Continuous hydrogen and methane production in a two-stage cheese whey fermentation system.

The feasibility of integrating biological hydrogen and methane production in a two-stage process using mixed cultures and cheese whey powder (CWP) as substrate was studied. The effect of operational parameters such as hydraulic retention time (HRT) and organic loading rate (OLR) on the volumetric hydrogen (VHPR) and methane (VMPR) production rates was assessed. The highest VHPR was 28 L H2/L/d, obtained during stable operation in a CSTR at HRT and OLR of 6 h and 142 g lactose/ L/d, respectively. Moreover, hydrogen (13 L/L/d) was produced even at HRT as low as 3.5 h and OLR of 163 g lactose/L/d, nonetheless, the reactor operation was not stable. Regarding methane production in an UASB reactor, the acidified effluent from the hydrogen-producing bioreactor was efficiently treated obtaining COD removals above 90% at OLR and HRT of 20 g COD/L/d and 6 h, respectively. The two-stage process for continuous production of hydrogen and methane recovered over 70% of the energy present in the substrate. This study demonstrated that hydrogen production can be efficiently coupled to methane production in a two-stage system and that CWP is an adequate substrate for energy production.

Catalytic effects of different redox mediators on the reductive decolorization of azo dyes.

The catalytic effects of redox mediators, with distinct standard redox potentials (E'0), were evaluated on the first-order rate constant of decolorization (Kd) of recalcitrant azo dyes by an anaerobic granular sludge. The dyes studied included mono-azo (Reactive Orange 14, RO14), di-azo (Direct Blue 53, DB53), and tri-azo (Direct Blue 71, DB71) compounds. Toxicity and auto-catalytic aspects seemed to play a role in determining the rate of decolorization. Addition of riboflavin, anthraquinone-2,6-disulphonate (AQDS) or lawsone as a redox mediator, increased the Kd value for all dyes studied, although their impact varied in every case. Kd values were increased from 1.1-fold up to 3.8-fold depending on the redox mediator applied. Moreover, catalysts with moderately similar E'0 value caused distinct stimulation on the rate of decolorization. These results should be considered for selecting the proper redox mediator to be applied during the anaerobic treatment of textile wastewaters and effluents containing electron-withdrawing pollutants, such as nitro-aromatic and polychlorinated compounds.

The role of sulphate reduction on the reductive S decolorization of the azo dye reactive orange 14.

The aim of this study was to investigate the impact of a broad range of sulphate concentrations (0-10g SO4(-2) L(-1)) on the reduction of an azo dye (reactive orange 14 (RO14)) by an anaerobic sludge. An increase in the sulphate concentration generally stimulated the reduction of RO14 by sludge incubations supplemented with glucose, acetate or propionate as electron donor. Sulphate and azo dye reductions took place simultaneously in all incubations. However, there was a decrease on the rate of decolorization when sulphate was supplied at 10g SO4(-2) L(-1). Abiotic incubations at different sulphide concentrations (0-2.5 g sulphide L(-1)) promoted very poor reduction of RO14. However, addition of riboflavin (20 microM), as a redox mediator, accelerated the reduction of RO14 up to 44-fold compared to a control lacking the catalyst. Our results indicate that sulphate-reduction may significantly contribute to the reduction of azo dyes both by biological mechanisms and by abiotic reductions implicating sulphide as an electron donor. The contribution of abiotic decolorization by sulphide, however, was only significant when a proper redox mediator was included. Our results also revealed that sulphate-reduction can out-compete with azo reduction at high sulphate concentrations leading to a poor decolorising performance when no sufficient reducing capacity is available.

Kinetic and metabolic study of benzene, toluene and m-xylene in nitrifying batch cultures.

