Archives of Microbiology

, Volume 186, Issue 3, pp 229–239 | Cite as

The acid tolerance response of Bacillus cereus ATCC14579 is dependent on culture pH, growth rate and intracellular pH

  • Séverine Thomassin
  • Michel P. Jobin
  • Philippe Schmitt
Original Paper

Abstract

The food pathogen Bacillus cereus is likely to encounter acidic environments (i) in food when organic acids are added for preservation purposes, and (ii) during the stomachal transit of aliments. In order to characterise the acid stress response of B. cereus ATCC14579, cells were grown in chemostat at different pH values (pHo from 9.0 to 5.5) and different growth rates (μ from 0.1 to 0.8 h−1), and were submitted to acid shock at pH 4.0. Cells grown at low pHo were adapted to acid media and induced a significant acid tolerance response (ATR). The ATR induced was modulated by both pHo and μ, and the μ effect was more marked at pHo 5.5. Intracellular pH (pHi) was affected by both pHo and μ. At a pHo above 6, the pHi decreased with the decrease of pHo and the increase of μ. At pHo 5.5, pHi was higher compared to pHo 6.0, suggesting that mechanisms of pHi homeostasis were induced. The acid survival of B. cereus required protein neo-synthesis and the capacity of cells to maintain their pHi and ΔpH (pHi - pHo). Haemolysin BL and non-haemolytic enterotoxin production were both influenced by pHo and μ.

Keywords

Bacillus cereus Chemostat Acid tolerance response Stress Intracellular pH 

Abbreviations

ATR

Acid tolerance response

cFDASE

Carboxyfluorescein diacetate succinimidyl ester

cFSE

Carboxyfluorescein succinimidyl ester

HBL

Haemolysin BL

JB

J broth

NHE

Non-haemolytic enterotoxin

pHi

Internal pH

pHo

External pH

ΔpH,

Delta pH

GLM

General linear model

HDS

Honest significant difference

Notes

Acknowledgements

We are grateful to Claire Dargaignaratz for her technical assistance. This work was supported by the Ministère de l’Education Nationale, de l’Enseignement Supérieur et de la Recherche (French Ministry for Education and Research).

