Partitioning of nutrient removal contribution between granules and flocs in a hybrid granular activated sludge system.

Sludge granulation in continuous-flow systems is an emerging technology to intensify existing activated sludge infrastructure for nutrient removal. In these systems, the nutrient removal contributions and partitioning of microbial functions between granules and flocs can offer insights into process implementations. To this end, a reactor system that simulates the continuous-flow environment using an equal amount of initial granule and floc biomass was investigated. The two operational strategies for maintaining granule growth in the continuous-flow system were (a) the higher solids retention time (SRT) for the granules versus flocs, as well as (b) selective feeding of carbon to the granules. The SRT of the large granule fractions (>425 µm, LG) and floc/small granule fractions (<425 µm, FSG) were controlled at 20 and 2.7-6.0 days, respectively. Long term operation of the hybrid granule/floc system achieved high PO43- and NH4+ removal efficiencies. Higher polyphosphate-accumulating organisms (PAO) activity was observed in the FSG than LG, while ammonia-oxidizing bacteria (AOB) activities were similar in the two biomass fractions. Nitrite shunt was observed in the FSG, possibly due to out-competition by the high NOB activity in LG. More importantly, washing out the FSG caused a reduction in LG's AOB and PAO activity, indicating a possible dependency of LG on FSG for maintaining its nutrient removal capacity. Our findings highlighted the partitioning and potential competition/cooperation of key microbial functional groups between LG and FSG, facilitating nutrient removal in a hybrid granular activated sludge system, as well as implications for practical application of the treatment platform.

An investigation into the optimal granular sludge size for simultaneous nitrogen and phosphate removal.

An aerobic granular sludge (AGS) pilot plant fed with a mixture of acetate amended centrate and secondary effluent was used to investigate the optimal granule size range for simultaneous nitrification and denitrification (SND) and ortho-phosphate removal. The anaerobic phase was mixed to understand how AGS will perform if integrated with a continuous flow activated sludge system that cannot feed the influent through the settled sludge bed. Five different granule size fractions were taken from the pilot (operated at DO setpoint of 2mgO2/L) and each size was subjected to activity tests in a well-controlled lab-scale AGS reactor at four dissolved oxygen (DO) concentrations of 1, 2, 3, and 4 mgO2/L. The size fractions were: 212 - 600 µm, 600 - 1000 µm, 1000 - 1400 µm, 1400 - 2000 µm, and >2000 µm. The smallest size range (212 - 600 µm) had the highest nitrification and phosphate removal rates at DO setpoints from 1 - 3 mgO2/L, which was attributed to the higher aerobic volume fraction in small granules and hence a higher abundance of phosphorus accumulating organisms (PAO) and ammonia oxidizing bacteria (AOB). In comparison, large granules (>1000 µm) had 1.4 - 4.7 times lower ammonia oxidation rates than the smallest size range, which aligned with their lower AOB abundance relative to granule biomass. The granules with the highest anoxic volume fraction had the highest abundance of nitrite reductase genes (nir gene) but did not show the highest specific nitrogen removal rate. Instead, smaller granules (212 - 600 and 600 - 1000 µm), which had a lower nir gene abundance, had the highest specific nitrogen removal rates (1.2 - 3.1 times higher than larger granules) across all DO values except at 4 mgO2/L. At a DO setpoint of 4 mgO2/L, nitrite production by ammonia oxidation (ammonia monooxygenase) exceeded nitrite reduction by nitrite reductase in granules smaller than 1000 µm, in addition, some denitrifying heterotrophs switched to oxygen utilization in deeper layers hence suppressing denitrification activity. At the DO range of 2 - 4 mg/L, granular size had a greater effect on nutrient removal than DO. Therefore, for AGS developed at an average DO setpoint of 2 mgO2/L, selecting for size fractions in the range of 212 - 1000 µm and avoiding DO values higher than 3 mgO2/L can achieve both a higher nitrogen removal capacity and energy savings. This study is the first to investigate the influence of different DO values on SND and biological phosphorus removal performance of different aerobic granular sludge sizes.

Aerobic granular sludge: Impact of size distribution on nitrification capacity.

