Decolorization and detoxification of different azo dyes by Phanerochaete chrysosporium ME-446 under submerged fermentation.

Azo dyes are widely used in the textile industry due to their resistance to light, moisture, and oxidants. They are also an important class of environmental contaminant because of the amount of dye that reaches natural water resources and because they can be toxic, mutagenic, and carcinogenic. Different technologies are used for the decolorization of wastewater containing dyes; among them, the biological processes are the most promising environmentally. The aim of this study was to evaluate the potential of Phanerochaete chrysosporium strain ME-446 to safely decolorize three azo dyes: Direct Yellow 27 (DY27), Reactive Black 5 (RB5), and Reactive Red 120 (RR120). Decolorization efficiency was determined by ultraviolet-visible spectrophotometry and the phytotoxicity of the solutions before and after the fungal treatment was analyzed using Lactuca sativa seeds. P. chrysosporium ME-446 was highly efficient in decolorizing DY27, RB5, and RR120 at 50 mg L-1, decreasing their colors by 82%, 89%, and 94% within 10 days. Removal of dyes was achieved through adsorption on the fungal mycelium as well as biodegradation, inferred by the changes in the dyes' spectral peaks. The intensive decolorization of DY27 and RB5 corresponded to a decrease in phytotoxicity. However, phytotoxicity increased during the removal of color for the dye RR120. The ecotoxicity tests showed that the absence of color does not necessarily translate to an absence of toxicity.

Activity of the endophytic fungi Phlebia sp. and Paecilomyces formosus in decolourisation and the reduction of reactive dyes' cytotoxicity in fish erythrocytes.

The current study investigates the potential for discolouration and degradation of Reactive Blue 19 and Reactive Black 5 textile dyes by endophytic fungi Phlebia sp. and Paecilomyces formosus as well as the potential cytotoxicity of products or by-products generated by the treatments in fish erythrocytes. It was observed at 30 days that both endophytes showed biodegradation activity with 0.1 g mL-1 of dyes. P. formosus showed highest extracellular and intracellular protein content levels after the 15th day, and Phlebia sp. stands out for production of extracellular laccase, indicating that this enzyme may be associated with the decolouration capacity. The dyes showed toxic effects in fishes at 0.01 g mL-1 concentration, resulting in the appearance of micronuclei in erythrocyte cells. When degraded dyes treated by endophytes were tested, the frequency of micronuclei reduced approximately 20%, indicating the effectiveness of these endophytic in the treatment of textile dyes with less environmental impact, thus indicating a potential for application of these fungi in bioremediation process.

Graphene oxide as electron shuttle for increased redox conversion of contaminants under methanogenic and sulfate-reducing conditions.

Graphene oxide (GO) is reported for the first time as electron shuttle to increase the redox conversion of the azo compound, reactive red 2 (RR2, 0.5mM), and the nitroaromatic, 3-chloronitrobenzene (3CNB, 0.5mM). GO (5mgL(-1)) increased 10-fold and 7.6-fold the reduction rate of RR2 and 3CNB, respectively, in abiotic incubations with sulfide (2.6mM) as electron donor. GO also increased by 2-fold and 3.6-fold, the microbial reduction rate of RR2 by anaerobic sludge under methanogenic and sulfate-reducing conditions, respectively. Deep characterization of GO showed that it has a proper size distribution (predominantly between 450 and 700nm) and redox potential (+50.8mV) to promote the reduction of RR2 and 3CNB. Further analysis revealed that biogenic sulfide plays a major role on the GO-mediated reduction of RR2. GO is proposed as an electron shuttle to accelerate the redox conversion of recalcitrant pollutants, such as nitro-benzenes and azo dyes.

Immobilized humic substances as redox mediator for the simultaneous removal of phenol and Reactive Red 2 in a UASB reactor.

