Decolorization kinetics of the azo dye Reactive Red 2 under methanogenic conditions: effect of long-term culture acclimation

The biological decolorization of the textile azo dye Reactive Red 2 was investigated using a mixed, mesophilic methanogenic culture, which was developed with mixed liquor obtained from a mesophilic, municipal anaerobic digester and enriched by feeding a mixture of dextrin/peptone as well as media containing salts, trace metals and vitamins. Batch decolorization assays were conducted with the unacclimated methanogenic culture and dye decolorization kinetics were determined as a function of initial dye, biomass, and carbon source concentrations. Dye decolorization was inhibited at initial dye concentrations higher than 100 mg l^-1 and decolorization kinetics were described based on the Haldane model. The effect of long-term culture exposure to the reactive dye on decolorization kinetics, culture acclimation, as well as possible dye mineralization was tested using two reactors fed weekly for two years with an initial dye concentration of 300 mg l^-1 and a mixture of dextrin/peptone. The maximum dye decolorization rate after a 2-year acclimation at an initial dye concentration of 300 mg l^-1 was more than 10-fold higher as compared to that obtained with the unacclimated culture. Aniline and the o-aminohydroxynaphthalene derivative resulting from the reductive azo bond cleavage of the dye were detected, but further transformation(s) leading to dye mineralization were not observed. Reactive Red 2 did not serve as the carbon and energy source for the mixed culture, and dye decolorization was sustained by the continuous addition of dextrin and peptone. Thus, biological decolorization of reactive azo dyes is feasible under conditions of low redox potential created and maintained in overall methanogenic systems, but supply of a biodegradable carbon source is necessary.     

Degradation of Remazol Red dye by <Emphasis Type="Italic">Galactomyces geotrichum</Emphasis> MTCC 1360 leading to increased iron uptake in <Emphasis Type="Italic">Sorghum vulgare</Emphasis> and <Emphasis Type="Italic">Phaseolus mungo</Emphasis> from soil

Removal of azo dyes from the effluent generated by textile industries is rather difficult. Azo dyes represent a major class of synthetic colorants that are both mutagenic and carcinogenic. Galactomyces geotrichum MTCC 1360, a yeast species, showed more than 96% decolorization of the azo dye Remazol Red (50 mg/L) within 36 h at 30°C and pH 11.0 under static condition with a significant reduction in the chemical oxygen demand (62%) and total organic carbon (41%). Peptone (5.0 g/L), rice husk (10 g/L extract), and ammonium chloride (5.0 g/L) were found to be more significant among the carbon and nitrogen sources used. The presence of tyrosinase, NADH-DCIP reductase, riboflavin reductase and induction in azo reductase and laccase activity during decolorization indicated their role in degradation. High performance thin layer chromatography analysis revealed the degradation of Remazol Red into different metabolites. Fourier transform infrared spectroscopy and high performance liquid chromatography analysis of samples before and after decolorization confirmed the biotransformation of dye. Atomic absorption spectroscopy analysis revealed a less toxic effect of the metabolites on iron uptake by Sorghum vulgare and Phaseolus mungo than Remazol Red dye. Remazol Red showed an inhibitory effect on iron uptake by chelation and an immobilization of iron, whereas its metabolites showed no chelation as well as immobilization of iron. Phytotoxicity study indicated the conversion of complex dye molecules into simpler oxidizable products which had a less toxic nature.     

The adsorption of Fusarium flocciferum spores on celite particles and their use in the degradation of phenol

Summary Spores of Fusarium flocciferum were inserted in porous celite beads. The effects of bead size, adsorption time course, washing cycle and spore concentration on spore loading were investigated. Cell loadings up to 50% (dry weight/beads) were obtained. The degradation of phenol using adsorbed cells was studied in batch experiments. The immobilized cell system was shown to efficiently degrade high concentrations of the substrate (up to 2.0 g/l) and to remain active for more than 2 motths. The oxygen uptake rate of free and immobilized cells was determined at various concentrations of phenol. The kinetic constants K _s=85 mg/l, K _i=345 mg/l and SMI=170 mg/l were estimated from the experimental data by linearization of the Haldane function for the free cells. The uptake rates exhibited by the confined cells were lower (30%) than those obtained for free cells and no significant differences were found for phenol concentrations between 150 and 1200 mg/l.     