The relationship between ammonia oxidation rate, nitrifiers population, and modelled aerobic zone volume in different granule sizes was investigated using aerobic granular sludge from a pilot-scale reactor. The pilot was fed with centrate and secondary effluent amended with acetate as the main carbon source. The maximum specific ammonia oxidation rates and community composition of different aerobic granular sludge size fractions were evaluated by batch tests, quantitative PCR, and genomic analysis. Small (331µm) granules had a 4.72 ± 0.09 times higher maximum specific ammonia oxidizing rate per 1 gVSS, and a 4.05 ± 0.17 times higher specific amoA gene copy number than large (2225µm) granules per 1 gram of wet biomass. However, when related to surface area, small granules had 1.43 ± 0.01 times lower maximum specific ammonia oxidation rate and a 1.66 ± 0.04 times lower specific amoA gene copy number per unit surface than large granules. Experimental results aligned with modeling results in which smaller granules had a higher specific aerobic zone volume to biomass and lower specific aerobic zone volume to surface area. Aerobic granular sludge reactors having the same average diameter of granules may have very different proportions of granule size fractions and hence possess different nitrification rates. Therefore, instead of the commonly reported average granule diameter, a new method was proposed to determine the aerobic volume density per sample, which correlated well with the nitrification rate. This work provides a roadmap to control nitrification capacity by two methods: (a) crushing larger granules into smaller fractions, or (b) increasing the mixed liquor suspended solid concentration to increase the total aerobic zone volume of the system.

Flocs in disguise? High granule abundance found in continuous-flow activated sludge treatment plants.

To date, high performance of full-scale aerobic granular sludge (AGS) technology has been demonstrated on a global scale. Its further integration with existing continuous flow activated sludge (CFAS) treatment plants is the next logical step. All granular sludge reactors operated in sequencing batch reactors (SBR) mode with anaerobic feeding conditions select for growth of phosphorus and glycogen accumulating organisms (PAO and GAO, respectively), which are known to enhance sludge settling characteristics. Therefore, we hypothesized that AGS are commonly present at full-scale CFAS processes with enhanced biological phosphorus removal (EBPR) and low sludge volume index (SVI). This hypothesis was confirmed at 13 EBPR plants, where granules were found present (at plants where SVI was lower than 100 ml/g) with a strong correlation between high granule abundance and low SVI. A wide range of granule abundance was found among the plants, ranging from 0.5% to as high as 80%. Evaluations of the EBPR plant process configurations showed that high granule abundances may be related to selector design features such as high anaerobic food to mass (F/M) ratios, unmixed in-line fermentation, and high influent soluble COD fraction. Granules were also observed at a non-EBPR plant with an aerobic selector receiving high F/M feeds. Quantitative PCR and 16S rRNA gene sequencing analyses revealed higher relative gene abundance of Accumulibacter PAO and Competibacter GAO in the granules over flocs, as well as a correlation between granule abundance and some possible EPS producers such as Flavobacterium and Competibacter. Our results indicated that process configurations that select for slow-growing or EPS-producing heterotrophs play an important role for granule formation in full-scale CFAS systems as previously shown in SBR configurations.

DNA-SIP based genome-centric metagenomics identifies key long-chain fatty acid-degrading populations in anaerobic digesters with different feeding frequencies.

Fats, oils and greases (FOG) are energy-dense wastes that can be added to anaerobic digesters to substantially increase biomethane recovery via their conversion through long-chain fatty acids (LCFAs). However, a better understanding of the ecophysiology of syntrophic LCFA-degrading microbial communities in anaerobic digesters is needed to develop operating strategies that mitigate inhibitory LCFA accumulation from FOG. In this research, DNA stable isotope probing (SIP) was coupled with metagenomic sequencing for a genome-centric comparison of oleate (C18:1)-degrading populations in two anaerobic codigesters operated with either a pulse feeding or continuous-feeding strategy. The pulse-fed codigester microcosms converted oleate into methane at over 20% higher rates than the continuous-fed codigester microcosms. Differential coverage binning was demonstrated for the first time to recover population genome bins (GBs) from DNA-SIP metagenomes. About 70% of the 13C-enriched GBs were taxonomically assigned to the Syntrophomonas genus, thus substantiating the importance of Syntrophomonas species to LCFA degradation in anaerobic digesters. Phylogenetic comparisons of 13C-enriched GBs showed that phylogenetically distinct Syntrophomonas GBs were unique to each codigester. Overall, these results suggest that syntrophic populations in anaerobic digesters can have different adaptive capacities, and that selection for divergent populations may be achieved by adjusting reactor operating conditions to maximize biomethane recovery.

