa en diferentes condiciones mediante su cuantificación en tres medios de cultivo a dos temperaturas diferentes. También se evaluó la cinética de crecimiento de ambas especies y se analizó la concentración de trehalosa durante la fase estacionaria temprana mediante un método enzimático. Los resultados mostraron que el medio 9K modificado con hierro ferroso a 28 °C tuvo la mayor producción de trehalosa, con A. ferrivorans CF27 con una mayor producción de 0.34 µmol/mg de proteína en comparación con A. ferrooxidans ATCC 23270 a 0.31 µmol/mg de proteína. Al utilizar CuS, la producción de trehalosa fue menor, con 0.02 y 0.03 µmol/mg de proteína para A. ferrivorans CF27 y A. ferrooxidans ATCC 23270, respectivamente, mientras que en presencia de zinc no se detectó trehalosa. A 15°C, el método enzimático no detectó trehalosa en los tres medios de cultivo, lo que indicaria que este carbohidrato no protege contra las bajas temperaturas en ninguna de las especies.

Bioleaching of metal sulfides is performed by diverse microorganisms. The dissolution of metal sulfides occurs via two chemical pathways, either the thiosulfate or the polysulfide pathway. These are determined by the metal sulfides' mineralogy and their acid solubility. The microbial cell enables metal sulfide dissolution via oxidation of iron(II) ions and inorganic sulfur compounds. Thereby, the metal sulfide attacking agents iron(III) ions and protons are generated. Cells are active either in a planktonic state or attached to the mineral surface, forming biofilms. This review, as an update of the previous one (Vera et al., 2013a), summarizes some recent discoveries relevant to bioleaching microorganisms, contributing to a better understanding of their lifestyle. These comprise phylogeny, chemical pathways, surface science, biochemistry of iron and sulfur metabolism, anaerobic metabolism, cell–cell communication, molecular biology, and biofilm lifestyle. Recent advances from genetic engineering applied to bioleaching microorganisms will allow in the future to better understand important aspects of their physiology, as well as to open new possibilities for synthetic biology applications of leaching microbial consortia. Key points: • Leaching of metal sulfides is strongly enhanced by microorganisms • Biofilm formation and extracellular polymer production influences bioleaching • Cell interactions in mixed bioleaching cultures are key for process optimization [ABSTRACT FROM AUTHOR]

The bioleaching process is carried out by aerobic acidophilic iron-oxidizing bacteria that are mainly mesophilic or moderately thermophilic. However, many mining sites are located in areas where the mean temperature is lower than the optimal growth temperature of these microorganisms. In this work, we report the obtaining and characterization of two psychrotolerant bioleaching bacterial strains from low-temperature sites that included an abandoned mine site in Chilean Patagonia (PG05) and an acid rock drainage in Marian Cove, King George Island in Antarctic (MC2.2). The PG05 and MC2.2 strains showed significant iron-oxidation activity and grew optimally at 20 °C. Genome sequence analyses showed chromosomes of 2.76 and 2.84 Mbp for PG05 and MC2.2, respectively, and an average nucleotide identity estimation indicated that both strains clustered with the acidophilic iron-oxidizing bacterium Acidithiobacillus ferrooxidans. The Patagonian PG05 strain had a high content of genes coding for tolerance to metals such as lead, zinc, and copper. Concordantly, electron microscopy revealed the intracellular presence of polyphosphate-like granules, likely involved in tolerance to metals and other stress conditions. The Antarctic MC2.2 strain showed a high dosage of genes for mercury resistance and low temperature adaptation. This report of cold-adapted cultures of the At. ferrooxidans species opens novel perspectives to satisfy the current challenges of the metal bioleaching industry. [ABSTRACT FROM AUTHOR]

Bioleaching of tannery sludge is an efficient and environmentally friendly way for chromium (Cr) removal, which supports the sustainable development of the leather industry. Acidithiobacillus thiooxidans has been reported effective in Cr bioleaching of tannery sludge. However, little is known about whether the presence of other benefiting species could further improve the Cr leaching efficiency of A. thiooxidans. Here, we studied the enhancing roles of four species namely Acidiphilium cryptum, Sulfobacillus acidophilus, Alicyclobacillus cycloheptanicus, and Rhodotorula mucilaginosa in chromium bioleaching of tannery sludge with A. thiooxidans by batch bioleaching experiments. We found that each of the four species facilitated the quick dominance of A. thiooxidans in the bioleaching process and significantly improved the bioleaching performance including bioleaching rate and efficiency. The bioleaching efficiency of Cr in the tannery sludge could reach 100% on the sixth day by co-inoculating A. thiooxidans and four auxiliary species. The achievements shed a light on the role of the community-level interactions on bioleaching and may also serve as guidance for managing bioleaching consortiums for better outcomes. [ABSTRACT FROM AUTHOR]

