Stability and performance of Xanthobacter autotrophicus GJ10 during 1,2-dichloroethane biodegradation

A nucleic acid-based approach was used to investigate the dynamics of a microbial community dominated by Xanthobacter autotrophicus GJ10 in the degradation of synthetic wastewater containing 1,2-dichloroethane (DCE). This study was performed over a 140-day period in a nonsterile continuous stirred-tank bioreactor (CSTB) subjected to different operational regimens: nutrient-limiting conditions, baseline operation, and the introduction of glucose as a cosubstrate. The microbial community was analyzed by a combination of fluorescence in situ hybridization (FISH) and denaturing gradient gel electrophoresis (DGGE). Under nutrient-limiting conditions, DCE degradation was restricted, but this did not affect the dominance of strain GJ10, determined by FISH to comprise 85% of the active population. During baseline operation, DCE degradation improved significantly to over 99.5% and then remained constant throughout the subsequent experimental period. DGGE profiles revealed a stable, complex community, while FISH indicated that strain GJ10 remained the dominant species. During the addition of glucose as a cosubstrate, DGGE profiles showed a proliferation of other species in the CSTB. The percentage of strain GJ10 dropped to 8% of the active population in just 5 days, although this did not affect the DCE biodegradation performance. The return to baseline conditions was accompanied by the reestablishment of strain GJ10 as the dominant species, suggesting that this system responds robustly to external perturbations, both at the functional biodegradation level and at the individual strain level.     

Biodegradation of 1,2-dichloroethane (1,2-DCA) by cometabolism in a nitrifying biofilm reactor

Biodegradation of 1,2-dichloroethane (1,2-DCA) by cometabolism was investigated in a continuous-flow nitrifying biofilm reactor over a time period of 218 days. The removal efficiency of 1,2-DCA ranged between 70 and 90%. Using the generation of chloride (Cl⁻) as an indicator of 1,2-DCA mineralization, it was shown that the cometabolic degradation of 1,2-DCA was initiated through oxidative dechlorination. However, Cl⁻ production rates were observed to be lower than the stoichiometric ones which indicated the partial mineralization of 1,2-DCA and the possibility of by-product formation due to incomplete dechlorination. At high 1,2-DCA removal rates, Cl⁻ release seemed to reach a saturation due to 1,2-DCA-dependent inactivation of NH₄–N oxidation. The cometabolic 1,2-DCA degradation capacity of nitrifiers was quite comparable to metabolic 1,2-DCA degradation capacities of pure cultures. A strong linear relationship was found between 1,2-DCA transformation yields and NH₄–N and 1,2-DCA loadings. The effect of 1,2-DCA loading on nitrifier population was monitored using molecular microbiological tools. Long-term input of 1,2-DCA to the biofilm reactor resulted in no significant changes in the quantities of Nitrosomonas, Nitrobacter and Nitrospira species and no shift in the diversities of ammonia oxidizing species. Those findings provide an insight into both the operation and the community structure in natural and managed nitrifying biofilm systems where cometabolic 1,2-DCA takes place.     

Aerobic biodegradation of 1,2-dichloroethane and 1,3-dichloropropene by bacteria isolated from a pulp mill wastewater effluent in South Africa

Large volumes of chlorinated aliphatic hydrocarbons are produced annually for a variety of industrial and commercial uses. They therefore constitute common contaminants of soil and groundwater causing serious environmental and human health problems. In this study, three bacteria were isolated from a pulp mill wastewater effluent in South Africa by culture enrichment technique and characterized for their ability to degrade 1,2-dichloroethane (1,2-DCE) and 1,3-dichloropropene (1,3-DCP). Specific growth rate constants of the organisms ranged between 0.864∼1.094 and 0.530∼0.585 d⁻¹ in 1.2-DCE and 1,3-DCP, respectively, while the degradation rate constant of the compounds ranged variously between 0.33 and 1.006 d⁻¹, with 1,2-DCE generally better utilized than 1,3-DCP. Gas chromatographic analysis revealed up to 75 and 80% removal of 1,2-DCE and 1,3-DCP, respectively, above that observed in the control bottles. These organisms also demonstrated high haloalkane dehalogenase activities with specific dehalogenase activities ranging between 0.25∼0.31 U (mg protein)⁻¹. Analysis of their 16S rRNA gene sequences revealed that they belong to the generaPaenibacillus, Bacillus, andMicrobacterium.     

