Optimal decolorization and kinetic modeling of synthetic dyes by Pseudomonas strains

Pseudomonas spp were isolated from an anaerobic-aerobic dyeing house wastewater treatment facility as the most active azo-dye degraders. Decolorization of azo dyes and non-azo dyes including anthraquinone, metal complex and indigo was compared with individual strains and a bacterial consortium consisting of the individual strain and municipal sludge (50 50wt). The consortium showed a significant improvement on decolorization of two recalcitrant non-azo dyes, but little effect on the dyes that the individual strains could degrade to a great or moderate extent. Decolorization of Acid violet 7 (monoazo) by a Pseudomonas strain GM3 was studied in detail under various conditions. The optimum decolorization activity was observed in a narrow pH range (7-8), a narrow temperature range (35-40 degrees C), and at the presence of organic and ammonium nitrogen. Nitrate had a severe inhibitory effect on azo dye decolorization: 10 mg/L led to 50% drop in decolorization activity and 1000 mg/L to complete activity depression. A kinetic model is established giving the dependence of decolorization rate on cell mass concentration (first-order) and dye concentration (half order). The rate increased with temperature from 10 to 35 C, which can be predicted by Arrhenius equation with the activation energy of 16.87 kcal/mol and the frequency factor of 1.49 x 10(11) (mg L)1/2/g DCM min.     

Biocalalyst effects of immobilized anthraquinone on the anaerobic reduction of azo dyes by the salt-tolerant bacteria

The accelerating effect of dissolved redox mediators has been studied in details in the bio-decolorization processes, but there are little literatures about the non-dissolved redox mediators. Here we describe the accelerating effect of anthraquinone as a redox mediator in the bio-decolorization. Decolorization of azo dyes was carried out experimentally using the salt-tolerant bacteria under immobilized anthraquinone and high salt conditions. Anthraquinone as a redox mediator was able to increase the decolorization rate of azo dyes, and was immobilized by entrapment in calcium alginate (CA), Polyvinyl alcohol (PVA)-H(3)BO(3) and agar, respectively. The effects of various operating conditions such as anthraquinone bead number, dissolved oxygen, temperature and pH on microbial decolorization were investigated experimentally. The reusability of the anthraquinone immobilization beads was evaluated with repeated-batch decolorization experiments. After four repeated experiments, the decolorization rate of CA immobilized anthraquinone retained over 90% of their original value. The experiments explored a great improvement of the redox mediator application and the new bio-treatment concept.     

Kinetic characteristics of bacterial azo-dye decolorization by Pseudomonas luteola

A Pseudomonas luteola strain expressing azoreductase activity was utilized to remove the color of an azo dye (reactive red 22) from contaminated solutions. The effects of substrate concentrations, medium compositions, and operation parameters (e.g., pH, temperature, dissolved oxygen, etc.) on decolorization of the azo dye by a P. luteola strain were systematically investigated to reveal the key factors that dominate the performance of azo-dye decolorization. The metabolites resulting from bacterial decolorization were analyzed by high-performance liquid chromatography (HPLC) and mass spectrometery (MS). The results show that the dissolved oxygen and glucose concentration retarded decolorization of reactive red 22 by P. luteola. The optimal azo-dye decolorization occurred at 37 degrees C, while more rapid decolorization took place over pH 7-9. Yeast extract and tryptone strongly enhanced the decolorization. The Michaelis-Menten model can satisfactorily describe the dependence of specific decolorization rate on the concentration of substrate (reactive red 22 or yeast extract). Decolorization of the azo dye by intact cells of P. luteola was essentially independent of the growth phase, whereas the azoreductase activity of the cell-free extract decreased in the order of late-stationary phase > early-stationary phase > mid-log phase. This suggests that mass transfer of the azo dye across the cell membrane may be the rate-limiting step. The HPLC and MS analyses suggest that both partial reduction and complete cleavage of the azo bond could contribute to decolorization of reactive red 22 by P. luteola.     

