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Critical Reviews in Environmental Science and Technology


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Removal of Dyes from the Effluent of Textile and Dyestuff Manufacturing Industry: A Review of Emerging Techniques With Reference to Biological Treatment
HARPREET SINGH RAI , MANI SHANKAR BHATTACHARYYA , JAGDEEP SINGH , T. K. BANSAL , PURVA VATS & U. C. BANERJEE
a c d e f a b

Department of Chemical Engineering, Thapar Institute of Engineering and Technology, Patiala, Punjab, India
b

Department of Pharmaceutical Technology, National Institute of Pharmaceutical Education and Research, Sector 67, S.A.S. Nagar, Punjab, India
c

Department of Civil Engineering, National Institute of Technology, Jalandhar, Punjab, India


d

Department of Chemical Engineering, Thapar Institute of Engineering and Technology, Patiala, Punjab, India
e f

Institute of Microbial Technology, Sector 39-A, Chandigarh, India

Department of Pharmaceutical Technology, National Institute of Pharmaceutical Education and Research, Sector 67, S.A.S. Nagar, Punjab, India Available online: 12 Jan 2007

To cite this article: HARPREET SINGH RAI, MANI SHANKAR BHATTACHARYYA, JAGDEEP SINGH, T. K. BANSAL , PURVA VATS & U. C. BANERJEE (2005): Removal of Dyes from the Effluent of Textile and Dyestuff Manufacturing Industry: A Review of Emerging Techniques With Reference to Biological Treatment, Critical Reviews in Environmental Science and Technology, 35:3, 219-238 To link to this article: http://dx.doi.org/10.1080/10643380590917932

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Critical Reviews in Environmental Science and Technology, 35:219238, 2005 Copyright Taylor & Francis Inc. ISSN: 1064-3389 print / 1547-6537 online DOI: 10.1080/10643380590917932

Removal of Dyes from the Efuent of Textile and Dyestuff Manufacturing Industry: A Review of Emerging Techniques With Reference to Biological Treatment
HARPREET SINGH RAI
Department of Chemical Engineering, Thapar Institute of Engineering and Technology, Patiala, Punjab, India

MANI SHANKAR BHATTACHARYYA


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Department of Pharmaceutical Technology, National Institute of Pharmaceutical Education and Research, Sector 67, S.A.S. Nagar, Punjab, India

JAGDEEP SINGH
Department of Civil Engineering, National Institute of Technology, Jalandhar, Punjab, India

T. K. BANSAL
Department of Chemical Engineering, Thapar Institute of Engineering and Technology, Patiala, Punjab, India

PURVA VATS
Institute of Microbial Technology, Sector 39-A, Chandigarh, India

U. C. BANERJEE
Department of Pharmaceutical Technology, National Institute of Pharmaceutical Education and Research, Sector 67, S.A.S. Nagar, Punjab, India

Biological removal of dyes from efuents of textile and dyestuff manufacturing industry offers some distinct advantages over the commonly used chemicals and physicochemical methods. These include possible mineralization of the dyes to harmless inorganic compounds like carbon dioxide and water, and formation of a lesser quantity of relatively harmless sludge. Removal of dyes from these wastewaters has been reviewed with respect to biological decolorization as well as complete biodegradation of the dye molecules. Emerging techniques with reference to biological treatment of these wastewaters have been discussed under aerobic, anaerobic, and combined anaerobicaerobic systems. Advantages and limitations

Address correspondence to U. C. Banerjee, Department of Pharmaceutical Technology, National Institute of Pharmaceutical Education and Research, Sector 67, SAS Nagar-160 062, Punjab, India. E-mail: ucbanerjee@niper.ac.in 219

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of different biological methods have been highlighted, and future studies to establish these techniques for their applications on industrial scale have been suggested. KEY WORDS: bioaugmentation, biodegradation, decolorization, synthetic efuent, textile and dyestuff efuent, textile dyes, triphenylmethane and azo dyes

