Microbial response and elimination capacity in biofilters subjected to high toluene loadings

Elimination capacity (EC) is frequently used as a performance and design criterion for vapor-phase biofilters without further verification of the microbial quantity and activity. This study was conducted to investigate how biofilters respond to high pollutant loadings and ultimately how this affects the EC of the biofilter. Two identical laboratory-scale biofilters were maintained at an initial toluene loading rate of 46 g m⁻³ h⁻¹ for a period of 24 days. After the initial biofilm development stage, the loading rates were increased to 91 g m⁻³ h⁻¹ and 137 g m⁻³ h⁻¹, respectively. Following a short period of pseudo-steady state, toluene removal efficiencies rapidly declined in both biofilters, with a concurrent decline in both critical and maximum ECs. The decline was mainly due to deterioration in the biodegradation activity of the biofilm and a decline in the toluene-degrading bacterial population within the biofilm phase. The findings imply that high toluene loadings accelerated the deterioration in overall performance due to a rapid accumulation of inactive biomass. As a result, care must be used when relying on EC values for biofilter design and operational purposes, since the values do not appropriately reflect the temporal changes in biodegradation activity and active biomass quantities that can occur in biofilters subjected to high inlet loadings.     

Influence of gaseous VOC concentration on the diversity and biodegradation performance of microbial communities

In this work, the influence of toluene gas concentration on the isolation of toluene degrading microbial communities from activated sludge was studied. Toluene biodegradation at gas phase concentration of 10?g?m^?3 (R1) resulted in process instability with removal efficiencies (RE) lesser than 33?%, while operation at toluene gas phase concentrations of 300?mg?m^?3 (R2) and 11?mg?m^?3 (R3) was stable with RE ranging from 74 to 94?%. The consortium isolated in R1 exhibited the highest tolerance toward toluene but the lowest biodegradation performance at trace level VOC concentrations. Despite R2 and R3 showed a similar sensitivity toward toluene toxicity, the microbial community from R2 supported the most efficient toluene biodegradation at trace level VOC concentrations. The Shannon-Wiener index showed an initial biodiversity decrease from 3.2 to 2.0, 1.9 and 2.7 in R1, R2 and R3, respectively. However, while R2 and R3 were able to recover their initial diversity levels by day 48, this loss in diversity was permanent in R1. These results showed that traditional inoculum isolation/acclimation techniques based on the exposure of the inoculum to high VOC concentrations, where toxicity tolerance plays a key role, may result in a poor abatement performance when the off-gas stream is diluted.     

Biotreatment of a gas-phase volatile mixture from fibreglass and composite manufacturing industries

Acetone, toluene and styrene (ATS) are representative air pollutants emanating during the production process in fibreglass and composite manufacturing industries. In this study, the performance of a steady-state biofilter inoculated with the fungus Sporothrix variecibatus was tested at different empty bed residence times (EBRTs), and at different inlet concentrations of ATS, corresponding to total pollutant loading rates ranging from 30 to 490 gm(-3)hour(-1). Styrene was somewhat better removed (47-100%) in the biofilter than acetone (34-100%) and toluene (42-100%), with maximum elimination capacities (EC(max)) of 108, 72 and 144 gm(-3)hour(-1), for ATS, respectively. Besides, it was observed that, although increasing the concentration of ATS decreased their removal, the presence of toluene also decreased the EC(max) of both acetone and toluene in the ternary mixture. During transient operations, the biofilter was subjected to intermittent shutdown and re-start operations where the gas-phase pollutant flow was stopped for either 5 or 16d. It was observed that, for longer shutdown periods (16d), the biofilter required nearly 8-10d to reach similar removal patterns to those observed before the shutdown phase. Batch biodegradation tests were conducted, using Sporothrix-like microorganisms present in the leachate of the biofilter, with a mixture of ATS as the sole carbon source. Complete removal of ATS was observed within the test period of 168 hours. Styrene was degraded faster, with a specific substrate utilization rate of 0.9 mg styrenemg biomass(-1)hour(-1), followed by toluene (0.6) and acetone (0.44). The effectiveness of the biofilter to reach high total EC (321.3 gm(-3)hour(-1)), and withstand transient operations shows the robustness of this fungal-bioreactor and its suitability to handle emissions from a fibreglass and composite manufacturing industry.     