Biodegradation of p-chlorophenol by a microalgae consortium

An aquatic community was recovered from a waste discharge container fed with several aromatic pollutants. After 3 months of selective enrichment with p-chlorophenol and p-nitrophenol, two microalgae species, Chlorella vulgaris and Coenochloris pyrenoidosa, were recovered from the microbial consortium. As an axenic culture, this microalgae consortium was able to remove p-chlorophenol under different photo-regimes. Cultures grown under a 24h light regime were capable of biodegrading 50mg l(-1) of p-chlorophenol within 5 days. Addition of zeolite, an adsorbing material, did not improve the p-chlorophenol removal. However, when p-chlorophenol at 150mgl(-1) was fed to the culture supplemented with zeolite, the growth rate of the consortium improved, but the lag phase was longer (16 against 14 days in the absence of zeolite).     

Bioremediation of crystal violet using air bubble bioreactor packed with Pseudomonas aeruginosa

Seven water and sediment samples were collected and tested for decolorizing crystal violet. Pseudomonas aeruginosa was the most effective isolate for dye decolorization. The LC₅₀ of the crystal violet (115 mg/l) was measured using Artemia salina as a biomarker. The effect of different heavy metals on crystal violet decolorization was investigated. Cd²⁺ and Fe³⁺ ions showed marginal enhancement of the decolorization process, the rate was 1.35 mg/l/h compared to (1.25 mg/l/h) for the control. Phenol and m-cresol showed no effect on crystal violet decolorization, meanwhile p-cresol and p-nitrophenol reduced the decolorization rate to 1.07 and 0.01 mg/l/h, respectively. P. aeruginosa cells were immobilized by entrapment in agar-alginate beads. The beads were cultivated and reused in Erlenmeyer flask and in an air bubble column bioreactor and they enhanced the crystal violet decolorization rate to 3.33 and 7.5 mg/l/h, respectively.     

Bioremediation of crystal violet using air bubble bioreactor packed with Pseudomonas aeruginosa

Seven water and sediment samples were collected and tested for decolorizing crystal violet. Pseudomonas aeruginosa was the most effective isolate for dye decolorization. The LC₅₀ of the crystal violet (115 mg/l) was measured using Artemia salina as a biomarker. The effect of different heavy metals on crystal violet decolorization was investigated. Cd²⁺ and Fe³⁺ ions showed marginal enhancement of the decolorization process, the rate was 1.35 mg/l/h compared to 1.25 mg/l/h for the control. Phenol and m-cresol showed no effect on crystal violet decolorization, meanwhile p-cresol and p-nitrophenol reduced the decolorization rate to 1.07 and 0.01 mg/l/h, respectively. P. aeruginosa cells were immobilized by entrapment in agar–alginate beads. The beads were cultivated and reused in Erlenmeyer flask and in an air bubble column bioreactor and they enhanced the crystal violet decolorization rate to 3.33 and 7.5 mg/l/h, respectively.     

Control of the sulfide (S2-) concentration for optimal zinc removal by sulfide precipitation in a continuously stirred tank reactor

Precipitation of Zn2+ with S2- was studied at room temperature in a continuously stirred tank reactor of 0.5l to which solutions of ZnSO4 (800-5800 mgl(-1) Zn2+) and Na2S were supplied. The pH was controlled at 6.5 and S2- concentration in the reactor was controlled at set point values ranging from 3.2x10(-19) to 3.2x10(-4) mgl(-1), making use of an ion-selective S2- electrode. In steady state, the mean particle size of the ZnS precipitate decreased linearly from 22 to 1 microm for S2- levels dropping from 3.2x10(-4) to 3.2x10(-18) mgl(-1). At 3.2x10(-11) mgl(-1) of S2-, the supplies of ZnSO4 and Na2S solutions were stoichiometric for ZnS precipitation. At this S2- level, removal of dissolved zinc was optimal with effluent zinc concentration <0.03 mgl(-1) while ZnS particles formed with a mean geometric diameter of about 10 microm. Below 3.2x10(-11) mgl(-1) of S2- insufficient sulfide was added for complete zinc precipitation. At S2- levels higher than 3.2x10(-11) mgl(-1) the effluent zinc concentration increased due to the formation of soluble zinc sulfide complexes as confirmed by chemical equilibrium model calculations.     

