Microbial diversity and abundance of Hg related genes from water, sediment and soil the Colombian amazon ecosystems impacted by artisanal and small-scale gold mining.

The Amazon region abounds in precious mineral resources including gold, copper, iron, and coltan. Artisanal and small-scale gold mining (ASGM) poses a severe risk in this area due to considerable mercury release into the surrounding ecosystems. Nonetheless, the impact of mercury on both the overall microbiota and the microbial populations involved in mercury transformation is not well understood. In this study we evaluated microbial diversity in samples of soil, sediment and water potentially associated with mercury contamination in two localities (Taraira and Tarapacá) in the Colombian Amazon Forest. To this end, we characterized the bacterial community structure and mercury-related functions in samples from sites with a chronic history of mercury contamination which today have different levels of total mercury content. We also determined mercury bioavailability and mobility in the samples with the highest THg and MeHg levels (up to 43.34 and 0.049 mg kg-1, respectively, in Taraira). Our analysis of mercury speciation showed that the immobile form of mercury predominated in soils and sediments, probably rendering it unavailable to microorganisms. Despite its long-term presence, mercury did not appear to alter the microbial community structure or composition, which was primarily shaped by environmental and physicochemical factors. However, an increase in the relative abundance of merA genes was detected in polluted sediments from Taraira. Several Hg-responsive taxa in soil and sediments were detected in sites with high levels of THg, including members of the Proteobacteria, Acidobacteria, Actinobacteria, Firmicutes and Chloroflexi phyla. The results suggest that mercury contamination at the two locations sampled may select mercury-adapted bacteria carrying the merA gene that could be used in bioremediation processes for the region.

Passive multi-unit field-pilot for acid mine drainage remediation: Performance and environmental assessment of post-treatment solid waste.

This study evaluated the performance of a passive multi-unit field-pilot operating for 16 months to treat acid mine drainage (AMD) from a coal mine in Colombia Andean Paramo. The multi-unit field-pilot involved a combination of a pre-treatment unit (550 L) filled with dispersed alkaline substrate (DAS), and six passive biochemical reactors (PBRs; 220 L) under two configurations: open (PBRs-A) and closed (PBRs-B) to the atmosphere. The AMD quality was 1200 ± 91 mg L-1 Fe, 38.0 ± 1.3 mg L-1 Mn, 8.5 ± 1.6 mg L-1 Zn, and 3200 ± 183.8 mg L-1 SO42-, at pH 2.8. The input and output effluents were monitored to establish AMD remediation. Physicochemical stability of the post-treatment solids, including metals (Fe2+, Zn2+, and Mn2+) and sulfates for environmental contamination from reactive mixture post-treatment, was also assessed. The passive multi-unit field-pilot achieved a total removal of 74% SO42-, 63% Fe2+, and 48% Mn2+ with the line of PBRs-A, and 91% SO42-, 80% Fe2+, and 66% Mn2+ with the line of PBRs-B, as well as 99% removal for Zn2+ without significant differences (p < 0.05) between the two lines. The study of the physicochemical stability of the post-treatment solids showed they can produce acidic leachates that could release large quantities of Fe and Mn, if they are disposed in oxidizing conditions; contact with water or any other leaching solutions must be avoided. Therefore, these post-treatment solids cannot be disposed of in a municipal landfill. The differences in configuration between PBRs, open or closed to the atmosphere, induced changes in the performance of the passive multi-unit field-pilot during AMD remediation.

Metagenomic Analysis of Biochemical Passive Reactors During Acid Mine Drainage Bioremediation Reveals Key Co-selected Metabolic Functions.