The bioleaching process is carried out by aerobic acidophilic iron-oxidizing bacteria that are mainly mesophilic or moderately thermophilic. However, many mining sites are located in areas where the mean temperature is lower than the optimal growth temperature of these microorganisms. In this work, we report the obtaining and characterization of two psychrotolerant bioleaching bacterial strains from low-temperature sites that included an abandoned mine site in Chilean Patagonia (PG05) and an acid rock drainage in Marian Cove, King George Island in Antarctic (MC2.2). The PG05 and MC2.2 strains showed significant iron-oxidation activity and grew optimally at 20°C. Genome sequence analyses showed chromosomes of 2.76 and 2.84 Mbp for PG05 and MC2.2, respectively, and an average nucleotide identity estimation indicated that both strains clustered with the acidophilic iron-oxidizing bacterium Acidithiobacillus ferrooxidans. The Patagonian PG05 strain had a high content of genes coding for tolerance to metals such as lead, zinc, and copper. Concordantly, electron microscopy revealed the intracellular presence of polyphosphate-like granules, likely involved in tolerance to metals and other stress conditions. The Antarctic MC2.2 strain showed a high dosage of genes for mercury resistance and low temperature adaptation. This report of cold-adapted cultures of the At. ferrooxidans species opens novel perspectives to satisfy the current challenges of the metal bioleaching industry.

Abstract Background Terrestrial hot spring settings span a broad spectrum of physicochemistries. Physicochemical parameters, such as pH and temperature, are key factors influencing differences in microbial composition across diverse geothermal areas. Nonetheless, analysis of hot spring pools from the Taupo Volcanic Zone (TVZ), New Zealand, revealed that some members of the bacterial genus, Acidithiobacillus, are prevalent across wide ranges of hot spring pHs and temperatures. To determine the genomic attributes of Acidithiobacillus that inhabit such diverse conditions, we assembled the genomes of 19 uncultivated hot spring Acidithiobacillus strains from six geothermal areas and compared these to 37 publicly available Acidithiobacillus genomes from various habitats. Results Analysis of 16S rRNA gene amplicons from 138 samples revealed that Acidithiobacillus comprised on average 11.4 ± 16.8% of hot spring prokaryotic communities, with three Acidithiobacillus amplicon sequence variants (ASVs) (TVZ_G1, TVZ_G2, TVZ_G3) accounting for > 90% of Acidithiobacillus in terms of relative abundance, and occurring in 126 out of 138 samples across wide ranges of temperature (17.5–92.9 °C) and pH (1.0–7.5). We recovered 19 environmental genomes belonging to each of these three ASVs, as well as a fourth related group (TVZ_G4). Based on genome average nucleotide identities, the four groups (TVZ_G1-TVZ_G4) constitute distinct species (ANI

The present work deals with the use of iron/sulfur-oxidizing bacterium Acidithiobacillus ferrooxidans for the desulfurization of a coal sample from Assam, India. The coal sample contains 3.87% w/w of organic sulfur, 1.4% w/w pyritic sulfur, and 1.29% w/w of sulfate sulfur which altogether are contributing to its total sulfur content of 6.56% w/w. The microbial process followed the oxidative-desulfurization mechanism for the removal of sulfur from the coal sample. The bacterium performed cleavage of the sulfide bonds in pyrite to release the inorganic sulfur. Mass spectroscopic analysis revealed that the bacterial strain A. ferrooxidans performed oxidative desulfurization of a model sulfur-containing organic compound dibenzothiophene to 2-hydroxy biphenyl through the formation of dioxy-dibenzothiophene. A. ferrooxidans cells played the catalytic role to release the sulfur present in organic and inorganic form in the coal sample. The microbial desulfurization process conducted in a shake flask scale (500 mL) removed about 79% of total sulfur from the coal sample in 14 days. [ABSTRACT FROM AUTHOR]

Acidithiobacillus ferrooxidans is an iron-oxidizing chemolithotroph used for bioleaching of precious metals and is also regarded as a potential host for bioelectrochemical production of value-added chemicals. Despite its industrial utility, however, it is difficult to genetically engineer A. ferrooxidans due to low transformation and recombination efficiencies. Here, we developed a clustered regularly interspaced short palindromic repeats interference (CRISPRi) system that can selectively repress the expression of a target gene in A. ferrooxidans. The mutated gene encoding a nuclease-deactivated Cas9 protein was cloned into the broad-host-range plasmid pBBR1-MCS2, and the applicability of the CRISPRi system was examined using the nitrogenase nifH gene as a knockdown target. Introduction of the CRISPRi plasmid into A. ferrooxidans resulted in decreased nifH transcription and retarded cell growth in the absence of nitrogen sources, demonstrating that the CRISPRi system altered the phenotype of this bacterium via selective gene knockdown. We suggest that the CRISPRi system developed in this study provides an efficient technique for constructing A. ferrooxidans knockdown mutants that are useful for the genetic dissection of this bacterium. [ABSTRACT FROM AUTHOR]

