Biodecolorization of azo dye mixture (Remazol Brilliant Violet 5R and Reactive Red 120) by indigenous bacterial consortium isolated from dye contaminated soil

Aims@#The present study investigated the biodegradation and removal of dye mixture (Remazol Brilliant Violet 5R and Reactive Red 120) using a new bacterial consortium isolated from dye-contaminated soil.@*Methodology and results@#Among the total 15 isolates screened, the two most efficient bacterial species (SS07 and SS09) were selected and identified as Enterobacter cloacae (MT573884) and Achromobacter pulmonis (MT573885). The removal efficiency of dye mixture by E. cloacae and A. pulmonis at an initial concentration of 100 mg/L was 82.78 and 84.96%, discretely. The bacterial consortium was developed using selected isolates and the optimum conditions for removing dyes were investigated. The maximum decolorization efficiency was achieved at pH 7; 35 °C; dye concentration, 100 mg/L; and initial inoculum concentration, 0.5 mL with mannitol and ammonium sulfate as carbon and nitrogen sources. The maximum removal efficiency of 91.3 ± 3.35% was achieved at the optimal conditions after 72 h of incubation.@*Conclusion, significance and impact of study@#Decolorization of azo dyestuff by the developed microbial consortia conforms to the zero-order reaction kinetics model. Consortia of E. cloacae and A. pulmonis was established as an effective decolorizer for the Remazol Brilliant violet 5R and Reactive Red 120 dye mixture with >90% color removal.

O estudo teórico do desempenho dos polímeros condutores dos corantes azoicos na detecção eletroquímica de indigo-carmim / The theoretical study of the function of conducting polymers of the azo-dyes in the electrochemical detection of indigo-carmine

RESUMO O desempenho dos polímeros condutores dos corantes azoicos durante a detecção eletroquímica de indigo-carmim foi investigado do ponto de vista teórico, sendo o modelo, correspondente ao caso, descrito e analisado mediante a teoria de estabilidade lineal e da análise de bifiircações. Foi mostrado que o sistema eletroanalítico depende fortemente do pH, pois as concentrações excessivas dos prótons levam à ineficiência eletroanalítica, haja vista o bloqueio dos centros ativos da reação. No entretanto, malgrado o supracitado, os polímeros dos corantes azoicos são modificadores eficientes para determinação do indigo-carmim. A possibilidade das instabilidades oscilatória e monotônica também foi verificada.

SUMMARY The function of the conducting polymers of azo-dyes during the indigo-carmine electrochemical detection has been investigated from the theoretical point of view. The correspondent model has been described and analyzed by means of linear stability theory and bifurcation analysis. It has been shown that the electroanalytical system depends strongly on pH, as the excessive protons concentrations drive the system to the electroanalytical inefficiency, as they block the reaction active sites. Nevertheless, despite of the mentioned, the azo-dyes conducting polymers are efficient modifiers for indigo-carmine electrochemical determination. The possibility of oscillatory and monotonic instabilities has also been verified.

Characterization of the biosorption of fast black azo dye K salt by the bacterium Rhodopseudomonas palustris 51ATA strain

BACKGROUND: Removal of dyes from wastewater by microorganisms through adsorption, degradation, or accumulation has been investigated. Biological methods used for dye treatment are generally always effective and environmentally friendly. In this study, biosorption of the Fast Black K salt azo dye by the bacterium Rhodopseudomonas palustris 51ATA was studied spectrophotometrically, at various pH (2­10), temperatures (25°C, 35°C, and 45°C) and dye concentrations (25­400 mg L-1). RESULTS: The bacterial strain showed extremely good dye-removing potential at various dye concentrations. IR studies at different temperatures showed that the dye was adsorbed on the bacterial surface at lower temperatures. Characteristics of the adsorption process were investigated by Scatchard analysis at 25°C and 35°C. Scatchard analysis of the equilibrium binding data for the dye on this bacterium gave rise to linear plots, indicating that the Langmuir model could be applied. The regression coefficients obtained for the dye from the Freundlich and Langmuir models were significant and divergence from the Scatchard plot was observed. CONCLUSION: The adsorption behavior of the dye on this bacterium was expressed by the Langmuir, Freundlich, and Temkin isotherms. The adsorption data with respect to various temperatures provided an excellent fit to the Freundlich isotherm. However, when the Langmuir and Temkin isotherm models were applied to these data, a good fit was only obtained for the dye at lower temperatures, thus indicating that the biosorption ability of R. palustris 51ATA is dependent on temperature, pH, and dye concentration.