I. INTRODUCTION
Highly colored substances, widely known as colorants, can be used to impart color to an innite variety of materials described technically as substrates. Colorants can be subdivided into dyes, which are soluble in the medium in which they are applied, and pigments, which are insoluble in the application medium. Dyes are dened as colored substances that when applied to bers give them a permanent color that is resistant to action of light, water, and soap. Practically every dyestuff is made from one or more of the compounds obtained by the distillation of coal tar. The chief of these are benzene (C6 H6 ), toluene (C6 H5 CH3 ), naphthalene (C10 H30 ), anthracene (C14 H10 ), phenol (C6 H5 OH), cresol (C7 H7 OH), acridine (C13 H9 N), and quinoline (C9 H7 N). These compounds are different from the actual dyestuffs, and they must rst be changed into other compounds called intermediates. These intermediates are hydrocarbons in which one or more of the hydrogen atoms are replaced by groups such as the nitro group (-NO2 ), amino group (-NH2 ), hydroxyl group (-OH), sulfonic acid group (-OSO3 H), and others. Examples of such compounds are nitrobenzene (C6 H5 NO2 ), aniline (C6 H5 NH2 ), -naphthol (C10 H7 OH), and -naphthalenesulfonic acid (C10 H7 SO3 H). There are two important conditions for a colored compound to act as a dye5,79 : 1) Presence of a chromophore: A chromophore is a group responsible for producing a color because of its capability to absorb light in the near ultraviolet region. Some important chromophores are N O, -NO2 , -N N, -C O, C S, -C N, and (CH-CH)n , and the compounds bearing chromophores are known as chromogens.5,79 2) Presence of auxochromes: Dye should get attached to the ber by means of stable chemical bonds. These chemical bonds are formed by groups that are either acidic or basic in nature. Such groups are known as auxochromes, and examples are -OH, -COOH, -SO3 H (all acidic), and -NH2 , -NHR, -NR2 (all basic). Some dyes require no mordant for xing them with the bers. These dyes are termed direct dyes. The color of direct dyes are duller than those provided by ber-reactive dyes, and the wash-fastness is poorexpect anything dyed with them to bleed forever. The one advantage is that direct dyes may be more lightfast, that is, resistant to

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fading in the light, than ber-reactive dyes. On the basis of the chemical structure of the chromophore group, dyes are classied as azo dyes, triphenylmethane dyes, anthroquinone dyes, phthalocyanine dyes, etc.5,79,112 Since 1856, when the rst synthetic dye was reported, to this day, the use of dyes in industries and households has increased remarkably. There are more than 10,000 dyes available commercially, and more than 7 105 tons of dyestuffs are produced annually.112 The main consumers of dyes are the textile, tannery, paper and pulp, and electroplating industries. Dyes are also used as additives in petroleum products. In addition, a number of dyes and dyestuffs are widely used in the food, pharmaceutical, and cosmetic industries. The huge growth in the textile dyeing and dyestuff manufacturing industries has resulted in an increase in the volume and complexity of the wastewater discharged to the environment. During textile processing, inefciencies in dyeing result in a large amount of dyestuff being directly lost in the wastewater, which ultimately nds way into the environment. It is estimated that 510% of the dyes is lost in the efuent during the dyeing process,91 while in the case of reactive dyes, as much as 50% of the initial dye load is present in the dye bath efuent.37,82 The release of textile and dye-house efuent may cause abnormal coloration of surface waters that captures the attention of both the public and the authorities. Apart from the aesthetic problems, the greatest environmental concern with the dyes is their absorption and reection of sunlight entering the water. This interferes with the growth of bacteria and plants, causing a disturbance of the ecology of the receiving water. Thus, the loss of dyes to the environment has become an environmental hazard. This is because of the low biodegradability and the toxic nature of the dyes. In addition, the human health impact of dyes has caused concern for a number of years. Some dyes, dye precursors, and their biotransformation products such as aromatic amines have been found to be toxic, mutagenic, and carcinogenic in nature, apart from having the potential of bioaccumulating in the food chain.6,25,57,61,77 As a result of this, legislation controlling the use of such substances is being developed in various countries.45 As the world has become more cautious about the environment and also due to ever-increasing and stringent laws, the textile industry around the world has begun using innovative methods for wastewater remediation, more so for the water containing the residual color from the dyes that remain almost unaffected by the conventional aerobic treatment systems.71,81 In light of these facts, attention has come to the efcient removal of dyes from the environment. However, as compared to the growth of the dye industries and dye products, a little growth has taken place towards their removal. Here we give an overview of the emerging techniques for the treatment of dye- waste from the dye industries and textile industries, giving a special emphasis on its biological treatment procedures.