Bioaugmentation with Nitrifying Granules in Low-SRT Flocculent Activated Sludge at Low Temperature.

Nitrifying granules were grown in a sidestream reactor fed municipal anaerobic digestion centrate and added in an initial slug dose and subsequent smaller daily doses to a non-nitrifying mainstream activated sludge system at 12 °C and 2.5-day aerobic solids retention time (SRT) to increase its nitrification capacity. Effluent NH3-N concentrations less than 1 mg/L were achieved with bioaugmentation, and nitrification was immediately lost when granules were removed after 30 days of bioaugmentation. Molecular microbial analyses indicated that nitrifying organisms remained attached to granules in the mainstream system with little loss to the flocculent sludge. Maximum specific nitrification activity of the bioaugmented granules decreased in mainstream treatment but the nitrification capacity remained due to new granule growth in the mainstream. This study demonstrated that bioaugmentation with sidestream nitrifying granules can intensify nitrification capacity in low-SRT, low-temperature flocculent activated sludge systems to achieve low effluent NH3-N concentrations and nitrogen removal.

Comparison of different aerobic granular sludge types for activated sludge nitrification bioaugmentation potential.

Three types of nitrifying granules were grown on media simulating anaerobic digestion dewatering reject water and compared for their potential to increase nitrification capacity when added to mainstream flocculent activated sludge treatment. An advantage of nitrification bioaugmentation with sidestream granules instead of flocculent biomass is that the granules can be selectively maintained at longer retention times than flocs and thus provide higher nitrification capacity from bioaugmentation. The three granule types and feeding conditions were: nitrifying granules with aerobic feeding, nitrifying-denitrifying granules with anoxic feeding, and nitrifying-denitrifying/phosphate-accumulating (NDN-PAO) granules with anaerobic feeding. NDN-PAO granular sludge showed the highest potential for nitrification bioaugmentation due to its better treatment performance, granule physical characteristics, and much greater production of granular mass and nitrification capacity. Dechloromonas-associated organisms were dominant in these granules; Candidatus Accumulibacter-related organisms were also present. Nitrosomonas was the dominant ammonia-oxidizing bacteria, while Candidatus Nitrotoga was an abundant nitrite-oxidizer in all granule types.

Long-chain fatty acid feeding frequency in anaerobic codigestion impacts syntrophic community structure and biokinetics.

This study investigated the impacts of long-chain fatty acid (LCFA) feeding frequencies on microbial community structure, bioconversion kinetics, and process stability during anaerobic codigestion. Parallel laboratory-scale anaerobic codigesters fed with dairy cattle manure were either pulse-fed every two days or continuously-fed daily, respectively, with oleate (C18:1) in incremental step increases over 200 days up to 64% of the influent chemical oxygen demand (COD). The effluent acetate concentration exceeded 3000 mg/L in the continuous-fed codigester at the highest oleate loading rate, but remained below 100 mg/L in the pulse-fed codigester at the end of its 48-hr oleate feed cycle. Maximum substrate conversion rates of oleate (qmax, oleate) and acetate (qmax, acetate) were significantly higher in the pulse-fed codigester compared to the continuous-fed codigester. 16S rRNA gene amplicon sequencing showed that Bacteria and Archaea community profiles diverged based on the codigester LCFA feeding pattern and loading rate. LCFA-degrading Syntrophomonas bacteria were significantly enriched in both LCFA codigesters relative to the control digester. The pulse-fed codigester had the highest community fraction of Syntrophomonas 16S rRNA genes by the end of the experiment with 43% of Bacteria amplicon sequences. qmax, oleate and qmax, acetate values were both significantly correlated to absolute concentrations of Syntrophomonas and Methanosaeta 16S rRNA genes, respectively. Multiple-linear regression models based on the absolute abundance of Syntrophomonas and Methanosaeta taxa provided improved predictions of oleate and acetate bioconversion kinetics, respectively. These results collectively suggest that pulse feeding rather than continuous feeding LCFA during anaerobic codigestion selected for higher microbial bioconversion kinetics and functional stability, which were related to changes in the physiological diversity and adaptive capacity of syntrophic and methanogenic communities.

