The microbial world in copper sulfide flotation plants (CSFP): Novel insights into bacterial communities and their application as potential pyrite bioreagents.

Operations in copper sulfide flotation plants (CSFP) are complex and governed by several variables such as available technologies, reagents, and environmental conditions. However, few investigations are related to studying the microbial communities. These aspects provide a reason to compare the bacterial communities of two CSFP operated with freshwater (FwFlo) and seawater (SwFlo), and study whether indigenous bacteria could be used as pyrite bioreagents. Analyses were determined through next-generation sequencing by Illumina MiSeq System and conducted throughout the entire process: (i) minerals before and after grinding; (ii) final concentrate and concentrate thickener overflow; (iii) final tailings and tailings thickener overflow; and (iv) intake water. Bacterial strains from both plants were tested as potential bioreagents, given their tendency to adhere to pyrite after 5 min. In both CSFP, Proteobacteria (relative abundance from 45.48% to 79.22%), followed by Bacteroidetes (9.37%-44.7%), were the most abundant phyla. Regarding species, Algoriphagus olei (11.35%-43.52%) was present exclusively in FwFlo samples in contact with process water and absent in the mineral before grinding, where Cupriavidus metallidurans (16.05%) and Pseudomonas_uc (11.79%) predominated. In SwFlo samples, Marinobacter flavimaris (3.47%-41.1%), and GU061212-s (10.92%-27.63%), were the most abundant microorganisms. All of them were also detected in intake seawater. The strains with the highest adhesion rate (from 29.84% ± 0.14-100%) were phylogenetically identified as species of the genera Marinobacter, Pseudomonas, Idiomarina, Halomonas, Bacillus, Aerocuccus, and Peribacillus. Our results reveal that bacterial communities are critically dependent on process waters during mining activities, and our data depicted that indigenous bacteria could be used as potential pyrite bioreagents, evidenced by a high adhesion rate. It is thus possible to propose that different indigenous bacterial strains could be considered as new bioreagents to reduce the impact of conventional flotation reagents on health from an environment friendly perspective.

Response to Static Magnetic Field-Induced Stress in Scenedesmus obliquus and Nannochloropsis gaditana.

Magnetic fields in biological systems is a promising research field; however, their application for microalgae has not been fully exploited. This work aims to measure the enzymatic activity and non-enzymatic activity of two microalgae species in terms of superoxide dismutase (SOD), catalase (CAT), and carotenoids, respectively, in response to static magnetic fields-induced stress. Two magnet configurations (north and south) and two exposure modes (continuous and pulse) were applied. Two microalgae species were considered, the Scenedesmus obliquus and Nannochloropsis gaditana. The SOD activity increased by up to 60% in S. obliquus under continuous exposure. This trend was also found for CAT in the continuous mode. Conversely, under the pulse mode, its response was hampered as the SOD and CAT were reduced. For N. gaditana, SOD increased by up to 62% with the south configuration under continuous exposure. In terms of CAT, there was a higher activity of up to 19%. Under the pulsed exposure, SOD activity was up to 115%. The CAT in this microalga was increased by up to 29%. For N. gaditana, a significant increase of over 40% in violaxanthin production was obtained compared to the control, when the microalgae were exposed to SMF as a pulse. Depending on the exposure mode and species, this methodology can be used to produce oxidative stress and obtain an inhibitory or enhanced response in addition to the significant increase in the production of antioxidant pigments.

Application of microbe-induced carbonate precipitation for copper removal from copper-enriched waters: Challenges to future industrial application.

Copper contamination in watercourses is a recent issue in countries where mining operations are prevalent. In this study, the application of copper precipitation through microbe-induced carbonate precipitation (MICP) was analyzed using urea hydrolysis by bacteria to evaluate precipitated copper carbonates. This article demonstrates the application of a copper precipitation assay involving Sporosarcina pasteurii (in 0.5 mM Cu2+ and 333 mM urea) and analyzes the resultant low removal (10%). The analysis indicates that the low removal was a consequence of Cu2+ complexation with the ammonia resulting from the hydrolysis of urea. However, the results indicate that there should be a positive correlation between the initial urea concentration and the bacterial tolerance to copper. This identifies a challenge in the industrial application of the process, wherein a minimum consumption of urea represents an economic advantage. Therefore, it is necessary to design a sequential process that decouples bacterial growth and copper precipitation, thereby decreasing the urea requirement.

Bioprospecting of Ureolytic Bacteria From Laguna Salada for Biomineralization Applications.