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II. BIOLOGICAL TREATMENT


Biological treatment, either aerobic or anaerobic, is generally considered to be the most effective means of removing the bulk of pollutants from complex and high-strength organic wastewater. Also, microorganisms are known to play a crucial role in the mineralization of biopolymers and xenobiotic compounds.59 Biological removal of the dyes from textile and dyestuff manufacturing industry can be broadly classied into three categories: aerobic treatment, anaerobic treatment, and combined anaerobicaerobic treatment. However, only the biological process of treatment of dye waste and textile efuent is not sufcient. The process requires the involvement of other physical, chemical, and physicochemical operations. Some of the physical processes are ltration and separation, dilution, and gamma irradiation. Physical methods are mainly used for the primary treatment of the dye wastewater. Physicochemical processes for the dye wastewater treatment include adsorption, occulation, ion-pair extraction, reverse osmosis, ion exchange, coagulation (by alum, ferric chloride, ferrous ammonium sulphate, lime, natural polymers like tamarind, seed extract, etc.), and clarication. Chemical treatments are oxidation, reduction, and chlorination. Activated carbon, charcoal, anhydrous sodium silicate, minerals, rice husk, tick wood bark, cotton waste, hair coal, y ash, ground nut shell powder, red soil, bauxite, gypsum, and clays were used for color removal from textile wastewater. Activated carbon is very effective in removing color, but it is capable of treating only small efuent volume, operates at slow speed, and has high capital cost.

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A. Aerobic Treatment
Most dyes have long been considered nonbiodegradable or nontransformable under aerobic conditions.21,71,106,107 In 1986, Pagga and Brown71 performed one of the most comprehensive studies on aerobic degradability of dyes. Eighty-seven dyestuffs were tested for their susceptibility to aerobic biodegradation. The dyestuffs chosen for the study were typical commercial products, and bacterial inocula were derived from a conventional (aerobic) efuent treatment plant. The results showed that the dyestuffs were most unlikely to show any signicant biodegradation under these conditions. However, efforts to isolate aerobic microorganisms capable of transforming dyes and dye related compounds have continued. Bacteria and fungi are the two microorganism groups that have been most widely studied for their ability to treat dye wastewaters.

1.

TREATMENT BY FUNGI

Fungal strains capable of decolorizing azo and triphenylmethane dyes have been studied in detail.19,73,78,79,94,103 Lignin-degrading white rot fungus, Phanerochaete chrysosporium, was investigated extensively during the

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last decade for its capability to degrade many recalcitrant pollutants like chlorophenols, nitrotoluenes, and polycyclic aromatic hydrocarbons.11,58 Furthermore, its ability to decolorize a wide range of dyes is now well established. Phanerochaete chrysosporium was reported to be able to decolorize three azo dyes, orange II, Congo red, and tropaeolin O, under aerobic conditions.31 Lignin peroxidase, an extracellular enzyme of P. chrysosporium, was found to be responsible for the decolorization process. Later studies revealed the potential of P. chrysosporium to decolorize direct dyes, reactive dyes, acidic dyes, and disperse dyes.23 Sani and Banerjee79 reported the decolorization of acid green 20 at 30 C by P. chrysosporium in a lowcost medium where the dye was initially converted to an unknown product, showing max at 522 nm and then transformed into another colorless compound. Ollikka et al.68 further established the potential of this fungus or its enzymes (such as lignin peroxidase) to decolorize various azo, heterocyclic, and polymeric dyes. Some other strains of fungi like Kurthia sp., Cyathus bulleri, Coriolus versicolor, Funalia trogii, Laetiporous sulphureus, and Streptomyces sp.73,79,94,103 have also been reported to decolorize some triphenylmethane and azo dyes. Phanerochaete chrysosporium, however, clearly holds an advantage over the other strains because of the nonspecic nature of the enzyme lignin peroxidase present in it; because of this, it can decolorize a diverse array of dyes. Lignin peroxidase has already been shown to be able to catalyze a wide variety of reactions, including benzylic oxidation, phenol dimerization, carboncarbon bond cleavage, hydroxylation, and O -demethylation.71,72,88 However, Wong and Yu98 identied two major problems in the application of P. chrysosporium to the treatment of real wastewater. First, lignin peroxidase, the key enzyme for dye degradation is released by fungal cells following a strict secondary metabolism under either carbon or nitrogen limitation. This means that the presence of carbon or nitrogen nutrient in the industrial efuent would prohibit the release of this enzyme by the fungal cells.111 Second, dye degradation by lignin peroxidase consumes considerable amounts of hydrogen peroxide and veratryl alcohol as reagents.104 Although veratryl alcohol is a metabolite released by the fungus, a large amount of hydrogen peroxide, veratryl alcohol, and lignin peroxidase may not be produced simultaneously in most industrial efuents. In the light of these facts, Wong and Yu98 decolorized azo and indigo dyes successfully using another fungus, Trametes versicolor. The responsible enzyme for dye decomposition was laccase, an extracellular oxidase, released by the fungus. Compared to P. chrysosporium, T. versicolor offered some distinct advantages. First, unlike P. chrysosporium, T. versicolor could produce the oxidative enzyme (laccase) even in the presence of nitrogen and carbon nutrients.62 More importantly, laccase is an oxidase, with a redox potential of 78 mV, and can catalyze the oxidation of organic pollutants even in the absence of hydrogen peroxide or other secondary metabolites.87