Microbial community adaptation influences long-chain fatty acid conversion during anaerobic codigestion of fats, oils, and grease with municipal sludge.

Codigesting fats, oils, and greases with municipal wastewater sludge can greatly improve biomethane recovery at wastewater treatment facilities. Process loading rates of fats, oils, and greases have been previously tested with little knowledge of the digester microbial community structure, and high transient fat loadings have led to long chain fatty acid (LCFA) accumulation and digester upsets. This study utilized recently-developed quantitative PCR assays for syntrophic LCFA-degrading bacteria along with 16S amplicon sequencing to relate changes in microbial community structure to LCFA accumulation during transient loading increases to an anaerobic codigester receiving waste restaurant oil and municipal wastewater sludge. The 16S rRNA gene concentration of the syntrophic ß-oxidizing genus Syntrophomonas increased to ∼15% of the Bacteria community in the codigester, but stayed below 3% in the control digester that was fed only wastewater sludge. Methanosaeta and Methanospirillum were the dominant methanogenic genera enriched in the codigester, and together comprised over 80% of the Archaea community by the end of the experimental period. Constrained ordination showed that changes in the codigester Bacteria and Archaea community structures were related to measures of digester performance. Notably, the effluent LCFA concentration in the codigester was positively correlated to the specific loading rate of waste oil normalized to the Syntrophomonas 16S rRNA concentration. Specific loading rates of 0-1.5 × 10(-12) g VS oil/16S gene copies-day resulted in LCFA concentrations below 30 mg/g TS, whereas LCFA accumulated up to 104 mg/g TS at higher transient loading rates. Based on the community-dependent loading limitations found, enhanced biomethane production from high loadings of fats, oils and greases can be achieved by promoting a higher biomass of slow-growing syntrophic consortia, such as with longer digester solids retention times. This work also demonstrates the potential for controlling the loading rate of fats, oils, and greases based on the analysis of the codigester community structure, such as with quantitative PCR measurements of syntrophic LCFA-degrading bacteria abundance.

Monitoring the dynamics of syntrophic ß-oxidizing bacteria during anaerobic degradation of oleic acid by quantitative PCR.

The ecophysiology of long-chain fatty acid-degrading syntrophic ß-oxidizing bacteria has been poorly understood due to a lack of quantitative abundance data. Here, TaqMan quantitative PCR (qPCR) assays targeting the 16S rRNA gene of the known mesophilic syntrophic ß-oxidizing bacterial genera Syntrophomonas and Syntrophus were developed and validated. Microbial community dynamics were followed using qPCR and Illumina-based high-throughput amplicon sequencing in triplicate methanogenic bioreactors subjected to five consecutive batch feedings of oleic acid. With repeated oleic acid feeding, the initial specific methane production rate significantly increased along with the relative abundances of Syntrophomonas and methanogenic archaea in the bioreactor communities. The novel qPCR assays showed that Syntrophomonas increased from 7 to 31% of the bacterial community 16S rRNA gene concentration, whereas that of Syntrophus decreased from 0.02 to less than 0.005%. High-throughput amplicon sequencing also revealed that Syntrophomonas became the dominant genus within the bioreactor microbiomes. These results suggest that increased specific mineralization rates of oleic acid were attributed to quantitative shifts within the microbial communities toward higher abundances of syntrophic ß-oxidizing bacteria and methanogenic archaea. The novel qPCR assays targeting syntrophic ß-oxidizing bacteria may thus serve as monitoring tools to indicate the fatty acid ß-oxidization potential of anaerobic digester communities.

