RESUMO

Sulfate-reducing bioreactors are a biotechnological alternative for the treatment of acid mine drainage (AMD). In this study, two separate bioreactors with pH and temperature-controlled (Bio I and II) were operated with two different acidophilic microbial consortia to determine their efficiencies in sulfate removal from a synthetic acidic mine water. The bioreactors were operated for 302 days in continuous flow mode under the same parameters: fed with a sulfate solution of ∼30 mM with a pH of 2.5, the temperature at 30°C, stirred gently at 40 rpm and using a continuous stream of nitrogen to help remove the H2S produced in the bioreactor. The glycerol consumption, acetate production, and sulfate removal were monitored throughout the course of the experiment. The community composition and potential metabolic functional groups were analyzed via 16S rRNA partial gene sequencing. Bio I consortium reduced the sulfate, achieving a range of sulfate concentration from 4.7 to 19 mM in the effluent liquor. The removal of sulfate in Bio II was between 5.6 and 18 mM. Both bioreactors' communities showed the presence of the genus De sulfosporosinus as the main sulfate-reducing bacteria (SRB). Despite differences in microbial composition, both bioreactors have similar potential metabolism, with a higher percentage of microorganisms that can use sulfate in respiration. Overall, both bioreactors showed similar performance in treating acidic mine water containing mostly sulfate using two different acidophilic sulfidogenic consortia obtained from different global locations.

RESUMO

In this study, bioleaching was carried out for the recovery of metals (copper, zinc, tin, lead, gold and silver) from printed circuit boards residues (PCBs), one of the most important wastes from electrical and electronic equipment, using an acidophilic iron-oxidizing bacterial consortium enriched with minerals from a gold mine in the Arequipa region, Peru. High-throughput sequencing and analysis of the 16S rRNA biomarker revealed that this consortium was predominantly composed of Tissierella, Acidiphilium and Leptospirillum bacteria, from which the latter is known to grow by chemolithotrophy through iron oxidation. After the enrichment process, the acidophilic iron-oxidizing consortium was first tested for its tolerance to different PCBs concentrations, showing best growth up to 10 g/L of PCBs and a tolerance index of 0.383. Based on these results, the bioleaching efficiency of the consortium was investigated for 10 g/L of PCBs in stirred tank reactors coupled to an aeration system, for 18 days. High bioleaching efficiencies were achieved for copper and zinc (69% and 91%, respectively), indicating that these two metals can be easily extracted in this leaching system. Lower extraction efficiencies were achieved for tin (16%) and gold (28%), while for lead and silver only a residual recovery (<0.25%) was detected. These results indicate that the enriched bacterial consortium originating from the Arequipa region, Peru, has a high capacity to recover different metals of economic importance.

RESUMO

In the actual mining scenario, copper bioleaching, mainly raw mined material known as run-of-mine (ROM) copper bioleaching, is the best alternative for the treatment of marginal resources that are not currently considered part of the profitable reserves because of the cost associated with leading technologies in copper extraction. It is foreseen that bioleaching will play a complementary role in either concentration-as it does in Minera Escondida Ltd. (MEL)-or chloride main leaching plants. In that way, it will be possible to maximize mines with installed solvent-extraction and electrowinning capacities that have not been operative since the depletion of their oxide ores. One of the main obstacles for widening bioleaching technology applications is the lack of knowledge about the key events and the attributes of the technology's critical events at the industrial level and mainly in ROM copper bioleaching industrial operations. It is relevant to assess the bed environment where the bacteria-mineral interaction occurs to learn about the limiting factors determining the leaching rate. Thus, due to inability to accurately determine in-situ key variables, their indirect assessment was evaluated by quantifying microbial metabolic-associated responses. Several candidate marker genes were selected to represent the predominant components of the microbial community inhabiting the industrial heap and the metabolisms involved in microbial responses to changes in the heap environment that affect the process performance. The microbial community's predominant components were Acidithiobacillus ferrooxidans, At. thiooxidans, Leptospirillum ferriphilum, and Sulfobacillus sp. Oxygen reduction, CO2 and N2 fixation/uptake, iron and sulfur oxidation, and response to osmotic stress were the metabolisms selected regarding research results previously reported in the system. After that, qPCR primers for each candidate gene were designed and validated. The expression profile of the selected genes vs. environmental key variables in pure cultures, column-leaching tests, and the industrial bioleaching heap was defined. We presented the results obtained from the industrial validation of the marker genes selected for assessing CO2 and N2 availability, osmotic stress response, as well as ferrous iron and sulfur oxidation activity in the bioleaching heap process of MEL. We demonstrated that molecular markers are useful for assessing limiting factors like nutrients and air supply, and the impact of the quality of recycled solutions. We also learned about the attributes of variables like CO2, ammonium, and sulfate levels that affect the industrial ROM-scale operation.

