Low-cost supports used to immobilize fungi and reliable technique for removal hexavalent chromium in wastewater

The main goal of this study was to exploit low-cost and efficient sorbents for the removal and recovery of Cr(VI) in wastewater. Three supports of sawdust, polyurethane and alginate were applied to immobilize living and dead R. cohnii cells, respectively. There was a distinct increase in the Cr(VI) removal efficiency before and after the HCl-pretreatment. Langmuir adsorption isotherm model was well used to describe the distribution of Cr(VI) between the liquid and solid phases in batch studies. The values of q0 predicted by Thomas model were near to experimental ones in the experiments of packed column. The breakthrough curves calculated with this model were consistent well with experimental ones at a largely extent. Desorption, regeneration and reuse of the packed column were studied. After 5 cycles, adsorption capacity was still kept at higher level, reaching to 91.4, 87.9, 91.4 and 93.3mg/l contrasted with the first cycle (94.1, 90.4, 94.8 and 98.5mg/l) and the desorption efficiency were 85.0%, 96.2%, 93.4% and 91.4% compared with the first cycle (87.6%, 95.4%, 96.7% and 94.3%), corresponding to living cells immobilized with sawdust, polyurethane, and dead cells immobilized with polyurethane and alginate, respectively. The results indicated that the packed columns with the immobilized living and dead R. cohnii cells were the better option to adsorb, desorb and recover Cr(VI) from wastewater.     

Phosphorus retention in the filter materials shellsand and Filtralite P®—Batch and column experiment with synthetic P solution and secondary wastewater

Filtralite P^® and shellsand as ideal constructed wetland substrates have been tested for their P sorption capacity, both with batch and column experiments. Two columns were filled with Filtralite P^® and one column with shellsand. The shellsand (SSPS) and one of the Filtralite P^® columns (FLSP) were loaded with a synthetic P solution, while the second Filtralite P^® column (FLWW) was loaded with secondary wastewater. Ca, Mg, pH, and the P concentrations were measured in the inlet and the seven outlets along the height of the three vertical upflow columns for up to 303 days. An overall P removal rate of 92, 91, and 54% was measured in the columns SSPS, FLWW, and FLPS, respectively, for the entire experimental period. The comparison of FLWW and FLPS showed that FLWW kept its high P removal efficiency (91%) throughout the experimental period while the removal efficiency of FLPS decreased fast after reaching the 1 ppm effluent P concentration. The competition of other negative ions and the development of biofilm did not have a negative effect on P removal from wastewater. The batch experiments showed a better sorption capacity of Filtralite P^® at low initial concentrations, while for high initial concentrations the shellsand sorbed more. Shellsand had, however, a higher sorption capacity in batch experiments with used column material and high initial P concentrations. The results from both the batch and the column experiment suggest that the shellsand has a more durable P sorption capacity than the Filtralite P^® material, possibly due to the persistent high concentrations of Ca in the shellsand.     

Next-Generation Pyrosequencing Analysis of Microbial Biofilm Communities on Granular Activated Carbon in Treatment of Oil Sands Process-Affected Water

The development of biodegradation treatment processes for oil sands process-affected water (OSPW) has been progressing in recent years with the promising potential of biofilm reactors. Previously, the granular activated carbon (GAC) biofilm process was successfully employed for treatment of a large variety of recalcitrant organic compounds in domestic and industrial wastewaters. In this study, GAC biofilm microbial development and degradation efficiency were investigated for OSPW treatment by monitoring the biofilm growth on the GAC surface in raw and ozonated OSPW in batch bioreactors. The GAC biofilm community was characterized using a next-generation 16S rRNA gene pyrosequencing technique that revealed that the phylum Proteobacteria was dominant in both OSPW and biofilms, with further in-depth analysis showing higher abundances of Alpha- and Gammaproteobacteria sequences. Interestingly, many known polyaromatic hydrocarbon degraders, namely, Burkholderiales, Pseudomonadales, Bdellovibrionales, and Sphingomonadales, were observed in the GAC biofilm. Ozonation decreased the microbial diversity in planktonic OSPW but increased the microbial diversity in the GAC biofilms. Quantitative real-time PCR revealed similar bacterial gene copy numbers (>10⁹ gene copies/g of GAC) for both raw and ozonated OSPW GAC biofilms. The observed rates of removal of naphthenic acids (NAs) over the 2-day experiments for the GAC biofilm treatments of raw and ozonated OSPW were 31% and 66%, respectively. Overall, a relatively low ozone dose (30 mg of O₃/liter utilized) combined with GAC biofilm treatment significantly increased NA removal rates. The treatment of OSPW in bioreactors using GAC biofilms is a promising technology for the reduction of recalcitrant OSPW organic compounds.     