Evaluating the role of point source discharges informs statewide nutrient control policy in Utah.

An evaluation of costs, rate, and environmental impacts of upgrading publically owned treatment works (POTWs) in the State of Utah to four levels of nutrient control allowed a variety of nutrient control policies to be assessed. Upgrade costs and rate impacts indicated that costs would be within a defined range for many POTWs, especially with design capacities greater than 40,000 m3/day (-10 mgd). However, costs were significantly higher for some POTWs with lower design capacities, and nutrient upgrades to the most stringent levels would not be affordable for these communities, representing about 15 percent of the service population. The resulting equity issues can be addressed through hardship grants program and/or regulations based on a trading scheme. Analysis demonstrated that trading offers advantages, including cost efficiency and flexibility to accommodate further nutrient reductions and population growth, and greater ability to interface with urban and rural nonpoint nutrient control. Currently, the State of Utah is establishing technology-based nutrient limits that can be affordably implemented at all POTWs in phases. Additionally, a multi-faceted approach is being evaluated that will consider prioritized watershed-scale strategies, point and nonpoint sources of pollution, ecological and socioeconomic implications, and stakeholder participation in nutrient reduction programs.

Influence of bioselector processes on 17α-ethinylestradiol biodegradation in activated sludge wastewater treatment systems.

The removal of the potent endocrine-disrupting estrogen hormone, 17α-ethinylestradiol (EE2), in municipal wastewater treatment plant (WWTP) activated sludge (AS) processes can occur through biodegradation by heterotrophic bacteria growing on other organic wastewater substrates. Different kinetic and metabolic substrate utilization conditions created with AS bioselector processes can affect the heterotrophic population composition in AS. The primary goal of this research was to determine if these changes also affect specific EE2 biodegradation kinetics. A series of experiments were conducted with parallel bench-scale AS reactors treating municipal wastewater with estrogens at 100-300 ng/L concentrations to evaluate the effect of bioselector designs on pseudo first-order EE2 biodegradation kinetics normalized to mixed liquor volatile suspended solids (VSS). Kinetic rate coefficient (kb) values for EE2 biodegradation ranged from 5.0 to 18.9 L/g VSS/d at temperatures of 18 °C to 24 °C. EE2 kb values for aerobic biomass growth at low initial food to mass ratio feeding conditions (F/Mf) were 1.4 to 2.2 times greater than that from growth at high initial F/Mf. Anoxic/aerobic and anaerobic/aerobic metabolic bioselector reactors achieving biological nutrient removal had similar EE2 kb values, which were lower than that in aerobic AS reactors with biomass growth at low initial F/Mf. These results provide evidence that population selection with growth at low organic substrate concentrations can lead to improved EE2 biodegradation kinetics in AS treatment.

Modeling organic nitrogen conversions in activated sludge bioreactors.

For biological nutrient removal (BNR) systems designed to maximize nitrogen removal, the effluent total nitrogen (TN) concentration may range from 2.0 to 4.0 g N/m(3) with about 25-50% in the form of organic nitrogen (ON). In this study, current approaches to modeling organic N conversions (separate processes vs. constant contents of organic fractions) were compared. A new conceptual model of ON conversions was developed and combined with Activated Sludge Model No. 2d (ASM2d). The model addresses a new insight into the processes of ammonification, biomass decay and hydrolysis of particulate and colloidal ON (PON and CON, respectively). Three major ON fractions incorporated are defined as dissolved (DON) (<0.1 µm), CON (0.1-1.2 µm) and PON (41.2 µm). Each major fraction was further divided into two sub-fractions - biodegradable and non-biodegradable. Experimental data were collected during field measurements and lab experiments conducted at the ''Wschod'' WWTP (570,000 PE) in Gdansk (Poland). The accurate steady-state predictions of DON and CON profiles were possible by varying ammonification and hydrolysis rates under different electron acceptor conditions. With the same model parameter set, the behaviors of both inorganic N forms (NH4-N, NOX-N) and ON forms (DON, CON) in the batch experiments were predicted. The challenges to accurately simulate and predict effluent ON levels from BNR systems are due to analytical methods of direct ON measurement (replacing TKN) and lack of large enough database (in-process measurements, dynamic variations of the ON concentrations) which can be used to determine parameter value ranges.

