Potential of acetic acid to restore methane production in anaerobic reactors critically intoxicated by ammonia as evidenced by metabolic and microbial monitoring.

BACKGROUND: Biogas and biomethane production from the on-farm anaerobic digestion (AD) of animal manure and agri-food wastes could play a key role in transforming Europe's energy system by mitigating its dependence on fossil fuels and tackling the climate crisis. Although ammonia is essential for microbial growth, it inhibits the AD process if present in high concentrations, especially under its free form, thus leading to economic losses. In this study, which includes both metabolic and microbial monitoring, we tested a strategy to restore substrate conversion to methane in AD reactors facing critical free ammonia intoxication. RESULTS: The AD process of three mesophilic semi-continuous 100L reactors critically intoxicated by free ammonia (> 3.5 g_N L-1; inhibited hydrolysis and heterotrophic acetogenesis; interrupted methanogenesis) was restored by applying a strategy that included reducing pH using acetic acid, washing out total ammonia with water, re-inoculation with active microbial flora and progressively re-introducing sugar beet pulp as a feed substrate. After 5 weeks, two reactors restarted to hydrolyse the pulp and produced CH4 from the methylotrophic methanogenesis pathway. The acetoclastic pathway remained inhibited due to the transient dominance of a strictly methylotrophic methanogen (Candidatus Methanoplasma genus) to the detriment of Methanosarcina. Concomitantly, the third reactor, in which Methanosarcina remained dominant, produced CH4 from the acetoclastic pathway but faced hydrolysis inhibition. After 11 weeks, the hydrolysis, the acetoclastic pathway and possibly the hydrogenotrophic pathway were functional in all reactors. The methylotrophic pathway was no longer favoured. Although syntrophic propionate oxidation remained suboptimal, the final pulp to CH4 conversion ratio (0.41 ± 0.10 LN_CH4 g_VS-1) was analogous to the pulp biochemical methane potential (0.38 ± 0.03 LN_CH4 g_VS-1). CONCLUSIONS: Despite an extreme free ammonia intoxication, the proposed process recovery strategy allowed CH4 production to be restored in three intoxicated reactors within 8 weeks, a period during which re-inoculation appeared to be crucial to sustain the process. Introducing acetic acid allowed substantial CH4 production during the recovery period. Furthermore, the initial pH reduction promoted ammonium capture in the slurry, which could allow the field application of the effluents produced by full-scale digesters recovering from ammonia intoxication.

Operation of a pilot-scale lipid accumulation technology employing parameters to select Microthrix parvicella for biodiesel production from wastewater.

Wastewater treatment plants (WWTPs) may play a crucial role in shifting to a zero-emission future by becoming more sustainable and contributing to the circular economy (CE). Recovered lipids from urban sewage can serve as a raw material for biofuel production contributing to a waste reduction, mitigation of natural resources depletion and reinforcing security and energy independence. A novel, pilot-scale lipid accumulation technology (LAT) employing parameters to select M. parvicella for the biofuel/biodiesel production was implemented on a side stream of an urban WWTP. The LAT proved its concept as the average amount of extracted lipids accumulated in the bioreactors was three-fold higher when compared to the lipids existing in activated sludge. The average transesterification of extracted lipids to biodiesel resulted in a 1.6 % yield, meaning that from 1 kg of dried sludge, 16 g of biodiesel could be formed. The biodiesel produced complies with European standard specifications (EN14214).

How microbial community composition, sorption and simultaneous application of six pharmaceuticals affect their dissipation in soils.

Pharmaceuticals may enter soils due to the application of treated wastewater or biosolids. Their leakage from soils towards the groundwater, and their uptake by plants is largely controlled by sorption and degradation of those compounds in soils. Standard laboratory batch degradation and sorption experiments were performed using soil samples obtained from the top horizons of seven different soil types and 6 pharmaceuticals (carbamazepine, irbesartan, fexofenadine, clindamycin and sulfamethoxazole), which were applied either as single-solute solutions or as mixtures (not for sorption). The highest dissipation half-lives were observed for citalopram (average DT50,S for a single compound of 152 ±â€¯53.5 days) followed by carbamazepine (106.0 ±â€¯17.5 days), irbesartan (24.4 ±â€¯3.5 days), fexofenadine (23.5 ±â€¯20.9 days), clindamycin (10.8 ±â€¯4.2 days) and sulfamethoxazole (9.6 ±â€¯2.0 days). The simultaneous application of all compounds increased the half-lives (DT50,M) of all compounds (particularly carbamazepine, citalopram, fexofenadine and irbesartan), which is likely explained by the negative impact of antibiotics (sulfamethoxazole and clindamycin) on soil microbial community. However, this trend was not consistent in all soils. In several cases, the DT50,S values were even higher than the DT50,M values. Principal component analyses showed that while knowledge of basic soil properties determines grouping of soils according sorption behavior, knowledge of the microbial community structure could be used to group soils according to the dissipation behavior of tested compounds in these soils. The derived multiple linear regression models for estimating dissipation half-lives (DT50,S) for citalopram, clindamycin, fexofenadine, irbesartan and sulfamethoxazole always included at least one microbial factor (either amount of phosphorus in microbial biomass or microbial biomarkers derived from phospholipid fatty acids) that deceased half-lives (i.e., enhanced dissipations). Equations for citalopram, clindamycin, fexofenadine and sulfamethoxazole included the Freundlich sorption coefficient, which likely increased half-lives (i.e., prolonged dissipations).