Acid mine drainage (AMD) is the major pollutant generated by the mining industry, and it is characterized by low pH and high concentration of metals and sulfate. The use of biochemical passive reactors (BPRs) is a promising strategy for its bioremediation. To date, there are various studies describing the taxonomical composition of BPR microbial communities, generally consisting of an assemblage of sulfate-reducing organisms inside Deltaproteobacteria, and a diverse set of anaerobic (ligno)cellulolytic bacteria; however, insights about its functional metagenomic content are still scarce. In previous studies, a laboratory-scale AMD bioremediation using biochemical passive reactors was designed and performed, tracking operation parameters, chemical composition, and changes, together with taxonomic composition of the microbiomes harbored in these systems. In order to reveal the main functional content of these communities, we used shotgun metagenomics analyses to explore genes of higher relative frequencies and their inferred functions during the AMD bioremediation from three BPRs representing the main microbiome compositions detected in the system. Remarkably, genes encoding for two-component regulatory systems and ABC transporters related to metal and inorganic ions, cellulose degradation enzymes, dicarboxylic acid production, and sulfite reduction complex were all detected at increased frequency. Our results evidenced that higher taxonomic diversity of the microbiome was arising together with a functional redundancy of the specific metabolic roles, indicating its co-selection and suggesting that its enrichment on BPRs may be implicated in the cumulative efficiency of these systems.

Acid mine drainage treatment using zero-valent iron nanoparticles in biochemical passive reactors.

Acid mine drainage (AMD) is the major effluent generated from metal and coal mines, causing serious ecological risks and degradation of aquatic habitats and surrounding soil quality. Biochemical passive reactors (BPRs) are an option for improving AMD affected water. This study investigates the effect of the size and concentration of zerovalent iron nanoparticles (nZVI) on the efficiency of batch BPRs during AMD remediation. Atomic force microscopy (AFM) and transmission electron microscopy (TEM) were also used as complementary techniques for the investigation of the changes in microbial cells and nZVI properties after the AMD remediation. The results from the batch experiment showed that the concentration of nZVI increases the pH and decreases ORP during AMD treatment, thus favouring the removal of metals. The results also suggest that metal sulfide precipitation occurred in all the batch with reactive mixture but was greater in reactors amended with nZVI of larger size. This study revealed that the presence of nZVI in the BPR leads to metal removal as well as the inhibition of sulfate-reducing bacteria (SRB) activity. Microscopy study indicated that the addition of nZVI creates a morphological change on certain microorganisms in which the cellular membrane was fully covered with nZVI, inducing cell lysis process. These results show that nZVI is a promising reactive material for the treatment of AMD in BPR systems.

Effect of hydraulic retention time on microbial community in biochemical passive reactors during treatment of acid mine drainage.

The effect of hydraulic retention time (HRT) on the microbial community during acid mine drainage (AMD) treatment was investigated. Physicochemical and molecular (illumina and qPCR) analyses were performed on reactive mixtures collected from seven bioreactors in three-operation period (8, 17 and 36weeks). Long HRT (4day) favored the relative abundance of SRB, causing the increase of residual sulfides and short HRT (1day) affected the anaerobic conditions of the bioreactors and favored the presence the acidophilic chemolithotrophic microorganisms. Besides qPCR indicated that genes related to cellulose degradation were present in low copy numbers and were affected by the HRT. Finally, environmental factors (pH, organic source, metal sulfides, and sulfate concentrations) had significant impact on relative abundance of the phylogenetic lineages, rather than the types of lineages present in the reactive mixture. The findings of this study indicate that HRT affects the stability of passive bioreactors and their microbial communities.

Prevención de drenajes ácidos de mina utilizando compost de champiñón como enmienda orgánica / Acid mine drainage prevention using mushroom compost as organic amendment

Resumen Los drenajes ácidos de mina (DAM) son vertimientos con bajo pH, alta concentración de metales y sulfato. Son considerados el mayor problema ambiental de la industria minera y prevenir su formación es la mejor alternativa ambiental y económica. En este estudio, se evaluó el compost de champiñón como enmienda de carbono orgánico para prevenir la formación de DAM. Se construyeron tres celdas en tubos de PVC (2,4 L), llenas con 300 g de mezcla de compost de champiñón y estéril de carbón en diferentes proporciones (40:60, 25:70, 60:40) y 400 mL de agua (18,5Ω). Los cambios químicos en el lixiviado, así como la actividad microbiana en las mezclas fueron monitoreados durante 6 semanas. En los lixiviados el oxígeno disuelto (< 2,0 mg L-1) y potencial de óxido reducción (< (100 mV) disminuyeron, mientras el pH (> 6,5) y la alcalinidad (> 1.500 mg CaCO3 L-1) incrementaron. Además, todas las mezclas fueron eficientes en precipitar los metales (Fe2+ > 95%; Mn2+ > 96%; Zn2+ > 52%) y remover sulfato (> 50%). Sin embargo, en la celda que contenía una proporción de compost y estéril de 25:75 se observó una producción significativa de sulfuro y una mayor actividad microbiana, indicando la presencia de bacterias sulfato-reductoras. Los resultados muestran que el compost de champiñón puede ser utilizado como enmienda orgánica de carbón para contrarrestar la formación de DAM y que la mezcla 25:75 puede ser una opción promisoria para usar en campo en el Distrito minero de Zipaquirá (Colombia).