含硝基苯、苯胺废水处理的工程实践

采用固定化微生物-曝气生物滤池与铁-炭微电解法联用的工艺方法处理含硝基苯、苯胺的废水.通过培养驯化微生物阶段、半负荷进水阶段、满负荷进水阶段的调试运行,表明:当进水CODCr＜1000 mg/L、硝基苯＜120 mg/L、苯胺＜30 mg/L时,出水可达到CODCr＜300 mg/L、硝基苯＜5 mg/L、苯胺＜5 mg/L的设计要求.铁-炭微电解法在pH值为3～4时,对废水有一定的脱色作用,但pH值升高后脱色效果不明显.     

Decolorization of textile dyes by laccases from a newly isolated strain of Trametes modesta

Four ligninolytic fungi, Trametes modesta, Trametes hirsuta, Trametes versicolor and Sclerotium rolfsii, were compared for their ability to produce laccases. The fungal laccases were screened for their ability to decolorize eight synthetic dyes (anthraquinone, azo, indigo and triarylmethane). The decolorization rate depended both on the source of the enzyme preparation and on the structure of the dye. Based on laccase production and dye decolorizing ability, T. modesta was selected for further studies. All the tested dyes were decolorized by the T. modesta laccase most efficiently under acid conditions (pH 3-6) but the optimum pH for decolorization of the individual dye varied. The decolorization rate of this laccase increased with the rise in temperature to 50-60 degrees C. The decolorization efficiency of T. modesta laccase was improved remarkably in the presence of mediators like 1-hydroxybenzotriazole and 2-methoxyphenothiazine.     

Laboratory column study for remediation of MTBE-contaminated groundwater using a biological two-layer permeable barrier

In this study, an in situ biological two-layer permeable reactive barrier system consisting of an oxygen-releasing material layer followed by a biodegradation layer was designed to evaluate the remediation effectiveness of MTBE-contaminated groundwater. The first layer containing calcium peroxide (CaO(2)) and other inorganic salts is to provide oxygen and nutrients for the immobilized microbes in the second layer in order to keep them in aerobic condition and maintain their normal metabolism. Furthermore, inorganic salts such as potassium dihydrogen phosphate (KH(2)PO(4)) and ammonium sulphate ((NH(4))(2)SO(4)) can also decrease the high pH caused by the alkali salt degraded from CaO(2). The second layer using granular expanded perlite as microbial carrier is able to biodegrade MTBE entering the barrier system. Batch experiments were conducted to identify the appropriate components of oxygen-releasing materials and the optimum pH value for the biodegradation of MTBE. At pH=8.0, the biodegradation efficiency of MTBE is the maximum and approximately 48.9%. A laboratory-scale experiment using two continuous upflow stainless-steel columns was then performed to evaluate the feasibility of this designed system. The fist column was filled with oxygen-releasing materials at certain ratio by weight. The second column was filled with expanded perlite granules immobilizing MTBE-degrading microbial consortium. Simulated MTBE-contaminated groundwater, in which dissolved oxygen (DO) content was 0mg/L, was pumped into this system at a flow rate of 500mL/d. Samples from the second column were analyzed for MTBE and its major degradation byproduct. Results showed that MTBE could be removed, and its metabolic intermediate, tert-butyl alcohol (TBA), could also be further degraded in this passive system.     

