Anna H. Kaksonen

Performance and ethanol oxidation kinetics of a sulfate-reducing fluidized-bed reactor treating acidic metal-containing wastewater.

The treatment of simulated acidic wastewater (pH 2.5–5)containing sulfate (1.0–2.2 g l-1), zinc (15–340 mg l -1) and iron (57 mg l -1) was studied in a sulfate-reducing fluidized-bed reactor (FBR) at 35 °C.The original lactate feed for enrichment and maintenance of the FBRculture was replaced stepwise with ethanol over 50 days. The robustnessof the process was studied by increasing stepwise the Zn, sulfate andethanol feed concentrations and decreasing the feed pH. The following precipitation rates were obtained: 360 mg l -1 d -1 for Zn and 86 mg l -1 d -1 for Fe, with over 99.8% Zn and Fe removal, with a hydraulic retention time of 16 h. Under these conditions, 77–95% of the electrons were accepted by sulfate reduction. The alkalinity produced from ethanol oxidation increased the wastewater pH from 2.5 to 7.5–8.5. Michaelis–Menten constants (Km) determined in batch FBR experiments, were 4.3–7.1 mg l -1 and 2.7–3.5 mg l -1 for ethanol and acetateoxidation, respectively. The maximum oxidation velocities (Vmax)were 0.19–0.22 mg gVS -1 min -1 and0.033–0.035 mg gVS -1 min -1, for ethanol and acetate, respectively. In summary, the FBR process produced a good quality effluent as indicated by its low organic content and Zn and Fe concentrations below0.1 mg l -1.

Sulfate Reduction Potential in Sediments in the Norilsk Mining Area, Northern Siberia

Abstract The purpose of this study was to characterize the distribution and activity of sulfate-reducing bacteria in tailings and sediments impacted by effluents from mining and smelting operations in the Norilsk area in northern Siberia. The Norilsk mining complex involves three smelter operations, a hydrometallurgical plant, and extensive tailings areas located in the permafrost zone. Sulfate reduction rates measured with a 35SO4 2− tracer technique under various in-situ conditions ranged from 0.05 to 30 nmol S cm−3 day−1. Acetate and glucose addition greatly stimulated sulfate reduction, whereas lactate had less effect. The most pronounced stimulation of sulfate reduction (6.5-fold) was observed with phosphate amendment. Most-probable-number (MPN) counts of sulfate-reducing bacteria in media with glucose, ethanol, lactate, and acetate as electron donors were generally highest at around 107 cells ml−1. The actual MPN counts varied with the sample, electron donor, and incubation conditions (pH 7.2 vs. pH 3.5; 28°C vs. 4°C). Enrichment cultures of sulfate-reducing bacteria were established from a sample that showed the highest rate of sulfate reduction. After multiple serial transfers, the dominant sulfate-reducers were identified by fluorescence in situ hybridization using genus and group-specific 16S rRNA-targeted oligonucleotide probes. Desulfobulbus spp. prevailed in ethanol and lactate enrichments and the Desulfosarcina-Desulfococcus group dominated in acetate and benzoate enrichments. Psychrophilic Desulfotalea-Desulfofustis and moderately psychrophilic Desulforhopalus spp. were identified in enrichments incubated at 4°C, but they were also found in mesophilic enrichments.

Culturable Diversity and Community Fatty Acid Profiling of Sulfate-Reducing Fluidized-Bed Reactors Treating Acidic, Metal-Containing Wastewater

Cultivable strains were identified from sulfate-reducing fluidized-bed reactors (FBR) treating acidic metal-containing wastewater. The FBR-communities were further characterized using culture-independent phenotypic markers, phospholipid fatty acid (PLFA) profiling. After morphological screening of 128 bacterial strains and partial sequencing of 55 strains, 17 distinct phylogenetic types were identified and characterized further. A total of 14 and 6 different bacterial strains were isolated from ethanol- and lactate-fed FBRs, respectively. Sequencing of the 16S rRNA gene showed that these strains were affiliated with members of the δ-Proteobacteria, Firmicutes and Bacteroidetes The strains were affiliated with members of the genera Desulfovibrio, Desulfotomaculum, Desulfobulbus, Desulfitobacterium, Clostridium, Caloramator, Oxobacter, and Bacteroides. Many of the strains were only distantly related to previously described species and, thus, may represent novel species or genera. A number of the strains were not detected in previously employed molecular analyses of the FBR communities, and the major component of each FBR as identified in the molecular analyses were not retrieved as cultures in this study. Most of the SRB, and two of the non-SRB utilized ethanol and lactate as a source of carbon and energy, but none of the isolates grew on acetate, an intermediate in the oxidation of ethanol and lactate. PLFA analysis revealed that the FBR community members contained large amounts of saturated fatty acids. Although the PLFA analysis showed some signatures consistent with sulfate-reducing communities, it did not show any substantial difference in the microbial communities between the reactors, an outcome that was quite contrary to the culture-independent phylogenetic analyses.

