Effect of didecyl dimethyl ammonium chloride on nitrate reduction in a mixed methanogenic culture.

The effect of the quaternary ammonium compound, didecyl dimethyl ammonium chloride (DDAC), on nitrate reduction was investigated at concentrations up to 100 mg/L in a batch assay using a mixed, mesophilic (35 degrees C) methanogenic culture. Glucose was used as the carbon and energy source and the initial nitrate concentration was 70 mg N/L. Dissimilatory nitrate reduction to ammonia (DNRA) and to dinitrogen (denitrification) were observed at DDAC concentrations up to 25 mg/L. At and above 50 mg DDAC/L, DNRA was inhibited and denitrification was incomplete resulting in accumulation of nitrous oxide. At DDAC concentrations above 10 mg/L, production of nitrous oxide, even transiently, resulted in complete, long-term inhibition of methanogenesis and accumulation of volatile fatty acids. Fermentation was inhibited at and above 75 mg DDAC/L. DDAC suppressed microbial growth and caused cell lysis at a concentration 50 mg/L or higher. Most of the added DDAC was adsorbed on the biomass. Over 96% of the added DDAC was recovered from all cultures at the end of the 100-days incubation period, indicating that DDAC did not degrade in the mixed methanogenic culture under the conditions of this study.     

Population dynamics of anaerobic microbial consortia in thermophilic methanogenic sludge treating paper-containing solid waste.

To maintain a stable thermophilic (55 degrees C) anaerobic digestion treating toilet paper-containing garbage, it is necessary to operate the digester at long hydraulic retention time (HRT) and low organic loading rate (OLR). Critical conditions of the digestion were investigated by operating the digester at HRT 23 days and OLR 3.4 gCOD(Cr)/L/d (R1) or HRT 14 days and OLR 5.6 gCOD/Cr)/L/d (R2) separately. Characteristics of methanogenesis of the two digesters were examined by measuring gas generation volume and volatile fatty acids (VFA) concentration, and the populations of four anaerobic acidogens and three methanogens were analyzed using quantitative PCR method. In digester R1, methanogenic activity was unstable but it could be recovered by stopping feeding as though VFA accumulation occurred. The population of acidogens and two methanogens were maintained at 10(11) - 10(13) copies/L, however, the population of Methanoculleus could not be recovered after methanogenesis recovering. In digester R2, the period of methanogenesis was significantly shorter than that in digester R1. Both the acidogens and the methanogens could not be maintained at a stable concentration. It is suggested that the critical HRT to sustain the population of acidogens in this process should be longer than 14 days and for all kinds of methanogens, HRT should be longer than 23 days.     

Biological wastewater treatment for removal of polymeric resins in UASB reactor: influence of oxygen.

The biological elimination of polymeric resins compounds (PRC) such as acrylic acid and their esters, vinyl acetate and styrene under methanogenic and oxygen-limited methanogenesis conditions was evaluated. Two UASB reactors (A and B) were used and the removal of the organic matter was studied in four stages. Reactor A was used as methanogenic control during the study. Initially both reactors were operated under methanogenic conditions. From the second stage reactor B was fed with 0.6 and 1 mg/L.d of oxygen (O2). Reactor A had diminution in chemical oxygen demand (COD) removal efficiency from 75+/-4% to 37+/-5%, by the increase of PRC loading rate from 750 to 1125 mg COD/L.d. In this reactor there was no styrene elimination. In reactor B the COD removal efficiency was between 73+/-5% and 80+/-2%, even with the addition of O2 and increase of the PRC loading rate, owing to oxygen being used in the partial oxidation of these compounds. In this reactor the yields were modified from 0.56 to 0.40 for CH4 and from 0.31 to 0.60 for CO2. The O2 in low concentrations increased 40.7% the consumption rates of acrylic acid, methyl acrylate and vinyl acetate, allowing styrene consumption with a rate of 0.103 g/L.d. Batch cultures demonstrated that under methanogenic and oxygen-limited methanogenesis conditions, the glucose was not used as an electron acceptor in the elimination of PRC.     

