Monitoring Microbial Growth by Bioluminescent ATP Assay

  • M.-R. Siro

Abstract

Quantitative determination of total biomass carbon as a function of time is often important both in general and applied microbiology. The classic techniques for detecting and monitoring microbes include total visual or electronic cell counting, viable cell counting using plating techniques, estimating cell number by measuring the turbidity, and dry weight determination. Both viable and total cell counting are somewhat time-consuming and open to serious criticism [1,2]. Although particle counting methods are faster, they suffer from inaccuracies caused by counting of any particulate matter, including air bubbles and dust particles. Moreover, direct counting includes both viable and nonviable cells. No rapid method for detecting or monitoring viable bacteria has thus far been available. Undoubtedly the determination of viable cells is important in all kinds of contamination studies and in processes that require viable cells. Because of the problems associated with conventional methods, there has been great interest in attempts to find a rapid routine procedure for determining the amount of microbes in different sources.

Keywords

Vortex Fermentation Adenosine Turbidity Microbe 

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References

  1. 1.
    Holm-Hansen O, Karl DM (1978) Biomass and adenylate energy charge determination in microbial cell extracts and environmental samples. Methods Enzymol 57:73–85CrossRefGoogle Scholar
  2. 2.
    Pringle JR, Mor JR (1975) Methods for monitoring the growth of yeast cultures and for dealing with the clumping problem. Methods Cell Biol XI:153–168Google Scholar
  3. 3.
    Holm-Hansen O, Booth CR (1966) The measurement of adenosine triphosphate in the ocean and its ecological significance. Limnol Oceanogr 11:510–519CrossRefGoogle Scholar
  4. 4.
    Karl DM (1980) Cellular nucleotide measurements and applications in microbial ecology. Microbiol Rev 44:739–796PubMedGoogle Scholar
  5. 5.
    De Luca M (1976) Firefly luciferase. In: Meister A (ed) Adv Enzymol 44:37–68Google Scholar
  6. 6.
    Strehler BL (1968) Bioluminescence assay: principles and practice. Methods Biochem Anal 16:99–181PubMedCrossRefGoogle Scholar
  7. 7.
    Lundin A, Rickardsson A, Thore A (1976) Continuous monitoring of ATP-converting reactions by purified firefly luciferase. Anal Biochem 75:611–620PubMedCrossRefGoogle Scholar
  8. 8.
    Lundin A, Thore A (1975) Analytical information obtainable by evaluation of the time course of firefly luminescence in the assay of ATP. Anal Biochem 66:47–63PubMedCrossRefGoogle Scholar
  9. 9.
    Lundin A, Thore A (1975) Comparison of methods for extraction of bacterial adenine nucleotides determined by firefly assay. Appl Microbiol 30:713–721PubMedGoogle Scholar
  10. 10.
    Holmsen H, Holmsen I, Bernhardsen A (1966) Microdetermination of adenosine diphosphate and adenosine triphosphate in plasma with the firefly luciferase system. Anal Biochem 17:456–473PubMedCrossRefGoogle Scholar
  11. 11.
    Rasmussen H, Nielsen R (1968) An improved analysis of adenosine triphosphate by the luciferase method. Acta Chem Scand 22:1745–1756PubMedCrossRefGoogle Scholar
  12. 12.
    Lin S, Cohen HP (1968) Measurement of adenosine triphosphate content of crayfish stretch receptor cell preparations. Anal Biochem 24:531–540PubMedCrossRefGoogle Scholar
  13. 13.
    Thore A (1979) Technical aspects of the bioluminescent firefly luciferase assay of ATP. Sci Tools 26:30–34Google Scholar
  14. 14.