The present study reports a novel treatment concept combining the redox-mediating capacity of immobilized humic substances with the biodegrading activity of anaerobic sludge for the simultaneous removal of two representative pollutants of textile wastewaters (e.g., phenol and Reactive Red 2 (RR2)) in a high-rate anaerobic reactor. The use of immobilized humic substances (1 g total organic carbon (TOC) L(-1), supported on an anion exchange resin) in an upflow anaerobic sludge blanket (UASB) reactor increased the decolorization efficiency of RR2 (~90 %), extent of phenol oxidation (~75 %), and stability as compared to a control UASB reactor operated without immobilized humic substances, which collapsed after 120 days of dye introduction (50-100 mg L(-1)). Increase in the concentration of immobilized humic substances (2 g TOC L(-1)) further enhanced the stability and efficiency of the UASB reactor. Detection of aniline in the effluent as RR2 reduction product confirmed that reduction of RR2 was the major mechanism of dye removal. This is the first demonstration of immobilized humic substances serving as effective redox mediators for the removal of recalcitrant pollutants from wastewater in a high-rate anaerobic bioreactor. The novel treatment concept could also be applicable to remove a wide variety of contaminants susceptible to redox conversion, which are commonly found in different industrial sectors.

Evaluation of the genotoxic potential of reactive black 5 solutions subjected to decolorizing treatments by three fungal strains.

The genotoxic potential of solutions of the textile dye "Reactive Black 5" that were subjected to decolorizing treatments with the fungal strains Coriolopsis polyzona MUCL33483, Penicillium sp. MUBA001 and Pycnoporus sp. MUBA002 was tested. The genotoxicity of the solutions was determined by evaluation of micronuclei formation in Vicia faba root cells and calculation of a damage index (MN(ID)). Non-treated Reactive Black 5 solutions (50-1000 ppm) caused a statistically significant increase in micronuclei formation and, by then, in damage index. Solutions of dye treated with C. polyzona MUCL33483 and Pycnoporus sp. MUBA002 showed color loss, probably due to enzymatic breakdown of the colorant, but maintenance or even an increase in genotoxicity. On the other hand, the Penicillium sp. strain MUBA001 caused decolorization of the dye, apparently by adsorption on mycelia, and, for solutions that initially contained 50 ppm of colorant, an elimination of the genotoxicity was observed after three weeks of treatment.

Kinetics during the redox biotransformation of pollutants mediated by immobilized and soluble humic acids.

The aim of this study was to elucidate the kinetic constraints during the redox biotransformation of the azo dye, Reactive Red 2 (RR2), and carbon tetrachloride (CT) mediated by soluble humic acids (HAs) and immobilized humic acids (HAi), as well as by the quinoid model compounds, anthraquinone-2,6-disulfonate (AQDS) and 1,2-naphthoquinone-4-sulfonate (NQS). The microbial reduction of both HAs and HAi by anaerobic granular sludge (AGS) was the rate-limiting step during decolorization of RR2 since the reduction of RR2 by reduced HAi proceeded at more than three orders of magnitute faster than the electron-transferring rate observed during the microbial reduction of HAi by AGS. Similarly, the reduction of RR2 by reduced AQDS proceeded 1.6- and 1.9-fold faster than the microbial reduction of AQDS by AGS when this redox mediator (RM) was supplied in soluble and immobilized form, respectively. In contrast, the reduction of NQS by AGS occurred 1.6- and 19.2-fold faster than the chemical reduction of RR2 by reduced NQS when this RM was supplied in soluble and immobilized form, respectively. The microbial reduction of HAs and HAi by a humus-reducing consortium proceeded 1,400- and 790-fold faster than the transfer of electrons from reduced HAs and HAi, respectively, to achieve the reductive dechlorination of CT to chloroform. Overall, the present study provides elucidation on the rate-limiting steps involved in the redox biotransformation of priority pollutants mediated by both HAs and HAi and offers technical suggestions to overcome the kinetic restrictions identified in the redox reactions evaluated.

Impact of the redox mediator sodium anthraquinone-2,6-disulphonate (AQDS) on the reductive decolourisation of the azo dye Reactive Red 2 (RR2) in one- and two-stage anaerobic systems.

This work assessed the impact of the redox mediator sodium anthraquinone-2,6-disulphonate (AQDS) on the reductive decolourisation of the azo dye Reactive Red 2 (RR2) in one- and two-stage anaerobic systems (R(1) and R(2), respectively). The two-stage system achieved better colour removal efficiencies (52-62%) than the single-stage system (23-33%) in the absence of AQDS. Addition of AQDS accelerated the electrons transfer from the substrate (ethanol) to the dye, which increased the colour removal efficiency of both anaerobic systems (≈ 85%). Finally, the impact of acidogenic and methanogenic phases separation was masked by AQDS supplementation.

