Birgitte Kiær Ahring

Effect of temperature increase from 55 to 65°C on performance and microbial population dynamics of an anaerobic reactor treating cattle manure

The effect of a temperature increase from 55 to 65 degrees C on process performance and microbial population dynamics were investigated in thermophilic, lab-scale, continuously stirred tank reactors. The reactors had a working volume of 31 and were fed with cattle manure at an organic loading rate of 3 g VS/l reactor volume/d. The hydraulic retention time in the reactors was 15 days. A stable reactor performance was obtained for periods of three retention times both at 55 degrees C and 65 degrees C. At 65 degrees C methane yield stabilized at approximately 165ml/g VS/d compared to 200 ml/g VS/d at 55 degrees C. Simultaneously, the level of total volatile fatty acids, VFA, increased from being below 0.3 g/l to 1.8-2.4 g acetate/l. The specific methanogenic activities (SMA) of biomass from the reactors were measured with acetate, propionate, butyrate, hydrogen, formate and glucose. At 65 degrees C. a decreased activity was found for glucose-, acetate-, butyrate- and formate-utilizers and no significant activity was measured with propionate. Only the hydrogen-consuming methanogens showed an enhanced activity at 65 degrees C. Numbers of cultivable methanogens, estimated by the most probable number (MPN) method, were significantly lower on glucose, acetate and butyrate at the increased operational temperature, while the numbers of hydrogenotrophic methanogens remained unchanged. No viable propionate-degrading bacteria were enriched at 65 C. Use of ribosomal oligonucleotide probes showed that an increase in temperature resulted in a decreased contribution of the rRNA of the domain bacteria from 74-79 to 57-62% of the universal probe, while the rRNA of the domain archaea, increased from 18-23 to 34-36%.

Anaerobic codigestion of municipal, farm, and industrial organic wastes : A survey of recent literature

Codigestion of organic wastes is a technology that is increasingly being applied for simultaneous treatment of several solid and liquid organic wastes. The main advantages of this technology are improved methane yield because of the supply of additional nutrients from the codigestates and more efficient use of equipment and cost-sharing by processing multiple waste streams in a single facility. Many municipal wastewater treatment plants (WWTPs) in industrialized countries currently process wastewater sludge in large digesters. Codigestion of organic wastes with municipal wastewater sludge can increase digester gas production and provide savings in the overall energy costs of plant operations. Methane recovery also helps to reduce the emission of greenhouse gases to the atmosphere. The goal of this literature survey was to summarize the research conducted in the last four years on anaerobic codigestion to identify applications of codigestion at WWTPs. Because the solids content in municipal wastewater sludge is low, this survey only focuses on codigestion processes operated at relative low solids content (slurry mode). Semi-solid or solid codigestion processes were not included. Municipal wastewater sludge, the organic fraction of municipal solid waste, and cattle manure (CAM) are the main wastes most often used in codigestion processes. Wastes that are codigested with these main wastes are wood wastes, industrial organic wastes, and farm wastes. These are referred to in this survey as codigestates. The literature provides many laboratory studies (batch assays and bench-scale digesters) that assess the digestibility of codigestates and evaluate the performance and monitoring of codigestion, inhibition of digestion by codigestates, the design of the process (e.g., single-stage or two-stage processes), and the operation temperature (e.g., mesophilic or thermophilic). Only a few reports on pilot- and full-scale studies were found. These evaluate general process performance and pretreatment of codigestates, energy production, and treatment costs.

Biodegradation of phthalate esters during the mesophilic anaerobic digestion of sludge

Phthalic acid esters (PAE) are commonly found in the sludge generated in the wastewater treatment plants. Anaerobic digestion followed by land application is a common treatment and disposal practice of sludge. To date, many studies exist on the anaerobic biodegradation rates of PAE, especially of the easily biodegradable ones, whereas the higher molecular weight PAE have reported to be non-biodegradable under methanogenic conditions. Furthermore, there is no information on the effect of the PAE on the performance of the anaerobic digesters treating sludge. In this study, the anaerobic biodegradation of di-n-butyl phthalate (DBP), di-ethyl phthalate (DEP) and di-ethylhexyl phthalate (DEHP) was investigated and their relative rates of anaerobic degradation were calculated. Also, the biological removal of PAE during the anaerobic digestion of sludge in bench-scale digesters was investigated using DBP and DEHP as model compounds of one biodegradable and one recalcitrant PAE respectively. The degradation of all the PAE tested in this study (DEP, DBP and DEHP) is adequately described by first-order kinetics. Batch and continuous experiments showed that DEP and DBP present in sludge are rapidly degraded under mesophilic anaerobic conditions (a first-order kinetic constant of 8.04 x 10(-2) and 13.69 x 10(-2)-4.35 day(-1) respectively) while DEHP is degraded at a rate between one to two orders of magnitude lower (0.35 x 10(-2)-3.59 x 10(-2) day(-1)). It is of high significance that experiments with anaerobic sludge of different origin (US and Europe) showed that degradation of DEHP occurs under methanogenic conditions. Accumulation of high levels of DEHP (more than 60 mg/l) in the anaerobic digester has a negative effect on DBP and DEHP removal rates as well as on the biogas production.