Isotope analysis as a natural reaction probe to determine mechanisms of biodegradation of 1,2-dichloroethane

1,2-Dichloroethane (1,2-DCA), a chlorinated aliphatic hydrocarbon, is a well-known groundwater contaminant. In this study, fractionation of stable carbon isotope values of 1,2-DCA during biodegradation was used as a novel reaction probe to provide information about the mechanism of 1,2-DCA biodegradation under both aerobic (O2-reducing) and anaerobic (NO3-reducing) conditions. Under O2-reducing conditions, an isotopic enrichment value (epsilon) of -25.8 +/- 1.1 per thousand (+/-95% confidence intervals) was measured for the enrichment culture. Under NO3-reducing conditions, an epsilon-value of -25.8 +/- 3.5 per thousand was measured. The microbial culture produced isotopic enrichment values (epsilon) that are not only large and reproducible, but also are the same whether O2 or NO3 was used as an electron acceptor. Combining data measured under both O2- and NO3-reducing conditions, an isotopic enrichment value (epsilon) of -25.8 +/- 1.6 per thousand is measured for the microbial culture during 1,2-DCA degradation. The epsilon-value can be converted into a kinetic isotope effect (KIE) value to relate the observed isotopic fractionation to the mechanism of degradation. This KIE value (1.05) is consistent with degradation via hydrolytic dehalogenation under both electron-accepting conditions. This study demonstrates the added value of compound-specific isotope analysis not only as a technique to verify the occurrence and extent of biodegradation in the field, but also as a natural reaction probe to provide insight into the enzymatic mechanism of contaminant degradation.     

Comparison between a moving bed membrane bioreactor and a conventional membrane bioreactor on organic carbon and nitrogen removal

A membrane bioreactor filled with carriers instead of activated sludge named a moving bed membrane bioreactor (MBMBR) was investigated for simultaneously removing organic carbon and nitrogen in wastewater. Its performance was compared with a conventional membrane bioreactor (CMBR) at various influent COD/TN ratios of 8.9–22.1. The operational parameters were optimized to increase the treatment efficiency. COD removal efficiency averaged at 95.6% and 96.2%, respectively, for MBMBR and CMBR during the 4 months experimental period. The MBMBR system demonstrated good performance on nitrogen removal at different COD/TN ratios. When COD/TN was 8.9 and the total nitrogen (TN) load was 7.58 mg/l h, the TN and ammonium nitrogen removal efficiencies of the MBMBR were maintained over 70.0% and 80.0%, respectively, and the removed total nitrogen (TN) load reached to 5.31 mg/l h. Multifunctional microbial reactions in the carrier, such as simultaneous nitrification and denitrification (SND), play important roles in nitrogen removal. In comparison, the CMBR did not perform so well. Its TN removal was not stable, and the removed total nitrogen (TN) load was only 1.02 mg/l h at COD/TN ratio 8.9. The specific oxygen utilization rate (SOUR) showed that the biofilm has a better microbial activity than an activated sludge. Nevertheless, the membrane fouling behavior was more severe in the MBMBR than in the CMBR due to a thick and dense cake layer formed on the membrane surface, which was speculated to be caused by the filamentous bacteria in the MBMBR.     

Demonstration of efficient trichloroethylene biodegradation in a hollow‐fiber membrane bioreactor

Rapid cometabolism of trichloroethylene (TCE) by pure cultures of Methylosinus trichosporium OB3b PP358 was demonstrated in a two‐stage hollow‐fiber membrane bioreactor over the course of 3 weeks. PP358 was grown in a continuous‐flow chemostat and circulated through the shell of a hollow‐fiber membrane module (HFMM), while TCE contaminated water (160 to 1450 μg/L) was pumped through the fiber lumen (fiber interior). In parallel‐flow HFMM biological experiments, 82% to 89% of the influent TCE was removed from the lumen (5.1‐min residence time) with 99% of the transferred TCE undergoing biodegradation. Biological experiments in a larger capacity baffled radial‐flow HFMM resulted in 66% to 99% TCE transferred and 93% to 96% TCE biodegradation at lumen residence times of between 1.5 and 3.7 min. Biodegradation was maintained throughout the experiments at pseudo‐first‐order biodegradation rate constants of 0.41 to 2.8 L/mg TSS/day. Best‐fit computer modeling of the baffled radial‐flow biological process estimated mass transfer coefficients as large as 2.7 × 10⁻² cm/min. The computer model was also shown to simulate the experimental results quite well. © 1999 John Wiley & Sons, Inc. Biotechnol Bioeng 62: 681–692, 1999.     