Assuntos

Acidithiobacillus/metabolismo , Ácidos/metabolismo , Bactérias/metabolismo , Biomarcadores/metabolismo , Cobre/metabolismo , Microbiologia Industrial/métodos , Laboratórios/normas , Acidithiobacillus/crescimento & desenvolvimento , Acidithiobacillus/isolamento & purificação , Bactérias/crescimento & desenvolvimento , Bactérias/isolamento & purificação , Biodiversidade , Cobre/isolamento & purificação

RESUMO

The Amarillo River in La Rioja, Argentina, is a natural acidic environment that is influenced by an abandoned mine. The river is characterized by extremely low pH and high concentrations of metals and metalloids. Fe(III)-bearing neoformed precipitated minerals are widespread along the hydrological basin. This work reports the presence of different species of iron-oxidizing bacteria and demonstrates that their action has a significant role in geochemical processes of the Amarillo River, mainly by catalyzing Fe2+ oxidation and intensifying the Fe(III)-bearing mineral precipitation. Various iron oxidizers (i.e. Acidithiobacillus ferrivorans, Leptospirillum ferrooxidans, Ferrimicrobium acidophilum, Alicyclobacillus cycloheptanicus) were detected in enrichment cultures at different temperatures. Moreover, this is the first report confirming that Acidithiobacillus ferrivorans is able to grow at 4 °C. Other acidophilic bacteria (i.e., Acidiphilium iwatensii) and fungi (e.g., Fodinomyces uranophilus, Coniochaeta fodinicola, Acidea extrema, Penicillium sp. and Cladosporium pseudocladosporioides) were also detected. In vitro laboratory studies recreating natural Fe(III)-bearing mineral formation showed that mineral precipitation rate was higher than 350 mg L-1 day-1 in the presence of microorganisms whereas it was about 15 mg L-1 day-1 under abiotic conditions. Jarosite was the only mineral detected in the precipitates generated by microbial action and it was also identified in the Amarillo River bed sediments. Biological Fe2+ oxidation rates depend on temperature which range from 8 to 32 mM day-1 at 4 and 30 °C, respectively. Finally, a conceptual model recognizing the significant microbial role is proposed to gain a better understanding of the biogeochemistry dynamics of the Amarillo River.

Assuntos

Compostos Férricos , Rios , Acidithiobacillus , Alicyclobacillus , Argentina , Ascomicetos , Bactérias , Cladosporium , Concentração de Íons de Hidrogênio , Oxirredução

RESUMO

The geothermal Copahue-Caviahue (GCC) system (Argentina) is an extreme acidic environment, dominated by the activity of Copahue volcano. Environments characterised by low pH values, such as volcanic areas, are of particular interest for the search of acidophilic microorganisms with application in biotechnological processes. In this work, sulfate-reducing microorganisms were investigated in geothermal acidic, anaerobic zones from GCC system. Sediment samples from Agua del Limón (AL1), Las Máquinas (LMa2), Las Maquinitas (LMi) and Baño 9 (B9-2, B9-3) were found to be acidic (pH values 2.1-3.0) to moderate acidic (5.1-5.2), containing small total organic carbon values, and ferric iron precipitates. The organic electron donor added to the enrichment was completely oxidised to CO2. Bacteria related to 'Desulfobacillus acidavidus' strain CL4 were found to be dominant (67-83% of the total number of clones) in the enrichment cultures, and their presence was confirmed by their isolation on overlay plates. Other bacteria were also detected with lower abundance (6-20% of the total number of clones), with representatives of the genera Acidithiobacillus, Sulfobacillus, Alicyclobacillus and Athalassotoga/Mesoaciditoga. These enrichment and isolates found at low pH confirm the presence of anaerobic activities in the acidic sediments from the geothermal Copahue-Caviahue system.