Using acidophilic bacteria to catalyze the reductive dissolution of oxidized minerals is an innovative process that facilitates the extraction of valuable base metals (principally cobalt and nickel) from limonites, which are otherwise often regarded as waste products of laterite mining. The most appropriate conditions required to optimize reductive mineral dissolution are unresolved, and the current work has reassessed the roles of Acidithiobacillus spp. in this process and identified novel facets. Aerobic bio-oxidation of zero-valent sulfur (ZVS) can generate sufficient acidity to counterbalance that consumed by the dissolution of oxidized iron and manganese minerals but precludes the development of low redox potentials that accelerate the reductive process, and although anaerobic oxidation of sulfur by iron-reducing species can achieve this, less acid is generated. Limited reduction of soluble iron (III) occurs in pure cultures of Acidithiobacillus spp. (Acidithiobacillus thiooxidans and Acidithiobacillus caldus) that do not grow by iron respiration. This phenomenon (“latent iron reduction”) was observed in aerated cultures and bioreactors and was independent of electron donor used (ZVS or hydrogen). Sufficient ferrous iron was generated in the presence of sterilized hydrophilic sulfur (bio-ZVS) to promote the effective reductive dissolution of Mn (IV) minerals in limonite and the solubilization of cobalt in the absence of viable acidophiles.

The copper-sensitive operon repressor (CsoR) family, which is the main Cu(I)-sensing family, is widely distributed and regulates regulons involved in detoxification in response to extreme copper stress (a general range of ≥3 g/liter copper ions). Here, we identified CsoR in hyper-copper-resistant Acidithiobacillus caldus (CsoRAc), an organism used in the bioleaching process of copper ores. CsoRAc possesses highly conserved Cu(I) ligands and structures within the CsoR family members. Transcriptional analysis assays indicated that the promoter (PIII) of csoR was active but weakly responsive to copper in Escherichia coli. Copper titration assays gave a stoichiometry of 0.8 mol Cu(I) per apo-CsoRAc monomer in vitro combined with atomic absorption spectroscopy analysis. CuI-CsoRAc and apo-CsoRAc share essentially identical secondary structures and assembly states, as demonstrated by circular dichroism spectra and size exclusion chromatography profiles. The average dissociation constants (KD = 2.26 × 10-18 M and 0.53 × 10-15 M) and Cu(I) binding affinity of apo-CsoRAc were estimated by bathocuproine disulfonate (BCS) and bicinchoninic acid (BCA) competition assays, respectively. Site-directed mutations of conserved Cu(I) ligands in CsoRAc did not significantly alter the secondary structure or assembly state. Competition assays showed that mutants had the same order of magnitude of Cu(I) binding affinity as apo-CsoRAc. Moreover, apo-CsoRAc could bind to the DNA fragment P08430 in vitro, although with low affinity. Finally, a working model was developed to illustrate putative copper resistance mechanisms in A. caldus. [ABSTRACT FROM AUTHOR]

Using acidophilic bacteria to catalyze the reductive dissolution of oxidized minerals is an innovative process that facilitates the extraction of valuable base metals (principally cobalt and nickel) from limonites, which are otherwise often regarded as waste products of laterite mining. The most appropriate conditions required to optimize reductive mineral dissolution are unresolved, and the current work has reassessed the roles of Acidithiobacillus spp. in this process and identified novel facets. Aerobic bio-oxidation of zero-valent sulfur (ZVS) can generate sufficient acidity to counterbalance that consumed by the dissolution of oxidized iron and manganese minerals but precludes the development of low redox potentials that accelerate the reductive process, and although anaerobic oxidation of sulfur by iron-reducing species can achieve this, less acid is generated. Limited reduction of soluble iron (III) occurs in pure cultures of Acidithiobacillus spp. (Acidithiobacillus thiooxidans and Acidithiobacillus caldus) that do not grow by iron respiration. This phenomenon ("latent iron reduction") was observed in aerated cultures and bioreactors and was independent of electron donor used (ZVS or hydrogen). Sufficient ferrous iron was generated in the presence of sterilized hydrophilic sulfur (bio-ZVS) to promote the effective reductive dissolution of Mn (IV) minerals in limonite and the solubilization of cobalt in the absence of viable acidophiles. [ABSTRACT FROM AUTHOR]