Biological hydrogen sulfide production in an ethanol–lactate fed fluidized-bed bioreactor

Sulfate-reducing fluidized-bed bioreactor (FBR) fed with ethanol-lactate mixture was operated at 35 degrees C for 540 days to assess mine wastewater treatment, biological hydrogen sulfide production capacity and acetate oxidation kinetics. During the mine wastewater treatment period with synthetic wastewater, the sulfate reduction rate was 62 mmol SO(4)(2-)L(-1)d(-1) and Fe and Zn precipitation rates were 11 mmol Fe L(-1)d(-1) and 1 mmol Zn L(-1)d(-1). After this, the hydrogen sulfide production was optimized, resulting in sulfate reduction rate of 100 mmol SO(4)(2-)L(-1)d(-1) and H(2)S production rate of 73.2 mmol H(2)SL(-1)d(-1). The limiting step in the H(2)S production was the rate of acetate oxidation, being 50 mmol acetate L(-1)d(-1). Therefore, FBR batch assays were designed to determine the acetate oxidation kinetics, and following kinetic parameters were obtained: K(m) of 63 micromol L(-1) and V(max) of 0.76 micromol acetate g VSS(-1)min(-1). The present study demonstrates high-rate hydrogen sulfide production and high-rate mine wastewater treatment with ethanol and lactate fed fluidized-bed bioreactor.

Metabolic and phylogenetic analysis of microbial communities during phytoremediation of soil contaminated with weathered hydrocarbons and heavy metals.

In the current study, the microbial ecology of weathered hydrocarbon and heavy metal contaminated soil undergoing phytoremediation was studied. The relationship of functional diversity, measured as carbon source utilisation in Biolog plates and extracellular enzymatic activities, and genetic diversity of bacteria was evaluated. Denaturing gradient gel electrophoresis was used for community analyses at the species level. Bulk soil and rhizosphere soil from pine and poplar plantations were analysed separately to determine if the plant rhizosphere impacted hydrocarbon degradation. Prevailing microbial communities in the field site were both genetically and metabolically diverse. Furthermore, both tree rhizosphere and fertilisation affected the compositions of these communities and increased activities of extracellular aminopeptidases. In addition, the abundance of alkane hydroxylase and naphthalene dioxygenase genes in the communities was low, but the prevalence of these genes was increased by the addition of bioavailable hydrocarbons. Tree rhizosphere communities had greater hydrocarbon degradation potential than those of bulk soil. Hydrocarbon utilising communities were dominated generally by the species Ralstonia eutropha and bacteria belonging to the genus Burkholderia. Despite the presence of viable hydrocarbon-degrading microbiota, decomposition of hydrocarbons from weathered hydrocarbon contaminated soil over four years, regardless of the presence of vegetation, was low in unfertilised soil. Compost addition enhanced the removal of hydrocarbons.

Low‐temperature (9°C) AMD treatment in a sulfidogenic bioreactor dominated by a mesophilic Desulfomicrobium species