Degradation kinetics of 4-amino naphthalene-1-sulfonic acid by a biofilm-forming bacterial consortium under carbon and nitrogen limitations.

By decolorization of azo dyes, caused by reductive cleavage of the azo linkage, toxic or recalcitrant amines are generated. The present study deals with the effect of the inflowing medium composition (C:N ratio) on the kinetic behavior of a bacterial biofilm-forming consortium, able to use as carbon, nitrogen and sulfur source, the molecule of 4-aminonaphthalene-1-sulfonic acid (4ANS), which is one of the most recalcitrant byproducts generated by decolorization of azo dyes. All the experiments were carried out at room temperature in a lab-scale packed-bed biofilm reactor. Because environmental conditions affect the bioreactor performance, two mineral salts media containing 4ANS, with distinct C:N ratios; 0.68 (carbon as the limiting nutrient) and 8.57 (nitrogen as the limiting nutrient) were used to evaluate their effect on 4ANS biodegradation. By HPLC and COD measurements, the 4ANS removal rates and removal efficiencies were determined. The cultivable bacterial strains that compose the consortium were identified by their 16S rDNA gene sequence. With the enrichment technique used, a microbial consortium able to use efficiently 4ANS as the sole carbon source and energy, nitrogen and sulfur, was selected. The bacterial strains that constitute the consortium were isolated and identified. They belong to the following genera: Bacillus, Arthrobacter, Microbacterium, Nocardioides, and Oleomonas. The results obtained with this consortium showed, under nitrogen limitation, a remarkable increase in the 4ANS removal efficiency η(ANS), and in the 4ANS volumetric removal rates R (V,4ANS), as compared to those obtained under carbon limitation. Differences observed in bioreactor performance after changing the nutrient limitation could be caused by changes in biofilm properties and structure.

Optimization of culture medium composition for manganese peroxidase and tyrosinase production during Reactive Black 5 decolourization by the yeast Trichosporon akiyoshidainum.

Decolourization and degradation of the diazo dye Reactive Black 5 was carried out by the yeast Trichosporon akiyoshidainum. A nine-factor Plackett-Burman design was employed for the study and optimization of the decolourization process and production of manganese peroxidase (MnP) and tyrosinase activities. In the present study, 26 individual experiments were conducted and three responses were evaluated. Raising yeast extract concentration significantly enhanced decolourization and MnP production. Carbon and nitrogen sources, glucose and (NH4)2 SO4, showed no significant effect on any response over the concentration range tested. Other culture medium components, such as CaCl2 or MgSO4, could be excluded from the medium formula, as they had no effect on the evaluated responses. Metal ions (Fe, Cu and Mn) showed different effects on decolourization and enzymatic activities. Addition of copper significantly enhanced MnP activity and decreased dye decolourization. On the contrary, iron had a positive effect on decolourization and no effect on enzyme production. Oddly, increasing manganese concentration had a positive effect on tyrosinase production without affecting decolourization or MnP activity. These results strongly suggest that dye decolourization should be regarded as a complex multi-enzymatic process, where optimal medium composition should arise as a compromise between those optimal for each implied enzyme production.

Sequential anaerobic/aerobic treatment of dye-containing wastewaters: colour and COD removals, and ecotoxicity tests.

Colour and COD removals of the azo dyes Congo Red (CR) and Reactive Black 5 (RB5) were individually evaluated in a sequential anaerobic/aerobic treatment system. Additionally, dye toxicity was assessed by using acute ecotoxicity tests with Daphnia magna as the indicator-organism. The anaerobic reactor was operated at approximately 27 °C and with hydraulic retention times of 12 and 24 h. The aerobic reactor was operated in batch mode with a total cycle of 24 h. During anaerobic step, high colour removals were obtained, 96.3% for CR (400 mg/L) and 75% for RB5 (200 mg/L). During the aerobic phase, COD effluent was considerably reduced, with an average removal efficiency of 52% for CR and 85% for RB5, which resulted in an overall COD removal of 88% for both dyes. Ecotoxicity tests with CR revealed that the anaerobic effluent presented a higher toxicity compared with the influent, and an aerobic post-treatment was not efficient in reducing toxicity. However, the results with RB5 showed that both anaerobic and aerobic steps could decrease dye toxicity, especially the aerobic phase, which removed completely the toxicity in D. magna. Therefore, the anaerobic/aerobic treatment is not always effective in detoxifying dye-containing wastewaters, sometimes even increasing dye toxicity.