Experiments were carried out to examine the oxido-reduction of manganese by extremely acidophilic Acidithiobacillus spp. grown with either elemental sulfur or molecular hydrogen as electron donor. While there was no evidence for manganese (II) oxidation, dissolution of solid phase manganese dioxide was observed in cultures grown aerobically on both electron donors, though this appeared not to involve reduction of the metal. Solubilization of MnO2 was much enhanced in cultures incubated anaerobically, even though biomass was smaller, and pH values increased significantly as a consequence of acid dissolution being accompanied by manganese (IV) reduction. Increases in cell numbers correlated with concentrations of soluble manganese in anaerobic cultures grown on hydrogen, demonstrating that iron-oxidizing/reducing Acidithiobacillus spp. can grow in the absence of oxygen using manganese (IV) as sole electron acceptor. Addition of ferric iron to anaerobic cultures further enhanced the reductive dissolution of MnO2 as a result of its reduction to ferrous iron which then reacted with the solid manganese phase, and confirming that Mn (IV) reduction by iron-reducing acidophiles can proceed both directly and indirectly, involving iron as a shuttle vector. The implications of these findings to developing technologies for bio-processing oxidized metal ore deposits are discussed. [ABSTRACT FROM AUTHOR]

Abstract Background Habitats colonized by acidophiles as an ideal physical barrier may induce genetic exchange of microbial members within the common communities, but little is known about how species in extremely acidic environments diverge and evolve. Results Using the acidophilic sulfur-oxidizer Acidithiobacillus as a case study, taxonomic reclassifications of many isolates provides novel insights into their phylogenetic lineage. Whole-genome-based comparisons were attempted to investigate the intra- and inter-species divergence. Recent studies clarified that functional and structural specificities of bacterial strains might provide opportunities for adaptive evolution responding to local environmental conditions. Acidophilic microorganisms play a key role in the acidification of natural waters and thus the formation of extremely acidic environments, and the feedbacks of the latter might confer the distinct evolutionary patterns of Acidithiobacillus spp. Varied horizontal gene transfer events occurred in different bacterial strains, probably resulting in the expansion of Acidithiobacillus genomes. Gene loss as another evolutionary force might cause the adaptive phenotypic diversity. A conceptual model for potential community-dependent evolutionary adaptation was thus proposed to illustrate the observed genome differentiation. Conclusions Collectively, the findings shed light on the phylogeny and divergent evolution of Acidithiobacillus strains, and provided a useful reference for evolutionary studies of other extremophiles.

Background: Terrestrial hot spring settings span a broad spectrum of physicochemistries. Physicochemical parameters, such as pH and temperature, are key factors influencing differences in microbial composition across diverse geothermal areas. Nonetheless, analysis of hot spring pools from the Taupo Volcanic Zone (TVZ), New Zealand, revealed that some members of the bacterial genus, Acidithiobacillus, are prevalent across wide ranges of hot spring pHs and temperatures. To determine the genomic attributes of Acidithiobacillus that inhabit such diverse conditions, we assembled the genomes of 19 uncultivated hot spring Acidithiobacillus strains from six geothermal areas and compared these to 37 publicly available Acidithiobacillus genomes from various habitats. Results: Analysis of 16S rRNA gene amplicons from 138 samples revealed that Acidithiobacillus comprised on average 11.4 ± 16.8% of hot spring prokaryotic communities, with three Acidithiobacillus amplicon sequence variants (ASVs) (TVZ_G1, TVZ_G2, TVZ_G3) accounting for > 90% of Acidithiobacillus in terms of relative abundance, and occurring in 126 out of 138 samples across wide ranges of temperature (17.5–92.9 °C) and pH (1.0–7.5). We recovered 19 environmental genomes belonging to each of these three ASVs, as well as a fourth related group (TVZ_G4). Based on genome average nucleotide identities, the four groups (TVZ_G1-TVZ_G4) constitute distinct species (ANI < 96.5%) of which three are novel Acidithiobacillus species (TVZ_G2-TVZ_G4) and one belongs to Acidithiobacillus caldus (TVZ_G1). All four TVZ Acidithiobacillus groups were found in hot springs with temperatures above the previously known limit for the genus (up to 40 °C higher), likely due to significantly higher proline and GC contents than other Acidithiobacillus species, which are known to increase thermostability. Results also indicate hot spring-associated Acidithiobacillus have undergone genome streamlining, likely due to thermal adaptation. Moreover, our data suggest that Acidithiobacillus prevalence across varied hot spring pHs is supported by distinct strategies, whereby TVZ_G2-TVZ_G4 regulate pH homeostasis mostly through Na+/H+ antiporters and proton-efflux ATPases, whereas TVZ_G1 mainly relies on amino acid decarboxylases. Conclusions: This study provides insights into the distribution of Acidithiobacillus species across diverse hot spring physichochemistries and determines genomic features and adaptations that potentially enable Acidithiobacillus species to colonize a broad range of temperatures and pHs in geothermal environments. BoTSrZ5f8ypScVzkmXt8zF Video Abstract [ABSTRACT FROM AUTHOR]