Abstract The Acid mine drainage (AMD) are discharges characterized by low pH and high concentrations of sulfate and metals. AMD is considered as a serious problem of the mining industry and preventing its formation is the best environmental and economical option. Mushroom compost was evaluated as organic carbon amendment to promote sulfate reduction and metal sulfide precipitation during AMD formation. Three PVC cells (2.4 L) were filled with 300 g of the mixture of mushroom compost and coal mining waste in different proportions (40:60, 25:70, 60:40 %) and 400 mL of water (18,5 Ω). The chemical change in the leachates and the microbial activity in the mixtures were evaluated for 6 weeks. In leachates, dissolved oxygen (< 2,0 mg L-1) and redox potential (< (100 mV) decreased while pH (> 6,5) and alkalinity (> 1500 mg CaCO3 L-1) increased. Besides, all mixtures were efficient for metals precipitation (Fe2+ > 95%; Mn2+ > 96%; Zn2+ > 52%) and sulfate reduction (> 50%). However, a significant production of sulfide and a greater microbial activity was observed in the mixture of mushroom compost and coal mining waste 25:75, indicating the presence of sulfate-reducing bacteria. The results showed that mushroom compost could be used as organic carbon amendment to prevent AMD generation and that the mixture 25:75 could be a promising option to be used in Zipaquirá Mining District (Colombia).

Biochemical passive reactors for treatment of acid mine drainage: Effect of hydraulic retention time on changes in efficiency, composition of reactive mixture, and microbial activity.

Biochemical passive treatment represents a promising option for the remediation of acid mine drainage. This study determined the effect of three hydraulic retention times (1, 2, and 4 days) on changes in system efficiency, reactive mixture, and microbial activity in bioreactors under upward flow conditions. Bioreactors were sacrificed in the weeks 8, 17 and 36, and the reactive mixture was sampled at the bottom, middle, and top layers. Physicochemical analyses were performed on reactive mixture post-treatment and correlated with sulfate-reducing bacteria and cellulolytic and dehydrogenase activity. All hydraulic retention times were efficient at increasing pH and alkalinity and removing sulfate (>60%) and metals (85-99% for Fe(2+) and 70-100% for Zn(2+)), except for Mn(2+). The longest hydraulic retention time (4 days) increased residual sulfides, deteriorated the quality of treated effluent and negatively impacted sulfate-reducing bacteria. Shortest hydraulic retention time (1 day) washed out biomass and increased input of dissolved oxygen in the reactors, leading to higher redox potential and decreasing metal removal efficiency. Concentrations of iron, zinc and metal sulfides were high in the bottom layer, especially with 2 day of hydraulic retention time. Sulfate-reducing bacteria, cellulolytic and dehydrogenase activity were higher in the middle layer at 4 days of hydraulic retention time. Hydraulic retention time had a strong influence on overall performance of passive reactors.

Temporal distribution of heavy metal concentrations in oysters Crassostrea rhizophorae from the central Venezuelan coast.

The oyster Crassostrea rhizophorae is a bivalve abundant in Venezuelan estuaries and consumed by local populations. No known values have been reported on trace metals in oysters from the central Venezuelan coast. We report the concentrations of Al, Cd, Co, Cr, Cu, Fe, Mn, Ni, Pb, Sr, V and Zn in the soft parts of C. rhizophorae, which were collected bimonthly between March 2008 and March 2009, at two sampling areas from the Central Venezuelan Coast: Buche estuary and Mochima estuary. Our results show that for each metal there is a similar temporal variation pattern. The concentrations of the heavy metals reported in this work are useful as reliable baselines and can be used for comparison in future environment studies. Concentrations in C. rhizophorae from the Buche estuary can be interpreted to be high on a global scale for Cd, Cu, Ni and Mn, indicating atypically raised bioavailabilities.