Detoxification of azo dyes mediated by cell-free supernatant culture with manganese-dependent peroxidase activity: effect of Mn2+ concentration and H2O2 dose.

White-rot fungi (WRF) are capable of degrading complex organic compounds such as lignin, and the enzymes that enable these processes can be used for the detoxification of recalcitrant organopollutants. The aim of this study is to evaluate a system based on the use of an in vitro ligninolytic enzyme for the detoxification of recalcitrant dye pollutants. The dyes selected for investigation were the anionic and cationic commercial azo dyes, basic blue 41 (BB41), acid black 1 (AB1), and reactive black 5 (RB5). A supernatant, cell-free culture of WRF with manganese peroxidase activity was used to investigate its degradative capacity under various conditions, and concentrations of cofactors, H(2)O(2) and Mn(2+). The assays were carried out using a 2(2) experimental designs whose variables were concentration of Mn(2+) (33 and 1,000 µM) and semicontinuous dosage of the H(2)O(2) (0.02 and 0.10 µmol) added at a frequency of 0.2 min(-1). The response variables analyzed were the efficiency and the initial rate of the decolorization process. The dye concentrations considered ranged from 10 to 200 mg L(-1). AB1 and RB5 were decolorized over the entire interval of concentrations studied; reaching efficiencies between 15 and 95%. Decolorization of up to 100 mg L(-1), BB41 had less than 30% efficiency. The decay of the concentration of AB1 was interpreted by two-stage kinetics model, with the exception of the condition of 33 µM Mn(2+)-0.02 µmol of H(2)O(2) in which only one stage was observed. For all assays performed with 33 µM Mn(2+), the initial rate of the decolorization process was found to be dependent on the dosage of H(2)O(2). The results of this study can be applied to the development bioreactors for the degradation of recalcitrant pollutants from the textile industry and may be used as a model for expanding the use of extracellular enzyme supernatants in bioremediation.

Immobilized humic substances on an anion exchange resin and their role on the redox biotransformation of contaminants.

A novel technique to immobilize humic substances (HS) on an anion exchange resin is presented. Immobilized HS were demonstrated as an effective solid-phase redox mediator (RM) during the reductive biotransformation of carbon tetrachloride (CT) and the azo model compound, Reactive Red 2 (RR2). Immobilized HS increased ∼4-fold the extent of CT reduction to chloroform by a humus-reducing consortium in comparison to incubations lacking HS. Immobilized HS also increased 2-fold the second-order rate constant of decolorization of RR2 as compared with sludge incubations lacking HS. To our knowledge, the present study constitutes the first demonstration of immobilized HS serving as an effective solid-phase RM during the reductive biotransformation of priority contaminants. The immobilizing technique developed could be appropriate for enhancing the redox biotransformation of recalcitrant pollutants in anaerobic wastewater treatment systems.

Anthraquinone-2,6-disulfonate (AQDS) as a catalyst to enhance the reductive decolourisation of the azo dyes Reactive Red 2 and Congo Red under anaerobic conditions.

In this study, we assessed the catalytic effect of anthraquinone-2,6-disulfonate (AQDS) to enhance the reductive decolourisation of the azo dyes Reactive Red 2 and Congo Red in batch and continuous-flow experiments. While testing the anaerobic sludge 1 in assays free of AQDS, the highest values for the first-order kinetic constant (k1) were found with co-substrates formate and glucose. In the assays that contained 50 microM of AQDS, the k1 values increased with all co-substrates tested, increasing by 3.5-fold when ethanol was the electron donor. The upflow anaerobic sludge blanket (UASB) reactors R1 (AQDS-free) and R2 (AQDS-supplemented) reached excellent decolourisation efficiencies (higher than 90%) even for the high Congo Red concentration tested (1.2 mM). However, electron donor depletion in the influent drastically decreased the colour removal capacity in both bioreactors. Reactor R2 presented higher stability and decolourisation efficiency compared to R1, indicating that the addition of a redox mediator can be valuable for treating dye-coloured wastewaters.