Estrogen biodegradation kinetics and estrogenic activity reduction for two biological wastewater treatment methods.

Estrogens from anthropogenic and livestock sources are a serious concern for aquatic ecosystems at concentrations less than 1 ng/L Fundamental process parameters to reduce estrogenic activity were investigated for two biotreatment methods: heterotrophic bacterial degradation in municipal activated sludge (AS) and a nitration process that is applicable to high NH4-N wastewaters. Batch tests with estrogen and nitro-estrogen compounds were conducted at nanogram per liter concentrations with mixed liquor from an AS wastewater treatment facility (WWTF) operating at a 3 day solids retention time (SRT) and a membrane bioreactor (MBR) WWTF operating at a 30-40 day SRT. The estrogenic activities of estrone (E1), 17beta-estradiol (E2), and 17alpha-ethinylestradiol (EE2) were reduced 80-97% following nitration. First-order biological degradation rate coefficients (kb) of the nitrated estrogens were 10-50% lower than the parent estrogen compounds. The kb values for EE2 in MBR and AS mixed liquors were similar, 1.67 and 1.63 L/gVSS-day respectively, indicating that the bacteria responsible for EE2 degradation were present at long and short SRTs. The kb values for E1 and E2 were 2 orders of magnitude greater than for EE2. EE2 degradation was 7.5 times faster in the presence of E1 and E2, and no effect was observed with other estrogen mixtures.

Estrogen nitration kinetics and implications for wastewater treatment.

Understanding estrogen-removal mechanisms in wastewater treatment is imperative, as estrogens have environmental effects at trace concentrations. Previous research investigating co-metabolic degradation of 17alpha-ethinylestradiol (EE2) by ammonia-oxidizing bacteria (AOB) revealed that, in batch tests where high nitrite-nitrogen (NO2-N) concentrations occurred as a result of ammonia-nitrogen (NH4-N) oxidation by AOB, an abiotic estrogen nitration reaction actually was occurring--not co-metabolic degradation. This paper addresses nitration kinetics. A first-order abiotic nitration model was developed that predicts nitration of EE2, 17beta-estradiol (E2), and estrone (El) as a function of temperature, pH, estrogen (EE2, E2, and E1), and NO2-N concentration. A contact time of 3.6 to 4.1 days is required for 90% estrogen nitration at 500 mg/L NO2-N and pH 6.4. At 20 degrees C and pH 6.4, the threshold NO2-N concentration for nitration to occur is 9 mg/L; therefore, estrogen nitration is not likely in activated sludge treatment of domestic wastewater, but has potential for high-NH4-N-strength wastewaters.

17alpha-ethinylestradiol transformation via abiotic nitration in the presence of ammonia oxidizing bacteria.

Impacts of trace concentrations of estrogens on aquatic ecosystems are a serious environmental concern, with their primary source being wastewater treatment facility effluents. Increased removal of 17alpha-ethinylestradiol (EE2) has been reported for activated sludge treatment with long enough solids retention time for nitrification. Previous work based on batch tests with Nitrosomonas europaea and nitrifying activated sludge at high EE2 concentrations (>300 000 ng/L) and high NH4-N concentrations (>200 mg/L) has led to the hypothesis that ammonia oxidizing bacteria cometabolically degrade EE2. This work investigated EE2 transformation with N. europaea and Nitrosospira multiformis at environmentally relevant EE2 concentrations and LC-MS-MS to observe transformation products. Degradation of EE2 was not observed in batch tests with no NH4-N addition or with 10 mg/L NH4-N fed daily. At increased NH4-N concentrations (200-500 mg/L) EE2 transformation was observed, but the only detected products were nitrated EE2. Abiotic assays with growth medium confirmed EE2 removal by nitration, which is enhanced at low pH and high NO2-N concentrations. These results suggest that EE2 removal at low concentrations found in municipal treatment activated sludge systems is not due to cometabolic degradation by ammonia oxidizing bacteria, or to abiotic nitration, but most likely due to heterotrophic bacteria.