An evaluation of waste gypsum-based precipitated calcium carbonate for acid mine drainage neutralization

Precipitated CaCO(3) compounds recovered from pulped waste gypsum using some carbonate and hydroxide-based reagents were evaluated for their utilization in acid mine drainage (AMD) neutralization. The neutralization potentials, acid neutralization capacities and compositions of the CaCO(3) compounds were determined and compared with some commercial CaCO(3). It was observed that CaCO(3) recovered from waste gypsum using Na(2)CO(3) significantly neutralized AMD compared with commercial CaCO(3) and that recovered using both (NH(4))(2)CO(3) or NH(4)OH-CO(2) reagents. Moreover, a higher acid neutralization capacity of 1,370 kg H(2)SO(4)/t was determined for CaCO(3) recovered from waste gypsum using Na(2)CO(3) compared with an average of 721 and 1,081 kg H(2)SO(4)/t for ammonium-based CaCO(3) and commercial CaCO(3) respectively. The inorganic carbon content for the CaCO(3) recovered using Na(2)CO(3) and ammonium-based reagents of 49 and 34% respectively confirmed their observed neutralization potentials and acid neutralization capacities, while energy dispersive X-ray fluorescence suggested absence of major oxide impurities, with the exception of residual SO(4)(2-) and Na(2)O which still requires further reduction in the respective compounds.     

An extension of the Anaerobic Digestion Model No. 1 to include the effect of nitrate reduction processes.

Nitrate reduction processes were incorporated into the IWA Anaerobic Digestion Model No. 1 (ADM1) in order to account for the effect of such processes on fermentation and methanogenesis. The general structure of the ADM1 was not changed except for modifications related to disintegration and hydrolysis of complex organic matter and decayed biomass. A fraction of butyrate/valerate and propionate degraders was assumed to be the fermentative denitrifiers carrying out fermentation in the absence of N-oxides. Nitrate reduction proceeded in a stepwise manner to nitrite, nitric oxide, nitrous oxide and nitrogen gas using four substrates as electron and/or carbon source. The utilization of the four substrates and N-oxides was based on stoichiometry and kinetics. The inhibitory effect of N-oxides on the methanogens was accounted for by the use of non-competitive inhibition functions. Model simulations were compared with experimental data obtained with a batch, mixed fermenting and methanogenic culture amended with various initial nitrate concentrations.     

Estimating evolution of δ(13)CH(4) during methanization of municipal solid waste based on chemical reactions, isotope accumulation in products and microbial ecology

Natural isotopic composition in substrate may be used to reveal the metabolic pathways of substrate transformation by microbial community. In this paper, a change in δ(13)CH(4) during methanization of reconstituted municipal solid waste was described using a mathematical model based on stoichiometric chemical reactions, equation for the (13)C isotope accumulation in products at the low natural C(13)/C(12) ratio and microbial ecology. A set of experimental data used in the model was taken from Qu et al. (2009a). According to the model, during mesophilic municipal solid waste methanization initially hydrogenotrophic and further aceticlastic methanogenesis dominated. At the final stage hydrogenotrophic methanogenesis followed by acetate oxidation dominated again. In spite of rather high measured values of δ(13)C for CO(2) above -21‰, a sharp decrease in δ(13)CH(4) from -20‰ to -60‰ at the final stage was explained by a larger fractionation against (13)C during methanogenesis from H(2)/H(2)CO(3) due to a kinetic isotope effect when hydrogenotrophic methanogens preferentially take down light (12)C. The model also confirmed that in thermophilic conditions a comparatively stable value of δ(13)CH(4) about -60‰ measured earlier (Qu et al. 2009b) was due to a dominance of hydrogenotrophic methanogenesis during all methanization process of cardboard waste.     

Investigation of the diversity of homoacetogenic bacteria in mesophilic and thermophilic anaerobic sludges using the formyltetrahydrofolate synthetase gene.