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Another interesting observation in this study was that while anthraquinone dye was directly oxidized by laccase, the decolorization of (nonsubstrate) azo and indigo dyes required mediation by some synthetic organic compounds or anthraquinone dye. This could be of considerable signicance in color removal from the actual industrial efuents, as they are likely to have the substrate dyes along with the nonsubstrate ones. Laccase needs O2 to catalyze the oxidation reaction, and various redox mediators have been reported to play an important role in the oxidation of phenolic compounds. Campos et al.20 reported that the pure fungal laccase, obtained from a commercial source formulation used in the textile industry, when applied without any redox mediator did not decolorize Remazol brilliant blue R (RBBR).79,80 But when a low-molecular-weight redox-mediator was added together with the laccase, decolorization was observed; violuric acid was found to be the most effective mediator, with almost complete decolorization within 20 min. Other mediators like 1-hydroxybenzonitriazole (HOBT) were found to be inefcient in comparision to the violuric acid, and a higher concentration of HOBT was inhibitory due to inactivation of laccase by the toxic (HOBT) radical. Other mediators like 2-methoxyphenothiaazine, acetosyringone, and 4-hydroxybenzenesulfonic acid (PHBS) were also found to be effective for the decolorization of various classes of dyes by laccase.

2.

TREATMENT BY BACTERIA

Similar efforts to identify and isolate aerobic bacteria capable of degrading various dyes have been going on since more than two decades ago. Ogawa et al.67 isolated a bacterium (from the sewerage system of a dyestuff factory) that could degrade various dye compounds. However, growth and respiration of the aerobic organisms involved were observed to be inhibited by the dyes used in the study. Idaka et al.50 reported the apparent aerobic reduction of simple azo compounds by Aeromonas hydrophilia. Subsequently, Kulla et al.56 described degradative pathways for sulfonated azo dyes by Pseudomonas strains previously adapted to carboxylated azo dyes. Later, Idaka et al.49 demonstrated the reductive cleavage of azo dyes by Pseudomonas cepacia. In 1991, Yatome et al.102 reported the degradation of ve triphenylmethane dyes by growing cells of Bacillus subtilis IFO 13719. A number of soil and water samples were screened for their ability to decolorize some of the triphenylmethane group of dyes, and one of the strains of Bacillus sp. MTCC B0006 decolorized 400 mg/L each of malachite green, magenta, crystal violet, brilliant green, ethyl violet, and pararosaniline.7,9 When the toxicity of dyes was assessed, it was found that crystal violet was toxic to all the organisms tested (E. coli DH5 , S. cerevisiae, and S . pombe); however, the dye decolorized by Bacillus sp. MTCC B0006 was found to be nontoxic.8 The process of decolorization of azo dyes in most studies highlighted the reduction of azo linkage via enzymatic reaction. However, the azo reductases isolated from Pseudomonas strains had a narrow substrate range.55,56,105

