Enhanced anaerobic digestion performance via combined solids- and leachate-based hydrolysis reactor inoculation.

Suboptimal conditions in anaerobic digesters (e.g., presence of common inhibitors ammonia and salinity) limit waste hydrolysis and lead to unstable performance and process failures. Application of inhibitor-tolerant inocula improves hydrolysis, but approaches are needed to establish and maintain these desired waste-hydrolyzing bacteria in high-solids reactors. Herein, performance was compared for leach bed reactors (LBRs) seeded with unacclimated or acclimated inoculum (0-60% by mass) at start-up and over long-term operation. High quantities of inoculum (∼60%) increase waste hydrolysis and are beneficial at start-up or when inhibitors are increasing. After start-up (∼112days) with high inoculum quantities, leachate recirculation leads to accumulation of inhibitor-tolerant hydrolyzing bacteria in leachate. During long-term operation, low inoculum quantities (∼10%) effectively increase waste hydrolysis relative to without solids-derived inoculum. Molecular analyses indicated that combining digested solids with leachate-based inoculum doubles quantities of Bacteria contacting waste over a batch and supplies additional desirable phylotypes Bacteriodes and Clostridia.

Temperature impacts on anaerobic biotransformation of LNAPL and concurrent shifts in microbial community structure.

Thermally-enhanced bioremediation is a promising treatment approach for petroleum contamination; however, studies examining temperature effects on anaerobic biodegradation in zones containing light non-aqueous phase liquids (LNAPLs) are lacking. Herein, laboratory microcosm studies were conducted for a former refinery to evaluate LNAPL transformation, sulfate reduction, and methane generation over a one-year period for temperatures ranging from 4 to 40 °C, and microbial community shifts were characterized. Temperatures of 22 and 30 °C significantly increased total biogas generation compared to lower (4 and 9 °C) and higher temperatures (35 and 40 °C; p < 0.1). Additionally, at 22 and 30 °C methane generation commenced ~6 months earlier than for 35 and 40 °C. Statistically significant biodegradation of benzene, toluene and xylenes was observed at elevated temperatures but not at lower temperatures (p < 0.1). Additionally, a novel differential chromatogram approach was developed to overcome challenges associated with resolving losses in complex mixtures of hydrocarbons, and application of this method revealed greater losses of hydrocarbons at 22 and 30 °C as compared to lower and higher temperatures. Finally, molecular biology assays revealed that the composition and activity of microbial communities shifted in a temperature-dependent manner. Collectively, results demonstrated that anaerobic biodegradation processes can be enhanced by increasing the temperature of LNAPL-containing soils, but biodegradation does not simply increase as temperature increases likely due to a lack of microorganisms that thrive at temperatures well above the historical high temperatures for a site. Rather, optimal degradation is achieved by holding soils at the high end of, or slightly higher than, their natural range.

Microbial community acclimation enhances waste hydrolysis rates under elevated ammonia and salinity conditions.

Hydrolysis rates under potentially inhibitory concentrations of ammonia and salinity were investigated for two model feedstocks (manure and food waste). Rates were determined under a range of ammonia and salinity concentrations (1.0-10.0 g TAN [total ammonia nitrogen] L(-1) and 3.9-20.0 g sodium L(-1)) with unacclimated and acclimated microbial inocula. Microbial community changes as a function of acclimation and feedstock were also investigated. Using unacclimated inocula, hydrolysis was found to be severely inhibited for elevated ammonia and salinity (~4 to 10-fold, respectively) or hydrolysis was not detected. However, for inocula acclimated over 2-4 months, statistically significant inhibition generally was not detectable. Molecular analyses demonstrated that microbial community composition changed during acclimation, and bacterial communities under elevated ammonia were distinct from communities under elevated salinity. Feedstock source also had a major influence on bacterial community structure.