Homoacetogenic bacteria are strict anaerobes capable of autotrophic growth on H(2)/CO(2) or CO, and of heterotrophic growth on a wide range of sugars, alcohols, methoxylated aromatic compounds and one carbon compounds, yielding acetate as their sole metabolic end-product. Batch activity tests on anaerobic granular sludge, using H(2)/CO(2) as a substrate and 2-bromoethanesulfonate (BES) as a specific methanogenic inhibitor revealed that H(2)/CO(2) conversion and concomitant acetate production commenced only after a lag period of 60-100 h. This finding suggests that the homoacetogenic population of digester sludge could be maintained by heterotrophic growth on sugars or other organic compounds, rather than by autotrophic growth on H(2)/CO(2). In the present study, two upflow anaerobic sludge bed (UASB) reactors were operated at 37 degrees C and 55 degrees C for two distinct trial periods, each characterised by the application of influents designed to enrich for homoacetogenic bacteria. Specific primers designed for the amplification of the functional gene encoding formyltetrahydrofolate synthetase (FTHFS), a key enzyme in the acetyl-CoA pathway of acetogenesis, were used as a specific probe for acetogenic bacteria. The diversity of acetogens in the granular sludge cultivated in each reactor was revealed by application of FTHFS targeted PCR. Results show that biomass acetogenic composition was dependent upon the operational temperature of the reactor and the substrate supplied as influent.     

Aeration tank odour by dimethyl sulphoxide (DMSO) waste in sewage.

Sewage plants can experience dimethyl sulphide (DMS) odour problems by at least one mg/L dimethylsulphoxide (DMSO) waste residue in plant influent, through a DMSO/DMS reduction mechanism. This bench-scale batch study simulates in bottles the role of poor aeration in wastewater treatment on the DMSO/DMS and sulphate/H2S reduction. The study compares headspace concentrations of sulphide odorants developed by activated sludge (closed bottles, half full) after six hours under anoxic versus anaerobic conditions, with 0 versus 2 mg/L DMSO addition. Anoxic sludge (0.1 - 2 mg/L dissolved oxygen, DO) with DMSO resulted in about 50 ppmv DMS and no other sulphide, while DMSO-free sludge was free of detectable sulphides. Anaerobic sludge (no measurable DO to the point of sulphate reduction) with DMSO resulted in 22/4/37 ppmv of H2S/methanethiol (MT)/DMS, while DMSO-free sludge resulted in 44/8/2 ppmv of H2S/MT/DMS. It is concluded that common "anoxic" aeration tank zones with measurable DO in bulk water but immeasurable DO inside sludge flocs (nitrate reducing) experience DMSO reduction to DMS that is oxidation resistant and becomes the most important odorant. Under anaerobic conditions, H2S from sulphate reduction becomes an additional important odorant. A strategy is developed that allows operators to determine from the quantity of different sulphides whether the DMSO/DMS mechanism is important at their wastewater plant.     

Calcium effect on fermentative hydrogen production in an anaerobic up-flow sludge blanket system.

The effects of calcium ions on a granular fermentative hydrogen production system were investigated in four lab-scale UASB reactors that fed on sucrose (20 g COD/L). The reactors were seeded with anaerobic sewage sludge microflora and operated at a temperature of 35 +/- 1 degrees, pH of 6.7 with hydraulic retention times (HRTs) of 24-6h. The experimental results indicated that calcium ion addition (75 - 150 mg/L) could enhance the granulation and elevate hydrogen production efficiency. However, an overly-high calcium concentration (300 mg-Ca(+2)/L) deteriorated the hydrogen productivity. A calcium concentration of 150 mg-Ca(+2)/L resulted in a peak HP of 3.6 mol H2/mol-sucrose and HPR of 807 mmol-H2/L-d at HRTs of 8 and 6 h, respectively. The EPS concentration of biohydrogenic biomass was higher than that of the aerobic or methanogenic biomass. The protein/carbon-ratio ranged from 0.17 to 0.26%. The multinomial regression analysis shows that the 75 - 150 mg-Ca(+2)/L calcium concentrations and HRT of 6 h were the optimal operating conditions to efficiently produce hydrogen.     

Enhanced degradation of paracetamol by UV-C supported photo-Fenton process over Fenton oxidation

For the treatment of paracetamol in water, the UV-C Fenton oxidation process and classic Fenton oxidation have been found to be the most effective. Paracetamol reduction and chemical oxygen demand (COD) removal are measured as the objective functions to be maximized. The experimental conditions of the degradation of paracetamol are optimized by the Fenton process. Influent pH 3, initial H(2)O(2) dosage 60 mg/L, [H(2)O(2)]/[Fe(2+)] ratio 60 : 1 are the optimum conditions observed for 20 mg/L initial paracetamol concentration. At the optimum conditions, for 20 mg/L of initial paracetamol concentration, 82% paracetamol reduction and 68% COD removal by Fenton oxidation, and 91% paracetamol reduction and 82% COD removal by UV-C Fenton process are observed in a 120 min reaction time. By HPLC analysis, 100% removal of paracetamol is observed at the above optimum conditions for the Fenton process in 240 min and for the UV-C photo-Fenton process in 120 min. The methods are effective and they may be used in the paracetamol industry.     