Levedura Pichia kudriavzevii SD5 como biocatalizador na descoloração do corante Preto Reativo 5 / Pichia kudriavzevii yeast as biocatalizer in Reactive Black 5

RESUMO As leveduras vêm apresentando bons resultados na biodegradação de corantes, tornando-se uma alternativa ambientalmente segura e de custo mais baixo para o tratamento de efluentes contendo corantes industriais. Assim, o objetivo deste trabalho foi estudar a descoloração do azo corante Preto Reativo 5 (PR5) pela levedura Pichia kudriavzevii SD5. Para otimização dos parâmetros de descoloração do Preto Reativo 5 foram realizados dois planejamentos experimentais do tipo Delineamento Composto Central Rotacional (DCCR) 23, sendo cada planejamento composto por 17 ensaios, a 150 rpm durante 24 h. As variáveis estudadas foram pH, temperatura e concentração do corante e tiveram como resposta a porcentagem de descoloração. Também foram realizados testes de toxicidade do sobrenadante após 16 h e 24 h de cultivo, utilizando-se sementes de alface (Lactuca sativa) e o microcrustáceo Artemia salina. Os resultados mostraram que a P. kudriavzevii SD5 é uma levedura tolerante a vários tipos de estresse, uma vez que apresentou capacidade de degradar elevadas concentrações do corante PR5 a 45° C. Entretanto, apenas a temperatura apresentou influência estatisticamente significativa (p < 0,05) na descoloração do PR5. Os bioensaios de toxicidade demonstraram que ocorreu diminuição da toxicidade após 24 h de cultivo, e o perfil de absorbância do sobrenadante apontou para um mecanismo degradativo de descoloração.

ABSTRACT Yeasts have been showing good results in the biodegradation of industrial dyes, becoming an environmentally safe and cost-effective alternative for the treatment of effluents containing industrial dyes. Thus, the objective of this work was to study the discoloration of the Reactive Black 5 (RB5) azo dye by the yeast Pichia kudriavzevii SD5. Two experimental designs were employed to optimize the discoloration parameters by means of Central Composite Design (CCD) 23, totalizing 17 trials each, at 150 rpm for 24 h. The studied independent variables were pH, temperature, and concentration of the dye and the outcome parameter was the rate of decolorization (%). Furthermore, the toxicity bioassays of the supernatant after 16 and 24 h of the culture were carried out using lettuce seeds (Lactuca sativa) and the microcrustacean Artemia salina. Results showed that P. kudriavzevii SD5 is a multi-stress tolerant yeast, being capable to degrade high concentrations of RB5 at 45° C. However, only the temperature showed statistical significance (p < 0.05) for dye discoloration. Toxicity bioassays demonstrated that toxicity decreased after 24 h of culture and the absorbance profile of the supernatant pointed to a degradative mechanism of discoloration.

Structural-functional analyses of textile dye degrading azoreductase, laccase and peroxidase: a comparative in silico study

Background: Textile industry not only plays a vital role in our daily life but also a prominent factor in improving global economy. One of the environmental concern is it releases huge quantities of toxic dyes in the water leading to severe environmental pollution. Bacterial laccase and azoreductase successfully oxidize complex chemical structure of nitrogen group-containing azo dyes. Additionally, the presence of textile dye infuriates bacterial peroxidase to act as a dye degrading enzyme. Our present study deals with three textile dye degrading enzymes laccase, azoreductase, and peroxidase through analyzing their structural and functional properties using standard computational tools. Result: According to the comparative analysis of physicochemical characteristics, it was clear that laccase was mostly made up of basic amino acids whereas azoreductase and peroxidase both comprised of acidic amino acids. Higher aliphatic index ascertained the thermostability of all these three enzymes. Negative GRAVY value of the enzymes confirmed better water interaction of the enzymes. Instability index depicted that compared to laccase and preoxidase, azoreductase was more stable in nature. It was also observed that the three model proteins had more than 90% of total amino acids in the favored region of Ramachandran plot. Functional analysis revealed laccase as multicopper oxidase type enzyme and azoreductase as FMN dependent enzyme, while peroxidase consisted of α-ß barrel with additional haem group. Conclusion: Present study aims to provide knowledge on industrial dye degrading enzymes, choosing the suitable enzyme for industrial set up and to help in understanding the experimental laboratory requirements as well.

