Enhancing iron biogeochemical cycling for canga ecosystem restoration: insights from microbial stimuli.

Introduction: The microbial-induced restoration of ferruginous crusts (canga), which partially cover iron deposits and host unique ecosystems, is a promising alternative for reducing the environmental impacts of the iron mining industry. Methods: To investigate the potential of microbial action to accelerate the reduction and oxidation of iron in substrates rich in hematite and goethite, four different microbial treatments (water only as a control - W; culture medium only - MO; medium + microbial consortium - MI; medium + microbial consortium + soluble iron - MIC) were periodically applied to induce iron dissolution and subsequent precipitation. Except for W, all the treatments resulted in the formation of biocemented blocks. Results: MO and MI treatments resulted in significant goethite dissolution, followed by precipitation of iron oxyhydroxides and an iron sulfate phase, due to iron oxidation, in addition to the preservation of microfossils. In the MIC treatment, biofilms were identified, but with few mineralogical changes in the iron-rich particles, indicating less iron cycling compared to the MO or MI treatment. Regarding microbial diversity, iron-reducing families, such as Enterobacteriaceae, were found in all microbially treated substrates. Discussion: However, the presence of Bacillaceae indicates the importance of fermentative bacteria in accelerating the dissolution of iron minerals. The acceleration of iron cycling was also promoted by microorganisms that couple nitrate reduction with Fe(II) oxidation. These findings demonstrate a sustainable and streamlined opportunity for restoration in mining areas.

Production and characterization of rhamnolipids by Pseudomonas aeruginosa isolated in the Amazon region, and potential antiviral, antitumor, and antimicrobial activity.

Biosurfactants encompass structurally and chemically diverse molecules with surface active properties, and a broad industrial deployment, including pharmaceuticals. The interest is growing mainly for the low toxicity, biodegradability, and production from renewable sources. In this work, the optimized biosurfactant production by Pseudomonas aeruginosa BM02, isolated from the soil of a mining area in the Brazilian Amazon region was assessed, in addition to its antiviral, antitumor, and antimicrobial activities. The optimal conditions for biosurfactant production were determined using a factorial design, which showed the best yield (2.28 mg/mL) at 25 °C, pH 5, and 1% glycerol. The biosurfactant obtained was characterized as a mixture of rhamnolipids with virucidal properties against Herpes Simplex Virus, Coronavirus, and Respiratory Syncytial Virus, in addition to antimicrobial properties against Gram-positive bacteria (Staphylococcus aureus and Enterococcus faecium), at 50 µg/mL. The antitumor activity of BS (12.5 µg/mL) was also demonstrated, with potential selectivity in reducing the proliferation of breast tumor cells, after 1 min of exposure. These results demonstrate the importance of studying the interconnection between cultivation conditions and properties of industrially important compounds, such as rhamnolipid-type biosurfactant from P. aeruginosa BM02, a promising and sustainable alternative in the development of new antiviral, antitumor, and antimicrobial prototypes.

Acquiring Iron-Reducing Enrichment Cultures: Environments, Methods and Quality Assessments.

Lateritic duricrusts cover iron ore deposits and form spatially restricted, unique canga ecosystems endangered by mining. Iron cycling, i.e., the dissolution and subsequent precipitation of iron, is able to restitute canga duricrusts, generating new habitats for endangered biota in post-mining landscapes. As iron-reducing bacteria can accelerate this iron cycling, we aim to retrieve microbial enrichment cultures suitable to mediate the large-scale restoration of cangas. For that, we collected water and sediment samples from the Carajás National Forest and cultivated the iron-reducing microorganisms therein using a specific medium. We measured the potential to reduce iron using ferrozine assays, growth rate and metabolic activity. Six out of seven enrichment cultures effectively reduced iron, showing that different environments harbor iron-reducing bacteria. The most promising enrichment cultures were obtained from environments with repeated flooding and drying cycles, i.e., periodically inundated grasslands and a plateau of an iron mining waste pile characterized by frequent soaking. Selected enrichment cultures contained iron-reducing and fermenting bacteria, such as Serratia and Enterobacter. We found higher iron-reducing potential in enrichment cultures with a higher cell density and microorganism diversity. The obtained enrichment cultures should be tested for canga restoration to generate benefits for biodiversity and contribute to more sustainable iron mining in the region.

Bioprospecting for Isoetes cangae Endophytes with Potential to Promote Plant Growth.

