Optimisation of process parameters for bioreduction of azo dyes using Bacillus firmus under batch anaerobic condition

A new dye decolourising bacterial strain was isolated from textile wastewater and identified as Bacillus firmus. The study indicated that the bacterium could efficiently decolourise different azo dyes under static culture conditions. Characterisation of the efficiency of azo dye reduction by this isolate using both spectral and HPLC analysis was found to be a function of process parameters which include dye concentration, culture broth pH, incubation temperature, aeration as well as nitrogen source. For decolourisation, the optimal pH and temperature were 7–8 and 20–35°C respectively, while remarkable dye degradation was obtained within 18 h for dye concentrations below 100 mg L^?1. With the addition of yeast extract and under optimal conditions, dye reduction was enhanced and complete colour removal was achieved within 12 h. Colour removal was shown to be due to biodegradation rather than adsorption of dyes on bacterial cells. This study confirms the ability of the new dye‐degrading strain, Bacillus firmus, to decolourise and degrade different azo dyes and highlights its high biotechnology potential for the eco‐friendly treatment of textile wastewater when optimal conditions are applied.     

Biodegradation and detoxification potential of rotating biological contactor (RBC) with Irpex lacteus for remediation of dye-containing wastewater

Use of fungal organisms in rotating biological contactors (RBC) for bioremediation of liquid industrial wastes has so far been limited in spite of their significant biodegradation potential. The purpose was to investigate the power of RBC using Irpex lacteus for decolorization and detoxification of industrial dyes and dyeing textile liquors. Recalcitrant dye Methylene Blue (150 mg L⁻¹) was decolorized within 70 days, its mutagenicity removed, and the biological toxicity decreased more than 10-fold. I. lacteus biofilm in the RBC completely decolorized within 26 and 47 days dyeing liquors containing disperse or reactive dyes adjusted to pH4.5 and 5-fold diluted with the growth medium, respectively. Their respective biological toxicity values were reduced 10- to 10⁴-fold in dependence of the test used. A battery of toxicity tests comprising Vibrio fisheri, Lemna minor and Sinapis alba was efficient to monitor the toxicity of textile dyes and wastewaters. Strong decolorization and detoxification power of RBC using I. lacteus biofilms was demonstrated.     

Application of the laccase,produced on coconut flesh by Pleurotus florida for dye decolorization

We investigated the ability of Pleurotus florida to produce laccase on coconut flesh as a solid substrate fermentation. The decolorization of two structurally different dyes such as azo (Reactive Blue 198) and triphenylmethane dye (Malachite Green) were analysed. The decolorization of Reactive blue 198 and Malachite Green at 8 hrs was 93% and 63% respectiely. The untreated and treated dye was characterized by UV-Vis spectral and fourier transform infrared (FTIR) Spectroscopy scan. FTIR analysis pointed out the involvement of alkene (C=C) and carboxylic (C-O) groups in the decolorization process. The toxicity with respect to Allium cepa root inhibition was measured to demonstrate the potential of laccase in the detoxication and bioremediation process.     

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.     

An approach to textile dye removal using sawdust from Aspidosperma polyneuron

The textile industry is responsible for discarding wastewater contaminated with dyes. The timber industry generates waste in the form of sawdust. The aim of the present study was to evaluate the adsorptive potential of sawdust obtained from the Aspidosperma polyneuron tree for the removal of the textile dye from wastewater. Sawdust was subjected to different pre-treatments (acid, alkaline and polyethyleneimine) in order to increase its adsorption capacity. Based on the results from the isotherms, treatment with polyethyleneimine (PEI) led to the greatest adsorption capacity and fits the Freundlich model, indicating cooperative adsorption. Other treatments with sawdust best fit the Langmuir model, but the untreated sawdust presented better results than the treated sawdust. These results were only surpassed by sawdust treated with PEI. A. polyneuron revealed good potential for use as an adsorbent to remove dyes, which is a novel result, since to date there is no study on its use as a sorbent material.     

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.     

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.     

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.     