Biological nitrogen removal using a modified oxic/anoxic reactor with zeolite circulation

A modified, laboratory-scale, oxic/anoxic process with zeolite circulation achieved a high efficiency (97%) of nitrogen removal from synthetic wastewater. This process lowered the turbidity and the suspended solid concentration in the effluent. The zeolite was effective as a biomedium due to the formation of biofilm on the surface of zeolite and the continuous biological regeneration.     

Removal of H(2)S by Thiobacillus denitrificans immobilized on different matrices

Biological removal of high concentrations of H(2)S was studied using the immobilized Thiobacillus denitrificans with peat moss, wood chip, ceramic and granular activated carbon (GAC) separately. Experiments on the physical adsorption capacity of matrix, retention time and pressure drop were carried out; the ability of bioreactor to buffer shock loading and the removal efficiency with different packing materials were also investigated. Besides, the kinetics of single-stage biodesulfuration was analyzed. The results showed that GAC provided higher bacteria adsorption capacity, showed a more resistance to shock loading and allowed better operational control with respect to pressure drop than other inert carriers. When the retention time was changed from 30 to 100 s at an influent concentration of 100 mg/L of H(2)S, the removal efficiencies were above 98%; when the inlet concentration of H(2)S were changed from 110 to 120 mg/L, an average 96.8% removal efficiency was achieved during the long-term operation for GAC bioreactor. Next to GAC, wood chip was found to be a good packing material; however, peat moss and ceramic had limited effectiveness and their removal efficiencies were less of 90%. The kinetic analysis showed that the maximum removal rate and the half-saturation constant of the GAC bioreactor were 666.7 mg (H(2)S)/(L.d) and 20.8 mg/L, respectively.     

Increased removal capacity for 1,2-dichloroethane by biological modification of the granular activated carbon process

The removal of 5 mg 1^–1 1,2-dichloroethane [(CH₂Cl)₂] was studied in two granular activated carbon (GAC) reactors run with hydraulic retention times of below 1 h. One reactor was operated abiotically. The other one was inoculated with microorganisms able to degrade (CH₂Cl)₂. While the (CH₂Cl)₂-adsorption capacity of the non-inoculated GAC reactor was exhausted after 20 days, it apparently did not exhaust for at least 170 experimental days in the biologically activated system because (CH₂Cl)₂ was removed to over 95% as a result of the microbial degradation. The biodegradation was quantified: during the passage through the biologically activated GAC reactor, (CH₂Cl)₂ (5± mg l^–1) disappeared, chloride ions (3.3±0.2 mg l^–1) were produced, and oxygen (4 to 6 mg l^–1) was consumed. Removal of 30% of GAC at the entrance of the reactor, which visibly carried most of the biomass, and its replacement by virgin GAC at the end of the column did not change the apparent (CH₂Cl)₂removal capacity of the GAC column, indicating that still enough biomass was available to degrade most of the chemical fed. After the addition of the virgin carbon, the effluent concentration fell for a short period of time from about 200 g l^–1 to below 100 g l^–1, indicating partial adsorption of the non-degraded (CH₂Cl)₂ at the end of the reactor by the virgin carbon. Thus, the modification of the adsorption process by inoculation and maintenance of bacteria with special degradation capabilities resulted in a lower consumption of GAC and thus led to an extended service life of the GAC columns.     

Characterization of microbial fuel cells enriched using Cr(VI)-containing sludge

Microbial fuel cells (MFCs) were successfully enriched using sludge contaminated with Cr(VI) and their characteristics were investigated. After enrichment, the charge of the final 10 peaks was 0.51 C +/- 1.16%, and the anodic electrode was found to be covered with a biofilm. The enriched MFCs removed 93% of 5 mg/l Cr(VI) and 61% of 25 mg/l Cr(VI). 16S rDNA DGGE profiles from the anodic electrode indicated that beta-Proteobacteria, Actinobacteria, and Acinetobacter sp. dominated. This study is the first to report that electrochemically active and Cr(VI)-reducing bacteria could be enriched in the anode compartment of MFCs using Cr(VI)-containing sludge and demonstrates the Cr(VI) removal capability of such MFCs.     