The processes of biomineralization, mediated by ureolytic bacteria, possess a wide range of technological applications, such as the formation of biocements and remediation of water and soil environments. For this reason, the bioprospecting of new ureolytic bacteria is interesting for its application to these technologies, particularly for water treatment. This study demonstrates the isolation, selection, and identification of halotolerant ureolytic bacteria from Laguna Salada (inland from Atacama Desert) and the evaluation of their ability to precipitate calcium carbonate crystals in freshwater in the presence of calcium ions, as well as the ability to induce the precipitation of crystals from different ions present in seawater. Twenty-four halotolerant ureolytic bacteria whose molecular identification gives between 99 and 100% identity with species of the genus Bacillus, Porphyrobacter, Pseudomonas, Salinivibrio, and Halomonas were isolated. When cultivated in freshwater, urea, and calcium chloride, all species are able to biomineralize calcium carbonate in different concentrations. In seawater, the strains that biomineralize the highest concentration of calcium carbonate correspond to Bacillus subtilis and Halomonas sp. SEM-EDX and XRD analyses determined that both bacteria induce the formation of 9-33% halite (NaCl), 31-66% monohydrocalcite (CaCO3 × H2O), and 24-27% struvite (MgNH4PO4 × 6H2O). Additionally, B.subtilis induces the formation of 7% anhydrite (CaSO4). In seawater, B.subtilis and Halomonas sp. were able to precipitate both calcium (96-97%) and magnesium (63-67%) ions over 14 days of testing. Ion removal assays with B.subtilis immobilized in beads indicate a direct relationship between the urea concentration and a greater removal of ions with similar rates to free cells. These results demonstrate that the biomineralization mediated by bacterial urea hydrolysis is feasible in both freshwater and seawater, and we propose its application as a new technology in improving water quality for industrial uses.

The chilean network of microbial culture collections: establishment and operation / Red chilena de colecciones de cultivos microbianos: establecimiento y operación

According to the data available at the World Data Center for Microorganism-WDCM from the World Federation for Culture Collection-WFCC, Chile has four registered culture collections that preserve 2777 microbial strains. At the global point of view, the culture collections in Chile are in different level of operation regarding its own infrastructure and compliancy with quality standards for preservation of strains and for services provide. The absence of funding to support the preservation of the Chilean microbial assets is a key issue for the development of the Chilean bioeconomy. Considering this, the Chilean culture collections started working together to establish the Chilean Network of Microbial Culture Collections (RCCCM, acronym in Spanish). In this note, the establishment and operation of the RCCCM is presented and discussed.

De acuerdo con los datos disponibles en el World Data Center for Microorganism-WDCM de la Federación Mundial para la Colección de Cultivos - WFCC, Chile tiene cuatro colecciones de cultivos registradas que preservan 2777 cepas microbianas. Desde el punto de vista global, las colecciones de cultivos en Chile se encuentran en diferentes niveles de operación con respecto a su propia infraestructura y cumplimiento con estándares de calidad para la preservación de las cepas y para los servicios que proporcionan. La ausencia de financiamiento para apoyar la preservación de los activos microbianos chilenos es un tema clave para el desarrollo de la bioeconomía chilena. Considerando esto, las colecciones chilenas de cultivos comenzaron a trabajar conjuntamente para establecer la Red Chilena de Colecciones de Cultivos Microbianos (RCCCM). En esta nota se presenta y discute el establecimiento y funcionamiento de la RCCCM.

Use of cellulolytic marine bacteria for enzymatic pretreatment in microalgal biogas production.

In this study, we designed and evaluated a microalgal pretreatment method using cellulolytic bacteria that naturally degrades microalgae in their native habitat. Bacterial strains were isolated from each of two mollusk species in a medium containing 1% carboxymethyl cellulose agar. We selected nine bacterial strains that had endoglucanase activity: five strains from Mytilus chilensis, a Chilean mussel, and four strains from Mesodesma donacium, a clam found in the Southern Pacific. These strains were identified phylogenetically as belonging to the genera Aeromonas, Pseudomonas, Chryseobacterium, and Raoultella. The cellulase-producing capacities of these strains were characterized, and the degradation of cell walls in Botryococcus braunii and Nannochloropsis gaditana was tested with "whole-cell" cellulolytic experiments. Aeromonas bivalvium MA2, Raoultella ornithinolytica MA5, and Aeromonas salmonicida MC25 degraded B. braunii, and R. ornithinolytica MC3 and MA5 degraded N. gaditana. In addition, N. gaditana was pretreated with R. ornithinolytica strains MC3 and MA5 and was then subjected to an anaerobic digestion process, which increased the yield of methane by 140.32% and 158.68%, respectively, over that from nonpretreated microalgae. Therefore, a "whole-cell" cellulolytic pretreatment can increase the performance and efficiency of biogas production.

Interactions of Botryococcus braunii cultures with bacterial biofilms.

