Effect of Organic Substrate Composition on Microbial Community Structure of Pilot-Scale Biochemical Reactors Treating Mining Influenced Water

Prieto, Maria Virginia; Hiibel, Sage R.; Pereyra, Luciana P.; Pruden, Amy; Reardon, Kenneth F.; Reisman, David

doi:10.21000/jasmr08010878

Cited by 4 publications

(3 citation statements)

References 20 publications

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“…Compared to the other inoculated columns, the EtOH columns contained the fewest mcrA genes (methanogens). This result supports a previous report by Prieto et al (2008), in which methanogens were lowest in concentration in ethanol-fed as compared to lignocellulose-fed BCRs. Suppressed methanogens may be advantageous considering that they compete with SRB for resources.…”

Section: Discussionsupporting

confidence: 93%

Response and Recovery of Sulfate-Reducing Biochemical Reactors From Aerobic Stress Events

Perrault¹,

Pereyra²,

Hiibel³

et al. 2009

JASMR

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Abstract. Microbially-mediated treatment of mining-influenced water (MIW) through the implementation of sulfate-reducing biochemical reactors (BCRs) is an attractive option for passive, in situ remediation with low operating costs and reduced maintenance requirements. However, BCRs can be unpredictable in terms of how they recover from environmental stresses such as oxygen exposure. Previous studies have demonstrated that the inoculum can impact performance positively, suggesting that engineered control of the microbial community structure could improve the resilience of BCRs to stress. The purpose of this study was to determine the effect of bioaugmentation and biostimulation on performance and recovery from oxygen stress. Twelve columns (six conditions in duplicate) were packed with a complex organic substrate (wood chips, etc.), inoculated with dairy manure, and fed simulated acid mine drainage containing Fe, Cd, and Zn at pH 5.5. The conditions tested were: 1.) bioaugmentation with cellulose degraders (CD); 2.) bioaugmentation with sulfate reducers (SRB); 3.) biostimulation with ethanol (EtOH); 4.) biostimulation with carboxymethyl cellulose (CMC); 5.) dairy manure only control (DM); 6.) un-inoculated control (CR). Once the columns reached steady state anaerobic performance, they were exposed to oxygen, allowed to reach steady state again, and then oxygen-exposed for a second, longer time. The results indicate that all columns performed well in terms of pH neutralization and removal of sulfate and heavy metals. However, the cellulose and ethanol biostimulated columns appeared to be the most resilient to oxygen exposure and performed best by the end of the experiment (171 days), while the bioaugmented columns performed similarly to the controls. The microbial communities were evaluated at each steady state using a suite of biomolecular tools, including active community profiling (ACP) and quantitative realtime PCR (Q-PCR) targeting key functional groups of sulfate-reducers, cellulosedegraders, fermenters, and methanogens. It was found that the active microbial community structure was distinct among the six column conditions and that bioaugmentation significantly impacted the kinds and diversity of bacteria present following stress events. Functional gene analysis supported these observations, but provided finer resolution regarding the response of each functional group to stress. Additional

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Section: Discussionsupporting

confidence: 93%

Response and Recovery of Sulfate-Reducing Biochemical Reactors From Aerobic Stress Events

Perrault¹,

Pereyra²,

Hiibel³

et al. 2009

JASMR

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“…For further primer validation, samples collected from two pilot-scale sulfate-reducing biochemical reactors that receive mine drainage from the National Tunnel in Black Hawk, CO (39) 58.8°C, and 60.1°C (mcrA). The optimized annealing temperature was selected based on the presence of one PCR product band of the expected size on a 1.2% agarose gel for the positive controls and the absence of any product in the pure-culture negative controls.…”

Section: Methodsmentioning

confidence: 99%

Detection and Quantification of Functional Genes of Cellulose- Degrading, Fermentative, and Sulfate-Reducing Bacteria and Methanogenic Archaea