BackgroundAcidithiobacillus caldus is a sulfur oxidizing extreme acidophile and the only known mesothermophile within the Acidithiobacillales. As such, it is one of the preferred microbes for mineral bioprocessing at moderately high temperatures. In this study, we explore the genomic diversity of A. caldus strains using a combination of bioinformatic and experimental techniques, thus contributing first insights into the elucidation of the species pangenome.Principal findingsComparative sequence analysis of A. caldus ATCC 51756 and SM-1 indicate that, despite sharing a conserved and highly syntenic genomic core, both strains have unique gene complements encompassing nearly 20% of their respective genomes. The differential gene complement of each strain is distributed between the chromosomal compartment, one megaplasmid and a variable number of smaller plasmids, and is directly associated to a diverse pool of mobile genetic elements (MGE). These include integrative conjugative and mobilizable elements, genomic islands and insertion sequences. Some of the accessory functions associated to these MGEs have been linked previously to the flexible gene pool in microorganisms inhabiting completely different econiches. Yet, others had not been unambiguously mapped to the flexible gene pool prior to this report and clearly reflect strain-specific adaption to local environmental conditions.SignificanceFor many years, and because of DNA instability at low pH and recurrent failure to genetically transform acidophilic bacteria, gene transfer in acidic environments was considered negligible. Findings presented herein imply that a more or less conserved pool of actively excising MGEs occurs in the A. caldus population and point to a greater frequency of gene exchange in this econiche than previously recognized. Also, the data suggest that these elements endow the species with capacities to withstand the diverse abiotic and biotic stresses of natural environments, in particular those associated with its extreme econiche.

Microorganisms of the genus Thiobacillus extreme acidophilus play an important role in biometallurgy, biological desulfurization and solid waste disposal, but the cell concentration of this type of microorganisms is very low during the culture process, which limits the wide application of this type of microorganisms. High-density cultivation is an effective means to improve the efficiency of microbial production. The application of high-density culture technology in acidophilic microorganisms can significantly reduce the production cost of microbial culture, shorten the production cycle, and the output rate of the microbial inoculants of Thiobacillus extreme acidophilus. This article summarizes the research status of high-density culture of Thiobacillus extreme acidophilus from strain selection, culture conditions and culture methods. [ABSTRACT FROM AUTHOR]