Efficiency of decolorization of different dyes using fungal biomass immobilized on different solid supports

Abstract Different technologies may be used for decolorization of wastewater containing dyes. Among them, biological processes are the most promising because they seem to be environmentally safe. The aim of this study was to determine the efficiency of decolorization of two dyes belonging to different classes (azo and triphenylmethane dyes) by immobilized biomass of strains of fungi (Pleurotus ostreatus - BWPH, Gleophyllum odoratum - DCa and Polyporus picipes - RWP17). Different solid supports were tested for biomass immobilization. The best growth of fungal strains was observed on the washer, brush, grid and sawdust supports. Based on the results of dye adsorption, the brush and the washer were selected for further study. These solid supports adsorbed dyes at a negligible level, while the sawdust adsorbed 82.5% of brilliant green and 19.1% of Evans blue. Immobilization of biomass improved dye removal. Almost complete decolorization of diazo dye Evans blue was reached after 24 h in samples of all strains immobilized on the washer. The process was slower when the brush was used for biomass immobilization. Comparable results were reached for brilliant green in samples with biomass of strains BWPH and RWP17. High decolorization effectiveness was reached in samples with dead fungal biomass. Intensive removal of the dyes by biomass immobilized on the washer corresponded to a significant decrease in phytotoxicity and a slight decrease in zootoxicity of the dye solutions. The best decolorization results as well as reduction in toxicity were observed for the strain P. picipes (RWP17).

Staining fungal structures with artificial dyes used in the industry of juices / Coloração de estruturas de fungos com corantes artificiais usados na fabricação de sucos

ABSTRACT: The objective of this research was to evaluate the efficiency of artificial dyes, sunset yellow and red bordeaux S, and the use of glycerol in different concentrations to consistently stain fungal structures in slides containing spores of Oidium sp., Albugo ipomoeae-panduratae, Pochonia chlamydosporia and hyphae of Phytopythium helicoides. Commercial product mixtures of the artificial dyes at 0.5, 1.0, 1.5, 2.0, 3.0 and 5.0% (w/v) added with glycerol at 0.25, 0.5 and 1.0% were evaluated. To stain chlamydospores, the suspension was placed in the staining solution or heated at 80ºC for 5 minutes. The slides were prepared by the wet mount slide method. Fungal spores were consistently stained starting at a concentration of 2% of the staining solution. The addition of glycerol to the staining solution improved the contrast of the sporangia, hyphae and chlamydospores. Higher intensity and uniformity of chlamydospore's staining was verified using 3% dye solution and 1% heated glycerol, when compared to the unheated and blue-cotton solution.

RESUMO: Neste trabalho, objetivou-se avaliar a eficiência dos corantes artificiais, amarelo crepúsculo e vermelho bordeaux S, e o uso do glicerol em diferentes concentrações, na montagem de lâminas com esporos de Oidium sp., Albugo ipomoeae-panduratae, Pochonia chlamydosporia e hifas de Phytopythium helicoides. Foram avaliadas as concentrações de 0,5, 1,0, 1,5, 2,0, 3,0 e 5,0% (p/v) do produto comercial da mistura dos corantes artificiais e adição de glicerol nas concentrações de 0,25, 0,5 e 1,0%. Para coloração de clamidósporos, a suspensão foi colocada na solução corante ou aquecida a 80ºC por 5 minutos e as lâminas preparadas com líquido de montagem. A partir da concentração de 2% da mistura dos corantes houve maior coloração dos esporos. A adição de glicerol na solução corante melhorou o contraste dos esporângios, hifas e clamidósporos. Maior intensidade e uniformidade de coloração de clamidósporos ocorreram na solução corante 3% e glicerol 1% aquecida, em comparação com a solução sem aquecimento e azul-de-algodão.