Assessment of the biodegradability of a monosulfonated azo dye and aromatic amines

Biodecolourisation of an azo dye by anaerobic cultures using a liposomal textile levelling agent as primary substrate was assessed. Liposomes seem to facilitate the uptake of the dye (Acid Orange 7) by anaerobic biomass, leading to a fast decolourisation (colour removal of 96% was achieved in the first sample port of the reactor profiles). On the other hand, the presence of dye (60–300 mg l⁻¹) caused a decrease in the chemical oxygen demand (COD) degradation rate (4.1–2.5 g COD removed l⁻¹ d⁻¹ for 60 and 300 mg l⁻¹ of dye, respectively), suggesting inhibitory effects.Aerobic degradation of aromatic amines was investigated in aerobic respirometric assays with different types of inocula. Sulfanilic acid and aniline were mineralised by inocula with a significant microbiological diversity, even with domestic effluent. These results were confirmed by a significant reduction of COD, total organic carbon (TOC) and a high oxygen consumption (biochemical oxygen demand/theoretical oxygen demand), 92±4%. Kinetic analysis showed that a sigmoid function describes quite well the experimental data, even better than the exponential model. Orthanilic and metanilic acids and 1-amino-2-naphtol were persistent under the tested conditions.     

Improved biodegradation of Congored by using Bacillus sp

The biodegradation of Congored, a toxic azo dye, was studied by using a hybrid technique involving sonolysis as pretreatment followed by biological treatment. The experiments were carried out with and without pretreatment using dye solution as a sole source of nutrition with an isolated and acclimatized strain of Bacillus sp. obtained from tannery industry effluent. The pretreatment time was varied as 30, 60, 90, 120, 150 and 180 min and then the pretreated dye solution was subjected to biological treatment. The effectiveness of pretreatment was compared with the results of biological degradation of non pretreated Congored and the results showed that the pretreatment improved the efficiency of the biodegradation of Congored. During the biological degradation, the increase in initial dye concentration decreased the decolorization rate and at high concentrations (1500 and 2000 mg/l), the inhibition was observed. The optimum pH and temperature were determined to be 7.0 and 37 degrees C, respectively. The data obtained through biodegradation experiments were fitted with five different kinetic models and the results were analyzed.     

Anaerobic degradation of azo dye Drimaren blue HFRL in UASB reactor in the presence of yeast extract a source of carbon and redox mediator

This paper presents results on anaerobic degradation of the azo dye blue HFRL in a bench scale Upflow anaerobic sludge blanket (UASB) reactor operated at ambient temperature. The results show that the addition of yeast extract (500 mg/L) increased color removal (P < 0.05) from 62 to 93% despite the low chemical oxygen demand (COD) removal (~35%) which happened due to volatile fatty acids (VFA) accumulation. There were no differences in color removal (~91%) when yeast extract (500 mg/L) was used in the presence or absence of glucose, suggesting that yeast extract acted as source of redox mediator (riboflavin) and carbon. The specific rate of dye removal increased along the operational phases and depended on the presence of yeast extract, suggesting progressive biomass acclimatization. Analysis of bacterial diversity by Polymerase Chain Reaction-Denaturing Gradient Gel Electrophoresis (PCR–DGGE) method showed there was biomass selection along the bioreactor operation and no evidence of azo dye degrading bacteria predominance. This strengthens the hypothesis that color removal happens extracellularly by the reduction of azo bond by reduced redox mediators, such as riboflavin, which is present in high amount in the yeast extract.     

Biological treatment of wastewater containing an azo dye using mixed culture in alternating anaerobic/aerobic sequencing batch reactors

The performance of one stage anaerobic/aerobic processes for the biological treatment of synthetic wastewaters containing Acid Red 18 was studied. In addition, a method for evaluating dye mineralization using lumped parameters was investigated. The selected initial dye concentrations were 0, 35, 70, 140, and 280 mg/L, in reactors R1 ～ R5, respectively. This study showed that average COD removal was not lower than 85% while the remaining COD originated from Acid Red 18 and its degradation products. The majority of the dye removal occurred in the anaerobic phases and the aerobic phase contributions were insignificant. The kinetics data of dye removal showed that the increase in initial dye concentration (35 ～ 280 mg/L) caused a decrease in first-order kinetic rate constants (0.0593 ～ 0.0384/h). The overall mineralization of AR 18 dye was at least 44, 35, 13, and 0% in R2 ～ R5, respectively. Increase in initial dye concentrations had no significant effect on sludge characteristics.     

Decolorizing kinetics of a recombinant Escherichia coli SS125 strain harboring azoreductase gene from Bacillus latrosporus RRK1

PCR amplified product containing gene responsible for dye decolorization was cloned and expressed in Escherichia coli. The resulting recombinant strain E. coli SS125 decolorized 200mg/l azo dye (Remazol Red) at 30 degrees C at 255 mg cell/l/h, while the host E. coli (DH5 alpha) had no color removal ability. The dependence of the decolorization rate on initial dye concentration and the maximum rate occurred with the dye at 100 mg l(-1). The decolorization rate of E. coli SS125 was optimal at 37-45 degrees C. Aeration strongly-inhibited the decolorization, but decolorization occurred effectively under static and anaerobic incubation conditions. The E. coli SS125 strain also exhibited excellent stability during reported batch operation.     