Ammonia, a selective agent for methane production by syntrophic acetate oxidation at mesophilic temperature.

In biogas processes, methane production from acetate proceeds by either aceticlastic methanogenesis or through syntrophic acetate oxidation (SAO). In the present study, the pathway for methane production from acetate was analysed; i) during a gradual increase of the ammonia concentration (final concentration 7 g NH(4)(+) -N/L) in a semi-continuous lab-scale anaerobic digester (4.3 L), operating at mesophilic temperature (37 degrees C) or ii) in diluted enrichment cultures (100 ml) experiencing a gradual increase in ammonia, sodium, potassium and propionic acid. The pathway for methane formation was determined by calculating the (14)CO(2)/(14)CH(4) ratio after incubating samples with (14)C-2-acetate. In the anaerobic digester, as well as in the enrichment cultures, the (14)CO(2)/(14)CH4 ratio clearly increased with increasing ammonium-nitrogen concentration, i.e. as the ammonia concentration increased, a shift from the aceticlastic mechanism to the syntrophic pathway occurred. The shift was very distinct and occurred as the NH(4)(+) -N concentration rose above 3 g/l. No shift in pathway was seen during increasing concentrations of sodium, potassium or propionic acid. The shift to SAO in the biogas digester resulted in a twofold decrease in the specific gas and methane yield.     

Advanced H2O2 oxidation for diethyl phthalate degradation in treated effluents: effect of nitrate on oxidation and a pilot-scale AOP operation

One of the objectives of this study was to delineate the effect of nitrate on diethyl phthalate (DEP) oxidation by conducting a bench-scale ultraviolet (UV)/H2O2 and O3/H2O2 operations as suggested in a previous study. We also aim to investigate DEP oxidation at various UV doses and H2O2 concentrations by performing a pilot-scale advanced oxidation processes (AOP) system, into which a portion of the effluent from a pilot-scale membrane bioreactor (MBR) plant was pumped. In the bench-scale AOP operation, the O3 oxidation alone as well as the UV irradiation without H2O2 addition could be among the desirable alternatives for the efficient removal of DEP dissolved in aqueous solutions at a low DEP concentration range of 85+/-15 microg/L. The adverse effect in the UV/H2O2 process was significantly greater than that in the UV oxidation alone, and its oxidation was almost halved by the nitrate. However, the nitrate clearly enhanced the DEP oxidation in the O3 oxidation and O3/H2O2 process. Especially, the addition of nitrate almost doubled the DEP oxidation efficiency in the O3/H2O2 process. The series of pilot-scale AOP operations confirmed that about 30-50% of DEP dissolved in the treated MBR effluent streams was, at least, oxidized by the O3 oxidation alone as well as the UV irradiation without H2O2 addition. The UV photolysis of H2O2 was most effective for DEP degradation with an H2O2 concentration of 40 mg/L at a UV dose of 500 mJ/cm2.     

Denitrification with epsilon-caprolactam by acclimated mixed culture and by pure culture of bacteria isolated from polyacrylonitrile fibre manufactured wastewater treatment system.

The aim of this study is to isolate denitrifying bacteria utilizing epsilon-caprolactam as the substrate, from a polyacrylonitrile fibre manufactured wastewater treatment system. The aim is also to compare the performance of PAN (polyacrylonitrile) mixed bacteria cultures acclimated to epsilon-caprolactam and isolated pure strain for treating different initial epsilon-caprolactam concentrations from synthetic wastewater under anoxic conditions. The result showed that the PAN mixed bacteria cultures acclimated to epsilon-caprolactam could utilize 1538.5 mg/l of epsilon-caprolactam as a substrate for denitrification. Sufficient time and about 2200 mg/l of nitrate were necessary for the complete epsilon-caprolactam removal. Paracoccus thiophilus was isolated from the polyacrylonitrile fibre manufactured wastewater treatment system and it could utilize 1722.5 mg/l of epsilon-caprolactam as a substrate for denitrification. About 3500 mg/l of nitrate was necessary for the complete removal of epsilon-caprolactam. When the initial epsilon-caprolactam concentration was below 784.3 mg/l, the removal efficiency of epsilon-caprolactam by Paracoccus thiophilus was better than that for the PAN mixed bacteria cultures. The growth of Paracoccus thiophilus was better. However, when the initial epsilon-caprolactam concentration was as high as 1445.8 mg/l, both the epsilon-caprolactam removal efficiency by Paracoccus thiophilus and Paracoccus thiophilus specific growth rate were similar to the PAN mixed bacteria cultures.     