Assuntos

Bactérias/classificação , Bactérias/isolamento & purificação , Sedimentos Geológicos/microbiologia , Fontes Termais/microbiologia , Ácidos , Argentina , Bactérias/genética , Meio Ambiente , Concentração de Íons de Hidrogênio , Ferro/metabolismo , Oxirredução , Filogenia , RNA Ribossômico 16S/genética , Sulfatos/metabolismo

RESUMO

Horizontal gene transfer is crucial for the adaptation of microorganisms to environmental cues. The acidophilic, bioleaching bacterium Acidithiobacillus ferrooxidans encodes an integrative-conjugative genetic element (ICEAfe1) inserted in the gene encoding a tRNAAla. This genetic element is actively excised from the chromosome upon induction of DNA damage. A similar genetic element (ICEAcaTY.2) is also found in an equivalent position in the genome of Acidithiobacillus caldus. The local genomic context of both mobile genetic elements is highly syntenous and the cognate integrases are well conserved. By means of site directed mutagenesis, target site deletions and in vivo integrations assays in the heterologous model Escherichia coli, we assessed the target sequence requirements for site-specific recombination to be catalyzed by these integrases. We determined that each enzyme recognizes a specific small DNA segment encoding the anticodon stem/loop of the tRNA as target site and that specific positions in these regions are well conserved in the target attB sites of orthologous integrases. Also, we demonstrate that the local genetic context of the target sequence is not relevant for the integration to take place. These findings shed new light on the mechanism of site-specific integration of integrative-conjugative elements in members of Acidithiobacillus genus.

Assuntos

Acidithiobacillus/genética , Elementos de DNA Transponíveis , DNA Bacteriano/genética , Transferência Genética Horizontal , RNA de Transferência de Alanina/genética , Acidithiobacillus/metabolismo , Anticódon/química , Anticódon/metabolismo , Sítios de Ligação Microbiológicos , Sequência de Bases , Mapeamento Cromossômico , Cromossomos Bacterianos/química , Cromossomos Bacterianos/metabolismo , Dano ao DNA , DNA Bacteriano/metabolismo , Escherichia coli/genética , Escherichia coli/metabolismo , Integrases/genética , Integrases/metabolismo , Mutagênese Sítio-Dirigida , Conformação de Ácido Nucleico , RNA de Transferência de Alanina/metabolismo , Recombinação Genética , Sintenia

RESUMO

The genome of the acidophilic, bioleaching bacterium Acidithiobacillus ferrooxidans, strain ATCC 23270, contains 95 predicted tRNA genes. Thirty-six of these genes (all 20 species) are clustered within an actively excising integrative-conjugative element (ICEAfe1). We speculated that these tRNA genes might have a role in adapting the bacterial tRNA pool to the codon usage of ICEAfe1 genes. To answer this question, we performed theoretical calculations of the global tRNA adaptation index to the entire A. ferrooxidans genome with and without the ICEAfe1 encoded tRNA genes. Based on these calculations, we observed that tRNAs encoded in ICEAfe1 negatively contribute to adapt the tRNA pool to the codon use in A. ferrooxidans. Although some of the tRNAs encoded in ICEAfe1 are functional in aminoacylation or protein synthesis, we found that they are expressed at low levels. These findings, along with the identification of a tRNA-like RNA encoded in the same cluster, led us to speculate that tRNA genes encoded in the mobile genetic element ICEAfe1 might have acquired mutations that would result in either inactivation or the acquisition of new functions.