Decolourization and Detoxification of Methyl Red by Aerobic Bacteria from A Wastewater Treatment Plant

Bacterial cultures from a wastewater treatment plant degraded a toxic azo dye (methyl red) by decolourization. Complete decolourization using a mixed-culture was achieved at pH 6, 30 °C within 6 h at 5 mg/l methyl red concentration, and 16 h at 20—30 mg/l. Four bacterial species were isolated that were capable of growth on methyl red as the sole carbon source, and two were identified, namely Vibrio logei and Pseudomonas nitroreducens. The Vibrio species showed the highest methyl red degradation activity at the optimum conditions of pH 6--7, and 30—35 °C. Analysis by NMR showed that previously reported degradation products 2-aminobenzoic acid and N,N-dimethyl-1,4-phenylenediamine were not observed. The decolourized dye was not toxic to a monkey kidney cell line (COS-7) at a concentration of 250 μM. This revised version was published online in July 2006 with corrections to the Cover Date.     

Effects of alkalinity and co-substrate on the performance of an upflow anaerobic sludge blanket (UASB) reactor through decolorization of Congo Red azo dye

The effect of substrate (glucose) concentrations and alkalinitiy (NaHCO3) on the decolorization of a synthetic wastewater containing Congo Red (CR) azo dye was performed in an upflow anaerobic sludge blanket (UASB). Color removal efficiencies approaching 100% were obtained at glucose-COD concentrations varying between 0 and 3000 mg/l. The methane production rate and total aromatic amine (TAA) removal efficiencies were found to be 120 ml per day and 43%, respectively, while the color was completely removed during glucose-COD free operation of the UASB reactor. The complete decolorization of CR dye under co-substrate free operation could be attributed to TAA metabolism which may provide the electrons required for the cleavage of azo bond in CR dye exist in the UASB reactor. No significant differences in pH levels (6.6-7.4), methane production rates (2000-2700 ml/day) and COD removal efficiencies (82-90%) were obtained for NAHCO3 concentrations ranging between 550 and 3000 mg/l. However, decolorization efficiency remained at 100% with decreasing NaHCO3 concentrations as low as 250 mg/l in the feed. An alkalinity/COD ratio of 0.163 in the feed was suggested for simultaneous optimum COD and color removal.     

Comparison of activated carbon and bottom ash for removal of reactive dye from aqueous solution

The adsorption of reactive dye from synthetic aqueous solution onto granular activated carbon (GAC) and coal-based bottom ash (CBBA) were studied under the same experimental conditions. As an alternative to GAC, CBBA was used as adsorbent for dye removal from aqueous solution. The amount of Vertigo Navy Marine (VNM) adsorbed onto CBBA was lower compared with GAC at equilibrium and dye adsorption capacity increased from 0.71 to 3.82 mg g(-1), and 0.73 to 6.35 mg g(-1) with the initial concentration of dye from 25 to 300 mg l(-1), respectively. The initial dye uptake of CBBA was not so rapid as in the case of GAC and the dye uptake was slow and gradually attained equilibrium.     

Biodegradation of azo dyes in a sequential anaerobic–aerobic system

A sequential anaerobic–aerobic treatment process based on mixed culture of bacteria isolated from textile dye effluent-contaminated soil was used to degrade sulfonated azo dyes Orange G (OG), Amido black 10B (AB), Direct red 4BS (DR) and Congo red (CR). Under anaerobic conditions in a fixed-bed column using glucose as co-substrate, the azo dyes were reduced and amines were released by the bacterial biomass. The amines were completely mineralized in a subsequent aerobic treatment using the same isolates. The maximum degradation rate observed in the treatment system for OG was 60.9 mg/l per day (16.99 mg/g glucose utilized), for AB 571.3 mg/l per day (14.46 mg/g glucose utilized), for DR 112.5 mg/l per day (32.02 mg/g glucose utilized) and for CR 134.9 mg/l per day (38.9 mg/g glucose utilized). Received: 6 August 1999 / Received revision: 20 December 1999 / Accepted: 24 December 1999     

The contribution of water soluble and water insoluble organic fractions to oxygen uptake rate during high rate composting