Panus tigrinus as a potential biomass source for Reactive Blue decolorization: isotherm and kinetic study

Background: Textile and dye industries pose a serious threat to the environment. Conventional methods used for dye treatment are generally not always effective and environmentally friendly. This drove attention of scores of researchers to investigate alternative methods for the biodegradation of dyes using fungal strains. In this work, white-rot fungus (Panus tigrinus) was used as a biosorbent for the decolorization of Reactive Blue 19. The process parameters that were varied were initial concentration (50­150 mg/L), contact time (30­90 min), and pH (2­6). In addition, to gain important data for the evaluation of a sorption process, the equilibrium and kinetics of the process were determined. Results: White-rot fungus showed great potential in decolorizing Azo dyes. The strain showed the maximum decolorization of 83.18% at pH 2, a contact time of 90 min, and an initial concentration of 50 mg/L. The Langmuir isotherm described the uptake of the Reactive Blue 19 dye better than the Freundlich isotherm. Analysis of the kinetic data showed that the dye uptake process followed the pseudo second-order rate expression. Conclusion: The biosorption process provided vital information on the process parameters required to obtain the optimum level of dye removal. The isotherm study indicated the homogeneous distribution of active sites on the biomass surface, and the kinetic study suggested that chemisorption is the rate-limiting step that controlled the biosorption process. According to the obtained results, P. tigrinus biomass can be used effectively to decolorize textile dyes and tackle the pollution problems in the environment.

Determination of Total Nitrogen in Seawater by Micro Sequential Injection-Cadmium Column Reduction Spectrophotometry / 分析化学

A cadmium column reduction-azo dyes spectrophotometric method based on micro sequential injection lab-on-valve was established for the determination of total nitrogen in seawater. The experimental parameters were optimized, and the interference experiment was carried out. The results showed that the interference of the main components and salinity in sea water could be eliminated by using a series of standard solution prepared by national standard seawater with certain salinity. The concentration of total nitrogen in seawater was linear with the absorbance in the range of 0 . 03-1 . 00 mg/L with a correlation coefficient of 0. 9993. When determining the national standard seawater at nitrogen concentration of 0. 20 mg/L, the relative standard deviation (RSD) was 4. 9%, the detection limit was 0. 010 mg/L, and the recoveries were 99. 5%-101 . 1%. There were not significance differences between the results of this method and national standard method in the t-test analysis. The method is suitable for the determination of total nitrogen in seawater.

Urinary Bladder Cancer with Focus on Occupational Dye Workers.

Benzidine based azo dyes are proven carcinogens, mutagens and have been linked to bladder cancer of human beings and laboratory animals. The textile and dyestuff manufacturing industry are the two major sources for releasing of azo dyes. Various research groups have started work on genotoxic effect of textile dyes in occupational workers of textile dye industry. Bladder cancer is the most common form of cancer in dye industries. Most of people between age 50 and 70 group of are diagnosed with bladder cancer. Men are more likely than the women to develop bladder cancer. Bladder cancer is a disease in which abnormal cells multiply without control in the bladder. The most common type of bladder cancer begins in cells lining the inside of the bladder and is called transitional cell carcinoma. Tumor markers are substances that can be found in the body when cancer is present. They are most often found in the blood or urine. The review deals about the impacts of the industry dyes on human health.

Effect of laccase from Hypsizygus ulmarius in decolorization of different dyes.

The objective of the study was to check the laccase (purified from Hypsizygus ulmarius) for decolorization of different dyes. The purified laccase from Hypsizygus ulmarius was studied for its decolorization of different dyes (Remazol brilliant blue R (RBBR), Alizarin red, Congo red, methyl orange and methyl violet). The results indicated that the percent of decolorization was increased when the time course and enzyme concentration was increased. The purified laccase showed maximum amount of decolorization in RBBR (85%) and followed by Methyl Orange (75%), Alizarin Red (73%), Methyl Violet (72%) and Congo Red (69%) without any additional redox mediator which suggest that this enzyme could be used in industries for effluent treatment.