Design of a biofilter packed with crab shell and operation of the biofilter fed with leaf mold solution as a nutrient

After measuring toluene adsorption (15.7 mg-toluene/g-material), water holding capacity (18.5%), organic content (53.8%), specific surface area (18.1 m²/g-material), and microbial attachment, crab shells were chosen as the main packing material for a biofilter design. The crab shells, cheap and abundant in the Gangneung area, also have relatively rigid structure, low density, and ability to neutralize acids generated during mineralization of toluene. Since towel scraps have water holding capacity as high as 301.2%, 10% of the total packing was supplemented with them to compensate for low water holding capacity of the crab shells. The biofilter fed with defined chemical medium under 0.8∼1.3 mg/L of inlet toluene concentration and 18 seconds of residence time showed satisfactory removal efficiency of over 97% and 72.8 g/h·m³ of removal capacity. For the purpose of deceasing operation costs, leaf mold solution was tried as an alternative nutrient instead of a defined chemical medium. The removal efficiency and removal capacity were 85% and 56.3 g/h·m³, respectively, using the same inlet toluene concentration and residence time. This research shows the possibility of recycling crab shell waste as packing material for biofilter. In addition, leaf mold was able to serve as an alternative nutrient, which remarkably decreased the operating cost of the biofilter.     

Evaluating the biodegradation of aromatic hydrocarbons by monitoring of several functional genes

Various microbial activities determine the effectiveness of bioremediation processes. In this work, we evaluated the feasibility of gene array hybridization for monitoring the efficiency of biodegradation processes. Biodegradation of ¹⁴C-labelled naphthalene and toluene by the aromatic hydrocarbon-degrading Pseudomonas putida F1, P. putida mt-2 and P. putida G7 was followed in mixed liquid culture microcosm by a preliminary, nylon membrane-based gene array. In the beginning of the study, toluene was degraded rapidly and increased amount of toluene degradation genes was detected by the preliminary gene array developed for the study. After toluene was degraded, naphthalene mineralization started and the amount of naphthalene degradation genes increased as biodegradation proceeded. The amount of toluene degradation genes decreased towards the end of the study. The hybridization signal intensities determined by preliminary gene array were in good agreement with mineralization of naphthalene and toluene and with the amount of naphthalene dioxygenase and toluene dioxygenase genes quantified by dot blot hybridization. The clear correlation between the results obtained by the preliminary array and the biodegradation process suggests that gene array methods can be considered as a promising tool for monitoring the efficiency of biodegradation processes.     

Shift of microbial community in gas-phase biofilters with different inocula,inlet loads and nitrogen sources

The research on gaseous VOCs biofilters has often concentrated on process optimization. However, the microbial community change upon operating conditions is not well understood. In this study, three lab-scale biofilters treating gaseous toluene were operated for 66 days with different inocula under changes in inlet loads and nitrogen sources. Three biofilters were inoculated with activated sludge, river sediment or microbial consortia, respectively. The microbial community differed a lot initially but gradually deviated toward similar structures with the same dominant microorganisms, i.e. Proteobacteria, Actinobacteria (phylum level) and Rhodococcus,Pseudomonas(genus level). Among three biofilters, the two biofilters inoculated with activated sludge and river sediment showed higher microbial diversity, better VOCs removal performance and higher metabolic activity. Higher relative abundance of Alcanivorax (3% compared with lower than 0.03%), Pimelobacte (0.05% compared with lower than 0.01%)were detected under low inlet load, and Zoogloea(0.1%), Alkaliphilus(0.2%) were detected when the inlet load was increased. the abundance of Pseudomonasdecreased from 14% to 2% when ammonia was used as nitrogen source instead of nitrate, meanwhile the abundance of Bacillus and Gordoniaincreased from 0.01% to 0.05% and 0.8% to 5.8% respectively. Some special organisms were observed i.e. the intestinal microorganism.     

