Applied Microbiology and Biotechnology

, Volume 75, Issue 5, pp 1191–1200 | Cite as

Bioaugmentation and coexistence of two functionally similar bacterial strains in aerobic granules

  • He-Long JiangEmail author
  • Abdul Majid Maszenan
  • Joo-Hwa Tay
Environmental Biotechnology


The survival of the inoculated microbial culture is critical for successful bioaugmentation but impossible to predict precisely. As an alternative strategy, bioaugmentation of a group of microorganisms may improve reliability of bioaugmentation. This study evaluated simultaneous bioaugmentation of two functionally similar bacterial strains in aerobic granules. The two strains, Pandoraea sp. PG-01 and Rhodococcus erythropolis PG-03, showed high phenol degradation and growth rates in phenol medium, but they were characterized as having a poor aggregation activity and weak bioflocculant-producing and biofilm-forming abilities. In the spatially homogeneous batch conditions, strain PG-01 with higher growth rates outcompeted strain PG-03. However, the two strains could stably coexist in the spatially heterogeneous conditions. Then the two strains were mixed and bioaugmented into activated sludge in two sequencing batch reactors, which were operated with the different settling times of 5 and 30 min, respectively. Aerobic granules were developed only in the reactor with a settling time of 5 min. Fluorescence in situ hybridization and denaturing gradient gel electrophoresis showed that the two strains could coexist in aerobic granules but not in activated sludge. These findings suggested that the compact structure of aerobic granules provided spatial isolation for coexistence of competitively superior and inferior strains with similar functions.


Aerobic granules Bioaugmentation Coexistence Competition Phenol degradation Spatial effect 



H.L.J. thanks Singapore Millennium Foundation for financial support.