Effect of anoxic selector configuration on sludge volume index control and bacterial population fingerprinting.

The effect of single-stage and 4-stage anoxic selectors and an anoxic/aerobic sequencing batch reactor (SBR) on the removal of readily biodegradable chemical oxygen demand (rbCOD) and slowly biodegradable COD (sbCOD) and on filamentous growth and sludge settleability was studied. Microbial community fingerprinting in the three selector configurations was done using automated ribosomal intergenic spacer analysis. Nostocoida limicola II and type 1851 filamentous bacteria were observed in all systems. The diluted sludge volume index (DSVI) decreased with increased selector staging. The rbCOD was almost completely removed in all selectors, and the lower DSVI values with increased selector staging were the result of greater sbCOD removal in the selector. The plug-flow kinetics in the SBR aerobic phase also improved DSVI. The bacteria community composition in the 4-stage selector system was found to be more similar to that for the SBR than for the single-stage selector.

Effects of oxygen exposure on anaerobic digester sludge.

Recuperative thickening of anaerobic digester sludge (thickening with solids return) yields increased digester capacity. Common thickening methods cause oxygen exposure to the digester sludge. This study evaluated the effects of various levels of oxygen exposure on the acetoclastic methanogens. Gravity belt thickening had no detrimental effect on the acetoclastic activity. From a 7-day batch test with continuous oxygen exposure of digester sludge, a 12% loss in acetoclastic activity was predicted for a digester with a 20-day solids retention time (SRT) and 100% recycle with recuperative thickening via dissolved air flotation thickening. However, a greater loss (27%) was found from a long-term, bench-scale digester operated under similar conditions. This loss did not affect the digester performance, as measured by volatile solids destruction. This research suggests that recuperative thickening may not affect digester performance at a long SRT with constant operation, but may change the reserve capacity of the anaerobic community.

Growth kinetics and competition between Methanosarcina and Methanosaeta in mesophilic anaerobic digestion.

Methanosarcina species with a high maximum specific growth rate (mumax) and high half-saturation coefficient (KS) and Methanosaeta species with a low mumax and low KS are the only known aceticlastic methanogens. Because of Methanosaeta's low KS, the low acetate concentrations in conventional, mesophilic anaerobic digestion yield Methanosaeta dominance. However, Methanosarcina absorbs increases in acetate more efficiently and thus promotes more stable digestion. This paper tests the hypothesis that decreasing digester feeding frequencies can increase Methanosarcina predominance. Two acetate-fed reactors were established at a 17-day solids retention time. One reactor was fed hourly, and one was fed once daily. Microscopic and molecular methods were used to verify that the hourly fed reactor enriched for Methanosaeta, while the daily fed reactor enriched for Methanosarcina. Growth and substrate-use kinetics were measured for each reactor. A digester overload condition was simulated, and the Methanosarcina-enriched reactor was found to perform better than the Methanosaeta-enriched reactor. These findings indicate that Methanosarcina dominance can be achieved with infrequent feedings, leading to more stable digestion.

Pseudonocardia chloroethenivorans sp. nov., a chloroethene-degrading actinomycete.

A bacterial strain, SL-1T, capable of degrading trichloroethene was isolated from a laboratory enrichment in the Department of Civil and Environmental Engineering, University of Washington, USA. The material in the enrichments was derived from a soil sample from Seattle, WA, USA. Strain SL-1T was capable of using phenol as a source of carbon and energy. Chemotaxonomic, morphological, physiological and phylogenetic analyses showed that strain SL-1T is a member of the genus Pseudonocardia. The ability of strain SL-1T to utilize phenol and degrade trichloroethene, as well as other phenotypic properties and the results from a 16S rRNA phylogenetic analysis, led to the proposal of a novel species, Pseudonocardia chloroethenivorans sp. nov. The type strain is SL-1T (=ATCC BAA-742T=DSM 44698T). Trichloroethene and other chloroethenes are major pollutants at many environmental sites, and P. chloroethenivorans has biodegradation properties that should be of interest to environmental microbiologists and engineers.