Degradation of BTEX compounds in liquid media and in peat biofilters

A mixed culture, enriched from Sphagnum peat moss, contaminated with gasoline vapours, degraded individual and mixed components of BTEX (benzene, toluene, ethylbenzene, xylene). Complete degradation of radiolabelled toluene by the mixed culture was observed in mineralisation studies. Individual isolates from a mixed culture containingPseudomonas maltophilia, P. testosteroni andP. putida biotype A exhibited contrasting BTEX degradation patterns. WhileP. putida biotype A degraded all of the BTEX compounds,P. maltophilia andP. testosteroni, appeared unable to degrade benzene and xylenes, respectively. When the peat, inoculated with the mixed culture, was used as a biofilter (6.2 cm diameter ×93 cm length) for degradation of toluene and ethylbenzene vapours, percentage removal efficiencies were 99 and 85, respectively. When the capacity of the biofilter to degrade a combination of BTEX compounds was evaluated, percentage removal efficiencies for toluene, ethylbenzene,p-xylene,o-xylene and benzene were 99, 85, 82, 80 and 78, respectively. The importance of using the mixed culture as an inoculum in the biofilter was established and also the relationship between contaminated vapour flow rate and percentage removal efficiency.     

Link between spatial structure of microbial communities and degradation of a complex mixture of volatile organic compounds in peat biofilters

AIMS: To investigate the relationships between the operation of the volatile organic compound (VOC) removal biofilter and the structure of microbial communities, and to study the impact on degradation activities and the structuring of microbial communities of biofilter malfunctions related to the qualitative composition of the polluted air. METHODS AND RESULTS: A microbiological study and a measurement of biodegradation activities were simultaneously carried out on two identical peat-packed columns, seeded with two different inocula, treating polluted air containing 11 VOCs. For both reactors, the spatial structure of the microbial communities was investigated by means of single-strand conformation polymorphism (SSCP) analysis. For both reactors, stratification of degradation activities in function of depth was observed. Oxygenated compounds were removed at the top of the column and aromatics at the bottom. Comparison of SSCP patterns clearly showed a shift in community structure in function of depth inside both biofilters. This distribution of biodegradation activities correlates with the spatialization of microbial density and diversity. Although the operating conditions of both reactors were identical and the biodegradation activities similar, the composition of microflora differed for biofilters A and B. Subdivision of biofilter B into two independent parts supplied with polluted air containing the complex VOC mixture showed that the microflora having colonized the bottom of biofilter B retained their potential for degrading oxygenated compounds. CONCLUSIONS: This work highlights the spatialization of biodegradation functions in a biofilter treating a complex mixture of VOCs. This distribution of biodegradation activities correlates with the spatialization of microbial density and diversity. SIGNIFICANCE AND IMPACT OF THE STUDY: This vertical structure of microbial communities must be taken into consideration when dealing with the malfunctioning of bioreactors. These results are also useful information about changes in microbial communities following natural or anthropogenic alterations in different ecosystems (soils and sediments) where structuring of microbial communities according to depth has been observed.     

Model based evaluation of a contaminant plume development under aerobic and anaerobic conditions in 2D bench-scale tank experiments

The influence of transverse mixing on competitive aerobic and anaerobic biodegradation of a hydrocarbon plume was investigated using a two-dimensional, bench-scale flow-through laboratory tank experiment. In the first part of the experiment aerobic degradation of increasing toluene concentrations was carried out by the aerobic strain Pseudomonas putida F1. Successively, ethylbenzene (injected as a mixture of unlabeled and fully deuterium-labeled isotopologues) substituted toluene; nitrate was added as additional electron acceptor and the anaerobic denitrifying strain Aromatoleum aromaticum EbN1 was inoculated to study competitive degradation under aerobic / anaerobic conditions. The spatial distribution of anaerobic degradation was resolved by measurements of compound-specific stable isotope fractionation induced by the anaerobic strain as well as compound concentrations. A fully transient numerical reactive transport model was employed and calibrated using measurements of electron donors, acceptors and isotope fractionation. The aerobic phases of the experiment were successfully reproduced using a double Monod kinetic growth model and assuming an initial homogeneous distribution of P. putida F1. Investigation of the competitive degradation phase shows that the observed isotopic pattern cannot be explained by transverse mixing driven biodegradation only, but also depends on the inoculation process of the anaerobic strain. Transient concentrations of electron acceptors and donors are well reproduced by the model, showing its ability to simulate transient competitive biodegradation.     