Decolorization of two azo dyes using marine Lysobacter sp. T312D9

Aims: Novel azo dye-degrading bacterium T312D9 strain has been isolated from Abou Quir Gulf, Alexandria, Egypt. Methodology and Results: The identification of the isolate by 16S rRNA gene sequencing revealed to be Lysobacter sp. This marine ecofriendly isolate was exploited for its ability to degrade two synthetic azo dyes considered as detrimental pollutants from industrial effluents: congo red and methyl red. Using different dye concentrations showed the highest metabolic activity for complete degradation obtained from 100 to 500 mg/L within 30 h under static condition, also, sustaining higher dye loading of 1 g/L was carried out. The significant induction of enzymes NADH - 2,6-dichloroindophenol (NADH-DCIP) reductase and tyrosinase indicated their prominent role in dye degradation. The biodegradation of two azo dyes were analyzed by gas chromatographicmass spectrum analysis (GC-MS) and Fourier transform infrared spectroscopy (FTIR) before and after treatment. Toxicity study revealed the much less toxic nature of the metabolites produced after complete decolorization. Conclusion, significance and impact of study: Lysobacter sp T312D9 represent an inexpensive and promising marine bacteria for removal of both methyl and congo red. High sustainable metabolic activity for biodegradation under static condition. NADHDCIP reductase and tyrosinase were significantly induced during biodegradation of dyes. The obtained metabolites revealed to be less toxic in nature which offers a practical biological treatment.

Mycoremediation of Congo red dye by filamentous fungi

Azo, anthroquinone and triphenylmethane dyes are the major classes of synthetic colourants, which are difficult to degrade and have received considerable attention. Congo red, a diazo dye, is considered as a xenobiotic compound, and is recalcitrant to biodegradative processes. Nevertheless, during the last few years it has been demonstrated that several fungi, under certain environmental conditions, are able to transfer azo dyes to non toxic products using laccases. The aim of this work was to study the factors influencing mycoremediation of Congo red. Several basidiomycetes and deuteromycetes species were tested for the decolourisation of Congo red (0.05 g/l) in a semi synthetic broth at static and shaking conditions. Poor decolourisation was observed when the dye acted as the sole source of nitrogen, whereas semi synthetic broth supplemented with fertilizer resulted in better decolourisation. Decolourisation of Congo red was checked in the presence of salts of heavy metals such as mercuric chloride, lead acetate and zinc sulphate. Decolourisation parameters such as temperature, pH, and rpm were optimized and the decolourisation obtained at optimized conditions varied between 29.25- 97.28 percent at static condition and 82.1- 100 percent at shaking condition. Sodium dodecyl sulphate polyacrylamide gel electrophoretic analysis revealed bands with molecular weights ranging between 66.5 to 71 kDa, a characteristic of the fungal laccases. High efficiency decolourisation of Congo red makes these fungal forms a promising choice in biological treatment of waste water containing Congo red.

Degradation and detoxification of three textile Azo dyes by mixed fungal cultures from semi-arid region of Brazilian Northeast

The aim of this work was to study the degradation and detoxification of three textile azo dyes (Reactive Red 198, Reactive Red 141 and Reactive Blue 214) by mixed fungal cultures from semi-arid region of Brazilian Northeast. Sediment samples of twenty water reservoirs in the surroundings of Serra da Capivara National Park, area of environmental preservation in the caatinga in the State of Piauí, with semi-arid climate, were evaluated in order to select the consortia of fungi capable to degrade and detoxify these dyes. The mixed fungal culture from Caldeirão Escuridão (CE) reservoir was the most efficient in the degradation and detoxification of the dyes tested.

Role of brown-rot fungi in the bioremoval of azo dyes under different conditions

The present study is vital to the understanding of bioremediation of structurally different azo dyes by some unusual Brown-rot fungi. Bioremoval of each dye (20 mg l-1) was tested in two different culture media under static and shaking conditions by taking inocula from different fungi. Fungal strains showed varying dyes removal abilities, though considerable high in case of Acid Red (AR) 151(di-azo) as compared to Orange (Or) II (mono-azo). With an exception of Aspergillus tereus SA3, all the fungal isolates showed higher removal of dyes in SDB. Under static condition, the maximum decolorizing fungal strains were; Aspergillus flavus SA2 (67 percent) and Alternaria spp. SA4 (57 percent) in AR 151, while Penicillium spp. (34 and 33 percent) in Orange II, in SDB and STE, respectively. Bioremoval of dyes was considerably increased when experiments were shifted from static to shaking mode. It was specifically increased ( percent) in; AR 151 (255) with Penicillium spp., Or II with A. flavus SA2 (112) and Alternaria spp. (111). The primary mechanism of dyes removal proved to be fungal biosorption. However, reduction of dyes (onto fungal) with formation of their products (α. naphthol, sulphalinic acid and aniline) furthermore revealed that dyes (specifically azo) were actually biodegraded.

