Microbial Ecology

, Volume 61, Issue 3, pp 557–567 | Cite as

Diversity of Dominant Bacterial Taxa in Activated Sludge Promotes Functional Resistance following Toxic Shock Loading

  • Pascal E. Saikaly
  • Daniel B. Oerther
Environmental Microbiology


Examining the relationship between biodiversity and functional stability (resistance and resilience) of activated sludge bacterial communities following disturbance is an important first step towards developing strategies for the design of robust biological wastewater treatment systems. This study investigates the relationship between functional resistance and biodiversity of dominant bacterial taxa by subjecting activated sludge samples, with different levels of biodiversity, to toxic shock loading with cupric sulfate (Cu[II]), 3,5-dichlorophenol (3,5-DCP), or 4-nitrophenol (4-NP). Respirometric batch experiments were performed to determine the functional resistance of activated sludge bacterial community to the three toxicants. Functional resistance was estimated as the 30 min IC50 or the concentration of toxicant that results in a 50% reduction in oxygen utilization rate compared to a referential state represented by a control receiving no toxicant. Biodiversity of dominant bacterial taxa was assessed using polymerase chain reaction-terminal restriction fragment length polymorphism (PCR-T-RFLP) targeting the 16S ribosomal RNA (16S rRNA) gene. Statistical analysis of 30 min IC50 values and PCR-T-RFLP data showed a significant positive correlation (P < 0.05) between functional resistance and microbial diversity for each of the three toxicants tested. To our knowledge, this is the first study showing a positive correlation between biodiversity of dominant bacterial taxa in activated sludge and functional resistance. In this system, activated sludge bacterial communities with higher biodiversity are functionally more resistant to disturbance caused by toxic shock loading.


Activate Sludge Terminal Restriction Fragment Length Polymorphism Oxygen Uptake Rate Ammonia Oxidize Bacterium Solid Retention Time 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.



The authors thank Brian Kinkle, Bruce Rittmann, Makram Suidan, and Jim Young for useful discussion. Financial support from the National Science Foundation to Daniel B. Oerther (BES 0238858) is gratefully acknowledged.