Isoetes cangae is a native plant found only in a permanent pond in Serra dos Carajás in the Amazon region. Plant-associated microbial communities are recognized to be responsible for biological processes essential for the health, growth, and even adaptation of plants to environmental stresses. In this sense, the aims of this work were to isolate, identify, and evaluate the properties of endophytic bacteria isolated from I. cangae. The bioprospecting of potentially growth-promoting endophytes required the following steps to be taken: isolation of endophytic colonies, molecular identification by 16S rDNA sequence analysis, and evaluation of the bacterial potential for nitrogen fixation, production of indole acetic acid and siderophores, as well as phosphate solubilization and mineralization. Bacillus sp., Rhizobium sp., Priestia sp., Acinetobacter sp., Rossellomorea sp., Herbaspirillum sp., Heyndrickxia sp., and Metabacillus sp., among other bacterial species, were identified. The isolates showed to be highly promising, evidencing the physiological importance for the plant and having the potential to promote plant growth.

Development of Rhizophora mangle (Rhizophoraceae) and Avicennia schaueriana (Avicenniaceae) in the presence of a hydrocarbon-degrading bacterial consortium and marine diesel oil.

The development of Rhizophora mangle and Avicennia schaueriana seedlings impacted by marine diesel oil (MDO) was evaluated in the presence or absence of a hydrocarbon-degrading bacterial consortium (HBC). The bioassays were conducted in a greenhouse during 6 months and consisted of three different treatments (control, MDO only and MDO + HBC). The bacterial consortium was mainly composed of Bacillus spp. (73%), but Rhizobium spp., Pseudomonas spp., Ochrobactrum spp., and Brevundimonas spp. were also present. After 6 months, A. schaueriana seedlings showed higher mortality compared to those of R. mangle; R. mangle exhibited 68% (control), 44% (MDO alone) and 50% (MDO + HBC) seedlings survivorship compared to 42% (control), 0% (MDO alone) and 4% (MDO + HBC) for A. schaueriana. This variability may be due to differences in species physiology. Stem growth, diameter and number of leaves remained constant during the 6 months of the experiment with marine diesel oil and hydrocarbon-degrading bacterial consortium (MDO + BBC). For both mangrove species, bacterial enzymatic activity in the sediments was sufficient to maintain cell counts of 107 cells cm-3 in the rhizospheric soil and possibly synthetize the extracellular polymeric substances (EPS) that may emulsify and solubilize oil products.

Comparison of two acidophilic sulfidogenic consortia for the treatment of acidic mine water.

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.

Accelerating microbial iron cycling promotes re-cementation of surface crusts in iron ore regions.

Accelerating microbial iron cycling is an innovative environmentally responsible strategy for mine remediation. In the present study, we extend the application of microbial iron cycling in environmental remediation, to include biocementation for the aggregation and stabilization of mine wastes. Microbial iron reduction was promoted monthly for 10 months in crushed canga (a by-product from iron ore mining, dominated by crystalline iron oxides) in 1 m3 containers. Ferrous iron concentrations reached 445 ppm in treatments and diverse lineages of the candidate phyla radiation dominated pore waters, implicating them in fermentation and/or metal cycling in this system. After a 6-month evaporation period, iron-rich cements had formed between grains and 20-cm aggregates were recoverable from treatments throughout the 1-m depth profile, while material from untreated and water-only controls remained unconsolidated. Canga-adapted plants seeded into one of the treatments germinated and grew well. Therefore, application of this geobiotechnology offers promise for stabilization of mine wastes, as well as re-formation of surface crusts that underpin unique and threatened plant ecosystems in iron ore regions.

Potential application of Pseudomonas stutzeri W228 for removal of copper and lead from marine environments.