Analogies and differences between photocatalytic oxidation of chemicals and photocatalytic inactivation of microorganisms

This study reports the analogies and differences found when comparing TiO₂ photocatalytic treatment for chemical oxidation and microorganisms inactivation, using methylene blue and Escherichia coli as references, respectively. In both processes the activation is based on the same physicochemical phenomena and consequently a good correlation between them is observed when analyzing the effect of operational variables such as catalyst concentration or incident radiation flux, both factors influencing common stages such radiation absorption and generation of reactive oxygen species. However, different microbiological aspects (osmotic stress, repairing mechanism, regrowth, bacterial adhesion to the titania surface, etc) makes disinfection kinetics significantly more complex than the first-order profiles usually observed for the oxidation of chemical pollutants. Moreover, bacterial inactivation reactions are found to be extremely sensitive to the composition of water and modifications of the catalysts in comparison with the decolorization of the dye solutions, showing opposite behaviors to the presence of chlorides, incorporation of silver to the catalysts or the use of different types of immobilized TiO₂ systems. Therefore, the activity observed for the photocatalytic oxidation of organics can not be always extrapolated to photocatalytic disinfection processes.     

The evaluation of low‐cost biosorbents for removal of an azo dye from aqueous solution

There is a need to develop innovative and alternative technologies that can remove dyes from wastewater. In this study, low‐cost and locally available two renewable biosorbents (cotton stalk and apricot seed) were investigated to remove of Astrazone Black from aqueous solution. The effects of various experimental parameters such as dye concentration, adsorbent amount, adsorbent particle size and initial pH were tested, and optimal experimental conditions were examined. The results showed that as the amount of adsorbent was increased, the percentage of dye removal increased accordingly. The ratios of dye sorbed increased as the adsorbent particle size decreased. In addition, antibacterial effect of untreated and treated (decolourized) dye on a soil bacterium, Pseudomonas aeruginosa, was determined. The removal of this dye with agricultural wastes reduced the toxic effect on P. aeruginosa. This reduction in toxic effect is important both in respect of environmental biotechnology and waste detoxification.     

Mutagenicity in Salmonella of fluorescent dye tablets used in water tracing

Fluorescent red dye tablets containing Rhodamine B induced dose-related increases of His⁺ revertant colonies in Salmonella typhimurium strains TA1538 and TA98 in the presence of S9. Fluorescent yellow/green tablets containing Uranine (Fluorescein Sodium) were not mutagenic. These fluorescent dyes are sold for public, commercial, and field use, in liquid and tablet forms, for sewage, plumbing, and water pollution tracing.     

The photocatalytic degradation of reactive black 5 using TiO2/UV in an annular photoreactor

The textile effluent is a major industrial polluter because it is highly colored, containing about 15% unfixed dyes as well as high levels of salts that can potentially be discharged into the environment. Photocatalytic oxidation using an thin gap annular UV reactor with TiO2 was used to break down the colour of a synthetic effluent ranging up to 400 ppm in dye concentration of Reactive Black 5 and up to 80 g/L in NaCl. Results show that the reaction kinetics was dominated by the TiO2 loading, the initial dye concentration, and the dissolved oxygen concentration; with the other parameters showing less significant effects. High rates of decolorization were found, with a linear fit to the Langmuir-Hinshelwood equation yielding a reaction rate constant (k) of 2.45 ppm/min, and an adsorption equilibrium constant (K) of 0.048 ppm(-1) based on color removal. The presence of the combination of high dissolved oxygen (15 ppm) and sodium chloride (up to 80 g/L) was found to enhance the decolorization and mineralization rates of the reactive dye. However, pH was found to not significantly affect the degradation rate. Since textile effluent is strongly alkaline, this result is significant, as no solution neutralisation is required and direct treatment of the effluent is possible.     

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.     

Application of 'waste' metal hydroxide sludge for adsorption of azo reactive dyes

The capacity and mechanism of metal hydroxide sludge in removing azo reactive dyes from aqueous solution was investigated with different parameters, such as charge amount of dyes, system pH, adsorbent particle size, and adsorbent dosage. The three anionic dyes used were CI Reactive Red 2, CI Reactive Red 120, and CI Reactive Red 141, increasing in number of sulfonic groups, respectively. Only 0.2% (w/v) of powdered sludge (<75microm) achieved color removal from 30 mg l(-1) reactive dye solutions within 5 min without pH adjustment. The larger the charge amount of the dyes, the greater the adsorption (>90%) on the metal hydroxide sludge. The system pH played a significant role in the adsorption on metal hydroxides and formation of dye-metal complexes. The optimum system pH for dye adsorption was 8-9 which was close to the pH(zpc) of the sludge while the precipitation of dye-metal complexes occurred at system pH 2. The maximum adsorption capacity (Q degrees ) of the sludge for the reactive dyes was 48-62 mg dye g(-1) adsorbent. The Langmuir and Freundlich models showed that the higher charged dyes had a higher affinity of adsorption. The smaller particle size and the greater amount of adsorbent showed the faster process, due to an increase in surface area of adsorbent. Desorption studies elucidated that metal hydroxide sludge had a tendency for ion exchange adsorption of sulfonated azo reactive dyes. Leaching data showed that the treated water was nontoxic at a system pH above 5 or a solution pH above 2.     