Characterization of compost biofiltration system degrading dichloromethane

The effects of acclimatization of microbial populations, compound concentration, and media pH on the biodegradation of low concentration dichloromethane emissions in biofiltration systems was evaluated. Greater than 98% removal efficiency was achieved for dichloromethane at superficial velocities from 1 to 1.5 m(3)/m(3). min (reactor residence times of 1 and 0.7 min, respectively) and inlet concentrations of 3 and 50 ppm Although acclimatization of microbial populations to toluene occurred within 2 weeks of operation start-up, initial dichloromethane acclimatization took place over a period of 10 weeks. This period was shortened to 10 days when a laboratory grown consortium of dichloromethane degrading organism, isolated from a previously acclimatized column, was introduced into fresh biofilter media. The mixed culture consisted to 12 members, which together were able to degrade dichloromethane at concentrations up to 500 mg/L. Only one member of the consortium was able to degrade dichloromethane were sustained for more than 4 months in a biofilter column receiving an inlet gas stream with 3 ppm(v) of dichloromethane acidification of the column and resulting decline in performance occurred when a 50-ppm(v) inlet concentration was used. A biofilm model incorporating first order biodegradation kinetics provided a good fit to observed concentration profiles, and may prove to be a useful tool for designing biofiltration systems for low concentration VOC emissions. (c) 1994 John Wiley & Sons, Inc.     

Application of biological activated carbon as a low pH biofilter medium for gas mixture treatment

Packing material is a crucial component of a bioreactor as it is the microbial population's habitat. This study assessed potential improvements to current biofiltration processes by investigating use of a novel support medium. Biological activated carbon (BAC) with microorganisms growing on granular activated carbon can produce a novel medium in which both adsorption and biodegradation contribute to pollutants removal. Investigation of carbon characteristics demonstrated that BAC was an ideal packing medium for biofiltration. The application of the novel packing medium for gas mixture treatment was evaluated in a low pH biofilter. Results demonstrated that BAC biofilter obtained high removal efficiency for both H(2)S and toluene. The removal mechanisms of BAC were investigated after the biofilter operation and it demonstrated that the performance of the BAC system was mainly controlled by the additive contributions of two removal mechanisms - adsorption and biodegradation. This study also indicated the potential for simultaneous treatment of hydrogen sulfide and toluene at low pH condition.     

In Situ Gene Expression in Mixed-Culture Biofilms: Evidence of Metabolic Interactions between Community Members

Microbial communities growing in laboratory-based flow chambers were investigated in order to study compartmentalization of specific gene expression. Among the community members studied, the focus was in particular on Pseudomonas putida and a strain of an Acinetobacter sp., and the genes studied are involved in the biodegradation of toluene and related aromatic compounds. The upper-pathway promoter (Pu) and the meta-pathway promoter (Pm) from the TOL plasmid were fused independently to the gene coding for the green fluorescent protein (GFP), and expression from these promoters was studied in P. putida, which was a dominant community member. Biofilms were cultured in flow chambers, which in combination with scanning confocal laser microscopy allowed direct monitoring of promoter activity with single-cell spatial resolution. Expression from the Pu promoter was homogeneously induced by benzyl alcohol in both community and pure-culture biofilms, while the Pm promoter was induced in the mixed community but not in a pure-culture biofilm. By sequentially adding community members, induction of Pm was shown to be a consequence of direct metabolic interactions between an Acinetobacter species and P. putida. Furthermore, in fixed biofilm samples organism identity was determined and gene expression was visualized at the same time by combining GFP expression with in situ hybridization with fluorescence-labeled 16S rRNA targeting probes. This combination of techniques is a powerful approach for investigating structure-function relationships in microbial communities.     