  1. 1.
    Briones AM, Daugherty BJ, Angenent LT, Rausch K, Tumbleson M, Raskin L (2007) Microbial diversity and dynamics in multi- and single-compartment anaerobic bioreactors processing sulfate-rich waste streams. Environ Microbiol 9:93–106PubMedCrossRefGoogle Scholar
  2. 2.
    Burmølle M, Web JS, Rao D, Hansen LH, Sørensen SJ, Kjelleberg S (2006) Enhanced biofilm formation and increased resistance to antimicrobial agents and bacterial invasion are caused by synergistic interactions in multispecies biofilms. Appl Environ Microbiol 72:3916–3923PubMedCrossRefGoogle Scholar
  3. 3.
    Cook KL, Garland JL, Layton AC, Dionisi HM, Levine LH, Sayler GS (2006) Effect of microbial species richness on community stability and community function in a model plant-based wastewater processing system. Microb Ecol 52:725–737PubMedCrossRefGoogle Scholar
  4. 4.
    Crosby LD, Criddle CS (2003) Understanding bias in microbial community analysis techniques due to rrn operon copy number heterogeneity. Biotechniques 34:790–799PubMedGoogle Scholar
  5. 5.
    Curtis TP, Head IM, Graham DW (2003) Theoretical ecology for engineering biology. Environ Sci Technol 64A–70AGoogle Scholar
  6. 6.
    Curtis TP, Sloan WT (2004) Prokaryotic diversity and its limits: microbial community structure in nature and implications for microbial ecology. Curr Opin Microbiol 7:221–226PubMedCrossRefGoogle Scholar
  7. 7.
    Daims H, Purkhold U, Bjerrum L, Arnold E, Wilderer PA, Wagner M (2001) Nitrification in sequencing biofilm batch reactors: lessons from molecular approaches. Water Sci Technol 43:9–18PubMedGoogle Scholar
  8. 8.
    Dalzell DJB, Alte S, Aspichueta E, de la Sota A, Etxebarria J, Gutierrez M, Hoffmann CC, Sales D, Obst U, Christofi N (2002) A comparison of five rapid direct toxicity assessment methods to determine toxicity of pollutants to activated sludge. Chemosphere 47:535–545PubMedCrossRefGoogle Scholar
  9. 9.
    Dunbar J, Ticknor LO, Kuske CR (2000) Assessment of microbial diversity in four southwestern United States soils by 16S rRNA gene terminal restriction fragment analysis. Appl Environ Microbiol 66:2943–2950PubMedCrossRefGoogle Scholar
  10. 10.
    Dunbar J, Ticknor LO, Kuske CR (2001) Phylogenetic specificity and reproducibility and new method for analysis of terminal restriction fragment profiles of 16S rRNA genes from bacterial communities. Appl Environ Microbiol 67:190–197PubMedCrossRefGoogle Scholar
  11. 11.
    Eichner CA, Erb RW, Timmis KN, Wagner-Dobler I (1999) Thermal gradient gel electrophoresis analysis of bioprotection from pollutant shocks in the activated sludge microbial community. Appl Environ Microbiol 65:102–109PubMedGoogle Scholar
  12. 12.
    Fernandez AS, Hashsham SA, Dollhope SL, Raskin L, Glagoleva O, Dazzo FB, Hickey RF, Criddle GS, Tiedje JM (2000) Flexible community structure correlates with stable community function in methanogenic bioreactor communities perturbed by glucose. Appl Environ Microbiol 66:4058–4067PubMedCrossRefGoogle Scholar
  13. 13.
    Forney LJ, Zhou X, Brown CJ (2004) Molecular microbial ecology: land of the one-eyed king. Curr Opin Microbiol 7:210–220PubMedCrossRefGoogle Scholar
  14. 14.
    Gentile M, Yan T, Tiquia SM, Fields MW, Nyman J, Zhou J, Criddle CS (2006) Stability in a denitrifying fluidized bed reactor. Microb Ecol 52:311–321PubMedCrossRefGoogle Scholar
  15. 15.
    Girvan MS, Campbell CD, Killham K, Prosser JI, Glover LA (2005) Bacterial diversity promotes community stability and functional resilience after perturbation. Environ Microbiol 7:301–313PubMedCrossRefGoogle Scholar
  16. 16.
    Grady CPL, Daigger GT, Lim HC (eds) (1999) Biological Wastewater Treatment, 2nd edn. Marcel Dekker, New YorkGoogle Scholar
  17. 17.
    Grady CPL, Smets BF, Barbeau DS (1996) Variability in kinetic parameter estimates: a review of possible causes and a proposed terminology. Water Res 30:742–748CrossRefGoogle Scholar
  18. 18.
    Graham DW, Smith VH (2004) Designed ecosystem services: application of ecological principles in wastewater treatment engineering. Front Ecol Environ 2:199–206CrossRefGoogle Scholar
  19. 19.