An anaerobic two-layer permeable reactive biobarrier for the remediation of nitrate-contaminated groundwater

In this paper, an anaerobic two-layer permeable reactive biobarrier system consisting of an oxygen-capturing layer followed by a biodegradation layer was designed firstly for evaluating the remediation effectiveness of nitrate-contaminated groundwater. The first layer filling with granular oxygen-capturing materials is used to capture dissolved oxygen (DO) in groundwater in order to create an anaerobic condition for the microbial denitrification. Furthermore, it can also provide nutrition, such as carbon and phosphorus, for the normal metabolism of immobilized denitrifying bacteria filled in the second layer. The second layer using granular activated carbon as microbial carrier is able to biodegrade nitrate entering the barrier system. Batch experiments were conducted to identify the effect of DO on microbial denitrification, oxygen-capturing performance of zero valent iron (ZVI) powder and the characteristics of the prepared oxygen-capturing materials used to stimulate growth of denitrifying bacteria. A laboratory-scale experiment using two continuous upflow stainless-steel columns was then performed to evaluate the feasibility of this designed system. The first column was filled with granular oxygen-capturing materials prepared by ZVI powder, sodium citrate as well as other inorganic salts, etc. The second column was filled with activated carbon immobilizing denitrifying microbial consortium. Simulated nitrate-contaminated groundwater (40 mg NO₃–N/L, pH 7.0) with 6 mg/L of DO content was pumped into this system at a flow rate of 235 mL/d. Samples from the second column were analyzed for nitrate and its major degradation byproduct. Results showed that nitrate could be removed more than 94%, and its metabolic intermediate, nitrite, could also be biodegraded further in this passive system. Further study is necessary in order to evaluate performance of its field application.     

Decolorization of an azo dye by unacclimated activated sludge under anaerobic conditions

Studies on the reductive decolorization of a complex azo dye, Reactive Red 3.1, were made as part of the development of a practical approach to better exploit the metabolic potential of biomass in wastewater treatment. Decolorization was achieved at low and variable rates by mixed microbial cultures under various environmental conditions, including low pH and high salt concentration. It was caused by reductive cleavage of the azo bond to yield two aromatic amines. More reliable and effective decolorization rates, of up to 20–30 mg l⁻¹ h⁻¹, were given by unadapted activated sludge, (6 g l⁻¹) incubated with 400 mg l⁻¹ of Reactive Red 3.1 under anaerobic conditions. Decolorization also occurred best in static conditions.     

Laccase-membrane reactors for decolorization of an acid azo dye in aqueous phase: Process optimization

In the present investigation, performance of various laccase-membrane reactor configurations including direct enzyme contact, enzyme impregnated, immobilized enzyme and a reactor system based on laccase immobilization in chitosan membranes for decolorization of azo dye (acid black 10 BX) were examined using laccase enzyme purified from white rot fungi Pleurotus ostreatus 1804. A five-step laccase purification procedure was employed, which improved the enzymatic activity by 8.27 folds. Laccase was confirmed by comparing with the standard marker using SDS-PAGE electrophoresis, which showed molecular weight of 63 kDa. Experimental data showed that laccase has great potential for color removal without addition of external redox mediators. Various process parameters viz. aqueous phase of pH 6.0, enzyme concentration of 1.75 U/ml, dye concentration of 20 mg/L, temperature of 30 °C and reaction time of 120 min were optimized to achieve maximum decolorization efficiencies. Moreover, different laccase-membrane reactor configurations were tested to determine the efficacy of repeated application of laccase on dye decolorization process. Among the different reactor configurations employed, laccase encapsulated in chitosan membrane showed advantages such as short-term contact period and reusability of enzyme for a number of cycles.     

Metabolism of azo dyes by Lactobacillus casei TISTR 1500 and effects of various factors on decolorization

Lactobacillus casei TISTR 1500 was isolated from soil of a dairy wastewater treatment plant and selected as the most active azo dye degrader of 19 isolates. Growing cells and freely suspended cells of this strain completely degraded methyl orange, thereby decolorizing the medium. The strain stoichiometrically converted methyl orange to N,N-dimethyl-p-phenylenediamine and 4-aminobenzenesulfonic acid, which were identified by HPLC, GC, and GC-MS analyses. The enzyme activity responsible for the cleavage of the azo bond of methyl orange was localized to the cytoplasm of cells grown on modified MRS medium containing methyl orange. The effect of sugars, oligosaccharides, organic acids, metal ions, pHs, oxygen and temperatures on methyl orange decolorization by freely suspended cells was investigated. The optimal conditions for the decolorization of methyl orange by the Lactobacillus casei TISTR 1500 are incubation at 35 degrees C and pH 6 with sucrose provided as the energy source.     