Biosorption of hexavalent chromium using biofilm of <Emphasis Type="Italic">E. coli</Emphasis> supported on granulated activated carbon

The optimization of hexavalent chromium biosorption has been studied by using three different biosorbents; biofilm of E. coli ASU 7 supported on granulated activated carbon (GAC), lyophilized cells of E. coli ASU 7 and granulated activated carbon. Supporting of bacteria on activated carbon decreased both the porosity and surface area of the GAC. Significant decrement of surface area was correlated to the blocking of microspores as a result of the various additional loads. The experimental data of adsorption was fitted towards the models postulated by Langmuir and Freundlich and their corresponding equations. The maximum biosorption capacity for hexavalent chromium using biofilm, GAC and E. coli ASU 7 were 97.70, 90.70, 64.36 mg metal/g at pH 2.0, respectively. Biosorption mechanism was related mainly to the ionic interaction and complex formation. Based on the experimental conditions, the presence of bacteria could be enhanced the capacity of activated carbon to adsorb hexavalent chromium ions from aqueous solutions.     

4-Chlorophenol degradation by a bacterial consortium: development of a granular activated carbon biofilm reactor

A bacterial consortium that can degrade chloro- and nitrophenols has been isolated from the rhizosphere of Phragmitis communis. Degradation of 4-chlorophenol (4-CP) by a consortium attached to granular activated carbon (GAC) in a biofilm reactor was evaluated during both open and closed modes of operation. During the operation of the biofilm reactor, 4-CP was not detected in the column effluent, being either adsorbed to the GAC or biodegraded by the consortium. When 4-CP at 100 mg l⁻¹ was fed to the column in open mode operation (20 mg g⁻¹ GAC total supply), up to 27% was immediately available for biodegradation, the rest being adsorbed to the GAC. Biodegradation continued after the system was returned to closed mode operation, indicating that GAC bound 4-CP became available to the consortium. Biofilm batch cultures supplied with 10–216 mg 4-CP g⁻¹ GAC suggested that a residual fraction of GAC-bound 4-CP was biologically unavailable. The consortium was able to metabolise 4-CP after perturbations by the addition of chromium (Cr VI) at 1–5 mg l⁻¹ and nitrate at concentrations up to 400 mg l⁻¹. The development of the biofilm structure was analysed by scanning electron microscopy and confocal laser scanning microscopy (CLSM) techniques. CLSM revealed a heterogeneous structure with a network of channels throughout the biofilm, partially occupied by microbial exopolymer structures. Received: 17 March 1999 / Received revision: 27 May 1999 / Accepted: 28 May 1999     

Treatment of chromium(VI) solutions in a pilot-scale bioreactor through a biofilm of Arthrobacter viscosus supported on GAC

The aim of this work is to evaluate the applicability of a biofilm to the removal of chromium in solution, at a pilot scale. The effect of the initial concentration of metal on the biosorption behavior of an Arthrobacter viscosus biofilm supported on granular activated carbon, in batch and column essays was also analyzed. Six isotherm equations have been tested in the present study. The best fit was obtained with the Freundlich model. It was observed that as the initial chromium concentration increases, the uptake increases too, but the removal percentage decreases, with values between 95.20% (C(0)=5mg/l) and 38.28% (C(0)=1000 mg/l). The batch adsorption studies were used to develop a pilot bioreactor able to remove chromium from aqueous solutions. Data obtained in a pilot-scale reactor showed an average removal percentage of 99.9%, during the first 30 days, for the initial concentration of 10mg/l and an average removal percentage of 72%, for the same period and for the initial concentration of 100mg/l. Uptake values of 11.35 mg/g and 14.55 mg/g were obtained, respectively, for the initial concentration of 10 and 100mg/l. The results obtained are very promising and encourage the utilization of this biofilm in environmental applications.     