The effect of benzene, toluene, and m-xylene (BTX) compounds on the nitrifying activity of a sludge produced in steady-state nitrification was evaluated in batch cultures. Benzene and m-xylene at 10 mg C/L decreased ammonium consumption efficiency by 57% and 26%, respectively, whereas toluene did not affect the ammonium oxidation process. The consumed NH4+-N was totally oxidized to NO3- -N. There was no significant effect at 5 mg C/L of each aromatic compound. BTX (5-20mg C/L) induced a significant decrease in the values for specific rates of NH4+ -N consumption (76-99%) and NO3- -N production (45-98%). At 10 mg C/L of BTX compounds, the inhibition order on nitrate production was: benzene > m-xylene > toluene while at 20 mg C/L, the sequence changed to m-xylene > toluene > benzene for both nitrification inhibition and BTX compounds persistence. At 5 mg C/L of BTX compounds, there was no toxic effect on the sludge whereas from 10 to 50 mgC/L, bacteria did not totally recover their nitrifying activity. At a concentration of 5 mg C/L, toluene was first oxidized to benzyl alcohol, which was later oxidized to butyrate while m-xylene was oxidized to acetate and butyrate.

Oxygen transfer and consumption in a thiosulfate oxidizing bioreactor with sulfur production.

AIMS: To evaluate the contribution of oxygen transfer and consumption in a sulfoxidizing system to increase the elemental sulfur yield from thiosulfate oxidation. METHODS AND RESULTS: A 10 l thiosulfate oxidizing bioreactor with suspended cells operating under microaerophilic conditions and a separated aerator with a variable volume of 0.8--1.7 l were operated with a consortium containing mainly Thiobacillus sp. that oxidizes several sulfide species to elemental sulfur and sulfate. From the gas-liquid oxygen balance, the k(L)a was estimated under different operation conditions. A k(L)a of around 200 h(-1) favoured elemental sulfur production and can serve as scale-up criterion. It was further shown that more than 50% of the oxygen fed to the system was consumed in the aerator. CONCLUSIONS: The performance of the sulfoxidizing system can be improved by controlling oxygen transfer. SIGNIFICANCE AND IMPACT OF THE STUDY: The proposed method for the k(L)a determination was based on the oxygen balance, which incorporates the oxygen concentrations measured in the liquid in steady state, reducing the interference of the response time in the traditional non-steady state methods. This approach can be used to optimize reactors where microaerophilic conditions are desirable.

Sulphide and oxygen inhibition over the anaerobic digestion of organic matter: influence of microbial immobilization type.

Two different types of microbial aggregates (granular sludge and biofilm onto a plastic matrix) were evaluated for their susceptibility to sulphide and dissolved oxygen. Their specific methanogenic and sulphate reducing activities were evaluated separately and simultaneously. Total sulphide concentrations that caused 50% loss of methanogenic activity were 800 and 1250 mg l(-1) and for sulphate reduction 750 and 860 mg l(-1) for the granular sludge and the attached biomass, respectively. Simultaneous methanogenesis and sulphate reduction resulted in an increased tolerance of the sulphate reducing process towards sulphide. Results suggest that methanogenesis in granular sludge is less resistant to sulphide than in the attached biomass structure, whereas in sulphate reduction the attached biomass exhibited a better tolerance to high concentrations of total sulphide than the granular sludge. The better sulphate reducing capacity in the attached biomass may suggest that biomass was selectively attached. The dissolved oxygen concentration that inhibited 50% the methanogenic activity was 4.9 and 6.4 mg l(-1) for the granular sludge and attached biomass, respectively. When methanogenesis and sulphate reduction were carried out simultaneously, the whole process was not affected by the supplied oxygen, as produced sulphide was used by sulphide oxidizing microorganisms thus scavenging oxygen. Results showed that the integration of anaerobic/aerobic conditions in a single bioreactor is quite possible and can be used as a good strategy for the complete transformation of sulphate to elemental sulphur.

Anaerobic biodegradation of BTEX using Mn(IV) and Fe(III) as alternative electron acceptors.