Assuntos

Acidithiobacillus/genética , Transferência Genética Horizontal , Genes Bacterianos , Genoma Bacteriano , Sequências Repetitivas Dispersas , RNA de Transferência/genética , Acidithiobacillus/classificação , Acidithiobacillus/metabolismo , Aminoacilação , Conjugação Genética , Mutação , Conformação de Ácido Nucleico , Filogenia , Biossíntese de Proteínas , RNA de Transferência/metabolismo

RESUMO

Background: Traditional methods of obtaining arsenic have disadvantages such as high cost and high energy consumption. Realgar is one of the most abundant arsenic sulphide minerals and usually treated as waste in industry. The aim of the present study was to screen an arsenic tolerant bacterium used for bioleaching arsenic from realgar. Results: An acidophilic iron-oxidizing bacterium BYQ-12 was isolated from Wudalianchi volcanic lake in northeast China. BYQ-12 was a motile, rod-shaped gram-negative bacterium with an optimum growth at 30°C and pH 2.5. 16S rDNA phylogeny showed that BYQ-12 was a new strain of Acidithiobacillus ferrooxidans. The inhibitory concentrations (ICs) of arsenite and arsenate were 32 and 64 mM, respectively. A significant second-order model was established using a Box­Behnken design of response surface methodology (BBD-RSM) and it estimated that a maximum arsenic bioleaching rate (73.97%) could be obtained when the pulp concentration, pH and initial ferrous ion concentration were set at optimized values of 0.95% w/v, 1.74 and 3.68 g/L, respectively. SEM, EDS and XRD analyses also revealed that there was direct bioleaching besides indirect electrochemical leaching in the arsenic bioleaching system. Conclusion: From this work we were successful in isolating an acidophilic, arsenic tolerant ferrous iron-oxidizing bacterium. The BBD-RSM analysis showed that maximum arsenic bioleaching rate obtained under optimum conditions, and the most effective factor for arsenic leaching was initial ferrous ion concentration. These revealed that BYQ-12 could be used for bioleaching of arsenic from arsenical minerals.

Assuntos

Arsênio/metabolismo , Arsênio/química , Acidithiobacillus/isolamento & purificação , Oxirredução , Concentração de Íons de Hidrogênio , Ferro/metabolismo

RESUMO

The understanding of biofilm formation by bioleaching microorganisms is of great importance for influencing mineral dissolution rates and to prevent acid mine drainage (AMD). Thermo-acidophilic archaea such as Acidianus, Sulfolobus and Metallosphaera are of special interest due to their ability to perform leaching at high temperatures, thereby enhancing leaching rates. In this work, leaching experiments and visualization by microscopy of cell attachment and biofilm formation patterns of the crenarchaeotes Sulfolobus metallicus DSM 6482(T) and the Acidianus isolates DSM 29038 and DSM 29099 in pure and mixed cultures on sulfur or pyrite were studied. Confocal laser scanning microscopy (CLSM) combined with fluorescent dyes as well as fluorescently labeled lectins were used to visualize different components (e.g. DNA, proteins or glycoconjugates) of the aforementioned species. The data indicate that cell attachment and the subsequently formed biofilms were species- and substrate-dependent. Pyrite leaching experiments coupled with pre-colonization and further inoculation with a second species suggest that both species may negatively influence each other during pyrite leaching with respect to initial attachment and pyrite dissolution rates. In addition, the investigation of binary biofilms on pyrite showed that both species were heterogeneously distributed on pyrite surfaces in the form of individual cells or microcolonies. Physical contact between the two species seems to occur, as revealed by specific lectins able to specifically bind single species within mixed cultures.

Assuntos

Acidianus/fisiologia , Biofilmes/crescimento & desenvolvimento , Microbiologia Ambiental , Interações Microbianas , Sulfolobus/fisiologia , Aderência Bacteriana , Ferro/metabolismo , Microscopia Confocal , Sulfetos/metabolismo , Enxofre/metabolismo

RESUMO

Leptospirillum ferriphilum Sp-Cl is a Gram negative, thermotolerant, curved, rod-shaped bacterium, isolated from an industrial bioleaching operation in northern Chile, where chalcocite is the major copper mineral and copper hydroxychloride atacamite is present in variable proportions in the ore. This strain has unique features as compared to the other members of the species, namely resistance to elevated concentrations of chloride, sulfate and metals. Basic microbiological features and genomic properties of this biotechnologically relevant strain are described in this work. The 2,475,669 bp draft genome is arranged into 74 scaffolds of 74 contigs. A total of 48 RNA genes and 2,834 protein coding genes were predicted from its annotation; 55 % of these were assigned a putative function. Release of the genome sequence of this strain will provide further understanding of the mechanisms used by acidophilic bacteria to endure high osmotic stress and high chloride levels and of the role of chloride-tolerant iron-oxidizers in industrial bioleaching operations.