Uranium bioremediation in continuously fed upflow sand columns inoculated with anaerobic granules

Reductive precipitation of soluble hexavalent uranium (U(VI)) to insoluble tetravalent uranium (U(IV)) containing minerals is one of the more promising approaches to uranium remediation. The objective of this study was to evaluate the long-term performance of methanogenic granules for the continuous treatment of U(VI). For this purpose, three sand-packed columns inoculated with anaerobic biofilm were operated with or without ethanol and one column was exposed to nitrate co-contamination. The columns were operated for 373 days and efficiently removed U (24 mg L(-1)) in excess of 99.8%. No long-term benefit of ethanol addition was observed, suggesting that endogenous substrates in the biofilm were sufficient to drive the reduction reactions. Nitrate addition was found to inhibit U(VI) reduction and cause re-oxidation of some U(IV) deposited in the column. Taken as a whole, the results indicate that methanogenic biofilms can be reliably applied in bioreactor technology for sustained U removal from groundwater.     

Reusability of avidin-biotinylated immunoglobulin Y columns in immunoaffinity chromatography

Reusability of avidin-biotinylated IgY columns for immunoaffinity chromatography was examined by repeated use and regeneration. An enzyme-linked immunosorbent assay-elution assay using CovaLink NH microtiter plates was used to find the optimal conditions for regeneration of columns. Actigel avidin-biotinylated IgY column retained about 90% of its initial IgG binding capacity after 50 cycles, with 0.1 M glycine-HCl buffer, pH 2.8, as eluent, requiring no regeneration. However, IgG binding capacity of UltraLink avidin-biotinylated IgY column gradually decreased to 75 and 65% after 10 and 20 cycles, respectively, with the commercial eluent, Actisep. Results from the CovaLink NH system agreed with those from UltraLink avidin-biotinylated IgY columns. The UltraLink avidin-biotinylated IgY column was regenerated twice, by applying 8 M guanidine-HCl, pH 1.6, to dissociate biotinylated IgY antibodies from the column. About 40 and 25% of IgG binding capacities remained after the first and second regeneration. By applying new biotinylated IgY to the treated columns, about 95 and 90% of the initial IgG binding capacity before any treatment were recovered. These results demonstrated that avidin-biotinylated IgY columns are reusable with or without regeneration depending on the avidin-immobilized matrix.     

Modeling hexavalent chromium removal in a Bacillus sp. fixed-film bioreactor

A one-dimensional diffusion-reaction model was developed to simulate Cr(VI) reduction in a Bacillus sp. pure culture biofilm reactor with glucose as a sole supplied carbon and energy source. Substrate utilization and Cr(VI) reduction in the biofilm was best represented by a system of (second-order) partial differential equations (PDEs). The PDE system was solved by the (fourth-order) Runge-Kutta method adjusted for mass transport resistance using the (second-order) Crank-Nicholson and Backward Euler finite difference methods. A heuristic procedure (genetic search algorithm) was used to find global optimum values of Cr(VI) reduction and substrate utilization rate kinetic parameters. The fixed-film bioreactor system yielded higher values of the maximum specific Cr(VI) reduction rate coefficient and Cr(VI) reduction capacity (kmc = 0.062 1/h, and Rc = 0.13 mg/mg, respectively) than previously determined in batch reactors (kmc = 0.022 1/h and Rc = 0.012 mg/mg). The model predicted effluent Cr(VI) concentration well with 98.9% confidence (sigmay2 = 2.37 mg2/L2, N = 119) and effluent glucose with 96.4 % confidence (sigmay(w)2 = 5402 mg2/L2, N = 121, w = 100) over a wide range of Cr(VI) loadings (10-498 mg Cr(VI)/L/d).     

Adsorptive removal of fermentation inhibitors from concentrated acid hydrolyzates of lignocellulosic biomass

Adsorptive purification of concentrated acid hydrolyzate of lignocellulose was investigated. Cation exchange resin (CS16GC), neutral polymer adsorbent (XAD-16), and granulated activated carbon (GAC) were studied to remove furfural, HMF, and acetic acid from a synthetic hydrolyzate containing 20 wt.% H₂SO₄. Adsorption isotherms were determined experimentally. Loading and regeneration were investigated in a laboratory scale column.GAC has the highest adsorption capacity, but regeneration with water was not feasible. XAD-16 and CS16GC had lower adsorption capacities but also shorter cycle times due to easier regeneration. Productivity increased when regenerating with 50 wt.% EtOH(aq) solution.To compare adsorbents, process performance was quantified by productivity and fraction of inhibitors removed. GAC yields highest performance when high purity is required and ethanol can be used in regeneration. For lower purities, XAD-16 and GAC yield approximately equal performance. When using ethanol must be avoided, CS16GC offers highest productivity.     