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More recently, Hu46 identied the bacterium Pseudomonas luteola, isolated from the sludge of an activated sludge treatment system treating dyeing wastewater, which removed the color of four reactive azo dyes: red G, RBP, RP2 B, and V2 RP. The sludge from activated sludge system was stabilized for 6 months prior to the isolation of P. luteola.48 The results showed that after undergoing shaking incubation for 48 h followed by static incubation for another 48 h, P . luteola gave color reduction of 37, 93, 93, and 88% for red G, RBP, RP2 B, and V2 RP, respectively. More recently, some successful studies on the color removal of certain azo dyes in aerobic condition have been reported.42,47,99,108 All the strains involved in these studies required an additional source of carbon and energy. Since the availability of this additional substrate could have led to the formation of anaerobic microniches, the occurrence of anaerobic reduction of azo dyes was very much suspected. Also, the possibility of anaerobic reduction of azo linkage could not be excluded even in the studies involving P. luteola,46 as the static incubation might have led to the formation of micro anaerobic zones within the aerobic system. In addition, a similar role of anaerobic microniches was also suspected in the degradation of azo dyes in anaerobic biolm reactors,28,51 especially when additional substrate was provided. On the other hand, very recent reports have demonstrated that some bacterial strains can mineralize various dyes under aerobic conditions.13,29,30,65,78 A sulfonated azo dye was decolorized and subsequently used as a carbon and energy source by a bacterial strain S5 derived from Hydrogenophaga palleronii S1.13 Also, bacterial strain MI2, isolated from an aerobic biolm reactor, was reported to mineralize acid orange 7 and acid orange 8.30 Furthermore, the ability of Sphingomonas sp. strain 1CX to mineralize acid orange 8, acid orange 10, acid red 4, and acid red 88 as sole carbon source was demonstrated.29 Most recently, successful degradation of acid 151 using an aerobic sequenced biolm reactor was reported.74 The dye served as the sole carbon source for the microorganisms, and 73% of the carbon was transformed into carbon dioxide.74 A number of triphenylmethane dyes, such as magenta, crystal violet, pararosaniline, brilliant green, malachite green and ethyl violet, were very efciently decolorized (96100%) by the cells of Kurthia sp.78 After biotransformation, the extent of COD reduction of the cell-free extract of triphenylmethane dyes was more than 88%, except in ethyl violet, where it was approximately 70%.78

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B. Anaerobic Treatment
Although some efforts in the recent past to decolorize dyes under aerobic conditions have met with success, the general perception of nonbiodegradability of most azo dyes in conventional aerobic systems still persists.21,71,81 On the other hand, the potential of anaerobic microorganisms to decolorize dyes is well documented and established.17,21,43,65,66,76,93

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The investigations on anaerobic decolorization of azo dyes were started way back in the early 1970s. Walker and Ryan95 reported decolorization of azo dyes using intestinal anaerobic bacteria. This potential of intestinal anaerobes to decolorize azo dyes was further established by other researchers.3,18,22 Later studies revealed that these compounds are readily decolorized by various other anaerobic cultures.17,22,26 However, as the phenomenon of azo dye reduction is not clearly understood yet, the term azo dye reduction may involve different mechanisms or locations, like enzymatic,43 nonenzymatic,41 mediated,93 intracellular,101 and extracellular.21 It has been established in these studies that the azo dye, instead of being biodegraded by the microorganisms, acts as an oxidizing agent for the reduced avin nucleotides of the microbial electron transport chain. Thus, the azo dye is reduced and decolorized concomitantly with reoxidation of the reduced avin nucleotides. An important feature of the reductive cleavage of these compounds is that the electrons required by the anaerobes for this process are derived from cosubstrates.41,43,66 It is thought that the presence of cosubstrates enhances the reduction of azo compounds by increasing the rate of formation of reducing equivalents.41,76 Many different cosubstrates have been found to suit as electron donors, such as glucose,21 hydrolyzed starch,97 tapioca,24 yeast extract,65 acetate, and a mixture of acetate, butyrate, and propionate.76 Along with the cosubstrate maintenance of proper environmental conditions in terms of pH, temperature, etc. also plays an important role in the decolorization process. The nature of the cosubstrate has considerable effect on the extent of decolorization of these dyes.66,76 In addition, the rate of azo dye reduction has also been observed to be dependent on the type of cosubstrates used, along with the chemical structure of the dye.93 Apart from electron donors, the role of mediators (compounds that facilitate the transport of electrons) to considerably enhance the azo reduction rate has also been observed.54,93 In a study conducted by Frank P. van der Zee et al.92 it was shown that redox-mediating compounds like anthraquinone disulfonate greatly enhanced both the chemical and biological dye reduction. It was also shown that redox-mediating enzyme cofactors released by cell lysis contribute as a stimulatory effect to the dye reduction.40 A recent report by Razo-Flores et al.,75 however, has gone a step beyond these studies. In this study, it was shown that it is possible not only to decolorize but also to completely mineralize the azo dye (mordant orange 1 MO1) in a lab-scale upow anaerobic sludge blanket (UASB) reactor even in the absence of a cosubstrate. The azo cleavage product of MO1 was identied as 5-azodisalicylate (5-ASA). It was seen that the presence of the cosubstrate was necessary only in the initial phase in order to establish an active methanogenic consortium during adaptation to the dye. The cosubstrate supplementation was obviated once 5-ASA-degrading bacteria developed in the consortium. This was because the reduction equivalents required for the azo cleavage were generated from metabolism of 5-ASA. This is contrary