Impact of culture media and sampling methods on Staphylococcus aureus aerosols

Staphylococcus aureus has been detected indoors and is associated with human infection. Reliable quantification of S. aureus using a sampling technique followed by culture assay helps in assessing the risks of human exposure. The efficiency of five culture media and eight sampling methods in recovering S. aureus aerosols were evaluated. Methods to extract cells from filters were also studied. Tryptic soy agar (TSA) presented greater bacterial recovery than mannitol salt agar (MSA), CHROMagar staph aureus, Chapman stone medium, and Baird–Park agarose (P < 0.05). Moreover, 93 ± 2%–95 ± 2% and 42 ± 1%–49 ± 2% of S. aureus were, respectively, recovered by a 15‐min heating of gelatin filters and 2‐min vortex of polycarbonate (PC) filters. Evaluation of two filtration (IOM with gelatin filter and cassette with PC filter), two impaction (Andersen 1‐STG loaded with TSA and MSA) and four impingement methods [AGI‐30 and BioSampler filled with Tween mixture (TM) and phosphate‐buffered saline (PBS)] revealed the BioSampler/TM performed best over 30 and 60 min of sampling (P < 0.05), while low recovery efficiencies were associated with the IOM/gelatin, cassette/PC, and AGI‐30/PBS combinations (P < 0.05). In addition to BioSampler/TM, collecting S. aureus onto TSA from the Andersen 1‐STG is also recommended, as it is the second best method at the 60‐min sampling (P < 0.05).     

Decolorization and toxicity of reactive anthraquinone textile dyes under methanogenic conditions

Reductive decolorization of two anthraquinone reactive dyes (Reactive Blue 4, RB4; Reactive Blue 19, RB19) under methanogenic conditions was performed using a mixed, methanogenic culture. Decolorization of the two anthraquinone dyes was investigated to evaluate the rate and extent of color removal as well as to assess possible toxic effects of the dyes and their decolorization product(s) on the methanogenic culture as a function of initial dye concentration ranging from 50 to 300 mg x L(-1). A dextrin/peptone mixture was used as the carbon and electron source. A high rate and extent of color removal was achieved ranging from 4.3 to 29.9 mg x L(-1)h(-1) and 73-91% for RB4, and 13.0-74.4 mg x L(-1)h(-1) and 90-95% for RB19. Initial RB4 concentrations up to 100 mg x L(-1) did not result in any significant inhibition. Both the 200 and 300 mg x L(-1) RB4-amended cultures, and all RB19-amended cultures resulted in severe inhibition of both acidogenesis and methanogenesis. Sequential dye addition at 300 mg x L(-1) for both RB4 and RB19 resulted in accumulation of volatile fatty acids (VFAs) and a very low methane production at the end of the first dye addition after 44 days of incubation. However, at the end of the second dye addition, after a relatively long incubation (384 days), recovery of methanogens in the RB4-amended culture was observed in contrast to the complete inhibition of methanogenesis in the RB19-amended culture. Therefore, RB19 resulted in a higher degree of inhibition of both acidogenesis and methanogenesis than RB4. Addition of dextrin/peptone to dye-inhibited cultures resulted in acidogenesis and a gradual recovery of methanogenesis (mainly aceticlastic methanogenesis) in the RB4-inhibited culture, and a slow recovery of acidogenesis but no recovery of methanogenesis in the RB19-inhibited culture. In contrast, addition of 80% H(2)-20% CO(2) gas to dye-inhibited cultures resulted in recovery of hydrogenotrophic methanogenesis in both the RB4- and RB19-inhibited cultures. In spite of the relatively severe inhibition of the two anthraquinone dyes on the mixed, methanogenic culture, a high extent of color removal was achieved.     