Bioremediation of toxic chromium from electroplating effluent by chromate-reducing Pseudomonas aeruginosa A2Chr in two bioreactors

The chromate-reducing ability of Pseudomonas aeruginosa A2Chr was compared in batch culture, with cells entrapped in a dialysis sac, and with cells immobilized in an agarose-alginate film in conjunction with a rotating biological contactor. In all three systems, the maximum Cr(VI) reduction occurred at 10 mg Cr(VI)/l. Whereas at 50 mg Cr(VI)/l concentration, only 16% of the total Cr(VI) was reduced, five spikings with 10 mg chromate/l at 2-h intervals led to 96% reduction of the total input of 50 mg Cr(VI)/l. Thus maximum Cr(VI) reduction was achieved by avoiding Cr(VI) toxicity to the cells by respiking with lower Cr(VI) concentrations. At 10 mg Cr(VI)/l, the pattern of chromate reduction in dialysis-entrapped cells was almost similar to that of batch culture and 86% of the bacterially reduced chromium was retained inside the dialysis sac. In electroplating effluent containing 100 mg Cr(VI)/l, however, the amount of Cr(VI) reduced by the cells immobilized in agarose-alginate biofilm was twice and thrice the amount reduced by batch culture and cells entrapped in a dialysis sac, respectively.     

A GAC biofilm reactor for the continuous degradation of 4-chlorophenol: treatment efficiency and microbial analysis

Using a continuous enrichment technique, a bacterial consortium capable of degrading 4-chlorophenol (4-CP) was obtained from the rhizosphere of Phragmites australis. A granular activated carbon (GAC) biofilm reactor was established using this consortium, and the degradation of 4-CP was investigated under continuous flow operation using a feed of 20-50 mg l(-1) with a hydraulic residence time of 17 min over a 6-month period. Chloride liberation occurred throughout the operation, and the reactor had 4-CP removal efficiencies of 69-100%. Periods of lower performance were attributed to clogging of the column with biomass and the formation of channels. Subsequently, the immobilized biofilm was subjected to a starvation period of 5 months, after which its degradative capacity was still maintained. The microbial consortium was characterized during the continuous flow experiment and dynamic population changes were observed throughout. One isolate recovered from the biofilm was shown to be capable of degrading 4-CP as a sole carbon and energy source.     

Leaching of boron through sewage sludge amended soil: the role of clinoptilolite

A laboratory experiment was conducted to determine the release of boron from soil-sewage sludge mixtures by leaching using a clinoptilolite type natural zeolite, before land application of the sewage sludge. Soil columns were filled up with the clinoptilolite soil after mixing with sewage sludge at a rate of 30 tons ha(-1) and with two different particle sizes (0.1-0.25 and 1.0-2.0 mm) of clinoptilolite each at the concentrations of 1% and 2%. The particle size and the application rate of clinoptilolite affected both boron leaching from soil compared to the control treatment (soil and sewage sludge mixture). The total soluble boron leached from a soil column varied from 66-92% depending on the applications of clinoptilolite and reached 96% for the control treatment, following application of 80 cm depth of water in all treatments. In the cases of the 1% application rate of 0.1-0.25 and 1-2 mm sized clinoptilolite 78% and 92% of the total boron leached, respectively. While at 2% application rate of 0.1-0.25 and 1-2 mm zeolite, 66% and 87% of total soluble boron leached, respectively. Boron concentrations in the soil layers increased as application rate increased and particle size of clinoptilolite decreased because of its high adsorption capacity. Adsorption isotherms indicated that clinoptilolite had a high adsorption capacity for boron compared to the sewage sludge and soil.     

