Journal of Applied & Environmental Microbiology
ISSN (Print): 2373-6747 ISSN (Online): 2373-6712 Website: Editor-in-chief: Sankar Narayan Sinha
Open Access
Journal Browser
Journal of Applied & Environmental Microbiology. 2014, 2(4), 143-154
DOI: 10.12691/jaem-2-4-8
Open AccessArticle

An Application of Mixed Consortium in Microbial Degradation of Reactive Red: Effective Strategy of Bioaugmentaiton

Maulin P Shah1,

1Industrial Waste Water Research Laboratory, Division of Applied & Environmental Microbiology, Enviro Technology Limited, Ankleshwar, Gujarat, India

Pub. Date: June 10, 2014

Cite this paper:
Maulin P Shah. An Application of Mixed Consortium in Microbial Degradation of Reactive Red: Effective Strategy of Bioaugmentaiton. Journal of Applied & Environmental Microbiology. 2014; 2(4):143-154. doi: 10.12691/jaem-2-4-8


In this paper, replacement-series method and contour analysis were applied to investigate optimal bioaugmentation strategies for the treatment of a dye-contaminated aquatic system using a constructed mixed-community for biodecolorization of a model azo dye Reactive Red. The novelty emphasizes that a species without essential target functions in a mixed culture could still play a crucial role in influencing the treatment performance. That is, although non-decolorizers (i.e., Escherichia coli DH5α) were considered metabolically ‘‘dormant’’ in this model binarybiosystem, their presence still significantly enhanced decolorization performance of the decolorizers (i.e., Pseudomonas spp.). In aerobic growth conditions, E. coli DH5α possessed a growth advantage to out-compete Pseudomonas spp. due to preferential growth rate of DH5α. However, in static decolorization conditions, DH5α seemed to produce decolorization-stimulating extracellular metabolites to help the major decolorizer (Pseudomonas spp.) decompose the toxic pollutant (i.e., the azo dye) in a short term for the benefit of total survival in the environment. The experimental results show that the presence of E. coli DH5α increased the decolorization efficiency of Pseudomonas spp. even though DH5α was an inefficient decolorizer in this microbial community. Thus, addition of DH5α into a mixed culture containing Pseudomonas spp. as a major decolorizer may lead to a bioaugmentation effect on decolorization activity. The optimal population ratio for bioaugmentation was determined by the contour analysis. The results indicate that the optimal community species ecology for maximum overall decolorization rate almost maintained at a ratio of one viable Pseudomonas spp. (0.78 x109 cells/mL) to one DH5α cell (0.70 x 109 cells/mL), representing a maximal diversity (i.e., Hmax 1.0).

Pseudomonas reactive red bioaugmentation decolorization

Creative CommonsThis work is licensed under a Creative Commons Attribution 4.0 International License. To view a copy of this license, visit