RESUMO

The present work aimed at determining the tolerance of Nile tilapia juveniles to highly acidic rearing waters and the effects of water acidity on the quality of tank effluents. The experimental design consisted of four treatments with different water pH values (4.12 ± 0.84; 5.13 ± 0.74; 6.14 ± 0.64 and 8.06 ± 0.48), with five replicates each. No exchange of water was performed throughout the study, only water replenishment to maintain the initial level. Variables of water quality, soil, growth performance, metabolism and effluents were monitored for eight weeks. Despite the lower total ammonia nitrogen (TAN) concentration in the pH 8 tanks, their levels of non-ionized ammonia (NH3) were the highest ones. At the end, the lowest body weight of fish was observed in the pH 8 tanks. There was a significant improvement in feed conversion ratio (FCR) and protein efficiency ratio (PER) due to to water acidification. There were reduced concentrations of NH3 in the acidified tanks effluents. It was concluded that the gradual water acidification up to pH 4 can improve the Nile tilapia juveniles growth performance.(AU)

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O presente trabalho teve por objetivo determinar a tolerância de juvenis de tilápia do Nilo à acidez elevada da água de cultivo e os efeitos da acidificação da água de cultivo na qualidade dos efluentes dos tanques. O delineamento experimental foi constituído por quatro tratamentos, águas de cultivo com diferentes valores de pH (4,12 ± 0,84; 5,13 ± 0,74; 6,14 ± 0,64 e 8,06 ± 0,48), com cinco repetições cada. Não houve troca da água de cultivo, apenas reposição para manter o nível inicial. Durante oito semanas, foram observadas variáveis de qualidade de água, solo, zootécnicas, metabólicas e efluentes. Embora a concentração de nitrogênio amoniacal total (NAT) na água tenha sido menor nos tanques pH8, sua concentração de amônia não-ionizada (NH3) foi maior que nos demais. Ao final, o menor peso corporal dos peixes foi observado nos tanques pH8. Houve significativa melhora nos resultados de conversão alimentar e na taxa de eficiência proteica com a acidificação do pH da água. As concentrações de NH3 nos efluentes dos tanques acidificados foram reduzidas. Concluiu-se que a acidificação gradual da água de cultivo de juvenis da tilápia do Nilo até pH 4 é benéfica ao crescimento corporal dos peixes.(AU)

Assuntos

Animais , Ciclídeos/anormalidades , Ciclídeos/crescimento & desenvolvimento , Aquicultura/organização & administração , Acidez da Água/efeitos adversos , Acidez da Água/análise , Concentração de Íons de Hidrogênio

RESUMO

The present work aimed at determining the tolerance of Nile tilapia juveniles to highly acidic rearing waters and the effects of water acidity on the quality of tank effluents. The experimental design consisted of four treatments with different water pH values (4.12 ± 0.84; 5.13 ± 0.74; 6.14 ± 0.64 and 8.06 ± 0.48), with five replicates each. No exchange of water was performed throughout the study, only water replenishment to maintain the initial level. Variables of water quality, soil, growth performance, metabolism and effluents were monitored for eight weeks. Despite the lower total ammonia nitrogen (TAN) concentration in the pH 8 tanks, their levels of non-ionized ammonia (NH3) were the highest ones. At the end, the lowest body weight of fish was observed in the pH 8 tanks. There was a significant improvement in feed conversion ratio (FCR) and protein efficiency ratio (PER) due to to water acidification. There were reduced concentrations of NH3 in the acidified tanks effluents. It was concluded that the gradual water acidification up to pH 4 can improve the Nile tilapia juveniles growth performance.