Uso de extrato de levedura como fonte de carbono e de mediadores redox, para a degradação anaeróbia de corante azo / Use of yeast extract as source of carbon and redox mediators for the anaerobic degradation of azo dyes

O trabalho investigou a influência do uso do extrato de levedura, fonte dos mediadores redox riboflavina e nicotinamida, na remoção de cor de solução de corante azo Drimaren Azul HF-RL em condições anaeróbias. O trabalho envolveu a execução de ensaios em batelada, em frascos-reatores mantidos a 25 ºC, incubados com o azo-corante e lodo anaeróbio na presença e ausência de fontes de carbono (extrato de levedura ou glicose) e de mediadores redox (riboflavina ou extrato de levedura). O monitoramento da variação temporal de cor, a demanda química de oxigênio (DQO) e ácidos graxos voláteis (AGV) mostraram que a adição de extrato de levedura (0,5 g/L) resultou em eficiências de remoção de cor de 80 a 85 por cento nas primeiras 24 horas de incubação, e que os produtos da degradação do azo-corante foram tóxicos para todo o consórcio anaeróbio, o que resultou em baixas eficiências de remoção de DQO na presença e ausência do extrato de levedura. Os resultados indicaram, ainda, que as eficiências de remoção de cor foram inferiores a 30 por cento na presença de apenas glicose (fonte de carbono) ou riboflavina (mediador redox), indicando que o extrato de levedura atuou simultaneamente como fonte de carbono e de mediadores redox.

This paper investigated the influence of using yeast extract, which is the source of redox mediators riboflavin and nicotinamide, in the decolorization of solutions containing the azo dye Drimaren Blue HF-RL in anaerobic conditions. It involved the incubation of serum bottles kept at 25 ºC and inoculated with the azo-dye, and anaerobic sludge in the presence and absence of carbon source (glucose or yeast extract) and redox mediators (riboflavin and yeast extract). The monitoring of color, chemical oxygen demand (COD) and volatile fatty acids (VFA) showed that the addition of yeast extract (0.5 g/L) resulted in 80 to 85 percent color removal in the first 24 hours of incubation; and that the metabolites of dye degradation were toxic to the anaerobic microorganisms, which led to low COD removal efficiencies either in the presence or absence of yeast extract. The results also showed that the efficiencies of color removal were below 30 percent in the presence of only glucose or riboflavin, indicating that the yeast extract acted simultaneously as source of carbon and redox mediators.

A simple method to screen for azo-dye-degrading bacteria.

A stab-culture method was adapted to screen for azo dyes-decolorizing bacteria from soil and water samples. Decolorized azo dye in the lower portion of the solid media indicates the presence of anaerobic azo dyes-decolorizing bacteria, while aerobic decolorizing bacteria decolorizes the surface portion of the solid media. Of twenty soil samples tested, one soil sample shows positive results for the decolourisation of two azo dyes; Biebrich scarlet (BS) and Direct blue 71 (DB) under anaerobic conditions. A gram negative and oxidase negative bacterial isolate was found to be the principal azo dyes degrader. The isolate was identified by using the BiologTM identification system as Serratia marcescens.

Review of Studies on Azo Dyes Decolorization / 微生物学通报

Biological process is an important approach to treat the dye wastewater. The azo dyes decolouration by special azoreductase of different aerobic bacteria and fungi was summarized. Under anaerobic condition, reductive decolourization of azo dyes was carried in the presence of redox mediators which act as electron shuttle. It was also pointed out that azo dye reduction occurred mainly under anaerobic condition. Different electron donor resulted in different decolourization rate. Problems of current biotechnology were analyzed and corresponding measures were discussed.