Purification and characterization of the 3‐chloro‐4‐hydroxy‐phenylacetate reductive dehalogenase of Desulfitobacterium hafniense

The membrane‐bound 3‐chloro‐4‐hydroxyphenylacetate (Cl‐OHPA) reductive dehalogenase from the chlorophenol‐reducing anaerobe Desulfitobacterium hafniense was purified 11.3‐fold to apparent homogeneity in the presence of the detergent CHAPS. The purified dehalogenase catalyzed the reductive dechlorination of Cl‐OHPA to 4‐hydroxyphenylacetate with reduced methyl viologen as the electron donor at a specific activity of 103.2 nkat/mg protein. SDS‐PAGE revealed a single protein band with an apparent molecular mass of 46.5 kDa. The enzyme contained 0.68±0.2 mol corrinoid, 12.0±0.7 mol iron, and 13.0±0.7 mol acid‐labile sulfur per mol subunit. The N‐terminal amino acid sequence of the enzyme was determined and no significant similarity was found to any protein present in the gene bank.

Toxicity of di-(2-ethylhexyl) phthalate on the anaerobic digestion of wastewater sludge

Previous studies on the microbial degradation of individual phthalic acid esters (PAEs) have demonstrated that the compounds with short ester hydrocarbon chains are easily biodegraded and mineralized, but PAEs with long ester chains are less susceptible to degradation and some of them are considered recalcitrant. Moreover, they inhibit methanogenesis. However, studies have not been made on the effect of feeding a combination of recalcitrant and biodegradable PAEs into anaerobic digesters treating wastewater sludge. The present study was conducted with wastewater sludge from the Los Angeles Bureau of Sanitations Hyperion Treatment Plant. Di (2-ethylhexyl) phthalate (DEHP), the most common persistent PAE found in wastewater, and di-n-butyl phthalate (DBP), a common PAE with short ester chains, were sorbed into the sludge fed to a bench-scale digester for a period of 12 weeks. DEHP degradation was always poor, and accumulation of DEHP was correlated with inhibition of the microbial degradation of DBP and with process instability of the test digester. Inhibition of the DBP removal was completely reversed after DEHP addition was discontinued, but biogas production never recovered to the level observed in a control digester. Other process parameters of digester performance were not affected by DEHP accumulation. These results are similar to the toxic effects of long chain fatty acids on sludge digestion, suggesting that DEHP or its degradation products affect all the microbial populations in the anaerobic bioreactor. Our results imply that high levels of DEHP or other recalcitrant PAEs in wastewater sludge are likely to compromise methanogenesis and removal of biodegradable PAEs in sludge digesters.

Hydrogen inhibition of acetate metabolism and kinetics of hydrogen consumption by Methanosarcina thermophila TM-1

Methanosarcina thermophila TM-1 produced hydrogen during growth on acetate, maintaining a concentration of approx. 0.3 μM in the culture, corresponding to a hydrogen partial pressure of approx. 40 Pa. Increasing the partial pressure of hydrogen to 250 Pa and more led to a gradually increasing inhibition of acetate metabolism. No growth was observed when the gas phase contained 2000 Pa or more and acetate metabolism did not occur even after prolonged incubation (more than 2 weeks). M. thermophila was capable of limited consumption of hydrogen. Consumption of low concentrations of hydrogen proceeded simultaneously with acetate utilization. The affinity for hydrogen (Km=5 μM) was within the range normally found for hydrogenutilizing methanogens, while the corresponding Vmax (1.2 μmol hydrogen consumed mg-1 · cells h-1) was orders of magnitude lower.

The genes coding for the hsp70(dnaK) molecular chaperone machine occur in the moderate thermophilic archaeon Methanosarcina thermophila TM-1

The hsp70(dnaK) locus of the moderate thermophilic archaeon Methanosarcina thermophila TM-1 was cloned, sequenced, and tested in vitro to measure gene induction by heat and ammonia, i.e., stressors pertinent to the biotechnological ecosystem of this methanogen that plays a key role in anaerobic bioconversions. The locus genes and organization, 5-grpE-hsp70(dnaK)-hsp40 (dnaJ)-trkA-3, are the same as those of the closely related mesophile Methanosarcina mazei S-6, but different from those of the only other archaeon for which comparable sequence data exist, the thermophile Methanobacterium thermoautotrophicum deltaH, from another genus, in which trkA is not part of the locus. The proteins encoded in the TM-1 genes are very similar to the S-6 homologs, but considerably less similar to the deltaH proteins. The TM-1 Hsp70(DnaK) protein has the 23-amino acid deletion--by comparison with homologs from gram-negative bacteria first described in the S-6 molecule and later found to be present in all homologs from archaea and gram positives. The genes responded to a temperature elevation in a manner that demonstrated that they are heat-shock genes, functionally active in vivo. Ammonia also induced a heat-shock type of response by hsp70(dnaK), and a similar response by trkA. The data suggest that the moderate thermophile TM-1 has an active Hsp70(DnaK)-chaperone machine in contrast to hyperthermophilic archaea, and that trkA is a stress gene, inasmuch as it responds like classic heat-shock genes to stressors that induce a typical heat-shock response.