Bacterial activity in plant (Schoenoplectus validus) biofilms of constructed wetlands

Biofilm-bacterial communities have been exploited in the treatment of wastewater in ‘fixed-film’ processes. Our understanding of biofilm dynamics requires a quantitative knowledge of bacterial growth-kinetics in these microenvironments. The aim of this paper was to apply the thymidine assay to quantify bacterial growth without disturbing the biofilm on the surfaces of emergent macrophytes (Schoenoplectus validus) of a constructed wetland. The isotope was rapidly and efficiently taken-up and incorporated into dividing biofilm-bacteria. Isotope diffusion into the biofilm did not limit the growth rate measurement. Isotope dilution was inhibited at >12 μM thymidine. Biofilm-bacterial biomass and growth rates were not correlated to the plant surface area (r² < 0.02). The measurements of in situ biofilm-bacterial growth rates both displayed, and accommodated, the inherent heterogeneity of the complex wetland ecosystem. Biofilm-bacterial respiratory activities, measured using the redox dye CTC, and growth rates were measured simultaneously. The dye did not interfere with bacterial growth. Biofilm-bacterial specific growth rates ranged from 1.4 ± 0.6 d⁻¹ to 3.3 ± 1.3 d⁻¹. In the constructed wetlands of this study biofilm-bacterial specific growth rates, compared to those of natural ecosystems, could be markedly improved through changes in wetland design that increased bacterial respiration while minimising biofilm growth.     

An approach using a novel phage mix for detecting Pseudomonas aeruginosa in water

The present study aims to integrate the benefits of plaque assay using a novel phage mix with phylogenetic and molecular analysis for detecting Pseudomonas aeruginosa in water. Three phages were isolated and the transmission electron microscope related their morphological resemblance to those of Siphoviridae and Podoviridae families, while molecular analysis showed different cp‐gene sizes. The Phage mix was highly specific (86.0%), and data misleading didn't exceed 14.0% compared to membrane filter assay (39.2%). Time elapsed for test completion required 24 h. Identified P. aeruginosa were verified using 16S‐rDNA. Nucleotide sequence data for both phages and bacteria were submitted to the NCBI GenBank database, USA and gained their accession numbers. Concluding remarks highlight the potential of plaque assay as specific, simple and rapid method. The study recommended future efforts to isolate and characterize new phages for detecting other bacterial pathogens of public health concern to control water pollution and maintain adequate hygiene.     

Bioaugmentation of floating treatment wetlands for the remediation of textile effluent

This pilot study evaluated the effects of bacterial augmentation on the efficiency of floating treatment wetlands (FTWs) to remediate textile wastewater. Two wetland plants, Phragmites australis and Typha domingensis, were used to develop FTWs, which were then augmented with a bacterial consortium of three strains (Acinetobacter junii, Pseudomonas indoloxydans, and Rhodococcus sp.). Results showed that both plant species removed colour, organic matter, toxicity, and heavy metals from textile wastewater and their removal efficiency was further enhanced by augmentation with bacteria. The maximum removal efficiencies of colour, COD and BOD after an 8‐day period were 97, 87 and 92%, respectively, by FTWs carrying P. australis inoculated with the bacterial consortium. Furthermore, the same combination showed 87–99% reduction of heavy metals in the textile wastewater as well. These results indicate that FTWs can be used for the treatment of textile effluent and their working efficiency can be improved by plant‐bacterial synergism.     

Submerged microfiltration membrane coupled with alum coagulation/powdered activated carbon adsorption for complete decolorization of reactive dyes

Even the presence of very low concentrations of dyes (1mgL(-1)) in the effluent is highly visible and is considered aesthetically undesirable. It must be removed from wastewater completely. This study systematically evaluates the performance of adsorption (three kinds of powdered activated carbons), coagulation (AlCl3.6H2O) and membrane (submerged hollow fiber microfiltration) processes individually in treating two kinds of reactive dyes (Orange 16 and Black 5) and then using a hybrid process with combined coagulation-adsorption-membrane treatment system. Adsorption capacity and kinetics of Orange 16 were much higher and faster than those of Black 5. The dye removal efficiency by coagulation was highly dependent on dye concentration and solution pH. The hybrid process performance was far more superior that individual process in removing both kinds of dyes. It was evident that the combined coagulation-adsorption-membrane process has a great potential application for complete reactive dye removal, production of high-quality treated water and allows the reduction in the use of coagulant and adsorbent.