Anaerobic BTEX biodegradation was tested in batch experiments using an anaerobic sediment as inoculum under Fe(III) and Mn(IV) reducing conditions. All BTEX were degraded under the conditions tested, specially under Mn(IV) reducing conditions, where benzene was degraded at a rate of 0.8 micromol l(-1) d(-1), significantly much faster than Fe(III) reducing conditions. Under Fe(III) reducing conditions, ethylbenzene was the compound that degraded at the faster rate of 0.19 micromol l(-1) d(-1). Mn(IV) reducing conditions are energetically more favourable than Fe(III), therefore, BTEX were more rapidly degraded under Mn(IV) reducing conditions. These results represent the first report of the degradation of benzene with Mn(IV) as the final electron acceptor. Amorphous manganese oxide is a natural widely distributed metal in groundwater, where it can be microbiologically reduced, leading to the degradation of monoaromatic compounds.

Simultaneous biodegradation of a phenol and 3,4-dimethylphenol mixture under denitrifying conditions.

Denitrification is a feasible alternative for the treatment of phenolic bearing-wastewaters. The aim of this study was to evaluate the biodegradability of phenolic compounds, as the only carbon and energy source in batch and continuous experiments, using nitrate as a final electron acceptor. Experiments in a continuous upward anaerobic sludge bed reactor demonstrated the possibility of biodegrading a mixture of phenol and 3,4-dimethylphenol at organic loads of 251.6 and 39.5 mg/L-d, respectively, at a COD/NO3(-)-N ratio of 2.57. A nitrogen production efficiency of 86% was obtained according to the nitrate consumption. GC-MS analyses demonstrated that m-cresol was an intermediate of 3,4-dimethylphenol degradation in batch conditions, and had an inhibitory effect on phenol degradation.

Inhibition of the acetoclastic methanogenic activity by phenol and alkyl phenols.

Chemical and petrochemical industries are important sources of aromatic pollutants. Petrochemical processes like caustic washing of middle distillates produce the spent caustic liquors highly concentrated in phenol and alkyl phenols. The anaerobic technology is considered a feasible strategy for petrochemical wastewater pre-treatment although high concentrations of phenol could limit its efficiency. The goal of this work was to determine the toxicity of both selected alkyl phenols and a synthetic "spent-caustic phenols mixture" on the acetoclastic Specific Methanogenic Activity (SMA) of unadapted and phenol-adapted granular sludge. Alkyl phenols were responsible for 50% (IC50) and 100% (IC100) inhibition of the SMA at concentrations ranging from 1.6 to 5.0 mM and from 4.1 to 27.5 mM, respectively, for un-adapted granular sludge. In the case of phenol-adapted granular sludge, the inhibitory concentrations ranged from 1.7 to 14.9 mM and from 4.0 to 83.0 for IC50 and IC100, respectively, highlighting the impact of sludge acclimation. The inhibition produced by 2-ethylphenol was more acute compared to phenol and was not reduced by the phenol acclimation process. The IC50 and IC100 values obtained for the synthetic "spent-caustic phenols mixture" were 9.5 mM and 88.4 mM, respectively. The inhibitory concentrations of phenol compounds were closely correlated with compound apolarity (log P), indicating that the lipophilic character of the tested compounds was responsible for their methanogenic toxicity. An inhibition model is confirmed to estimate the IC50 and IC100.

Toxicity and kinetic parameters of the aerobic biodegradation of the phenol and alkylphenols by a mixed culture.

A mixed culture aerobically metabolized phenol, cresol isomers (o-,m-,p-), 2-ethylphenol and xylenol isomers (2,5-DMP and 3,4-DMP) as the sole carbon and energy source. This culture had a high tolerance towards phenol with values of maximum degradation rate (Vmax) of 47 microM phenol mg-1 protein h-1 and inhibition substrate constant (Ki) of 10 mM. These kinetic parameters were considerably diminished and the toxicity increased with the alkylphenols. For example with 2,5-xylenol, Vmax and Ki values of 0.8 microM 2,5-xylenol mg-1 protein h-1 and 1.3 mM, respectively, were obtained. The cresols were 5-fold more toxic than phenol, whereas 2-ethylphenol and 3,4-xylenol were 11-fold more toxic, and 2,5-xylenol was 34-fold more toxic than phenol.