Unicellular microalgae generally grow in the presence of bacteria, particularly when they are farmed massively. This study analyzes the bacteria associated with mass culture of Botryococcus braunii: both the planktonic bacteria in the water column and those forming biofilms adhered to the surface of the microalgal cells (∼107-108 culturable cells per gram microalgae). Furthermore, we identified the culturable bacteria forming a biofilm in the microalgal cells by 16S rDNA sequencing. At least eight different culturable species of bacteria were detected in the biofilm and were evaluated for the presence of quorum-sensing signals in these bacteria. Few studies have considered the implications of this phenomenon as regards the interaction between bacteria and microalgae. Production of C4-AHL and C6-AHL were detected in two species, Pseudomonas sp. and Rhizobium sp., which are present in the bacterial biofilm associated with B. braunii. This type of signal was not detected in the planktonic bacteria isolated from the water. We also noted that the bacterium, Rhizobium sp., acted as a probiotic bacterium and significantly encouraged the growth of B. braunii. A direct application of these beneficial bacteria associated with B. braunii could be, to use them like inoculants for large-scale microalgal cultures. They could optimize biomass production by enhancing growth, particularly in this microalga that has a low growth rate.

Second acyl homoserine lactone production system in the extreme acidophile Acidithiobacillus ferrooxidans.

The acidophilic proteobacterium Acidithiobacillus ferrooxidans is involved in the industrial biorecovery of copper. It is found in acidic environments in biofilms and is important in the biogeochemical cycling of metals and nutrients. Its genome contains a cluster of four genes, glyQ, glysS, gph, and act, that are predicted to encode the alpha and beta subunits of glycine tRNA synthetase, a phosphatase, and an acyltransferase, respectively (GenBank accession no. DQ149607). act, cloned and expressed in Escherichia coli, produces acyl homoserine lactones (AHLs) principally of chain length C14 according to gas chromatography and mass spectrometry measurements. The AHLs have biological activity as shown by in vivo studies using the reporter strain Sinorhizobium meliloti Rm41 SinI-. Reverse transcription-PCR (RT-PCR) experiments indicate that the four genes are expressed as a single transcript, demonstrating that they constitute an operon. According to semiquantitative RT-PCR results, act is expressed more highly when A. ferrooxidans is grown in medium containing iron than when it is grown in medium containing sulfur. Since AHLs are important intercellular signaling molecules used by many bacteria to monitor their population density in quorum-sensing control of gene expression, this result suggests that A. ferrooxidans has two quorum-sensing systems, one based on Act, as described herein, and the other based on a Lux-like quorum-sensing system, reported previously. The latter system was shown to be upregulated in A. ferrooxidans grown in sulfur medium, suggesting that the two quorum-sensing systems respond to different environmental signals that may be related to their abilities to colonize and use different solid sulfur- and iron-containing minerals.

A Lux-like quorum sensing system in the extreme acidophile Acidithiobacillus ferrooxidans.

The genome of the acidophilic, proteobacterium Acidithiobacillusferrooxidans, contains linked but divergently oriented genes, termed afel and afeR, whose predicted protein products are significantly similar to the LuxI and LuxR families of proteins. A possible promoter and Lux box are predicted upstream of afel. A cloned copy of afel, expressed in E. coli, encodes an enzyme that catalyzes the production of a diffusible compound identified by gas chromatography and mass spectrometry as an unsubstituted N-acyl homoserine lactone (AHL) of chain length C14. This AHL can be detected by a reporter strain of Sinorhizobium meliloti Rm41 suggesting that it is biologically active. The reporter strain also responds to extracts of the supernatant of A. ferrooxidans grown to early stationary phase in sulfur medium indicating that a diffusible AHL is produced by this microorganism. Semi-quantitative RT-PCR experiments indicate that afeI and afeR are expressed maximally in early stationary phase and are more expressed when A. ferrooxidans is grown in sulfur--rather than iron-containing medium. Given the predicted amino acid sequence and functional properties of AfeI and AfeR it is proposed that A. ferrooxidans has a quorum sensing system similar to the LuxI-LuxR paradigm.

A Lux-like Quorum Sensing System in the Extreme Acidophile Acidithiobacillus ferrooxidans

The genome of the acidophilic, proteobacterium Acidithiobacillus ferrooxidans, contains linked but divergently oriented genes, termed afeI and afeR, whose predicted protein products are significantly similar to the LuxI and LuxR families of proteins. A possible promoter and Lux box are predicted upstream of afeI. A cloned copy of afeI, expressed in E. coli, encodes an enzyme that catalyzes the production of a diffusible compound identified by gas chromatography and mass spectrometry as an unsubstituted N-acyl homoserine lactone (AHL) of chain length C14. This AHL can be detected by a reporter strain of Sinorhizobium meliloti Rm41 suggesting that it is biologically active. The reporter strain also responds to extracts of the supernatant of A. ferrooxidans grown to early stationary phase in sulfur medium indicating that a diffusible AHL is produced by this microorganism. Semi-quantitative RT-PCR experiments indicate that afeI and afeR are expressed maximally in early stationary phase and are more expressed when A. ferrooxidans is grown in sulfur- rather than iron-containing medium. Given the predicted amino acid sequence and functional properties of AfeI and AfeR it is proposed that A. ferrooxidans has a quorum sensing system similar to the LuxI-LuxR paradigm.