    Thore A, Ånsehn S, Lundin A, Bergman S (1975) Detection of bacteriuria by luciferase assay of adenosine triphosphate. J Clin Bacteriol 1:1–8Google Scholar
  15. 15.
    Chappelle EW, Levin GV (1968) Use of the firefly bioluminescent reaction for rapid detection and counting of bacteria. Biochem Med 2:41–52CrossRefGoogle Scholar
  16. 16.
    Bagnara AS, Finch LR (1972) Quantitative extraction and estimation of intracellular nucleoside triphosphates of Escherichia coli. Anal Biochem 45:24–34PubMedCrossRefGoogle Scholar
  17. 17.
    Guinn G, Eidenbock MP (1972) Extraction, purification and estimation of ATP from leaves, floral buds and immature fruits of cotton. Anal Biochem 50:89–97PubMedCrossRefGoogle Scholar
  18. 18.
    Sharpe AN, Woodrow MN, Jackson AK (1970) Adenosine triphosphate (ATP) levels in foods contaminated by bacteria. J Appl Bacteriol 33:758–767PubMedCrossRefGoogle Scholar
  19. 19.
    Siro M-R, Romar H, Lövgren T (1982) Continuous flow method for extraction and bioluminescence assay of ATP in baker’s yeast. Eur J Appl Microbiol Biotechnol 15:258–264CrossRefGoogle Scholar
  20. 20.
    Hysert DW, Kovecses F, Morrison NM (1976) A firefly bioluminescence ATP assay method for rapid detection and enumeration of brewery microorganisms. Am Soc Brew Chem J 34:145–150Google Scholar
  21. 21.
    Stannard CJ, Wood JM (1982) The estimation of viable bacteria in raw meat by measurement of microbial ATP. The British Food Manufacturing Industries Research Association (Research reports no 379)Google Scholar
  22. 22.
    Siro M-R, Lövgren T (1979) Influence of glucose on the a-glucoside permease activity of yeast. Eur J Appl Microbiol Biotechnol 7:59–66CrossRefGoogle Scholar
  23. 23.
    Levin BV, Usdin E, Slonim AR (1968) Rapid detection of microorganisms in aerospace water systems. Aerosp Med 14–16Google Scholar
  24. 24.
    Forsberg CW, Lam K (1977) Use of adenosine 5′-triphosphate as an indicator of the microbiota biomass in rumen contents. Appl Environ Microbiol 33/3:528–537PubMedGoogle Scholar
  25. 25.
    Thore A, Lundin A, Ånsehn S (1983) Firefly luciferase ATP assay as a screening method for bacteriuria. J Clin Microbiol 17:218–224PubMedGoogle Scholar
  26. 26.
    Molin Ö, Nilsson L, Ånsehn S (1983) Rapid detection of bacterial growth in blood cultures by bioluminescent assays of bacterial ATP. J Clin Microbiol 18:521–525PubMedGoogle Scholar
  27. 27.
    Hysert DW, Morrison NM (1977) Studies on ATP, ADP and AMP concentrations in yeast and beer. Am Soc Brew Chem J 35:160–167Google Scholar
  28. 28.
    Lehtokari M, Nikkola P, Paatero J (1983) Determination of ATP from compost using the firefly bioluminescence technique. Eur J Appl Microbiol Biotechnol 17:187–190CrossRefGoogle Scholar
  29. 29.
    Thore A, Nilsson L, Hojer H, Ånsehn S, Bröte L (1977) Effects of ampicillin on intracellular levels of adenosine triphosphate in bacterial cultures related to antibiotic susceptibility. Acta Pathol Microbiol Scand [B] 85:161Google Scholar
  30. 30.
    Nilsson L (1982) Rapid bioluminescent assay of serum amikacin. J Antimicrob Chemother 10: 125–130PubMedCrossRefGoogle Scholar
  31. 31.
    Nilsson L, Höjer H, Ånsehn S, Thore A (1977) A rapid semiautomated bioassay of gentamicin based on luciferase assay of bacterial adenosine triphosphate. Scand J Infect Dis 9:232–236PubMedGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 1985

Authors and Affiliations

  • M.-R. Siro
    • 1
  1. 1.Department of Biochemistry and PharmacyÅbo AkademiÅboFinland

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