This study aims to establish the contribution of the water soluble and water insoluble organic fractions to total oxygen uptake rate during high rate composting process of a mixture of organic fraction of municipal solid waste and lignocellulosic material. This mixture was composted using a 20 l self-heating pilot scale composter for 250 h. The composter was fully equipped to record both the biomass-temperature and oxygen uptake rate. Representative compost samples were taken at 0, 70, 100, 110, 160, and 250 h from starting time. Compost samples were fractionated in water soluble and water insoluble fractions. The water soluble fraction was then fractionated in hydrophilic, hydrophobic, and neutral hydrophobic fractions. Each fraction was then studied using quantitative (total organic carbon) and qualitative analysis (diffuse reflectance infrared spectroscopy and biodegradability test). Oxygen uptake rates were high during the initial stages of the process due to rapid degradation of the soluble degradable organic fraction (hydrophilic plus hydrophobic fractions). Once this fraction was depleted, polymer hydrolysis accounted for most of the oxygen uptake rate. Finally, oxygen uptake rate could be modeled using a two term kinetic. The first term provides the oxygen uptake rate resulting from the microbial growth kinetic type on easily available, no-limiting substrate (soluble fraction), while the second term considers the oxygen uptake rate caused by the degradation of substrate produced by polymer hydrolysis.     

Characteristics and purification of an oxygen insensitive azoreductase from Caulobacter subvibrioides strain C7-D

An azo dye-degrading bacterium, Caulobacter subvibrioides strain C7-D, semi-constitutively produces an azoreductase that reduced the azo bond of the dyes Acid Orange (AO) 6, AO7, AO8, AO12, Acid Red (AR) 88, AR151, and Methyl Red (MR). This activity was oxygen insensitive. Of the dyes tested, AO7 was the best inducer and the most rapidly reduced substrate suggesting that dye AO7 most closely mimics the natural physiological substrate for this enzyme. The K _m for AO7 was 1 μM. Purification of the azoreductase from C. subvibrioides strain C7-D was achieved through dye-ligand affinity chromatography using the dye Orange-A covalently coupled to an agarose support. The azoreductase is approximately 30 kDa and enzyme studies indicate a single azoreductase. The optimal activity, pH, cofactor usage, substrate specificity, molecular weight and K _m characteristics of the enzyme set it apart from other known oxygen-insensitive azoreductases. Received 18 May 1999/ Accepted in revised form 13 July 1999     

Accumulation of Acid Orange 7, Acid Red 18 and Reactive Black 5 by growing Schizophyllum commune

The effect of Acid Orange 7, Acid Red 18 and Reactive Black 5 on the growth and decolorization properties of Schizophyllum commune was studied with respect to the initial pH varying from 1 to 6 and initial dye concentration (10-100 mg/L). The optimum pH value was found to be 2 for both growth and color removal of these azo dyes. Increasing the concentration of azo dyes inhibited the growth of S. commune. It was observed that S. commune was capable of removing Acid Orange 7, Acid Red 18 and Reactive Black 5 with a maximum specific uptake capacity of 44.23, 127.53 and 180.17 (mg/g) respectively for an initial concentration of 100 mg/L of the dye. Higher decolorization was observed at lower concentrations for all the dyes. Finally it was found that the percentage decolorization was more in the case of Reactive Black 5 dye compared to the other two dyes used in the present investigation.     

Fed-batch bioreactor strategies for microbial decolorization of azo dye using a Pseudomonas luteola strain

A Pseudomonas luteola strain possessing azoreductase activity was utilized to decolorize a reactive azo dye (C. I. Reactive Red 22) with fed-batch processes consisting of an aerobic cell growth stage and an anaerobic fed-batch decolorization stage. The fed-batch decolorization was conducted with different agitation and aeration rates, initial culture volumes, dye loading strategies, and yeast extract to dye (Y/D) ratios, and the effect of those operation parameters on azo dye decolorization was evaluated. Dissolved oxygen strongly inhibited the azo reduction activity; thus aeration should be avoided during decolorization but slight agitation (around 50 rpm) was needed. With the periodical feeding strategy, the specific decolorization rate (v(dye)) and overall decolorization efficiency (eta(dye)) tended to increase with increasing feeding concentrations of dye, whereas substrate inhibition seems to arise when the feeding concentration exceeded 600 mg dye/L. In the continuous feeding mode, higher initial culture volume resulted in better eta(dye) due to higher biomass loading, but lower v(dye) due to lower dye concentration in the bioreactor. With a volumetric flow rate (F) of 25 mL/h, both v(dye) and eta(dye) increased almost linearly with the increase in the loading rate of dye (F(dye)) over the range of 50-200 mg/h, while further increase in F(dye) (400 mg/h) gave rise to a decline in v(dye) and eta(dye). As the F was doubled (50 mL/h), the v(dye) and eta(dye) increased with F(dye) only for F(dye) < 80 mg/h. The best v(dye) (113.7 mg dye g cell(-)(1) h(-)(1)) and eta(dye) (86.3 mg dye L(-)(1) h(-)(1)) were achieved at F(dye) = 200 mg/h and F = 25 mL/h. The yield coefficient representing the relation between dye decolorized and yeast extract consumed was estimated as 0.8 g/g. With F(dye) = 75 mg/h, the Y/D ratio should be higher than 0.5 to ensure sufficient supply of yeast extract for stable fed-batch operations. However, performance of the fed-batch decolorization process was not appreciably improved by raising the Y/D ratio from 0.5 to 1.875 but was more sensitive to the changes in the dye loading rate.     