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to the popular perception that the azo cleavage products are recalcitrant to anaerobic degradation.15,16,43 A recent report by Bromley-Challenor et al.14 adds another interesting aspect to the perspective just described. In this study, it was found that unadapted activated sludge could quite effectively decolorize an azo dye under anaerobic conditions in the absence of any external cosubstrate. From the linear correlation between the dye decolorization rate and its concentration, without any external source of carbon and electrons, it was concluded that the sludge utilized its endogenous energy reserves, and these were sufcient for decolorizing even higher concentration (up to 400 mg L1 ) of azo dye. This offers a distinct advantage in cost and exibility over processes using dened bacterial cultures or anaerobic digester sludge, which requires addition of a carbon source.21,66 Furthermore, the same sludge decolorized three other structurally dissimilar azo dyes, suggesting that decolorization was not a dye-specic process. It was concluded that this gratuitous anaerobic reduction (process in which microbes obtain no benet from reduction) may be applicable to other xenobiotic compounds as well. On an industrial scale, an advantage of this normally hidden metabolic potential of the microora may be found in cycling of activated sludge or in the use of submerged biological lters between aerobic and anaerobic conditions.14

C. Treatment in Combined AnaerobicAerobic System


The general perception that has emerged over the years is that most dyes are generally recalcitrant to aerobic degradation21,71,106 but can be reductively decolorized under anaerobic conditions.17,22,35 The biotransformation products generated in this process however, are not further susceptible to anaerobic attack, but are readily biodegraded under aerobic conditions.53,60,84 Hence, anaerobic decolorization followed by aerobic posttreatment is generally recommended for treating colored wastewater from textile and dyestuff manufacturing industries.16,38 This condition can be implemented either by spatial separation of the anaerobic and aerobic sludge using a sequential anaerobicaerobic reactor system110 or within one reactor, commonly termed an integrated anaerobicaerobic reactor system.38

1.

SEQUENTIAL ANAEROBIC--AEROBIC TREATMENT

This treatment system involves spatially separated sequential exposure of dye wastewater to anaerobic condition followed by aerobic environment. Numerous researchers have studied this system during the last decade.4,39,62,69 For example, in the study by An et al.,4 three dye solutions consisting of CI acid yellow 17(AY 17), CI basic blue 3 (BB 3), and CI basic red 2 (BR 2) were treated in a UASB reactor followed by a semicontinuous activated sludge tank. It was observed that in the anaerobic stage, the dyes such as AY 17,

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BB 3, and BR 2 were decolorized by 20, 72, and 78%, respectively, with no signicant color removal in the aerobic stage. Later on, an actual efuent from a dye-manufacturing factory was used for the experiment. The efuent had COD and color concentration of 1200 mg/L and 500 degrees (dilution factor) respectively. The COD and color removal of 83 and 90%, respectively, were achieved in the combined anaerobicaerobic system. It was concluded from these studies that the anaerobic stage of the combined system removes both color and COD. In addition, it also improves the biodegradability of dyes for further aerobic treatment. Likewise, most of these studies report appreciable color and organic matter removal. However, in most cases clear evidence for complete biodegradation of dyes is missing. This is primarily because there is no concrete proof of mineralization of the biotranformed products of the dyes. Only in one case43 is a clear proof of mineralization of an azo dye by bacterial coculture provided.