Influence of magnetic field on Cr(VI) adsorption capability of given anaerobic sludge

To provide beneficial guide for the application of the magnetic field in the bio-treatment of the Cr(VI)-contained wastewater, sludge samples from the control bio-system A (absent of magnetic field) and the contrast bio-system B (present of magnetic field) were used to adsorb the synthetic wastewater with 100 mg l⁻¹ Cr(VI). Influences of two adsorption modes, single adsorption and once continuous adsorption, on the Cr(VI) adsorption capacities of both sludge samples were compared. And the influence of regeneration on the Cr(VI) adsorption capacities were also studied. The results of adsorption experiments showed that the Cr(VI) adsorption capacities of the first single adsorption for sludge sample A and B were pretty nearly, which were 9.79 and 9.93 mg, respectively. And after 5 single adsorption periods, the total Cr(VI) adsorption capacity and efficiency of the sample B were 25.88 and 55.66 mg Cr(VI) g⁻¹VSS, while those of the control were 14.95 and 33.98 mg Cr(VI) g⁻¹VSS, respectively. For the sludge sample A and B after a single adsorption, both functions of regeneration were remarkable. But after 13 cycles of the single adsorption-regeneration, the Cr(VI) adsorption capacity and efficiency of the sample B were 110.15 and 189.91 mg Cr(VI) g⁻¹VSS, while those of the control were 70.89 and 140.38 mg Cr(VI) g⁻¹VSS, respectively. Though the Cr(VI) adsorption capacity of a once continuous adsorption period was more than that of a single adsorption period obviously, the Cr(VI) removal rates of the sludge sample A and B in the third period of once continuous adsorption-regeneration were only 8.12 and 33.51%, respectively. It was concluded that the weak magnetic field did improve the Cr(VI) bio-removal efficiency and the sludge stability, the batch treatment was an ideal operation mode for the bio-treatment of the Cr(VI)-contained wastewater, as compared with the continuous operation mode, but regeneration and enough sludge content were two necessary conditions to ensure the efficiency of batch treatment.     

Ammonium exchange and bioregeneration of bio-flocculated zeolite in a sequencing batch reactor

Natural zeolite was added to the sequencing batch reactors to assess its role in ammonium exchange. Even though the biofilm was fully developed on the zeolite surface, ammonium removal and the biological regeneration of the zeolite occurred constantly during the anoxic-fill phase and the reaction phase, respectively. However, the specific nitrification rate of the bio-flocculated zeolite was lower than that observed in the control on account of the limited ammonium release to the liquid phase.     

Reduction of Cr(VI) by immobilized cells of Desulfovibrio vulgaris NCIMB 8303 and Microbacterium sp. NCIMB 13776

Hexavalent chromium, a carcinogen and mutagen, can be reduced to Cr(III) by Desulfovibrio vulgaris NCIMB 8303 and Microbacterium sp. NCIMB 13776. This study examined Cr(VI) reduction by immobilized cells of the two strains in a common solution matrix using various entrapment matrices. Chitosan and PVA-borate beads did not retain integrity and supported low or no reduction of Cr(VI) by the cells. A commercial preparation (Lentikats) was stable but also did not support Cr(VI) reduction. K-carrageenan beads were stable in batch suspensions but gel integrity was lost after only 5 h in a flow-through system in the presence of 100 microM Cr(VI). The best immobilization matrices were agar and agarose, where the initial rates of reduction of Cr(VI) (from 500 microM solution) for D. vulgaris NCIMB 8303 and Microbacterium sp. NCIMB 13776 were 127 (agar) and 130 (agarose), and 15 (agar) and 12 (agarose) nmol h(-1) mg dry cell wt(-1), respectively. The higher removal of Cr(VI) by D. vulgaris was also seen in 14-mL packed-bed flow-through columns, where, at a flow rate of 2.4 mL h(-1), the percentage removal of Cr(VI) was approximately 95% and 60% for D. vulgaris and Microbacterium sp., respectively (agar-immobilized cells). The Cr(VI) reducing activities of D. vulgaris and Microbacterium sp. were lost after 159 and 140 h, respectively. Examination of the beads for structural integrity within the columns in situ using magnetic resonance imaging after 24 and 100 h of continuous operation against Cr(VI) (with negligible Cr retained within the columns) showed that agar beads were more stable with time. The most appropriate system for development of a continuous bioprocess is thus the use of D. vulgaris NCIMB 8303 immobilized in an agar gel matrix.