High concentrations of metals in the environment alter bacterial diversity, selecting resistant and tolerant species. The study evaluated the selection of a potential bacterial strain from Sepetiba Bay-Rio de Janeiro, Brazil marine sediments to remove Cu and Pb. The bacterial strain isolated from the sediments was used in three different bioassays: (1) Cu at concentrations of 0 (control), 6 and 50 µg.mL-1; (2) Pb at concentrations of 0 (control), 6 and 50 µg.mL-1; (3) Cu + Pb in concentrations of 3 µg.mL-1 Cu + 3 µg.mL-1 Pb (6 µg.mL-1) and 25 µg.mL-1 Cu + 25 µg.mL-1 Pb (50 µg.mL-1). The number of cells and the enzymatic activities of dehydrogenases and esterases were quantified. Results of taxonomic identification indicated the selection of the Pseudomonas stutzeri W228 strain, showing a greater degree of similarity (±73%) with the database used. There was no significant variation in the number of cells, 108 cells.mL-1, which represents a high biomass production in the presence of stressors. However, we observed a reduction in dehydrogenase activity at all tested concentrations of Cu, Pb and Cu + Pb. The activity of esterase increased, indicating a higher energy demand to complete the bacterial life cycle. The study showed significant results for the absorption of Pb by the extracellular polymeric substances (EPS) and the efflux of Cu. The capacity of Pb absorption by EPS can be considered a resistance mechanism, as well as the efflux of Cu, so that the available EPS sites could be occupied by the most toxic ions demonstrating that Pseudomonas stutzeri is resistant to Pb and Cu.

Inhibitory effect of sildenafil on the human isolated seminal vesicle.

OBJECTIVE: To investigate the effects of sildenafil on noradrenaline- and potassium-induced contractions of isolated human seminal vesicles (SVs), as premature ejaculation is a relatively common male sexual dysfunction that currently lacks an adequate therapy, and recent in vitro tests showed that sildenafil induces relaxation of rodent isolated SVs, but it is not known whether it also inhibits isolated human SV. MATERIAL AND METHODS: Isolated strips of human SVs were suspended in an organ bath and cumulative concentration-response curves to either noradrenaline or KCl were constructed in the presence and absence of sildenafil to evaluate its inhibitory actions. RESULTS: Sildenafil (25-100 microm) induced concentration-dependent inhibitory effects on the human SV contracted by either noradrenaline or KCl. These actions of sildenafil do not therefore appear to be mediated via a competitive antagonism of alpha-adrenoceptors and are consistent with its recognized ability to inhibit phosphodiesterase-5. CONCLUSIONS: The present results show an important inhibitory action of sildenafil in the isolated human SV, supporting the therapeutic indication of this drug for treating premature ejaculation.

Copper and lead removal from aqueous solutions by bacterial consortia acting as biosorbents.

A bacterial consortium was selected in the presence of Cu from sediment samples taken from Sepetiba Bay, Brazil, which is a site historically contaminated by metals. Bacteria were exposed to 0, 1, 6, 12.5, 25 and 50µg·mL(-1) Cu, Pb and Cu+Pb for 11days of bioassay. Results showed Alcanivorax dominance (81%) and cell counts of 10(8)cells·mL(-1). However, a reduction in dehydrogenase activity was observed from the fifth day of exposure for all Cu, Pb, and Cu+Pb concentrations tested. Esterase activity tended to increase, indicating higher energy demand to complete the bacterial lifecycle. Pb concentrations in the filtered culture medium (0.2µm) were below the detection limit, indicating biosorption, whereas concentrations of Cu were close to the tested concentrations, indicative of efflux. Results suggest the need for biomarkers, such as esterase and dehydrogenase enzymatic activity, in the assessment of resistance and tolerance of communities previously exposed to stressors.

Evaluation of the sensitivity to zinc of ciliates Euplotes vannus and Euplotes crassus and their naturally associated bacteria isolated from a polluted tropical bay.

The aim of this study was to evaluate the Zn sensitivity of Euplotes vannus, Euplotes crassus, and their naturally associated bacteria sampled from sediments in the northwest and east regions of Guanabara Bay. The unexposed ciliates and bacteria did not appear to be negatively affected by 96 h of assay. In the control group, E. vannus exhibited an increase in the biomass content from 2.3 × 10(2) to 2.3 × 10(3) µg C cm(-3) between 0 and 96 h, and E. crassus increased up to 7.07 × 10(2) µg C cm(-3) at 48 h. The maximum biomass was pointed by E. crassus (1.33 × 10(3) µg C cm(-3)) in the presence of 0.005 mg Zn L(-1) and E. vannus was naturally associated bacteria (2.40 × 10(-1) µg C cm(-3)) in the presence of 1.0 mg Zn L(-1) (96 h). The growth of E. vannus from the northwest region showed concentration-dependent manners, and it is more sensitive to zinc than E. crassus from the southeast. Naturally associated bacteria showed better adaptation to increasing concentrations of Zn, and the Dunnett test showed that previous environmental selection is important. These results show that new bioremediation tools are necessary.