Using volatile organic compounds to enhance atrazine biodegradation in a biobed system

The effect of the terpenes α-pinene, eucalyptol, and limonene, individually and as mixtures, on atrazine (ATZ) biodegradation and on biological activity in a biobed biomixture was evaluated. Additionally, terpenes emitted from the biomixture were captured using solid-phase microextraction. Terpenes added individually at relatively low concentrations (50 μg kg^?1) significantly enhanced ATZ degradation and biological activity during the first incubation days. No significant effect on ATZ degradation was found from adding the terpene mixture, and, interestingly, an inhibitory effect on phenoloxidase activity was found during the first 20 days of incubation when mixed terpenes were present at 100 μg kg^?1. Capturing terpenes demonstrated that during the first hour of incubation a significant fraction of the terpenes was volatilized. These results are the first to demonstrate the feasibility of using terpenes to enhance the degradation of a pesticide. However, successive applications of terpenes or the addition of materials that slowly release terpenes could sustain the ATZ degradation enhancement.     

Microbial community related to volatile organic compound (VOC) emission in household biowaste

Malodorous emissions and potentially pathogenic microorganisms which develop during domestic organic waste collection are not only a nuisance but may also pose health risks. The aim of the present study was to determine whether the presence of specific microorganisms in biowastes is directly related to the composition of the emitted volatile organic compounds (VOCs). The succession of microbial communities during 16 days of storage in organic waste collection bins was studied by denaturing gradient gel electrophoresis (DGGE) of amplified 16S ribosomal DNA in parallel with a classical cultivation and isolation approach. Approximately 60 different bacterial species and 20 different fungal species were isolated. Additionally, some bacterial species were identified through sequencing of excised DGGE bands. Proton transfer reaction mass spectrometry (PTR-MS) was used to detect VOCs over the sampling periods, and co-inertia analyses of VOC concentrations with DGGE band intensities were conducted. Positive correlations, indicating production of the respective VOC or enhancement of microbial growth, and negative correlations, indicating the use of, or microbial inhibition by the respective compound, were found for the different VOCs. Measurement of the VOC emission pattern from a pure culture of Lactococcus lactis confirmed the positive correlations for the protonated masses 89 (tentatively identified as butyric acid), 63 (tentatively identified as dimethylsulfide), 69 (likely isoprene) and 73 (likely butanone).     

A review of the genotoxicity of 1,2-dichloroethane (EDC)

1,2-Dichloroethane (EDC, CAS#107-06-2) is a high production volume halogenated aliphatic hydrocarbon that is used mainly in the manufacture of vinyl chloride. EDC has been found in ambient and residential air samples, as well as in groundwater, surface water and drinking water. EDC has been well-studied in a variety of genotoxicity assays, and appears to involve the metabolic activation of the parent compound. We critically evaluated the genotoxicity data of EDC and its metabolites as part of an evaluation of carcinogenic mechanisms of action of EDC. EDC is genotoxic in multiple test systems via multiple routes of exposure. EDC has been shown to induce DNA adduct formation, gene mutations and chromosomal aberrations in the presence of key activation enzymes (including CYP450s and/or GSTs) in laboratory animal and in vitro studies. EDC was negative for clastogenesis as measured by the micronucleus assay in mice. In general, an increased level of DNA damage is observed related to the GSH-dependent bioactivation of EDC. Increased chromosomal aberrations with increased CYP450 expression were suggestive of a role for the oxidative metabolites of EDC in inducing chromosomal damage. Taken together, these studies demonstrate that EDC exposure, in the presence of key enzymes (including CYP450s and/or GSTs), leads to DNA adduct formation, gene mutations and chromosomal aberrations.     