Pressure-drops control strategy in a fixed-bed reactor.

This paper presents a strategy to control pressure-drops (head loss) in a biofilter designed according to the "Mist-Foam" concept. This concept is based on the mixing of the gaseous substrate and a liquid nutrient solution with an atomization nozzle to generate a mist passing subsequently through a synthetic polyurethane foam. In this type of bioreactor, the microbial growth reduces progressively the empty bed volume of the biofilter and causes an increase in the pressure-drops. This phenomenon can result in a complete clogging of the biofilter. The strategy of pressure-drops control presented here consists of successive interruption of the liquid flow, automatically controlled, resulting in a drying effect of the biomass. Tested during a 160 days experiment, this system has permitted to reduce and stabilize the pressure-drops in a biofilter in which the carrier exhibited a high likelihood of clogging.

Anaerobic biodegradability of alkylphenols and fuel oxygenates in the presence of alternative electron acceptors.

Alkylphenols and fuel oxygenates are important environmental pollutants produced by the petrochemical industry. A batch biodegradability test was conducted with selected ortho-substituted alkylphenols (2-cresol, 2,6-dimethylphenol and 2-ethylphenol), fuel oxygenates (methyl tert-butyl ether, ethyl tert-butyl ether and tert-amylmethyl ether) and tert-butyl alcohol (TBA) as model compounds. The ortho-substituted alkylphenols were not biodegraded after 100 days of incubation under methanogenic, sulfate-, or nitrate-reducing conditions. However, biodegradation of 2-cresol and 2-ethylphenol (150 mg l(-1)) was observed in the presence of Mn (IV) as electron acceptor. The biodegradation of these two compounds took place in less than 15 days and more than 90% removal was observed for both compounds. Mineralization was indicated since no UV-absorbing metabolites accumulated after 23 days of incubation. These alkylphenols were also slowly chemically oxidized by Mn (IV). No biodegradation of fuel oxygenates or TBA (1 g l(-1)) was observed after 80 or more days of incubation under methanogenic, Fe (III)-, or Mn (IV)-reducing conditions, suggesting that these compounds are recalcitrant under anaerobic conditions. The fuel oxygenates caused no toxicity towards acetoclastic methanogens activity in anaerobic granular sludge.

Biotransformation and biodegradation of selected nitroaromatics under anaerobic conditions

The fate of four nitroaromatic compounds (5-nitrosalicylate, 5NSA; 4-nitrobenzoate, 4NBc; 2,4-dinitrotoluene, 2,4DNT; nitrobenzene, NB) was studied in 160 mL laboratory-scale upward-flow anaerobic sludge bed reactors supplied with a mixture of volatile fatty acids and/or glucose as electron donors. All the nitroaromatics were transformed stoichiometrically to their corresponding aromatic amines. After prolonged reactor operation, 5NSA and 4NBc were completely mineralized to CH4 and CO2, whereas 2,4DNT was partially transformed to a nonidentified and nondegradable metabolite. Batch nitro-reduction experiments indicated that the position of the nitro group in relation to the other substituents in the aromatic ring plays a key role in the rate of the nitro-group reduction. The results obtained indicate that certain nitroaromatic compounds can be completely mineralized and serve as a carbon and energy source for anaerobic bacteria.

Biotransformation and biodegradation of N-substituted aromatics in methanogenic granular sludge.

N-Substituted aromatic compounds are environmental contaminants associated with the production and use of dyes, explosives, pesticides and pharmaceuticals. In this article, we examine the potential of anaerobic granular sludge from anaerobic treatment systems towards the detoxification, transformation, and mineralization of nitroaromatic and azo compounds. Nitroaromatics and azo dyes with strong electron withdrawing are highly inhibitory to acetoclastic methanogenic bacteria. However, nitro and azo substituted aromatics are readily reductively detoxified in methanogenic consortia to their respective aromatic amines, which are several orders of magnitude less toxic. This reductive detoxification has allowed the successful operation of anaerobic reactors for the treatment of highly toxic aromatic compounds. In the course of the experiments it was discovered that some aromatic amines were mineralized. These results indicate that some N-substituted aromatic compounds can be completely mineralized and serve as a carbon and energy source for anaerobic bacteria.