References

  1. Agata N, Ohta M, Mori M, Isobe M (1995) A novel dodecadepsipeptide, cereulide, is an emetic toxin of Bacillus cereus. FEMS Microbiol Lett 129:17–20PubMedGoogle Scholar
  2. Belli WA, Marquis RE (1991) Adaptation of Streptococcus mutans and Enterococcus hirae to acid stress in continuous culture. Appl Environ Microbiol 57:1134–1138PubMedGoogle Scholar
  3. Breeuwer P, Drocourt J, Rombouts F, Abee T (1996) A novel method for continuous determination of the intracellular pH in bacteria with the internally conjugated fluorescent probe 5 (and 6-)-carboxyfluorescein succinimidyl ester. Appl Environ Microbiol 62:178–183PubMedGoogle Scholar
  4. Browne N, Dowds BC (2002) Acid stress in the food pathogen Bacillus cereus. J Appl Microbiol 92:404–414PubMedCrossRefGoogle Scholar
  5. Carlin F, Guinbretière MH, Choma CD, Pasqualini R, Braconnier A, Nguyen-The C (2000) Spore-forming bacteria in commercial cooked, pasteurized and chilled vegetable purées. Food Microbiol 17:153–165CrossRefGoogle Scholar
  6. Choma C et al (2000) Prevalence, characterization and growth of Bacillus cereus in commercial cooked chilled foods containing vegetables. J Appl Microbiol 88:617–625PubMedCrossRefGoogle Scholar
  7. Claus D, Berkeley RCW (1986) Endospore-forming gram-positive rods and cocci. In: Sneath PHA, Mair NS, Sharpe ME, Holt JG (eds) Bergey’s manual of systematic bacteriology. Williams & Wilkins, Baltimore, pp 1104–1139Google Scholar
  8. Clavel T, Carlin F, Lairon D, Nguyen-The C, Schmitt P (2004) Survival of Bacillus cereus spores and vegetative cells in acid media simulating human stomach. J Appl Microbiol 97:214–219PubMedCrossRefGoogle Scholar
  9. Cotter PD, Hill C (2003) Surviving the acid test: responses of gram-positive bacteria to low pH. Microbiol Mol Biol Rev 67:429–453PubMedCrossRefGoogle Scholar
  10. Datta AR, Benjamin MM (1997) Factors controlling acid tolerance of Listeria monocytogenes: effects of nisin and other ionophores. Appl Environ Microbiol 63:4123–4126PubMedGoogle Scholar
  11. Davis MJ, Coote PJ, O’Byrne CP (1996) Acid tolerance in Listeria monocytogenes: the adaptive acid tolerance response (ATR) and growth-phase-dependent acid resistance. Microbiology 142:2975–2982PubMedGoogle Scholar
  12. Duport C, Thomassin S, Bourel G, Schmitt P (2004) Anaerobiosis and low specific growth rates enhance hemolysin BL production by Bacillus cereus F4430/73. Arch Microbiol 182:90–95PubMedCrossRefGoogle Scholar
  13. Jobin MP, Clavel T, Carlin F, Schmitt P (2002) Acid tolerance response is low-pH and late-stationary growth phase inducible in Bacillus cereus TZ415. Int J Food Microbiol 79:65–73PubMedCrossRefGoogle Scholar
  14. Karem KL, Foster JW, Bej AK (1994) Adaptive acid tolerance response (ATR) in Aeromonas hydrophila. Microbiology 140:1731–1736PubMedGoogle Scholar
  15. Kramer JM, Gilbert RJ (1989) Bacillus cereus and other Bacillus species. In: Doyle MP (ed) Foofborn bacterial pathogens. Dekker M, New York, pp 21–70Google Scholar
  16. Lund T, Granum PE (1997) Comparison of biological effect of the two different enterotoxin complexes isolated from three different strains of Bacillus cereus. Microbiology 143:3329–3336PubMedGoogle Scholar
  17. Lund T, De Buyser ML, Granum PE (2000) A new cytotoxin from Bacillus cereus that may cause necrotic enteritis. Mol Microbiol 38:254–261PubMedCrossRefGoogle Scholar
  18. Minton KW, Karmin P, Hahn GM, Minton AP (1982) Nonspecific stabilization of stress-susceptible proteins by stress-resistant proteins: a model for the biological role of heat shock proteins. Proc Natl Acad Sci USA 79:7107–7111PubMedCrossRefGoogle Scholar
  19. Nascimento MM, Lemos JAC, Abranches J, Goncalves RB, Burne RA (2004) Adaptive acid tolerance response of Streptococcus sobrinus. J Bacteriol 186:6383–6390PubMedCrossRefGoogle Scholar
  20. O’Driscoll B, Gahan CG, Hill C (1996) Adaptive acid tolerance response in Listeria monocytogenes: isolation of an acid-tolerant mutant which demonstrates increased virulence. Appl Environ Microbiol 62:1693–1698PubMedGoogle Scholar
  21. O’Hara GW, Glenn AR (1994) The adaptive acid tolerance response in root nodule bacteria and Escherichia coli. Arch Microbiol 161:286–292PubMedCrossRefGoogle Scholar
  22. O’Sullivan E, Condon S (1999) Relationship between acid tolerance, cytoplasmic pH, and ATP and H + -ATPase levels in chemostat cultures of Lactococcus lactis. Appl Environ Microbiol 65:2287–2293PubMedGoogle Scholar
  23. Rao M, Streur TL, Aldwell FE, Cook GM (2001) Intracellular pH regulation by Mycobacterium smegmatis and Mycobacterium bovis BCG. Microbiology 147:1017–1024PubMedGoogle Scholar
  24. Roberts D, Watson GN, Gilbert RJ (1982) Contamination of food plants and plant products with bacteria of public health significance. In: Rhodes-Roberts ME, Skinner FA (eds) Bacteria and Plants. Academic, London, pp 169–195Google Scholar
  25. Siegumfeldt H, Rechinger KB, Jakobsen M (1999) Use of fluorescence ratio imaging for intracellular pH determination of individual bacterial cells in mixed cultures. Microbiology 145:1703–1709PubMedCrossRefGoogle Scholar
  26. Siegumfeldt H, Rechinger KB, Jakobsen M (2000) Dynamic changes of intracellular pH in individual lactic acid bacterium cells in response to a rapid drop in extracellular pH. Appl Environ Microbiol 66:2330–2335PubMedCrossRefGoogle Scholar
  27. Sutherland AD, Limond AM (1993) Influence of pH and sugars on the growth and production of diarrhoeagenic toxin by Bacillus cereus. J Dairy Res 60:575–580PubMedGoogle Scholar
  28. Tiwari RP, Sachdeva N, Hoondal GS, Grewal JS (2004) Adaptive acid tolerance response in Salmonella enterica serovar Typhimurium and Salmonella enterica serovar Typhi. J Basic Microbiol 44:137–146PubMedCrossRefGoogle Scholar
  29. Ultee A, Kets EPW, Smid EJ (1999) Mechanisms of action of carvacrol on the food-borne pathogen Bacillus cereus. Appl Environ Microbiol 65:4606–4610PubMedGoogle Scholar
  30. Valero M, Fernandez PS, Salmeron MC (2003) Influence of pH and temperature on growth of Bacillus cereus in vegetable substrates. Int J Food Microbiol 82:71–79PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2006

Authors and Affiliations

  • Séverine Thomassin
    • 1
    • 2
  • Michel P. Jobin
    • 1
    • 2
  • Philippe Schmitt
    • 1
    • 2
  1. 1.IUT Génie BiologiqueUniv Avignon, UMR 408, Sécurité et Qualité des Produits d’Origine VégétaleAvignonFrance
  2. 2.INRAUMR 408Avignon cedexFrance

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