Pereyra¹,

Hiibel²,

Riquelme

et al. 2010

Appl Environ Microbiol

134

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Cellulose degradation, fermentation, sulfate reduction, and methanogenesis are microbial processes that coexist in a variety of natural and engineered anaerobic environments. Compared to the study of 16S rRNA genes, the study of the genes encoding the enzymes responsible for these phylogenetically diverse functions is advantageous because it provides direct functional information. However, no methods are available for the broad quantification of these genes from uncultured microbes characteristic of complex environments. In this study, consensus degenerate hybrid oligonucleotide primers were designed and validated to amplify both sequenced and unsequenced glycoside hydrolase genes of cellulose-degrading bacteria, hydA genes of fermentative bacteria, dsrA genes of sulfate-reducing bacteria, and mcrA genes of methanogenic archaea. Specificity was verified in silico and by cloning and sequencing of PCR products obtained from an environmental sample characterized by the target functions. The primer pairs were further adapted to quantitative PCR (Q-PCR), and the method was demonstrated on samples obtained from two sulfate-reducing bioreactors treating mine drainage, one lignocellulose based and the other ethanol fed. As expected, the Q-PCR analysis revealed that the lignocellulose-based bioreactor contained higher numbers of cellulose degraders, fermenters, and methanogens, while the ethanol-fed bioreactor was enriched in sulfate reducers. The suite of primers developed represents a significant advance over prior work, which, for the most part, has targeted only pure cultures or has suffered from low specificity. Furthermore, ensuring the suitability of the primers for Q-PCR provided broad quantitative access to genes that drive critical anaerobic catalytic processes.

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“…These elevated metal concentrations and low pH contribute directly to conditions toxic for aquatic life and exceed the EPA ambient aquatic standards (Table 1, 6.0-6.6 4.8-6.4 6.5-8.5 Temperature 6.2-17.1 4.6-9.2 N/G 2 1 Current National Ambient Water Quality Criteria (U.S. EPA, 2004) 2 Not a goal for this study The BCRs supported active sulfate reducing bacterial communities (Prieto et al 2008) and demonstrated the removal of metals including Fe, Cu, and Zn from National Tunnel MIW over the course of six months in 2006 (Buccambuso et al 2007). Biogenic sulfide and elevated alkalinity were presumed to be responsible for metal removal in the BCRs, primarily by raising MIW pH to circum-neutral values and subsequent precipitation of metals in carbonate and sulfide forms.…”

Section: Introductionmentioning

confidence: 99%

Pilot-Scale Evaluation of Solid and Liquid Phase Organic Substrates Used in Biochemical Reactors for the Treatment of Mining Influenced Water

Venot

Figueroa²,

Wildeman³

et al. 2008

JASMR

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Sulfate-reducing biochemical reactors (BCRs) were installed to provide a basis for substrate selection for a final treatment remedy for mining influenced water (MIW) from the National Tunnel adit. Black Hawk, CO. The MIW was characterized by elevated metal and SO 4 2concentrations as well as pH fluctuations from 4.8-6.4. Three pairs of BCRs were installed in 2006, each of which contained a different mixture of solid or liquid-phase substrates. The above-ground BCRs partially froze during the first winter, prompting a redesign of the hydraulic system in May 2007 and the installation of heating and insulation in preparation for the following winter in October 2007. During the hydraulic system re-design, a fourth pair of reactors that contained CHITOREM ® SC-20 chitin complex was added to provide data on an alternative solid-phase substrate.Results from 2007 suggest that nearly all of the reactors removed greater than 95% of the Fe and Zn and 50-95% of the sulfate from the MIW during September through December 2007. Copper removal was typically above 95%. In addition, chitin complex reactors demonstrated high Mn removal (average removal was 80%) and high alkalinity (average of 4200 mg/L as CaCO 3 ) during the same period. The high alkalinity was partially due to high concentrations of ammonium (up to 450 mg/L as nitrogen). Operational challenges due to winter conditions in November and December 2007 caused significant temperature and flow fluctuations; however, effective removal of metals was still observed. The BCRs will be monitored through summer 2008 and the data will be evaluated in late-summer 2008 to determine which substrates (CHITOREM ® SC-20, ethanol or solid phase organic mixture) are best suited for long-term treatment of the National Tunnel MIW by BCRs.

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Effect of Organic Substrate Composition on Microbial Community Structure of Pilot-Scale Biochemical Reactors Treating Mining Influenced Water

Cited by 4 publications

References 20 publications

Response and Recovery of Sulfate-Reducing Biochemical Reactors From Aerobic Stress Events

Response and Recovery of Sulfate-Reducing Biochemical Reactors From Aerobic Stress Events

Detection and Quantification of Functional Genes of Cellulose- Degrading, Fermentative, and Sulfate-Reducing Bacteria and Methanogenic Archaea

Pilot-Scale Evaluation of Solid and Liquid Phase Organic Substrates Used in Biochemical Reactors for the Treatment of Mining Influenced Water

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