Biodecolorization screening of synthetic dyes by four white-rot fungi in a solid medium: possible role of siderophores.

AIMS: Four selected fungi were screened for their ability to decolourize a textile effluent and commercial reactive dyes in a solid medium. METHODS AND RESULTS: Ligninolytic enzymes activities (lignin peroxidase, manganese peroxidase and laccase) and siderophores presence were monitored in decolourized plates. RESULTS: The results showed low lignin peroxidase activity and no manganese peroxidase activity was detected for all fungi. Laccase activity was observed in Reactive Blue 19 decolourized plates by Trametes versicolor and Trametes villosa. Siderophores presence was observed in Trametes versicolor, Phanerochaete chrysosporium and Lentinus edodes decolourized plates. CONCLUSION: Lentinus edodes displayed the greatest decolourization ability both in terms of extent and rapidity of decolourization. SIGNIFICANCE AND IMPACT OF THE STUDY: The transformation observed for dyes open the possibility to study siderophores to treat dyes and textile effluents.

Acidic pH modulates the interaction between human granulocyte-macrophage colony-stimulating factor and glycosaminoglycans.

Granulocyte-macrophage colony-stimulating factor (GM-CSF) controls growth and differentiation of hematopoietic cells. Previous reports have indicated that the mitogenic activity of GM-CSF may be modulated by the glycosidic moiety of proteoglycans associated with the membrane of stromal cells. In this work, we have performed in vitro studies of the interaction between GM-CSF and glycosaminoglycans. The addition of heparin promoted a marked blue shift in the fluorescence emission spectrum of GM-CSF as well as a 30-fold increase in the intensity of light scattering, which indicates formation of large molecular weight complexes between the two molecules. Interestingly, heparin-induced changes in the spectral properties of GM-CSF were only observed at acidic pH. The dependence on acidic pH, together with a strict dependence on glycosaminoglycan sulfation and the fact that high ionic strength destabilized the interaction, indicates that the association between GM-CSF and glycosaminoglycans is mediated by electrostatic interactions. These interactions probably involve sulfate groups in the glycosaminoglycans and positively charged histidine residues in GM-CSF. We propose that negatively charged glycolipids present on the plasma membrane of the hematopoietic and/or the stromal cell could promote an acidic microenvironment capable of triggering interaction between GM-CSF and membrane-bound proteoglycans in vivo.

[Degradation of naphthalene-2-sulfonate by strains of Micromonospora]. / Degradación de ácido naftalén-2-sulfónico por cepas de Micromonospora.

It was possible to isolate several strains of Micromonospora sp. from waters of Rio Reconquista. They all were filamentous bacteria with lateral spores, highly resistant to Cr(III) and another heavy metal cations. All these strains were able to grow on naphthalene-2-sulphonate as sole carbon source in a mineral medium. The biodegradation of the xenobiotic proceeds via the formation of salicylate and gentisate. These compounds have been isolated and mass spectrometry identified.

[2-Naphthalene-sulphonic acid degrading microorganisms]. / Microorganismos degradadores de ácido naftalen-2-sulfónico.

Derivatives of anionic surfactants, specially naphthalene sulphonates, when discharged into natural waters are accumulated in waters and sediments because of their poor biodegradability. A four-membered bacterial community, able to degrade the sodium salt of 2-naphthalene-sulphonic acid (2NS), was isolated from Río Reconquista. All the isolated strains consisted of gram-negative, strictly aerobic rods, with a strong proteolytic activity and resistance to high levels of cations like Cr (III), Hg (II), Cd (II) and Pb (II). Some of them were resistant to gentamicin, amikacin and chloramphenicol. These strains appeared to be related to the genera Pseudomonas and Alcaligenes. When isolated, if growing on 2NS, a brown-dark pigment is formed by three of them, resulting in an inhibition of growth. The presence of a Pseudomonas aeruginosa strain avoided the production of pigment and resulting in a complete consume of 2NS.