Performance and macrokinetic analysis of biofiltration of toluene and p-xylene mixtures in a conventional biofilter packed with inert material

Interactions of toluene and p-xylene in air treatment biofilters packed with an inert filter media were studied. The effect of the inlet load of toluene, p-xylene and mixtures of both compounds on the biodegradation rate was analyzed in three lab-scale biofilters. A maximum elimination capacity (EC) of 26.5 and 40.3 g C m⁻³ h⁻¹ for an inlet load (IL) of 65.6 and 57.8 g C m⁻³ h⁻¹ was obtained for p-xylene and toluene biofilters, respectively. Inhibition of p-xylene biodegradation by the presence of toluene took place when the mixture was treated, whereas the presence of p-xylene had an enhancing effect on the toluene removal efficiency. Specific growth rates (μ) from 0.019 to 0.068 h⁻¹ were calculated in the mixed biofilter, where the highest values were similar to mixtures with lower p-xylene levels (IL_p-Xyl 8.84 ± 0.29 g C m⁻³ h⁻¹). Michaelis-Menten and Haldane type models were fitted to experimental EC for p-xylene and toluene biofilters, respectively.     

Benzene, toluene, and o-xylene degradation by free and immobilized P. putida F1 of postconsumer agave-fiber/polymer foamed composites

Benzene, toluene, and o-xylene (BTX) degradation by immobilized Pseudomonas putida F1 of postconsumer agave-fiber/polymer foamed-composites (AFPFC) and suspended cultures was studied under controlled conditions. Analyses using FTIR-ATR and SEM showed that P. putida F1 adhered onto the composite surface and developed a biofilm. In this sense, the AFPFC were successfully used as a support for bacterial immobilization. Both systems, immobilized and suspended cells of P. putida F1, were able to completely degrade benzene and toluene from initial concentrations of 15, 30, 60, and 90 mg l⁻¹. An inhibitory effect of the intermediary catechol from benzene degradation was observed in suspended cultures but it was not presented in the immobilized system. The degradation of o-xylene was partially accomplished in both systems. The Monod equation was used to model the experimental data obtained from the biodegradation kinetics, and they were adequately described with this model.     

Development and operation of a trickling biofilter system for continuous treatment of gas-phase trichloroethylene

A parallel trickling biofilter (TBF) system that consists of two TBFs units in parallel, one for biodegradation of trichloroethylene (TCE) and the other for reactivation of an inactivated biofilm, was developed and operated for continuous treatment of gas-phase TCE by Burkholderia cepacia G4. For inlet loadings below 8.6 mg TCE l^–1 d^–1, complete removal of TCE was achieved. The maximal TCE elimination capacity was 17 mg l^–1 d^–1.     

Pyrosequencing analysis of bacterial communities in drinking water biofilters receiving influents of different types

Biological activated carbon (BAC) filters are commonly used in the world for improvement of drinking water quality. The indigenous microbiota in BAC filters can play a crucial role in reduction or biotransformation of contaminants. Molecular analysis can enhance our understanding of ecological functions of the microbial communities in drinking water BAC filters. In this study, three laboratory-scale drinking water BAC filters receiving influents of different types were constructed. Differences of bacterial communities in the three BAC filters were characterized using 454 pyrosequencing analysis. Pyrosequencing analysis illustrated the usefulness in elucidating the bacterial community structure in drinking water biofilter. High bacterial diversity in granular activated carbon (GAC) samples from each BAC biofilter was observed. Proteobacteria was the largest bacterial phylum in each GAC sample, with a marked shift of the proportions of Alpha-, Beta-, and Gammaproteobacteria. The levels of dissolved organic carbon and ammonia nitrogen in the influents could affect the bacterial diversity and community composition in the BAC biofilters. This work might add some new insights into microbial community and its influential factors in drinking water biofilters.     

Biological oxidation of hydrogen sulfide under steady and transient state conditions in an immobilized cell biofilter

The removal of hydrogen sulfide (H(2)S) was investigated in a lab scale biofilter packed with biomedia, encapsulated by sodium alginate and polyvinyl alcohol (PVA). The main H(2)S oxidation products were SO(4)(2-), SO(3)(2-), S(2-) and S(0). The immobilized cell biofilter required no separate acclimatization period and showed high removal efficiencies (RE) within the first few days of experiments. The removal efficiencies in the biofilter were consistently greater than 99% even when H(2)S loading was 6 g m(-3)h(-1). The maximum elimination capacity achieved in this study is 8 g H(2)Sm(-3)h(-1) at a loading rate of 13 g H(2)Sm(-3) h(-1). The response of the immobilized cells to fluctuations in inlet concentration and flow rate was determined by subjecting the biofilter to inlet loads of up to 10 g H(2)Sm(-3)h(-1). The biofilter responded effectively to these shock loading conditions and convalesced rapidly within 4-8h. Pressure drop values were consistently less throughout the operational period. The results from this study suggest that an immobilized cell biofilter is effective in treating H(2)S under steady and transient operating conditions.     