Comparison of the performance of submerged membrane bioreactor (SMBR) and submerged membrane adsorption bioreactor (SMABR)

This study focuses on comparing the performance of submerged membrane bioreactor (SMBR) and submerged membrane adsorption bioreactor (SMABR) over a period of 20 days at a hydraulic retention time (HRT) of 3.1h. The effects of PAC on critical flux and membrane fouling were also investigated. The SMABR exhibited better results in terms of mixed liquor suspended solids (MLSS) growth, DOC removal (over 96%), COD removal (over 95%), transmembrane pressure (TMP) and oxygen uptake rate. Nearly 100% of bacteria and 100% of total coliforms were removed in both systems. The addition of PAC could maintain the critical flux at a lower TMP value (7.5 kPa), while irreversible fouling caused by PAC occurred when the filtration flux exceeded critical flux.     

Removal of microbial indicators from municipal wastewater by a membrane bioreactor (MBR)

The impact of removable and irremovable fouling on the retention of viral and bacterial indicators by the submerged microfiltration membrane in an MBR pilot plant was evaluated. Escherichia coli, sulphite-reducing Clostridium spores, somatic coliphages and F-specific RNA bacteriophages were used as indicators. The membrane demonstrated almost complete removal of E. coli and sulphite-reducing Clostridium spores. However, there was no correlation with membrane fouling. The phage removal varied in accordance with the irremovable fouling, rising from 2.6 to 5.6 log₁₀ units as the irremovable fouling increased (measured by the change in the transmembrane pressure). In contrast, removable fouling did not have any effect on the retention of viruses by the membrane. These results indicate that irremovable membrane fouling may affect the removal efficiency of MBRs and, therefore, their capacity to ensure the required microbiological standards for the permeate achieved.     

Resolution of 1,2-epoxyhexane by Rhodotorula glutinis using a two-phase membrane bioreactor

Large-scale resolution of epoxides by the yeast Rhodotorula glutinis was demonstrated in an aqueous/organic two-phase cascade membrane bioreactor. Due to the chemical instability and low solubility of epoxides in aqueous phases, an organic solvent was introduced into the reaction mixture in order to enhance the resolution of epoxide. A cascade hollow-fiber membrane bioreactor was used (1) to minimize the toxicity of organic solvents towards the epoxide hydrolase of R. glutinis, and (2) to remove inhibitory amounts of formed diol from the yeast cell containing aqueous phase. Dodecane was selected as a suitable solvent and 1,2-epoxyhexane as a model substrate. By use of this membrane bioreactor, highly concentrated (0.9 M in dodecane) enantiopure (> 98% ee) (S)-1,2-epoxyhexane (6.5 g, 30% yield) was obtained from the racemic mixture. Received: 28 June 1999 / Received revision: 26 August 1999 / Accepted: 3 September 1999     

Kinetics of organic carbon removal by a mixed culture in a membrane bioreactor

The aim of this work was to model the biological activity and anticipate the kinetic behaviour of microorganisms and the overall performance of the process according to a specific model and running parameters. The bacterial inoculum used in these experiments was a mixture of cultures taken from the wastewater treatment plant in Montpellier. The fermentor, used in association with an ultrafiltration separation stage (with a filtration area of 0.2 m²) had a working volume of 15.8 l. For various working conditions (different solid retention times, different hydraulic retention times and substrate concentrations), the biomass concentration and the residual substrate concentration, expressed in terms of dry weight and chemical oxygen demand, respectively, were measured. The basic idea of modelling was related to the concept of maintenance. The coefficient of maintenance, E, and the theoretical conversion yield, y, were therefore calculated. The values of E and y, measured for total cell recycling experiments and for experiments with various solid retention times, remained similar and were found to equal 0.040 mgCOD mgVSS h⁻¹ and 0.36 mgVSS mgCOD⁻¹, respectively. Determining these two constants and modelling the treatment process made it possible to anticipate the optimal biomass concentration for a defined removal efficiency under different steady-state operating conditions.     