Chemisorption of oxygen onto activated carbon can enhance the stability of biological perchlorate reduction in fixed bed biofilm reactors

Fixed bed biofilm reactors with granular activated carbon (GAC) or glass beads as support media were used to evaluate the influence of short-term (12h) and long-term (23 days) increases of influent dissolved oxygen (DO) concentrations on biological perchlorate removal. The goal was to evaluate the extent by which chemisorption of oxygen to GAC can enhance the stability of biological perchlorate reduction. Baseline influent concentrations were 50 microg/L of perchlorate, 2 mg/L of acetate as C, and 1mg/L of DO. Perchlorate removal in the glass bead reactor seized immediately after increasing influent DO concentrations from 1 to 4 mg/L since glass beads have no sorptive capacity. In the biologically active carbon (BAC) reactor, chemisorption of oxygen to GAC removed a substantial fraction of the influent DO, and perchlorate removal was maintained during short-term increases of influent DO levels up to 8 mg/L. During long-term exposure to influent DO concentrations of 8.5mg/L, effluent perchlorate and DO concentrations increased slowly. Subsequent exposure of the BAC reactor bed to low DO concentrations partially regenerated the capacity for oxygen chemisorption. Microbial analyses indicated similar microbial communities in both reactors, which confirmed that the differences in reactor performance during dynamic loading conditions could be attributed to the sorptive properties of GAC. Using a sorptive biofilm support medium can enhance biological perchlorate removal under dynamic loading conditions.     

Comparative studies on potential of consortium and constituent pure bacterial isolates to decolorize azo dyes

The decolorization potential of the consortium HM-4 constituted by mixing four laboratory isolates identified as Bacillus cereus (BN-7), Pseudomonas putida (BN-4), Pseudomonas fluorescens (BN-5) and Stenotrophomonas acidaminiphila (BN-3) was compared with that of individual isolates. Six different azo dyes viz., C.I. Acid Red 88 (AR-88), C.I. Acid Red 119 (AR-119), C.I. Acid Red 97 (AR-97), C.I. Reactive Red 120 (RR-120), C.I. Acid Blue 113 (AB-113) and C.I. Acid Brown 100 (AB-100) were used in this study. The individual bacterial isolates were not able to completely decolorize these dyes, except for dyes AR-119 and AB-113. The consortium HM-4 was able to decolorize all the dyes used at an initial dye concentration of 20 mg L⁻¹ at a significantly higher rate as compared to that achieved by individual isolates.     

天然粉末二氧化锰处理染料废水的试验研究

分别以罗丹明B和甲基橙为目标化合物配制染料废水，研究了粉末态二氧化锰（PMN）的脱色性能，考察了pH值和接触时间对其脱色效果的影响。结果表明，pH值是影响脱色效果的关键因素，较低的pH值和适宜的接触时间有利于废水脱色。pH值为1．2时，PMN对甲基橙的脱色率为92．8％，对罗丹明B的脱色率为81．9％；pH值为1．5时，达到最大脱色率所需的时间为15min。酸性条件下，PMN对染料的去除为吸附和氧化综合作用的结果。     

Stability of the bacterial communities supported by a seven-stage biological process treating pharmaceutical wastewater as revealed by PCR-DGGE