The possibilities for the treatment of low‐temperature mine waste waters have not been widely studied. The amenability of low‐temperature sulfate reduction for mine waste water treatment at 9°C was studied in a bench‐scale fluidized‐bed bioreactor (FBR). Formate was used as the electron and carbon source. The first influent for the FBR was acidic, synthetic waste water containing iron, nutrients, and sulfate, followed by diluted barren bioleaching solution (DBBS). The average sulfate reduction rates were 8u2009mmolu2009L−1u2009day−1 and 6u2009mmolu2009L−1u2009day−1 with synthetic waste water and DBBS, respectively. The corresponding specific activities were 2.4 and 1.6u2009mmol SO u200942− u2009g VSS−1 day−1, respectively. The composition of the microbial community and the active species of the FBR was analyzed by extracting the DNA and RNA, followed by PCR‐DGGE with the universal bacterial 16S rRNA gene primers and dsrB‐primers specific for sulfate‐reducing bacteria. The FBR microbial community was simple and stable and the dominant and active species belonged to the genus Desulfomicrobium. In summary, long‐term operation of a low‐temperature bioreactor resulted in enrichment of formate‐utilizing, psychrotolerant mesophilic sulfate reducing bacteria. Biotechnol. Bioeng. 2009; 104: 740–751

Precipitation of Cu-sulfides by copper-tolerant Desulfovibrio isolates

The purpose of this work was to isolate Cu-tolerant sulfate-reducers that could be used to produce copper sulfides under pure culture conditions. Three sulfate-reducing bacteria were isolated from wastewater effluents of a zinc-smelter in the Urals and their tolerance to copper varied between 325 and 2600 mg Cu l −1 . Analysis of 16S rRNA gene sequences placed the isolates in the genus Desulfovibrio. The isolates showed pronounced saccharolytic growth. Growing cultures precipitated Cu 2+ as covellite (CuS) within less than a week. Extended incubation for 1 month lead to the formation of chalcocite (Cu2S) and chalcopyrite (CuFeS2).

Mine wastewater treatment using Phalaris arundinacea plant material hydrolyzate as substrate for sulfate-reducing bioreactor.

A low-cost substrate, Phalaris arundinacea was acid hydrolyzed (Reed Canary Grass hydrolyzate, RCGH) and used to support sulfate reduction. The experiments included batch bottle assays (35 degrees C) and a fluidized-bed bioreactor (FBR) experiment (35 degrees C) treating synthetic mine wastewater. Dry plant material was also tested as substrate in batch bottle assays. The batch assays showed sulfate reduction with the studied substrates, producing 540 and 350mgL(-1) dissolved sulfide with RCGH and dry plant material, respectively. The soluble sugars of the RCGH presumably fermented into volatile fatty acids and hydrogen, which served as electron donors for sulfate reducing bacteria. A sulfate reduction rate of 2.2-3.3gL(-1)d(-1) was obtained in the FBR experiment. The acidic influent was neutralized and the highest metal precipitation rates were 0.84g FeL(-1)d(-1) and 15mg ZnL(-1)d(-1). The sulfate reduction rate in the FBR was limited by the acetate oxidation rate of the sulfate-reducing bacteria.

Biological iron oxidation and sulfate reduction in the treatment of acid mine drainage at low temperatures

Biological iron oxidation and sulfate reduction in the treatment of acid mine drainage at low temperatures

Characterization of a thermophilic sulfur oxidizing enrichment culture dominated by a Sulfolobus sp. obtained from an underground hot spring for use in extreme bioleaching conditions

A thermoacidophilic elemental sulfur and chalcopyrite oxidizing enrichment culture VS2 was obtained from hot spring run-off sediments of an underground mine. It contained only archaeal species, namely a Sulfolobus metallicus-related organism (96% similarity in partial 16S rRNA gene) and Thermoplasma acidophilum (98% similarity in partial 16S rRNA gene). The VS2 culture grew in a temperature range of 35–76°C. Sulfur oxidation by VS2 was optimal at 70°C, with the highest oxidation rate being 99xa0mg S0xa0l−1xa0day−1. At 50°C, the highest sulfur oxidation rate was 89xa0mg l−1xa0day−1 (in the presence of 5xa0gxa0Cl−xa0l−1). Sulfur oxidation was not significantly affected by 0.02–0.1xa0gxa0l−1 yeast extract or saline water (total salinity of 0.6xa0M) that simulated mine water at field application sites with availability of only saline water. Chloride ions at a concentration above 10xa0gxa0l−1 inhibited sulfur oxidation. Both granular and powdered forms of sulfur were bioavailable, but the oxidation rate of granular sulfur was less than 50% of the powdered form. Chalcopyrite concentrate oxidation (1% w/v) by the VS2 resulted in a 90% Cu yield in 30xa0days.