Ores serve as energy and nutrient sources for microorganisms. Through complex biochemical processes, microorganisms disrupt the surface structure of ores and release metal elements. However, there is limited research on the mechanisms by which bacteria with different nutritional modes act during the leaching process of different crystal structure ores. This study evaluated the leaching efficiency of two types of bacteria with different nutritional modes, heterotrophic bacterium Bacillus mucilaginosus (BM) and autotrophic bacterium Acidithiobacillus ferrooxidans (AF), on different crystal structure lithium silicate ores (chain spodumene, layered lepidolite and ring elbaite). The aim was to understand the behavioral differences and decomposition mechanisms of bacteria with different nutritional modes in the process of breaking down distorted crystal lattices of ores. The results revealed that heterotrophic bacterium BM primarily relied on passive processes such as bacterial adsorption, organic acid corrosion, and the complexation of small organic acids and large molecular polymers with metal ions. Autotrophic bacterium AF, in addition to exhibiting stronger passive processes such as organic acid corrosion and complexation, also utilized an active transfer process on the cell surface to oxidize Fe 2+ in the ores for energy maintenance and intensified the destruction of ore lattices. As a result, strain AF exhibited a greater leaching effect on the ores compared to strain BM. Regarding the three crystal structure ores, their different stacking modes and proportions of elements led to significant differences in structural stability, with the leaching effect being highest for layered structure, followed by chain structure, and then ring structure. These findings indicate that bacteria with different nutritional modes exhibit distinct physiological behaviors related to their nutritional and energy requirements, ultimately resulting in different sequences and mechanisms of metal ion release from ores after lattice damage., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Mineral processing encounters the challenge of separating chalcopyrite and pyrite, with the conventional high alkali process characterized by issues such as large dosages of reagents, complex procedures, and environmental pollution. This study addresses this challenge by isolating and enriching Thiobacillus ferrooxidans (T·f) from acidic mine drainage, employing it as a biosurfactant. The modification mechanism of T·f was thoroughly analyzed. Fe dissolution through biological oxidation formed a passivation layer (jarosite [KFe 3 (SO 4 ) 2 (OH) 6 ], elemental sulfur (S 0 ), and metal sulfides (Cu/Fe-S) on the surface of minerals. Metal oxides, hydroxides, and sulfates were detected on the surface of two minerals, but the difference was that elemental sulfur (S 0 ) and copper sulfide (Cu-S) were detected on the surface of chalcopyrite. elucidating the fundamental reason for the significant difference in surface hydrophobicity between chalcopyrite and pyrite. T·f has been successfully used as a biosurfactant to achieve copper-sulfur separation., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Strain S30A2 T , isolated from the acid mine drainage sediment of Mengzi Copper Mine, Yunnan, is proposed to represent a novel species of the sulphur-oxidizing genus Acidithiobacillus . Cells were Gram-stain-negative, non-endospore forming, highly motile with one or two monopolar flagella and rod-shaped. The strain was mesophilic, growing at 30-50 °C (optimum, 38 °C), acidophilic, growing at pH 2.0-4.5 (optimum, pH 2.5), and tolerant of 0-4 % (w/v; 684 mol l -1 ) NaCl. The 16S rRNA gene-based sequence analysis showed that strain S30A2 T belongs to the genus Acidithiobacillus and shows the largest similarity of 96.6 % to the type strain Acidithiobacillus caldus KU T . The genomic DNA G+C content of strain S30A2 T was 59.25 mol%. The average nucleotide identity ANIb and ANIm values between strain S30A2 T and A. caldus KU T were 70.95 and 89.78 %, respectively and the digital DNA-DNA hybridization value was 24.9 %. Strain S30A2 T was strictly aerobic and could utilize elementary sulphur and tetrathionate to support chemolithotrophic growth. The major cellular fatty acid of S30A2 T was C 19 : 1 ω7 c . The respiratory quinones were ubiquinone-8 and ubiquinone-7. Based upon its phylogenetic, genetic, phenotypic, physiologic and chemotaxonomic characteristics, strain S30A2 T is considered to represent a novel species of the genus Acidithiobacillus , for which the name Acidithiobacillus acidisediminis sp. nov. is proposed. The type strain is S30A2 T (=CGMCC 1.17059 T =KCTC 72580 T ).

In this study, Acidithiobacillus thiooxidans was used for the bioleaching of copper (Cu) from sewage sludge. In order to find optimization conditions, three factors including solid-to-liquid ratio (S/L) (0.01-0.2 %(w/v)), initial element sulfur (S 0 ) (1-10 g/L), and initial pH (1-3) have been investigated. Based on response surface methodology (RSM) determined a significant reduced quadratic model with a p-value of 0.0022 (<0.05 significant level). The maximum Cu recovery was 85.3% in the optimum condition of S/L = 0.16% (w/v), S 0  = 8.2 g/L, and pH = 1.4. Furthermore, a kinetic study based on a shrinking core model was performed and the result showed that chemical reaction was rate limiting in the extraction. Toxicity Characteristic Leaching Procedure (TCLP) results after bioleaching showed the bioleaching process detoxified sludge and the bioleached sludge residue was well within the regulatory limits for disposal. The germination seed with adding bioleached and unbioleached sludge to the soil was determined. Various parameters such as Germination Index (GI), Tolerance Index (TI), Vigor Index (VI), and stem length showed that the sewage sludge indices significantly increased than the sample soil with unbioleached sludge., Competing Interests: Declaration of competing interest Authors confirm that this manuscript has not been published elsewhere and is not under consideration by another journal for publication. All authors have approved the manuscript and agreed with its submission to this journal. The authors have no conflicts of interest to declare., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Hydrogen can serve as an electron donor for chemolithotrophic acidophiles, especially in the deep terrestrial subsurface and geothermal ecosystems. Nevertheless, the current knowledge of hydrogen utilization by mesophilic acidophiles is minimal. A multi-omics analysis was applied on Acidithiobacillus ferrooxidans growing on hydrogen, and a respiratory model was proposed. In the model, [NiFe] hydrogenases oxidize hydrogen to two protons and two electrons. The electrons are used to reduce membrane-soluble ubiquinone to ubiquinol. Genetically associated iron-sulfur proteins mediate electron relay from the hydrogenases to the ubiquinone pool. Under aerobic conditions, reduced ubiquinol transfers electrons to either cytochrome aa3 oxidase via cytochrome bc1 complex and cytochrome c4 or the alternate directly to cytochrome bd oxidase, resulting in proton efflux and reduction of oxygen. Under anaerobic conditions, reduced ubiquinol transfers electrons to outer membrane cytochrome c (ferrireductase) via cytochrome bc1 complex and a cascade of electron transporters (cytochrome c4, cytochrome c552, rusticyanin, and high potential iron-sulfur protein), resulting in proton efflux and reduction of ferric iron. The proton gradient generated by hydrogen oxidation maintains the membrane potential and allows the generation of ATP and NADH. These results further clarify the role of extremophiles in biogeochemical processes and their impact on the composition of the deep terrestrial subsurface.