The stabilities of the bacterial community structures supported by seven full-scale biological reactors treating pharmaceutical wastewater were investigated by denaturing gradient gel electrophoresis (DGGE) of polymerase chain reaction (PCR) amplified 16S rRNA gene fragments. Effluent quality from this treatment process was consistently high with respect to BOD5 (<30 mgl(-1)), soluble COD (<500 mgl(-1)), and total ammonia (< 5 mgl(-1) as N) concentrations. Long-term community structure stability was studied by comparing the similarity of PCR-DGGE fingerprints from samples collected 87 days apart between which the influent wastewater characteristics were relatively stable. The Dice index (Cs) of similarity was moderately high for the first four reactors (Cs = 0.61-0.77) and very high for the last three reactors (Cs = 0.89-0.91). Short-term community structure stability was studied by comparing PCR-DGGE fingerprints from samples collected 15 days apart between which the influent wastewater characteristics changed significantly, while the effluent quality remained consistently high. The bacterial community composition of each of the seven bioreactors showed a moderate community shift (Cs = 0.70-0.76). Short-term variability in influent wastewater composition, therefore, affected a greater community shift than did long-term operation treating a wastewater of relatively consistent composition. These results indicate that functionally stable wastewater treatment bioreactors have stable microbial community structures under normal operating conditions but are able to adapt in response to perturbations to sustain high effluent quality.     

Sulphate reduction and the removal of carbon and ammonia in a laboratory-scale constructed wetland

Sulphate is a normal constituent of domestic wastewater and reduced sulphur compounds are known to be potent inhibitors of plant growth and certain microbial activities. However, the knowledge about sulphate reduction and the effect on the removal of C and N in constructed wetlands is still limited. Investigations in laboratory-scale constructed wetland reactors were performed to evaluate the interrelation of carbon and nitrogen removal with the sulphate reduction by use of artificial domestic wastewater. Carbon removal was found to be only slightly affected and remained at high levels of efficiency (75-90%). Only at sulphate reduction intensities above 75 mgl(-1) (50% removal), a decrease of carbon removal of up to 20% was observed. A highly contrary behaviour of ammonia removal was found in general, which decreased exponentially from 75% to 35% related to a linear increase of sulphate reduction up to 75 mgl(-1) (50% removal). Since sulphate removal is considered to be dependant on the load of electron donors, the carbon load of the system was varied. Variation of the load changed the intensities of sulphate reduction immediately, but did not influence the carbon removal effectiveness. Doubling of the carbon concentration of 200 mgl(-1) BOD(5) for domestic wastewater usually led to sulphate reduction of up to 150 mgl(-1) (100% removal). The findings show that, particularly in constructed wetland systems, the sulphur cycle in the rhizosphere is of high importance for performance of the waste water treatment and may initiate a reconsideration of the amount of sulphate present in the tap water systems.     

Monitoring and kinetic study of ammonia oxidation using dissolved oxygen electrode and NAD(P) H fluorometer

The ammonia oxidation of a mixed culture enriched from a wastewater treatment plant sludge was monitored by a DO probe and a nicotinamide adenine dinucleotide (phosphate) (NAD(P)H) fluorometer. Under fixed aeration, DO reflected ammonia oxidation effectively. According to the DO profiles and the total ammonia concentrations analyzed experimentally, the ammonia oxidation kinetics of the enriched culture was determined. The ammonia oxidation rate was found sensitive to pH, especially at low total ammonia concentrations. At high concentrations of total ammonia, the maximum ammonia oxidation rate occurred at pH 7.6+/-0.1. At low concentrations, the pH sensitivity intensified significantly and the apparent optimal pH shifted higher with decreasing total ammonia concentrations, because NH3 molecules were the true substrate for ammonia oxidation and more NH4+ ions were converted to NH3 molecules at higher pH. The ammonia oxidation kinetics was therefore developed according to the concentration of NH3 molecules, instead of the commonly used total ammonia concentration. The kinetics followed Michaelis-Menten behavior for both DO and NH3 concentration: the maximum rate was 16.7+/-0.7 mg NH3-N/(g TSS-h) and the Michaelis constants for DO and NH3 were (14+/-2)% air saturation and (4.4+/-0.4) x 10(-2) (mg NH3-N/L), respectively. It was also concluded from the study that with or without exogenous organic substances, the NAD(P)H fluorescence of the enriched nitrifying culture was undetectable. The fluorescence did not respond to addition or depletion of substrate (ammonia, glucose, or acetate), change between aerobic and anaerobic conditions, or even KCN addition to kill the culture.