Continuous detoxification, transformation, and degradation of nitrophenols in upflow anaerobic sludge blanket (UASB) reactors.

The anaerobic transformation and degradation of nitrophenols by granular sludge was investigated in upflow anaerobic sludge blanket (UASB) reactors continuously fed with a volatile fatty acid (VFA) mixture as the primary substrate. During the start-up, subtoxic concentrations of 2-nitrophenol (2-NP), 4-nitrophenol (4-NP), and 2, 4-dinitrophenol (2, 4-DNP) were utilized. 4-NP and 2, 4-DNP were readily converted to the corresponding aromatic amine; whereas 2-NP was converted to nonaromatic products via intermediate formation of 2-aminophenol (2-AP). These conversions led to a dramatic detoxification of the mononitrophenols because the reactors treated the nitrophenolics at the concentrations which were over 25 times higher than those that caused severe inhibition. VFA removal efficiencies greater than 99% were achieved in both reactors at loading rates greater than 11.4 g COD per liter of reactor volume per day even at volumetric loading of mononitrophenols up to 910 mg/L . d.The sludges obtained from each of the reactors at the end of the continuous experiments were assayed for their specific nitrophenol reducing activity in the presence of different primary substrates. Reduction rates of 45 and 26 mg/g volatile suspended solids per day were observed for 2-NP and 4-NP, respectively, when utilizing the VFA mixture as primary substrate. Hydrogen, an interspecies-reduced compound, and substrates that provide interspecies-reducing equivalents-such as butyrate, propionate, and ethanol stimulated nitrophenol reduction, whereas acetate and methanol did not. Anaerobic batch biodegradability tests with the 2-NP-adapted sludge revealed that its corresponding aromatic amine, 2-AP, was degraded to methane at a specific rate of 14.5 mg/g VSS . d. Acetate was observed to be the major intermediate during 2-AP degradation in the presence of a specific methanogenic inhibitor 2-bromoethanesulfonate. The results of this study indicate that UASB reactors can be applied to rapidly detoxify and, under certain circumstances, degrade nitroaromatic compounds. (c) 1996 John Wiley & Sons, Inc.

Toxicity of N-substituted aromatics to acetoclastic methanogenic activity in granular sludge.

N-substituted aromatics are important priority pollutants entering the environment primarily through anthropogenic activities associated with the industrial production of dyes, explosives, pesticides, and pharmaceuticals. Anaerobic treatment of wastewaters discharged by these industries could potentially be problematical as a result of the high toxicity of N-substituted aromatics. The objective of this study was to examine the structure-toxicity relationships of N-substituted aromatic compounds to acetoclastic methanogenic bacteria. The toxicity was assayed in serum flasks by measuring methane production in granular sludge. Unacclimated cultures were used to minimize the biotransformation of the toxic organic chemicals during the test. The nature and the degree of the aromatic substitution were observed to have a profound effect on the toxicity of the test compound. Nitroaromatic compounds were, on the average, over 500-fold more toxic than their corresponding aromatic amines. Considering the facile reduction of nitro groups by anaerobic microorganisms, a dramatic detoxification of nitroaromatics towards methanogens can be expected to occur during anaerobic wastewater treatment. While the toxicity exerted by the N-substituted aromatic compounds was closely correlated with compound apolarity (log P), it was observed that at any given log P, N-substituted phenols had a toxicity that was 2 orders of magnitude higher than that of chlorophenols and alkylphenols. This indicates that toxicity due to the chemical reactivity of nitroaromatics is much more important than partitioning effects in bacterial membranes.