Pilot-scale two-stage process: a combination of acidogenic hydrogenesis and methanogenesis.

This study was performed to optimize both acidogenic hydrogenesis and methanogenesis, and then to develop a pilot-scale two-stage process producing not only CH4 but also H2. Firstly, acidogenic hydrogenesis of food waste was examined in pilot-scale leaching-bed reactors using dilution rate (D) as a tool to improve the environmental conditions. The maximum efficiency of 71.4% was obtained by adjusting D from 4.5 to 2.5 d(-1) depending on the state of degradation. Secondly, the wastewater from acidogenic hydrogenesis was converted to CH4 in a pilot-scale UASB reactor. The COD removal efficiency exceeded 95% up to the loading rates of 13.1 g COD/Ld, which corresponded to HRT of 0.25 d (6 h). Lastly, a pilot-scale two-stage process was devised based on a combination of acidogenic hydrogenesis and methanogenesis. Over 120 days, the pilot-scale process resulted in large VS reduction of 70.9% at the high loading rate of 12.5 kg VS/m3/d in a short SRT of 8 days.     

Leachate pre-treatment strategies before recirculation in landfill bioreactors.

Nitrified leachate recirculation represents a promising strategy for a more sustainable landfill management. Our objective was to determine the reactions involved in nitrate reduction in municipal solid waste batch biodegradation tests. Anaerobic digestion of waste in the three control reactors showed a good reproducibility. In two test reactors, nitrate was added at various moments of the waste degradation process. We observed that: (1) H2S concentration controlled the nitrate reduction pathway: above a certain threshold of H2S, dissimilatory nitrate reduction to ammonium (DNRA) replaced denitrification. (2) N2O/N2 ratio varied with the organic carbon concentration: the lower the easily biodegradable carbon concentration, the higher the N2O/N2 ratio. (3) N2 was consumed after denitrification. The possibility of a nitrogen fixation reaction in the presence of NH4 is discussed. Nitrified leachate recirculation during acidogenesis should be avoided because of higher H2S production which could induce DNRA.     

Simulation of denitrification in groundwater from Chaohu Lake Catchment,China

The eutrophication of Chaohu Lake in China is mainly attributed to nitrate inflow from non-point sources in the lake catchment. In this study,biological nitrate reduction from groundwater in the Chaohu Lake Catchment was investigated under laboratory conditions in a continuous upflow reactor. Sodium acetate served as the carbon source and electron donor. Results showed that a carbon-to-nitrogen(C/N) molar ratio of 3:1 and hydraulic retention time(HRT) of 8 d could achieve the most rapid nitrate nitrogen(NO_3~--N) depletion(from 100 mg/L to 1 mg/L within120 h). This rate was confirmed when field groundwater was tested in the reactor, in which a NO_3~--N removal rate of 97.71% was achieved(from60.35 mg/L to 1.38 mg/L within 120 h). Different levels of the initial NO_3~--N concentration(30, 50, 70, and 100 mg/L) showed observable influence on the denitrification rates, with an overall average NO_3~--N removal efficiency of 98.25% at 120 h. Nitrite nitrogen(NO_2~--N)accumulated in the initial 12 h, and then kept decreasing, until it reached 0.0254 mg/L at 120 h. Compared with the initial value, there was a slight accumulation of 0.04 mg/L for the ammonia nitrogen(NH4-N) concentration in the effluent, which is, however, less than the limit value.These results can provide a reference for evaluating performance of denitrification in situ.     

Characteristics of granular sludge in a single upflow sludge blanket reactor treating high levels of nitrate and simple organic compounds.