Role of Thiobacillus thioparus in the biodegradation of carbon disulfide in a biofilter packed with a recycled organic pelletized material

This study reports the biodegradation of carbon disulfide (CS₂) in air biofilters packed with a pelletized mixture of composted manure and sawdust. Experiments were carried out in two lab-scale (1.2 L) biofiltration units. Biofilter B was seeded with activated sludge enriched previously on CS₂-degrading biomass under batch conditions, while biofilter A was left as a negative inoculation control. This inoculum was characterized by an acidic pH and sulfate accumulation, and contained Achromobacter xylosoxidans as the main putative CS₂ biodegrading bacterium. Biofilter operation start-up was unsuccessfully attempted under xerophilic conditions and significant CS₂ elimination was only achieved in biofilter A upon the implementation of an intermittent irrigation regime. Sustained removal efficiencies of 90–100 % at an inlet load of up to 12 g CS₂ m^?3 h^?1 were reached. The CS₂ removal in this biofilter was linked to the presence of the chemolithoautotrophic bacterium Thiobacillus thioparus, known among the relatively small number of species with a reported capacity of growing on CS₂ as the sole energy source. DGGE molecular profiles confirmed that this microbe had become dominant in biofilter A while it was not detected in samples from biofilter B. Conventional biofilters packed with inexpensive organic materials are suited for the treatment of low-strength CS₂ polluted gases (IL <12 g CS₂ m^?3 h^?1), provided that the development of the adequate microorganisms is favored, either upon enrichment or by inoculation. The importance of applying culture-independent techniques for microbial community analysis as a diagnostic tool in the biofiltration of recalcitrant compounds has been highlighted.     

Characterization of a biofilter treating toluene contaminated air

The removal of toluene from an experimental gas-stream was studied in an industrial biofilter filled with poplar wood bark. Toluene degradation, approximately 85% through the operating period, resulted in low levels of toluene in the off-gas effluent. For a toluene load of 6.7 g m^-3 h^-1 the elimination capacity of the biofilter was found to be 6.0 g m^-3 h^-1. Toluene removal was due to biodegradative activity of microorganisms in the filter bed; the most probable number counts of toluene degraders increased from 2.4×10² to 6.4×10⁷ MPN/g dry packing material in about seven months of air-toluene supply. The degradative capacity of a Burkholderia (Pseudomonas) cepacia strain, isolated from the biofilter material, as an example of the effectiveness of microbial toluence removal was tested in batch culture. The microorganism degraded completely 250 ppm of toluence supplied as sole carbon source in 24 hours. The high performance demonstrated for a long period and the mechanical and physico-chemical stability of the biofilter favour its use in industrial full-scale off-gas control.     

Biofiltration of toluene and acetone mixtures by a trickle-bed air biofilter

Toluene and acetone mixtures are commonly encountered from the manufacture of semi-conductor or opto-electronic apparatus. This study attempts to employ a trickle-bed air biofilter (TBAB) for treating toluene and acetone mixtures under different gas flow rates and influent concentrations. In the pseudo-steady-states, the elimination capacities of toluene and acetone increased but the removal efficiencies decreased with the increase of influent carbon loading. The removal efficiencies of toluene were higher than those of acetone, indicating that toluene is a preferred substrate in the mixtures. Greater than 90% removal efficiencies were achieved with influent carbon loadings of toluene and acetone below 125 and 15 g/m³ h, respectively. The TBAB appears efficient for controlling toluene and acetone mixture with medium toluene and low acetone loadings. Applicable operating conditions of TBAB for treating mixed toluene and acetone emission are suggested.