2.
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INTEGRATED ANAEROBIC--AEROBIC TREATMENT

The basis of these systems lies in the fact that anaerobic and aerobic microorganisms can coexist benecially in a single biolm.52,109 Supplying oxygen to an oxygen-tolerant anaerobic consortium86 can create an integrated anaerobicaerobic system. Alternatively, exposing a biolm to low concentration of oxygen along with a cosubstrate28,38 can also create such conditions. Study conducted by Tan et al.86 demonstrated that two azo dyes, 4-phenylazophenol (4-PAP) and mordant yellow 10 (MY10), were appreciably degraded in an integrated anaerobicaerobic treatment system. Exposing anaerobic granular sludge to oxygen implemented the integration of anaerobic and aerobic conditions. The system observed temporary accumulation of aromatic amines, that is, 4-aminophenol (4-AP) and aniline from 4-PAP, and 5-aminosalicylic acid (5-ASA) and sulfanilic acid (SA) from MY10, resulting from the reduction of the dye molecules. These compounds were subsequently mineralized either by facultative aerobic bacteria present in the anaerobic sludge or by addition of inocula from aerobic enrichment culture (developed from aromatic amines) to the anaerobic sludge. The study concluded that aerobic enrichment culture developed from aromatic amines, combined with oxygen tolerant anaerobic granular sludge, could potentially be used to completely biodegrade azo dyes under integrated anaerobic aerobic conditions. Anaerobic digestion was enhanced in efciency by appropriate bioaugmentation. This was followed by aerobic posttreatment. The cosubstrate ethanol was successfully applied as an electron donor for azo dye reduction. It also created anaerobic microniches to facilitate anaerobic azo dye reduction in the presence of oxygen. A recent study by Tan et al.85 aimed at studying the mineralization of three azo dyes, mordant orange 1 (MO1), 4-phenylazophenol (4-PAP), and mordant yellow 10 (MY10), under integrated as well as sequential anaerobic aerobic conditions. The results of this study demonstrated that azo dyes were

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mineralized in both integrated and sequential anaerobicaerobic conditions. However, the sequential anaerobicaerobic system was recommended for two reasons. Firstly, faster azo dye reduction rates were observed under sequential anaerobicaerobic conditions as compared to the integrated ones. This was because of the fact that in the anaerobic stage of the sequential system, the cosubstrate is used only for donating electrons for azo dye reduction, whereas in the case of an integrated system, part of the cosubstrate is used in creating anaerobic microniches. Second, aerobic degradation of the cosubstrates under integrated conditions usually gives rise to deciency of either oxygen or cosubstrate, which eventually is unfavorable for decolorization and mineralization of azo dyes. Where an excess of cosubstrate caused shortage of oxygen, resulting in reduced rate of aerobic degradation of aromatic amines, excess of oxygen resulted in a severe decline in the azo dye reduction efciency in the bioreactor. These negative effects obviously do not occur in the sequential anaerobic aerobic system as observed in this study. Thus, a good balance between cosubstrate and oxygen is very much required, which was not easily achieved in the integrated anaerobicaerobic conditions. Another study conducted by Van der Zee93 focused on optimizing the anaerobic phase of an integrated anaerobicaerobic treatment system when used for azo dye degradation. It states that the system, though effective, is sluggish due to slowness of the anaerobic phase. The aim was to speed up the anaerobic phase of the treatment. It was shown that the anaerobic phase can be optimized by using redox mediators, either by continuous dosing of soluble quinones or by incorporation of AC (activated carbon) in the sludge blanket. Thus, an efcient as well as fast integrated anaerobic aerobic treatment system can also now be developed.

III. FUTURE WORK A. Other Important Classes of Dyes


Most of the reports published in literature are either on azo or reactive azo dyes. This is because azo dyes account for 50% of all the dyestuffs produced and the azo bond is the most common chromospheres of reactive textile dye.96 Furthermore, the toxicity, mutagenicity, and carcinogenicity of these dyes and their precursors and biotransformation products are widely reported.25,57,61,77 Reports are also available on other important classes of dyes like triphenylmethane, anthraquinone, and indigoid, and their degradation is generally mediated by the enzyme laccase.1,10,20,27,62,83 Some dyes from the triphenylmethane group have also been reported to be toxic, mutagenic, and carcinogenic to biota.12,44 Therefore, more studies are required to evaluate and establish biological methods for degradation and mineralization of these and other important classes of dyes.