A multitude of microbes are involved in the solubilisation of minerals and metals as this approach offers numerous advantages over traditional methods. This strategy is preferred as it is eco-friendly and economical, thus overcoming the drawbacks of the traditional approach of pyrometallurgy. Many different types of bacteria are employed in the process of Bioleaching, which are collectively grouped under chemolithotrophs, as they derive their energy from inorganic compounds. Bioleaching is the mobilization of metal cations from insoluble ores by microorganisms. All chemolithotropic bacteria are extremophiles since they have the ability to survive in extreme conditions. They carry out the process of Bioleaching through three mechanisms: Indirect, contact/ direct and cooperative bioleaching. This review gives a sneak peek into the different strains of chemolithotrophs which are used in bioleaching, and some recent work in the field. It also gives an insight into the general process and mechanism of Bioleaching, the study of which will pave way for developing better and efficient industrial bioleaching operations.

Hydrogen can serve as an electron donor for chemolithotrophic acidophiles, especially in the deep terrestrial subsurface and geothermal ecosystems. Nevertheless, the current knowledge of hydrogen utilization by mesophilic acidophiles is minimal. A multi-omics analysis was applied on Acidithiobacillus ferrooxidans growing on hydrogen, and a respiratory model was proposed. In the model, [NiFe] hydrogenases oxidize hydrogen to two protons and two electrons. The electrons are used to reduce membrane-soluble ubiquinone to ubiquinol. Genetically associated iron-sulfur proteins mediate electron relay from the hydrogenases to the ubiquinone pool. Under aerobic conditions, reduced ubiquinol transfers electrons to either cytochrome aa 3 oxidase via cytochrome bc 1 complex and cytochrome c 4 or the alternate directly to cytochrome bd oxidase, resulting in proton efflux and reduction of oxygen. Under anaerobic conditions, reduced ubiquinol transfers electrons to outer membrane cytochrome c (ferrireductase) via cytochrome bc 1 complex and a cascade of electron transporters (cytochrome c 4, cytochrome c 552, rusticyanin, and high potential iron-sulfur protein), resulting in proton efflux and reduction of ferric iron. The proton gradient generated by hydrogen oxidation maintains the membrane potential and allows the generation of ATP and NADH. These results further clarify the role of extremophiles in biogeochemical processes and their impact on the composition of the deep terrestrial subsurface. [ABSTRACT FROM AUTHOR]

Sulfur-oxidizing bacteria that are halophilic and acidophilic have gained interest because of their potential use in bioleaching operations in salt-containing environments. Acidithiobacillus sp. strain SH, which was previously identified as Acidithiobacillus thiooxidans based on its 16S rRNA gene sequence, is a chemolithoautotrophic marine bacterium exhibiting sodium chloride-stimulated thiosulfate-oxidizing activities. A novel thiosulfate:quinone oxidoreductase from strain SH (SH-TQO) has been purified from its solubilized membrane fraction. The gene for SH-TQO was determined from the draft genome sequence of the strain SH. Amino acid sequences of peptides generated by the in-gel trypsin digestion of SH-TQO were found in a protein encoded by locus tag B1757_09800 of the genome of the strain SH. The gene encoded 444 amino acids with a signal peptide of 29 amino acids and was annotated to encode a porin. The gene was located in a unique genomic region, not found in A. thiooxidans strains, suggesting that the strain SH acquired this region through a horizontal gene transfer. A protein–protein basic local alignment search revealed that sulfur-oxidizing bacteria, such as Acidithiobacillus species have proteins homologous to SH-TQO, though the degree of homologies was relatively low. The protein, DoxXA, which is homologous to TQO from Acidianus amvibalens , was also found in the genomic region. [ABSTRACT FROM AUTHOR]