[1]  Banat IM, Nigam P, Singh D, Marchant R. Microbial decolorization of textile-dye-containing effluents: a review. Bioresour Technol 1996; 58: 217-27.
[2]  Chen B-Y. Understanding decolorization characteristics of reactive azo dyes by Pseudomonas luteola: toxicity and kinetics. Process Biochem 2002; 38: 437-46.
[3]  Keck A, Klein J, Kudlich M, Stolz A, Knackmuss H-J, Mattes R. Reduction of azo dyes by redox mediators originating in the naphthale- nesulfonic acid degradation pathway Sphingomonas sp. Strain BN6. Appl Environ Microbiol 1997; 63: 3684-90.
[4]  Maymard Jr CW. In: Kent JA, editor. Riegel’s handbook of industrial chemistry. New York: Van Nostrand Reinhold; 1983. p. 809-61.
[5]  Zollinger H. Color chemistry—syntheses, properties and applications of organic dyes and pigments New York: VCH Publishers; 1987. pp. 92-102.
[6]  Heiss GS, Gowan B, Dabbs ER. Cloning of DNA from a Rhodococcus strain conferring the ability to decolorize sulfonated azo dyes. FEMS Microbiol Lett 1992; 99: 221-6.
[7]  Chung K-T, Stevens Jr SE. Degradation of azo dyes by environmental microorganisms and helminthes. Environ Toxicol Chem 1993; 12: 2121-32.
[8]  Stolz A. Basic and applied aspects in the microbial degradation of azo dyes. Appl Microbiol Biotechnol 2001; 56: 69-80.
[9]  Zimmermann T, Kulla HG, Leisinger T. Properties of purified orange II azoreductase, the enzyme initiating azo dye degradation by Pseudomonas KF46. Eur J Biochem 1982; 129: 197-203.
[10]  Hu TL. Decolourization of reactive azo dyes by transformation with Pseudomonas luteola. Bioresour Technol 1994; 49: 47-51.
[11]  Chang J-S, Chou C, Lin Y-C, Lin P-J, Ho J-Y, Hu T-L. Kinetic characteristics of bacterial azo-dye decolorization by Pseudomonas luteola. Water Res 2001; 35: 2841-50.
[12]  Chang J-S, Lin Y-C. Fed-batch bioreactor strategies for microbial decolorization of azo dyes using a Pseudomonas luteola strain. Biotechnol Prog 2000; 16: 979-85.
[13]  HaugW, Schmidt A, Nortemann B, Hempel DC, Stolz A, Knackmuss H-J. Mineralization of the sulfonated azo dye mordant yellow 3 by 6-aminonaphthalene-2-sulfonate-degrading bacterial consortium. Appl Environ Microbiol 1991; 57: 3144-9.
[14]  Seshadri S, Bishop PL, Agha AM. Anaerobic/aerobic treatment of selected azo dyes in wastewater. Waste Manage 1994; 14: 127-37.
[15]  Flores ER, Luijten M, Donlon BA, Lettinga G, Field JA. Complete biodegradation of the azo dye azodisalicylate under anaerobic conditions. Environ Sci Technol 1997; 31: 2098-103.
[16]  Glenn JK, Gold MH. Decolorization of several polymeric dyes by the lignin-degrading Basidiomycete Phanerochaete chrysosporium. Appl Environ Microbiol 1983; 45: 1741-7.
[17]  Yang F, Yu J. Development of a bioreactor system using an immobilized white rot fungus for decolourization, Part II: Continuous decolourization tests. Bioprocess Eng 1996; 16: 9-11.
[18]  Palleria S, Chambers RP. Characterization of a Ca-alginate-immobilized Trametes versicolor bioreactor for decolourization and AOX reduction of paper mill effluents. Bioresour Technol 1997; 60: 1-8.
[19]  Zhang F-M, Knapp JS, Tapley KN. Development of bioreactor systems for decolorization of orange II using white rot fungus. Enzyme Microb Technol 1999; 24: 48-53.
[20]  Coughlin MF, Kinkle BK, Tepper A, Bishop PL. Characterization of aerobic azo dye-degrading bacteria and their activity in biofilms. Water Sci Technol 1997; 36: 215-20.
[21]  Stolz A. Basic and applied aspects in the microbial degradation of azo dyes. Appl Microbiol Biotechnol 2001; 56: 69-80.
[22]  Russ R, Rau J, Stolz A. The function of cytoplasmic flavin reductases in the bacterial reduction of azo dyes. Appl Environ Microbiol 2000; 66: 1429-34.
[23]  Morin PL. Communities. In: Community ecology. Malden, MA, USA: Blackwell Science Inc.; 1999. pp. 3-28 [chapter 1].
[24]  Mills JS, Soule ME, Doak DF. The Keystone-species concept in ecology and conservation. BioScience 1993; 43: 219-24.
[25]  Williams AC, McCarthy BC. A new index of interspecific competition for replacement and additive designs. Ecol Res 2001; 16: 29-40.
[26]  Akey WC, Jurik TW, Dekker J. A replacement series evaluation of competition between velvetleaf (Abutilon theophrasti) and soybean (Glycine max). Weed Res 1991; 31: 63-72.
[27]  Mark W, Lindow SE. Ecological similarity and coexistence of epiphytic Ice-nucleating (Ice+) Pseudomonas syringae strain and a Non-Ice-nucleating (Ice-) biological control agent. Appl Environ Microbiol 1994; 60: 3128-37.
[28]  Pianka ER. Competition and the ecological niche. In: Evolutionary ecology4th ed., New York: Harper Collins Publication Inc.; 1988. pp. 213-65 [chapter 11].
[29]  Edwards C. Some problems posed by natural environments for monitoring microorganisms.In: Environmental monitoring of bacteria. Edwards C, editor. Methods in biotechnology, vol. 12. Totowa, NJ: Human Press; 1999. p. 1-13 [chapter 1].
[30]  Duetz WA, deJong C, Williams PA, Van Andel JG. Competition in chemostat culture between Pseudomonas strains that use different pathways for the degradation of toluene. Appl Environ Microb 1994; 60:2858-63.
[31]  Chen B-Y, Chang J-S, Chen S-Y. Bacterial species diversity and dye decolorization of a two-species mixed consortium. Environ Eng Sci 2003; 20: 337-45.
[32]  Chen B-Y, Chang J-S, Chen S-Y. Bacterial decolourization enhancement using a constructed mixed consortium. J Chin Inst Chem Eng 2003; 34: 513-24.
[33]  Chen B-Y, Chen Y-W, Wu D-J, Cheng Y-C. Metal toxicity assessment upon indigenous Thiobacillus thiooxidans BC1. Environ Eng Sci 2003; 20: 375-85.
[34]  McMullan G, Meehan C, Conneely A, Kirby N, Robinson T, Nigam P, Banat IM, Marchant R, Smyth WF. Microbial decolourisation and degradation of textile dyes. Appl Microbiol Bitechnol 2001; 56: 81-7.
[35]  Chang J-S, Chen B-Y, Lin Y-S. Stimulation of bacterial decolorization of an azo dye by extracellular metabolites from Escherichia coli strain NO3. Bioresour Technol 2004; 91: 243-8.
[36]  Chen B-Y, Lim HC. Bioreactor studies on temperature induction of the Qmutant of bacteriophage l in Escherichia coli. J Biotechnol 1996; 51: 1-20.
[37]  Sen R, Swaminathan T. Application of response-surface methodology to evaluate the optimum environmental conditions for the enhanced production of surfactin. Appl Microbiol Biotechnol 1997; 47: 358-63.
[38]  Shannon CE, Weaver W. The mathematical theory of communication Urbana: University of Illinois Press; 1949.
[39]  Lin S-K. Molecular diversity assesment: logarithmic relations of information and species diversity and logarithmic relations of entropy and indistinguishability after rejection of Gibbs paradox entropy of mixing. Molecules 1996; 1: 57-67.
[40]  Chen B-Y, Induction Evolution. Entropy of bacteriophage lQ_ mutant in Escherichia coli. J Chin Inst Chem Eng 2001; 32: 81-7.
[41]  Williams GC. Design for what?In: Plan and purpose in nature—the limits of Darwinian evolution, science masters series. London, Great Britain: Phoenix; 1996. pp. 55-82.
[42]  Dover G. The ignorant gene. In: Dear Mr Darwin—letters on the evolution of life and human nature. London, Great Britain:Weidenfeld & Nicolson; 2000. pp. 49-66.