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B. Anaerobic High-Rate Reactors


In a study conducted by Donlon et al.,33 degradation of nitrophenols (contaminants originating from manufacturing of dyes) was investigated in a UASB reactor, which was continuously fed with a volatile fatty acid (VFA) mixture as a primary substrate. The results showed dramatic detoxication of the nitrophenols in the reactor where the concentration of nitrophenol was 25 times higher than the normal values. The overall results indicated that a UASB reactor can be used to rapidly detoxify and under certain conditions completely degrade nitroaromatic compounds. The successful operation of continuous anaerobic reactors based on reductive detoxication of other similar organic compounds has also been cited in the literature.64,65,100 The main cause of effective treatment of these xenobiotics under anaerobic conditions in high rate reactors is the rapid facile reduction of these compounds to products of lower toxicity.34,83 Furthermore, the immobilization of anaerobic bacteria and maintenance of a high concentration of biomass in the high rate reactors are factors that improve the tolerance of the anaerobic system to toxic substances.36,70 Also, the use of VFAs, glucose, ethanol, etc. as primary substrates not only builds up a high concentration of biomass in the reactor, but also increases the number of reducing equivalents (electron donors), which leads to rapid reduction of these compounds to lesser toxic metabolites.76,89,90 In the light of this, anaerobic high-rate reactors should be given special attention for future studies on biological treatment of dye wastewaters.

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C. Bioaugmentation
Bioaugmentation is the process of obtaining maximum biodegradability by the addition of nonindigenous microbial cultures to have optimum bacterial biomass. This offers considerable advantage in dealing with the problems of bacterial acclimatization, toxicity of compounds, and restart of the system. It was successfully employed in the anaerobic phase of anaerobicaerobic integrated treatment of dyes.86 Also, for example, in the study conducted by Razo-ores et al.76 the introduction of 5-ASA-degrading bacteria at the very beginning would have obviated the need for the cosubstrate in the early stages of the experiment. In addition, the time lost in developing the 5-ASA-degrading bacteria in the reactor would have been avoided.

D. Mineralization of Dyes
Decolorization of the dyes often results from the initial degradation of the dye molecules to some intermediate organic compounds. These compounds contribute to biochemical oxygen demand (BOD) and chemical oxygen demand

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(COD) of the wastewater, This means that dye wastewater even if effectively decolorized, may more often fall short of the required consent standard for its disposal into the receiving bodies. It may be necessary to reveal the biological mechanisms responsible for the decolorization and degradation of dyes. Besides, the biotranformation products of the dyes may some times be even more toxic than the dyes themselves. Therefore, mineralization of these compounds should be an integral part of the studies on biological decolorization of dye wastewater.72 Enrichment studies to isolate organisms responsible for utilizing various dyes as the sole source of carbon and energy would go a long way in effective decolorization and mineralization of these pollutants.

IV. CONCLUSION
Accumulation of dyestuff and dye wastewater creates not only environmental pollution but also medical and aesthetic problems associated with human health and society. Though biological means of dye degradation and removal from the efuents of dye and textile industries hold promises for meaningful addressing the problem, the method relies on success of nding out a suitable organism and designing of condition for the process. At the same time, biological means of dye degradation alone cannot tackle the problem successfully. Therefore, along with biological processes, other preand posttreatment methods are required. Pretreatment of dyeing efuent by advanced oxidation processes (AOP) catalyzed by a source of ultraviolet (UV) light and a powerful oxidant is a promising alternative for the effective removal of color and refractory organics from the efuent.2 Metal oxides, mainly TiO2 , in combination with a strongly photoreactivating agent like UV rays are used for this process. Zinc oxide appears to be a suitable alternative to TiO2 for water treatment.32 A crucial feature in designing such system is the optimization of operating conditions (such as UV and oxidant dosages), to yield maximum removal at acceptable costs. On the other hand, to date, dye degradation and removal of dye waste from the environment are somewhat at an immature stage from the microbiological point of view, and a better understanding of the processes is required to apply a suitable combination of aerobic, anaerobic, pure culture, and mixed culture methodologies for effective treatment of dye waste. Processes of mineralization of dye waste also have to be integrated with the biodegradation so that the products originate from the biodegradation processes do not contaminate the environment further more. An understanding of the enzymology of dye degradation will further enhance the efciencies of these processes towards its successful application. Application of modern molecular biology techniques for cloning and overexpression is expected to enter the eld of dye degradation by biological means, with signicant impact on it.

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