Simultaneous denitrification and methanogenesis were accomplished in a single upflow sludge blanket (USB) reactor. More than 99% and 95% of nitrate and chemical oxygen demand (COD) removal rates were obtained at a loading of 600 mg NO3-N/L x d and 3,300 mg COD/L x d, respectively. The specific denitrification rate (SDR) increased as COD/NO3-N ratios decreased. Maximum SDR with acetate could reach 1.05 g NO3-N/gVSS x d. Significant sludge flotation was observed at the top of the reactor due to the change of microbial composition and the formation of hollow granules. Granules became fluffy and buoyant due to the growth of denitrifiers. Microscopic examination showed that granules exhibited layered structure and they were mainly composed of Methanosarcina sp., Pseudomonas sp., and rod-shaped bacteria.     

Microbial characteristics of a methanogenic phenol-degrading sludge.

Microbial properties of a methanogenic granular phenol-degrading sludge were characterized using the 16S rRNA/DNA-based techniques, including polymerase chain reaction (PCR) amplification, cloning, DNA sequencing, and fluorescence in situ hybridization (FISH). The sludge was sampled from an upflow anaerobic sludge blanket reactor, which removed 98% of phenol (up to 1260 mg/l) in wastewater at 26 degrees C with 12 hours of hydraulic retention. Based on DNA analysis, the Eubacteria in the sludge was composed of 13 operational taxonomy units (OTUs). Two OTUs, one resembling Clostridium and the other remotely resembling Desulfotomaculum, were likely responsible for the conversion of phenol to benzoate, which was further degraded by five Syntrophus-resembling OTUs to acetate and H2/CO2; methanogens lastly converted acetate and H2/CO2 into methane. The role of six remaining OTUs remains unclear. Overall, the sludge was composed of 26 +/- 6% Eubacteria and 74 +/- 9% methanogens, of which 54 +/- 6% were acetotrophic Methanosaetaceae, 14 +/- 3% and 3 +/- 2% were hydrogenotrophic Methanomicrobiales and Methanobacteriaceae, respectively.     

Anaerobic biodegradability of Tween surfactants used as a carbon source for the microbial reductive dechlorination of hexachlorobenzene.

Three structurally-related, nonionic, polysorbate surfactants (Tween 60, 61, and 65) were used as the sole carbon source to sustain the microbial, sequential reductive dechlorination of hexachlorobenzene (HCB) in a mixed, methanogenic culture derived from a contaminated estuarine sediment. The surfactants were partially degraded and fermented to methane with no measurable accumulation of volatile fatty acids, indicating that methanogenesis was rapid relative to the rates of hydrolysis and acidogenesis. Addition of the methanogenesis inhibitor 2-bromoethanesulfonic acid resulted in acetate accumulation without impact on the sequential dechlorination of HCB. An anaerobic biodegradability assay was performed and the following data were obtained for the Tween 60, 61, and 65, respectively: 53, 62, and 62% COD destruction; 35, 57, and 48% COD to methane conversion; and 38, 38, and 45% COD to acetate conversion. These data suggest that the hydrophobic moiety (stearate) of the surfactants was preferentially degraded, most likely through beta-oxidation, to acetate and ultimately to methane and carbon dioxide. Between 38 and 47% of the initial surfactant COD remained after 46 d incubation, which most likely corresponds to the hydrophilic polyoxyethylene moiety. An anaerobic biodegradation pathway of the Tween surfactants is proposed.     

Effect of pH on nitrate and selenate reduction in flue gas desulfurization brine using the H2-based membrane biofilm reactor (MBfR)

Increased tightening of air regulations is leading more electric utilities to install flue gas desulfurization (FGD) systems. These systems produce brine containing high concentrations of nitrate, nitrite, and selenate which must be removed before discharge. The H2-based membrane biofilm reactor (MBfR) was shown to consistently remove nitrate, nitrite, and selenate at high efficiencies. The maximum selenate removal flux reached 362 mgSe m(-2)d(-1) and was higher than that observed in earlier research, which shows continual improvement of the biofilm for selenate reduction. A low pH of 6.8 inhibited precipitation when treating actual FGD brine, yet did not inhibit removal. SO4(2-) was not removed and therefore did not compete with nitrate, nitrite, and selenate reduction for the available H2.