The goal of the present work was to study the oxidation of sulfide mineral pyrrhotite by members of microbial groups predominant in biohydrometallurgical processes (mixotrophic iron- and sulfur-oxidizing bacterium Sulfobacillus thermosulfidooxidans strain VKMV 1269T, autotrophic sulfur-oxidizing bacterium Acidithiobacillus caldus strain MBC-1, and heterotrophic iron- and sulfur-oxidizing archaeon Acidiplasma sp. strain MBA-1) to evaluate the role of microorganisms with different physiological properties in the leaching of this mineral. It was shown that pyrrhotite was most actively leached by the sulfur-oxidizing strain Acidithiobacillus caldus MBC-1, as well as by mixed cultures in which this strain was present: S. thermosulfidooxidans VKMV 1269T + A. caldus MBC-1 and Acidiplasma sp. MBA-1 + A. caldus MBC-1. These cultures leached 70, 43, and 60% of pyrrhotite, respectively, after 30 days of the experiment. In experiments with pure cultures S. thermosulfidooxidans VKMV 1269T, Acidiplasma sp. MBA-1 and a mixed culture S. thermosulfidooxidans VKMV 1269T + Acidiplasma sp. MBA-1, the rates of pyrrhotite leaching were significantly lower: 29, 37 and 30%, respectively. An experiment on sulfur biooxidation by pure cultures of the studied microorganisms demonstrated that A. caldus strain MBC-1 was the most active sulfur oxidizer. Thus, it was shown that the bioleaching of pyrrhotite was carried out mainly by sulfur-oxidizing microorganisms, with sulfur biooxidation as the most important process. Active biooxidation of the Fe2+ ion by iron-oxidizing microorganisms did not increase activity of pyrrhotite oxidation, but, on the contrary, led to inhibition of pyrrhotite leaching. Thus, active biooxidation of sulfur, which is formed during the leaching of pyrrhotite, was a factor determining the leaching rate for this sulfide mineral. [ABSTRACT FROM AUTHOR]

Ferrous iron- and sulfur-oxidizing Acidihalobacter species and similar so far unclassified bacteria have been isolated from the islands of Vulcano (Italy) and Milos (Greece), specifically from where seawater was acidified at sulfide-rich geothermal sites. Acidithiobacillus species which tolerated concentrations of chloride that inhibit most Acidithiobacillus spp. were also isolated from sites on both islands: these were At. thiooxidans strains and an unclassified species, Acidithiobacillus sp. strain V1. The potential of salt-tolerant acidophiles for industrial application in promoting copper extraction from mineral sulfides where chloride is naturally present at concentrations which would inhibit most acidophiles, or where seawater rather than fresh water is available, appears to be limited by the sensitivity of ferrous-iron oxidizing Acidihalobacter spp. to copper. However, tolerance of copper and chloride shown by At. thiooxidans strain A7 suggests it could oxidize sulfur and benefit acid leaching if ferric iron or copper was provided as the primary oxidant of sulfide ores. [ABSTRACT FROM AUTHOR]

Acidophiles play a dominant role in driving elemental cycling in natural acid mine drainage (AMD) habitats and exhibit important application value in bioleaching and bioremediation. Acidity is an inevitable environmental stress and a key factor that affects the survival of acidophiles in their acidified natural habitats; however, the regulatory strategies applied by acidophilic bacteria to withstand low pH are unclear. We identified the significance of the ferric uptake regulator (Fur) in acidophiles adapting to acidic environments and discovered that Fur is ubiquitous as well as highly conserved in acidophilic bacteria. Mutagenesis of the fur gene of Acidithiobacillus caldus, a prototypical acidophilic sulfur-oxidizing bacterium found in AMD, revealed that Fur is required for the acid resistance of this acidophilic bacterium. Phenotypic characterization, transcriptome sequencing (RNA-seq), mutagenesis, and biochemical assays indicated that the Acidithiobacillus caldus ferric uptake regulator (AcFur) is involved in extreme acid resistance by regulating the expression of several key genes of certain cellular activities, such as iron transport, biofilm formation, sulfur metabolism, chemotaxis, and flagellar biosynthesis. Finally, a Furdependent acid resistance regulatory strategy in A. caldus was proposed to illustrate the ecological behavior of acidophilic bacteria under low pH. This study provides new insights into the adaptation strategies of acidophiles to AMD ecosystems and will promote the design and development of engineered biological systems for the environmental adaptation of acidophiles. IMPORTANCE This study advances our understanding of the acid tolerance mechanism of A. caldus, identifies the key fur gene responsible for acid resistance, and elucidates the correlation between fur and acid resistance, thus contributing to an understanding of the ecological behavior of acidophilic bacteria. These findings provide new insights into the acid resistance process in Acidithiobacillus species, thereby promoting the study of the environmental adaptation of acidophilic bacteria and the design of engineered biological systems. [ABSTRACT FROM AUTHOR]