Degradation and enantiomeric fractionation of mecoprop in soil previously exposed to phenoxy acid herbicides - New insights for bioremediation.

Phenoxy acid-contaminated subsoils are common as a result of irregular disposal of residues and production wastes in the past. For enhancing in situ biodegradation at reducing conditions, biostimulation may be an effective option. Some phenoxy acids were marketed in racemic mixtures, and biodegradation rates may differ between enantiomers. Therefore, enantio-preferred degradation of mecoprop (MCPP) in soil was measured to get in-depth information on whether amendment with glucose (BOD equivalents as substrate for microbial growth) and nitrate (redox equivalents for oxidation) can stimulate bioremediation. The degradation processes were studied in soil sampled at different depths (3, 4.5 and 6m) at a Danish urban site with a history of phenoxy acid contamination. We observed preferential degradation of the R-enantiomer only under aerobic conditions in the soil samples from 3- and 6-m depth at environmentally relevant (nM) MCPP concentrations: enantiomer fraction (EF)<0.5. On the other hand, we observed preferential degradation of the S-enantiomer in all samples and treatments at elevated (µM) MCPP concentrations: EF>0.5. Three different microbial communities were discriminated by enantioselective degradation of MCPP: 1) aerobic microorganisms with little enantioselectivity, 2) aerobic microorganisms with R-selectivity and 3) anaerobic denitrifying organisms with S-selectivity. Glucose-amendment did not enhance MCPP degradation, while nitrate amendment enhanced the degradation of high concentrations of the herbicide.

Large-scale bioreactor production of the herbicide-degrading Aminobacter sp. strain MSH1.

The Aminobacter sp. strain MSH1 has potential for pesticide bioremediation because it degrades the herbicide metabolite 2,6-dichlorobenzamide (BAM). Production of the BAM-degrading bacterium using aerobic bioreactor fermentation was investigated. A mineral salt medium limited for carbon and with an element composition similar to the strain was generated. The optimal pH and temperature for strain growth were determined using shaker flasks and verified in bioreactors. Glucose, fructose, and glycerol were suitable carbon sources for MSH1 (µ = 0.1 h(-1)); slower growth was observed on succinate and acetic acid (µ = 0.01 h(-1)). Standard conditions for growth of the MSH1 strain were defined at pH 7 and 25 °C, with glucose as the carbon source. In bioreactors (1 and 5 L), the specific growth rate of MSH1 increased from µ = 0.1 h(-1) on traditional mineral salt medium to µ = 0.18 h(-1) on the optimized mineral salt medium. The biomass yield under standard conditions was 0.47 g dry weight biomass/g glucose consumed. An investigation of the catabolic capacity of MSH1 cells harvested in exponential and stationary growth phases showed a degradation activity per cell of about 3 × 10(-9) µg BAM h(-1). Thus, fast, efficient, large-scale production of herbicide-degrading Aminobacter was possible, bringing the use of this bacterium in bioaugmentation field remediation closer to reality.

Degradation of three benzonitrile herbicides by Aminobacter MSH1 versus soil microbial communities: pathways and kinetics.

BACKGROUND: The herbicide dichlobenil was banned in the European Union after its metabolite 2,6-dichlorobenzamide (BAM) was encountered in groundwater. Owing to structural similarities, bromoxynil and ioxynil might be converted to persistent metabolites in a similar manner. To examine this, we used an indigenous soil bacterium Aminobacter sp. MSH1 which is capable of mineralizing dichlobenil via BAM and 2,6-dichlorobenzoic acid (2,6-DCBA). RESULTS: Strain MSH1 converted bromoxynil and ioxynil to the corresponding aromatic metabolites, 3,5-dibromo-4-hydroxybenzoic acid (BrAC) and 3,5-diiodo-4-hydroxybenzoic acid (IAC) following Michaelis-Menten kinetics (adjusted R(2) between 0.907 and 0.999). However, in contrast to 2,6-DCBA, degradation of these metabolites was not detected in the pure-culture studies, suggesting that they might pose an environmental risk if similar partial degradation occurred in soil. By contrast, experiments with natural soils indicated 20-30% mineralization of ioxynil and bromoxynil within the first week. CONCLUSION: The degradation pathway of the three benzonitriles is initially driven by similar enzymes, after which more specific enzymes are responsible for further degradation. Ioxynil and bromoxynil mineralization in soil is not dependent on previous benzonitrile exposure. The accumulation of dead-end metabolites, as seen for dichlobenil, is not a major problem.