Aerobic degradation of a mixture of azo dyes in a packed bed reactor having bacteria-coated laterite pebbles

A microbial consortium capable of aerobic degradation of a mixture of azo dyes consisting of two isolated strains (RRL,TVM) and one known strain of Pseudomonas putida (MTCC 1194) was immobilized on laterite stones. The amount of bacterial biomass attached to the laterite stones was 8.64 g per 100 g of the stone on a dry weight basis. The packed bed reactor was filled with these stones and had a total capacity of 850 mL and a void volume of 210 mL. The feed consisted of an equal mixture of seven azo dyes both in water as well as in a simulated textile effluent, at a pH of 9.0 and a salinity of 900 mg/L. The dye concentrations of influent were 25, 50, and 100 microg/mL.The residence time was varied between 0.78 and 6.23 h. It was found that at the lowest residence time 23.55, 45.73, and 79.95 microg of dye was degraded per hour at an initial dye concentration of 25, 50, and 100 microg, respectively. The pH was reduced from 9.0 to 7.0. Simulated textile effluent containing 50 microg/mL dye was degraded by 61.7%. Analysis of degradation products by TLC and HPLC showed that the dye mixture was degraded to nontoxic smaller molecules. The bacteria-coated pebbles were stable, there was no washout even after 2 months, and the reactor was found to be suitable for the aerobic degradation of azo dyes.     

氧气对混合菌群脱色降解偶氮染料效果的影响

【背景】偶氮染料及其中间产物具有一定的环境毒性,利用混合菌群降解偶氮染料是一种环境友好型方法,但降解过程中氧气的存在起到至关重要的作用,可以促进或抑制偶氮染料的微生物降解作用。【目的】探讨氧气对偶氮染料微生物脱色液的影响,分析氧气对混合菌群脱色降解偶氮染料效果的影响。【方法】利用混合菌群DDMY1在3种培养条件(好氧、厌氧、兼氧)下,对7种偶氮染料进行脱色降解,探讨偶氮染料脱色液对氧气的响应情况,利用紫外可见分光光度法(ultraviolet visible spectrophotometry,UV-vis)和傅里叶变换红外光谱法(Fourier transform infrared spectroscopy,FTIR)对脱色产物进行分析。【结果】在兼氧和厌氧条件下反应48 h后的染料脱色液,与氧气充分接触后,部分偶氮染料微生物脱色液发生较为明显的复色现象,如活性黑5、直接黑38;UV-vis分析结果表明,这种复色现象是由于脱色液与氧气接触之后产生新物质所致;FTIR分析结果表明,混合菌群对发生复色反应的偶氮染料仍然具有一定脱色降解效果,但是脱色尚不够完全。【结论】兼氧和厌氧条件下,氧气对部分偶氮染料微生物脱色液具有较为明显的影响,从而影响混合菌群对偶氮染料的整体脱色效果,这可为今后研究偶氮染料彻底生物降解提供理论基础。     

海洋产电菌Shewanella marisflavi EP1的脱色特性

以一株新筛选得到的海洋产电菌Shewanella marisflavi EP1作为实验材料,研究了该菌株关于偶氮、蒽醌、三苯基甲烷等染料的脱色能力及脱色机制。结果表明,该菌株对这些染料均具有较好的脱色能力,最高脱色容量达到925 mg染料/(g细胞干重.d)。EP1能利用葡萄糖、蔗糖、木糖、乳酸、甲酸、柠檬酸等多种碳源将单偶氮染料丽春红2R脱色。脱色的pH、温度和NaCl浓度范围分别是:pH 6-10、15°C-40°C、0-8%。最优脱色条件:乳酸,pH 8、35°C、1%-2%NaCl,10 h内脱色率高达99.95%。分光光谱结果表明,在0-8%NaCl浓度范围内EP1脱色机制为降解脱色。