Contribution of sludge adsorption and biodegradation to the removal of five pharmaceuticals in a submerged membrane bioreactor

A submerged membrane bioreactor was set up to investigate the removal efficiencies of five pharmaceuticals from synthetic domestic wastewater. Batch experiments were conducted with sterilized sludge and activated sludge to explore the contributions of sludge adsorption and biodegradation for those pharmaceuticals. Notable difference of those pharmaceuticals removal efficiencies was observed, at about 92.2, 90.0, 55.4, 38.5 and 3.2% for acetaminophen, 17β-estradiol, naproxen, diclofenac sodium, and carbamazepine, respectively. Results of batch adsorption experiments via sterilized sludge showed that the removal efficiencies of five pharmaceuticals by sludge adsorption were 7.9, 68.2, 60.1, 40.1 and 71.5%, respectively, which were positively correlated with their octanol–water partition coefficients. Results of batch experiments via activated sludge showed that 83.4% of acetaminophen, 98.0% of 17β-estradiol, and 46.8% of naproxen were removed through the combination of sludge adsorption and biodegradation, while adsorption accumulation in sludge phase was only 1.8, 1.3 and 7.0%. This implies that the removals of these three drugs were mainly achieved by biodegradation. The total removal efficiency of diclofenac sodium was 19.7%, and the contributions of sludge adsorption and biodegradation were 14.9 and 4.8%, which indicated that the removal of diclofenac sodium was mainly achieved by sludge adsorption. The total removal efficiency of carbamazepine was only 8.9% and this implies that neither sludge adsorption nor biodegradation is effective for its removal.     

Origin of volatile organic compound emissions from subarctic tundra under global warming

Warming occurs in the Arctic twice as fast as the global average, which in turn leads to a large enhancement in terpenoid emissions from vegetation. Volatile terpenoids are the main class of biogenic volatile organic compounds (VOCs) that play crucial roles in atmospheric chemistry and climate. However, the biochemical mechanisms behind the temperature‐dependent increase in VOC emissions from subarctic ecosystems are largely unexplored. Using ¹³CO₂‐labeling, we studied the origin of VOCs and the carbon (C) allocation under global warming in the soil–plant–atmosphere system of contrasting subarctic heath tundra vegetation communities characterized by dwarf shrubs of the genera Salix or Betula. The projected temperature rise of the subarctic summer by 5°C was realistically simulated in sophisticated climate chambers. VOC emissions strongly depended on the plant species composition of the heath tundra. Warming caused increased VOC emissions and significant changes in the pattern of volatiles toward more reactive hydrocarbons. The ¹³C was incorporated to varying degrees in different monoterpene and sesquiterpene isomers. We found that de novo monoterpene biosynthesis contributed to 40%–44% (Salix) and 60%–68% (Betula) of total monoterpene emissions under the current climate, and that warming increased the contribution to 50%–58% (Salix) and 87%–95% (Betula). Analyses of above‐ and belowground ^12/13C showed shifts of C allocation in the plant–soil systems and negative effects of warming on C sequestration by lowering net ecosystem exchange of CO₂ and increasing C loss as VOCs. This comprehensive analysis provides the scientific basis for mechanistically understanding the processes controlling terpenoid emissions, required for modeling VOC emissions from terrestrial ecosystems and predicting the future chemistry of the arctic atmosphere. By changing the chemical composition and loads of VOCs into the atmosphere, the current data indicate that global warming in the Arctic may have implications for regional and global climate and for the delicate tundra ecosystems.     

Extraction of volatile fatty acids from diluted aqueous solution using a supported liquid membrane

Experimental and analytical studies on the extraction of volatile fatty acids (VFAs) from an aqueous solution using a supported liquid membrane (SLM) were carried out. Teflon and 20% (w/w) tri-n-octyl phosphine oxide (TOPO) in kerosene were used as the supporting membrane and liquid, respectively. The extraction rate of VFAs transferred from the source across the SLM to the sink was measured. It was observed that only undissociated forms of VFAs can penetrate into the SLM and that the complex formation between TOPO and VFA (1/1 molar ratio) inside the membrane enhanced the transfer rate of VFA across the membrane. This phenomenon was explained by mathematical models based on mass transfer and the chemical reactions occurring inside the membrane, suggesting that diffusion of the VFA-TOPO complex inside the membrane may be the rate-limiting step in this experiment.