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O presente trabalho teve por objetivo determinar a tolerância de juvenis de tilápia do Nilo à acidez elevada da água de cultivo e os efeitos da acidificação da água de cultivo na qualidade dos efluentes dos tanques. O delineamento experimental foi constituído por quatro tratamentos, águas de cultivo com diferentes valores de pH (4,12 ± 0,84; 5,13 ± 0,74; 6,14 ± 0,64 e 8,06 ± 0,48), com cinco repetições cada. Não houve troca da água de cultivo, apenas reposição para manter o nível inicial. Durante oito semanas, foram observadas variáveis de qualidade de água, solo, zootécnicas, metabólicas e efluentes. Embora a concentração de nitrogênio amoniacal total (NAT) na água tenha sido menor nos tanques pH8, sua concentração de amônia não-ionizada (NH3) foi maior que nos demais. Ao final, o menor peso corporal dos peixes foi observado nos tanques pH8. Houve significativa melhora nos resultados de conversão alimentar e na taxa de eficiência proteica com a acidificação do pH da água. As concentrações de NH3 nos efluentes dos tanques acidificados foram reduzidas. Concluiu-se que a acidificação gradual da água de cultivo de juvenis da tilápia do Nilo até pH 4 é benéfica ao crescimento corporal dos peixes.

Assuntos

Animais , Acidez da Água/análise , Acidez da Água/efeitos adversos , Aquicultura/organização & administração , Ciclídeos/anormalidades , Ciclídeos/crescimento & desenvolvimento , Concentração de Íons de Hidrogênio

RESUMO

Background: Inulinases have been extracted and characterized from inulin-storing tissues; however, production of microbial inulinases have recently draw much attention as they offer several industrial advantages. Many microorganisms, including filamentous fungi, yeast and bacteria have been claimed as inulinase producers. These hydrolases are usually inducible and their exo-acting forms may hydrolyze fructose polymers (inulin) and oligosaccharides such as sucrose and raffinose. Fungal inulinase extracts are often produced as stable mixture of highly active fructanhydrolases. From a practical prospective, the best known inulinases to date are those produced by species of Penicillium, Aspergillus and Kluyveromyces. Results: The production of extracellular inulinase by A. kawachii in liquid cultures, using either inulin or yacon derived materials as CES as well as inulinase inducers, is reported. In addition, a partial characterization of the enzyme activity is included. Conclusions: Yacon derived products, particularly yacon juice, added to the culture medium proved to be a good CES for fungal growth as well as an inducer of enzyme synthesis. Partial characterization of the enzyme revealed that it is quite stable in a wide range of pH and temperature. In addition, characterization of the reaction products revealed that this enzyme corresponds to an exo-type. These facts are promising considering its potential application in inulin hydrolysis for the production of high fructose syrups.

Assuntos

Aspergillus/enzimologia , Glicosídeo Hidrolases/metabolismo , Temperatura , Estabilidade Enzimática , Reatores Biológicos , Asteraceae , Técnicas de Cultura Celular por Lotes , Concentração de Íons de Hidrogênio , Hidrólise , Íons

RESUMO

The coal mining activity is characterized by the generation of large amount of by-products. One of them is pyrite, which tends to acidify the water, solubilizing heavy metals. As a consequence the environment becomes acid and rich in heavy metals, selecting microorganisms able to survive in this condition, which are of great interest as bioremediation agents. This work describes the isolation and characterization of microorganisms from a coal mining area in Santa Catarina. These microorganisms comprised bacteria, fungi and yeasts resistant to zinc, nickel and cadmium.

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A atividade de mineração do carvão é responsável pela geração de diferentes sub-produtos. Entre esses, está a pirita que acidifica a água e acelera o processo de solubilização de metais. Como conseqüência, o ambiente torna-se ácido e rico em metais pesados, os quais selecionam os microrganismos capazes de sobreviver nestas condições. Esses microrganismos podem, por sua vez, serem empregados como agentes para a biorremediação de áreas contaminadas com metais pesados. No presente trabalho é descrito o isolamento e a caracterização de bactérias, fungos e leveduras resistentes aos metais zinco, níquel e cádmio.