    Griffiths BS, Kuan HL, Ritz K, Glover LA, McCaig AE, Fenwick C (2004) The relationship between microbial community structure and functional stability, tested experimentally in an upland pasture soil. Microb Ecol 47:104–113PubMedCrossRefGoogle Scholar
  20. 20.
    Griffiths BS, Ritz K, Bardgett RD, Cook R, Christensen S, Ekelund F, Sorensen SJ, Baath E, Bloem J, de Ruiter PC, Dolfing J, Nicolardot B (2000) Ecosystem response of pasture soil communities to fumigation-induced microbial diversity reductions: an examination of the biodiversity-ecosystem function relationship. Oikos 90:279–294CrossRefGoogle Scholar
  21. 21.
    Gutierrez M, Etxebarria J, de las Fuentes L (2002) Evaluation of wastewater toxicity: comparative study between Microtox and activated sludge oxygen uptake inhibition. Water Res 36:919–924PubMedCrossRefGoogle Scholar
  22. 22.
    Hashsham SA, Fernandez AS, Dollhope SL, Dazzo FB, Hickey RF, Tiedje JM, Criddle GS (2000) Parallel processing of substrate correlates with greater functional stability in mathanogenic bioreactor communities perturbed by glucose. Appl Environ Microbiol 66:4050–4057PubMedCrossRefGoogle Scholar
  23. 23.
    Hughes JB, Hellmann JJ, Ricketts TH, Bohannan BJM (2001) Counting the uncountable: statistical approaches to estimating microbial diversity. Appl Environ Microbiol 67:4399–4406PubMedCrossRefGoogle Scholar
  24. 24.
    Juretschko S, Timmermann G, Schmid M, Scleifer KH, Pommerening-Roser A, Koops HP, Wagner M (1998) Combined molecular and conventional analyses of nitrifying bacterium diversity in activated sludge; Nitrosococcus mobilis and Nitrospira-like bacteria as dominant populations. Appl Environ Microbiol 64:3042–3051PubMedGoogle Scholar
  25. 25.
    Kitts CL (2001) Terminal restriction fragment patterns: a tool for comparing microbial communities and assessing community dynamics. Curr Issues Intest Microbiol 2:17–25PubMedGoogle Scholar
  26. 26.
    LaPara TM, Nakatsu CH, Pantea LM, Alleman JE (2002) Stability of the bacterial communities supported by a seven-stage biological process treating pharmaceutical wastewater as revealed by PCR-DGGE. Water Res 36:638–646PubMedCrossRefGoogle Scholar
  27. 27.
    Liu WT, Marsh TL, Cheng H, Forney LJ (1997) Characterization of microbial diversity by determining terminal restriction fragment length polymorphisms of genes encoding 16S rRNA. Appl Environ Microbiol 63:4516–4522PubMedGoogle Scholar
  28. 28.
    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) Ecology: biodiversity and ecosystem functioning: current knowledge and future challenges. Science 294:804–808PubMedCrossRefGoogle Scholar
  29. 29.
    Love NG, Bott CB (2000) A review and needs survey of upset early warning devices. Water Environment Research Foundation; Report No. 99-WWF-2. Alexandria, VA.Google Scholar
  30. 30.
    Love NG, Bott CB (2002) Evaluating the role of microbial stress response mechanisms in causing biological treatment system upset. Water Sci Technol 46:11–18PubMedGoogle Scholar
  31. 31.
    Marquardt DW (1963) An algorithm for least squares estimation of parameters. J Soc Ind Appl Math 11:431–441CrossRefGoogle Scholar
  32. 32.
    McCann KS (2000) The diversity–stability debate. Nature 405:228–233PubMedCrossRefGoogle Scholar
  33. 33.
    McGrady-Steed J, Harris PM, Morin PJ (1997) Biodiversity regulates ecosystem predictability. Nature 390:162–165CrossRefGoogle Scholar
  34. 34.
    Muyzer G, Smalla K (1998) Application of denaturing gradient gel electrophoresis (DGGE) and temperature gradient gel electrophoresis (TGGE) in microbial ecology. Antonie Leeuwenhoek 73:127–141PubMedCrossRefGoogle Scholar
  35. 35.
    Osborn AM, Moore ERB, Timmis KN (2000) An evaluation of terminal-restriction fragment length polymorphism (T-RFLP) analysis for the study of microbial community structure and dynamics. Environ Microbiol 2:39–50PubMedCrossRefGoogle Scholar
  36. 36.
    Oviedo MDC, Marquez DS, Alonso JMQ (2002) Toxic effects of metals on microbial activity in the activated sludge process. Chem Biochem Eng Q 16:139–144Google Scholar
  37. 37.
    Pedros-Alio C (2006) Marine microbial diversity: can it be determined? Trends Microbiol 14:257–263PubMedCrossRefGoogle Scholar
  38. 38.