  1. Abed RMM, Safi NMD, Koster J, de Beer D, El-Nahhal Y, Rullkotter J, Garcia-Pichel F (2002) Microbial diversity of a heavily polluted microbial mat and its community changes following degradation of petroleum compounds. Appl Environ Microbiol 68:1674–1683CrossRefGoogle Scholar
  2. Alm EW, Oerther DB, Larsen N, Stahl DA, Raskin L (1996) The oligonucleotide probe database. Appl Environ Microbiol 62:3557–3559CrossRefGoogle Scholar
  3. American Public Health Association (APHA) (1998) Standard methods for the examination of water and wastewater, APHA, Washington, DCGoogle Scholar
  4. Ayala-del-Rio HL, Callister SJ, Criddle CS, Tiedje JM (2004) Correspondence between community structure and function during succession in phenol-and phenol-plus-trichloroethene-fed sequencing batch reactors. Appl Environ Microbiol 70:4950–4960CrossRefGoogle Scholar
  5. Beun JJ, Hendriks A, van Loosdrecht MCM, Morgenroth E, Wilderer PA, Heijnen JJ (1999) Aerobic granulation in a sequencing batch reactor. Water Res 33:2283–2290CrossRefGoogle Scholar
  6. Boon N, de Gelder L, Lievens H, Siciliano SD, Top EM, Verstraete W (2002) Bioaugmenting bioreactors for the continuous removal of 3-chloroaniline by a slow release approach. Environ Sci Technol 36:4698–4704CrossRefGoogle Scholar
  7. Bouchez T, Patureau D, Dabert P, Juretschko S, Dore J, Delgenes P, Moletta R, Wagner M (2000) Ecological study of a bioaugmentation failure. Environ Microbiol 2:179–190CrossRefGoogle Scholar
  8. Bruns MA, Hanson JR, Mefford J, Scow KM (2001) Isolate PM1 populations are dominant and novel methyl tert-butyl ether-degrading bacteria in compost biofilter enrichments. Environ Microbiol 3:220–225CrossRefGoogle Scholar
  9. Chen JQ, Weimer PJ (2001) Competition among three predominant ruminal cellulolytic bacteria in the absence or presence of non-cellulolytic bacteria. Microbiology 147:21–30CrossRefGoogle Scholar
  10. Davenport RJ, Curtis TP, Goodfellow M, Stainsby FM, Bingley M (2000) Quantitative use of fluorescent in situ hybridization to examine relationships between mycolic acid-containing actinomycetes and foaming in activated sludge plants. Appl Environ Microbiol 66:1158–1166CrossRefGoogle Scholar
  11. Ellis RJ, Thompson IP, Bailey MJ (1999) Temporal fluctuations in the pseudomonad population associated with sugar beet leaves. FEMS Microbiol Ecol 28:345–356CrossRefGoogle Scholar
  12. Fernandez A, Huang SY, Seston S, Xing J, Hickey R, Criddle C, Tiedje J (1999) How stable is stable? Function versus community composition. Appl Environ Microbiol 65:3697–3704CrossRefGoogle Scholar
  13. Fernandez AS, Hashsham SA, Dollhopf SL, Raskin L, Glagoleva O, Dazzo FB, Hickey RF, Criddle CS, Tiedje JM (2000) Flexible community structure correlates with stable community function in methanogenic bioreactor communities perturbed by glucose. Appl Environ Microbiol 66:4058–4067CrossRefGoogle Scholar
  14. Jiang HL, Tay JH, Tay STL (2002) Aggregation of immobilized activated sludge cells into aerobically grown microbial granules for the aerobic biodegradation of phenol. Lett Appl Microbiol 35:439–445CrossRefGoogle Scholar
  15. Jiang HL, Tay JH, Tay STL (2004a) Changes in structure, activity and metabolism of aerobic granules as a microbial response to high phenol loading. Appl Microbiol Biotechnol 63:602–608CrossRefGoogle Scholar
  16. Jiang HL, Tay JH, Maszenan AM, Tay STL (2004b) Bacterial diversity and function of aerobic granules engineered in a sequencing batch reactor for phenol degradation. Appl Environ Microbiol 70:6767–6775CrossRefGoogle Scholar
  17. Jiang HL, Tay JH, Maszenan AM, Tay STL (2006) Physiological traits of bacterial strains isolated from phenol-degrading aerobic granules. FEMS Microbiol Ecol 57:182–191CrossRefGoogle Scholar
  18. Kurane R, Hatamochi K, Kakuno T, Kiyohara M, Kawaguchi K, Mizuno Y, Hirano M, Tamihuchi Y (1994) Purification and characterization of lipid bioflocculant produced by Rhodococcus erythropolis. Biosci Biotechnol Biochem 58:1977–1982CrossRefGoogle Scholar
  19. Liu Y, Tay JH (2004) State of the art of biogranulation technology for wastewater treatment. Biotechnol Advances 22:533–563CrossRefGoogle Scholar
  20. Loreau M, Naeem S, Inchausti P, Bengtsson J, Grime JP, Hector A, Hooper DU, Huston MA, Raffaelli D, Schmid B, Tilman D, Wardle DA (2001) Biodiversity and ecosystem functioning: current knowledge and future challenges. Science 294:804–808CrossRefGoogle Scholar
  21. Maidak BL, Cole JR, Lilburn TG, Parker CT, Saxman PR, Farris RJ, Garrity GM, Olsen GJ, Schmidt TM, Tiedje JM (2001) The RDP-II (Ribosomal Database Project). Nucleic Acids Res 29:173–174CrossRefGoogle Scholar
  22. Morgenroth E, Sherden T, van Loosdrecht MCM, Heijnen JJ, Wilderer PA (1997) Aerobic granular sludge in a sequencing batch reactor. Water Res 31:3191–3194CrossRefGoogle Scholar
  23. Muyzer G, Dewaal EC, Uitterlinden AG (1993) Profiling of complex microbial-populations by denaturing gradient gel-electrophoresis analysis of polymerase chain reaction-amplified genes-coding for 16S ribosomal-RNA. Appl Environ Microbiol 59:695–700CrossRefGoogle Scholar
  24. Naeem S, Li SB (1997) Biodiversity enhances ecosystem reliability. Nature 390:507–509CrossRefGoogle Scholar
  25. Norberg J, Swaney DP, Dushoff J, Lin J, Casagrandi R, Levin SA (2001) Phenotypic diversity and ecosystem functioning in changing environments: a theoretical framework. Proc Natl Acad Sci USA 98:11376–11381CrossRefGoogle Scholar
  26. O’Toole GA, Kolter R (1998) Initiation of biofilm formation in Pseudomonas fluorescens WCS365 proceeds via multiple, convergent signalling pathways: a genetic analysis. Mol Microbiol 28:449–461CrossRefGoogle Scholar
  27. Rainey PB, Travisano M (1998) Adaptive radiation in a heterogeneous environment. Nature 394:69–72CrossRefGoogle Scholar
  28. Rosenberg M, Gutnick D, Rosenberg E (1980) Adherence of bacteria to hydrocarbons: a simple method from measuring cell-surface hydrophobicity. FEMS Microbiol Lett 9:29–33CrossRefGoogle Scholar
  29. Tay STL, Moy BYP, Maszenan AM, Tay JH (2005) Comparing activated sludge and aerobic granules as microbial inocula for phenol biodegradation. Appl Microbiol Biotechnol 115:387–395Google Scholar
  30. Thompson IP, van der Gast CJ, Ciric L, Singer AC (2005) Bioaugmentation for bioremediation: the challenge of strain selection. Environ Microbiol 7:909–915CrossRefGoogle Scholar
  31. Treves DS, Xia B, Zhou J, Tiedje JM (2003) A two-species test of the hypothesis that spatial isolation influences microbial diversity in soil. Microb Ecol 45:20–28CrossRefGoogle Scholar
  32. van der Gast CJ, Whiteley AS, Thompson IP (2004) Temporal dynamics and degradation activity of an bacterial inoculum for treating waste metal-working fluid. Environ Microbiol 6:254–263CrossRefGoogle Scholar
  33. vanVeen JA, vanOverbeek LS, vanElsas JD (1997) Fate and activity of microorganisms introduced into soil. Microbiol Mol Biol Rev 61:121–135CrossRefGoogle Scholar
  34. von Canstein H, Kelly S, Li Y, Wagner-Dobler I (2002) Species diversity improves the efficiency of mercury-reducing biofilms under changing environmental conditions. Appl Environ Microbiol 68:2829–2837CrossRefGoogle Scholar
  35. Walker B, Kinzig A, Langridge J (1999) Plant attribute diversity, resilience, and ecosystem function: the nature and significance of dominant and minor species. Ecosystems 2:95–113CrossRefGoogle Scholar
  36. Zhou JZ, Xia BC, Treves DS, Wu LY, Marsh TL, O’Neill RV, Palumbo AV, Tiedje JM (2002) Spatial and resource factors influencing high microbial diversity in soil. Appl Environ Microbiol 68:326–334CrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2007

Authors and Affiliations

  • He-Long Jiang
    • 1
    • 2
    Email author
  • Abdul Majid Maszenan
    • 1
  • Joo-Hwa Tay
    • 1
  1. 1.Environmental Engineering Research Centre, School of Civil and Environmental EngineeringNanyang Technological UniversitySingaporeSingapore
  2. 2.Department of Botany and MicrobiologyUniversity of OklahomaNormanUSA

Personalised recommendations