    Prosser JI, Bohannan BJM, Curtis TP, Ellis RJ, Firestone MK, Freckleton RP, Green JL, Green LE, Killham K, Lennon JL, Osborn MA, Solan M, van der Gast CJ, Young JP (2007) The role of ecological theory in microbial ecology. Nat Rev Microbiol 5:384–392PubMedCrossRefGoogle Scholar
  39. 39.
    Ricco G, Tomei MC, Ramadori R, Laera G (2004) Toxicity assessment of common xenobiotic compounds on municipal activated sludge: comparison between respirometry and Microtox. Water Res 38:2103–2110PubMedCrossRefGoogle Scholar
  40. 40.
    Rosello-Mora R, Amann R (2001) The species concept for prokaryotes. FEMS Microbiol Rev 25:39–67CrossRefGoogle Scholar
  41. 41.
    Rowan AK, Snape JR, Fearnside D, Barer MR, Curtis TP, Head IM (2003) Composition and diversity of ammonia-oxidizing bacterial communities in wastewater treatment reactors of different design treating identical wastewater. FEMS Microbiol Ecol 43:195–206PubMedCrossRefGoogle Scholar
  42. 42.
    Saikaly PE, Oerther DB (2004) Bacterial competition in activated sludge: theoretical analysis of varying solids retention times on diversity. Microb Ecol 48:274–284PubMedCrossRefGoogle Scholar
  43. 43.
    Saikaly PE, Stroot PG, Oerther DB (2005) Assessing the impact of solids retention time on activated sludge bacterial diversity by 16S rRNA gene terminal restriction fragment length polymorphism. Appl Environ Microbiol 71:5814–5822PubMedCrossRefGoogle Scholar
  44. 44.
    Schwartz MW, Brigham CA, Hoeksema JD, Lyons KG, Mills MH, van Mantgem PJ (2000) Linking biodiversity to ecosystem function: implications for conservation ecology. Oecologia 122:297–305CrossRefGoogle Scholar
  45. 45.
    Shannon CE, Weaver W (1963) The mathematical theory of communication. University of Illinois Press, UrbanaGoogle Scholar
  46. 46.
    Simpson EH (1949) Measurement of diversity. Nature 163:688CrossRefGoogle Scholar
  47. 47.
    Sogin ML, Morrison HG, Huber JA, Welch DM, Huse SM, Neal PR, Arrieta JM, Herndl GJ (2006) Microbial diversity in the deep sea and the underexplored ‘rare biosphere’. Proc Natl Acad Sci 103:12115–12120PubMedCrossRefGoogle Scholar
  48. 48.
    Solow AR (1993) A simple test for change in community structure. J Anim Ecol 62:191–193CrossRefGoogle Scholar
  49. 49.
    Tilman D (1999) The ecological consequences of changes in biodiversity: a search for general principles. Ecology 80:1455–1474Google Scholar
  50. 50.
    Tilman D (2002) Biodiversity: population versus ecosystem stability. Ecology 77:350–363CrossRefGoogle Scholar
  51. 51.
    Tilman D, Knops J, Wedin D, Reich P, Ritchie M, Siemann E (1997) The influence of functional diversity and composition on ecosystem processes. Science 277:1300–1302CrossRefGoogle Scholar
  52. 52.
    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
  53. 53.
    von Wintzingerode F, Gobel UB, Stackebrandt E (1997) Determination of microbial diversity in environmental samples: pitfalls of PCR-based rRNA analysis. FEMS Microbiol Rev 21:213–229CrossRefGoogle Scholar
  54. 54.
    Ward JH (1963) Hierarchical grouping to optimize an objective function. J Am Stat Assoc 58:236–244CrossRefGoogle Scholar
  55. 55.
    Wertz S, Degrange V, Prosser JI, Poly F, Commeaux C, Freitag T, Guillaumaud N, Roux XL (2007) Decline of soil microbial diversity does not influence the resistance and resilience of key soil microbial functional groups following a model disturbance. Environ Microbiol 9:2211–2219PubMedCrossRefGoogle Scholar
  56. 56.
    Wong KY, Zhang MQ, Li XM, Lo W (1997) A luminescence-based scanning respirometer for heavy metal toxicity monitoring. Biosens Bioelectron 12:125–133CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2010

Authors and Affiliations

  1. 1.Water Desalination and Reuse Research CenterKing Abdullah University of Science and Technology (KAUST)ThuwalKingdom of Saudi Arabia
  2. 2.Division of Chemical and Life Sciences and EngineeringKing Abdullah University of Science and TechnologyThuwalSaudi Arabia
  3. 3.Department of Civil and Environmental EngineeringAmerican University of BeirutBeirutLebanon
  4. 4.Department of Civil, Architectural, and Environmental EngineeringMissouri University of Science and TechnologyRollaUSA
  5. 5.Environmental Research CenterMissouri University of Science and TechnologyRollaUSA

Personalised recommendations