Current Microbiology

, Volume 76, Issue 12, pp 1495–1502 | Cite as

Comparison of Starvation-Induced Persister Cells with Antibiotic-Induced Persister Cells

  • Shridhar S. Paranjape
  • Ravindranath ShashidharEmail author


The phenotypic heterogeneity in a large population arises because of fluctuation in microenvironments and stochastic gene expressions. In this report, we isolated two types of persistent sub-populations of Vibrio cholerae, one triggered by starvation and another by antibiotics. We characterised starvation-induced (E-cells) and antibiotic-induced (P-cell) persister cells for stress tolerance, colony morphology and toxin gene expressions. Both the sub-populations differ with respect to morphology, temperature tolerance and oxidative stress tolerance. The E-cells were smaller than the P-cells and formed tiny colonies (1–2 mm). The E-cells were more sensitive to heat and oxidative stress compared with P-cells. The up-regulated genes of P-cells include, genes of antioxidant enzymes (>5 fold), cholera toxin (>26 fold) and toxin: antitoxin protein hipA (>100 fold). Upon nutrient up-shift, the E-cells recovered after lag time of 6 h. However, such lag extension was not visible during P-cell recovery, suggesting that P-cell physiology is more akin to normal cells than E-cells. This is the first comparative report on the two different persister sub-populations of V. cholerae. The E-cells and P-cells are similar regarding antibiotic tolerance. However, the sub-populations differ significantly in stress tolerance and other phenotypes studied.



Funding was provided by Department of Atomic Energy, India.

Supplementary material

284_2019_1777_MOESM1_ESM.docx (131 kb)
Supplementary file1 (DOCX 131 kb)


  1. 1.
    Gefen O, Balaban NQ (2009) The importance of being persistent: heterogeneity of bacterial populations under antibiotic stress. FEMS Microbiol Rev 33:704–717CrossRefGoogle Scholar
  2. 2.
    Jõers A, Tenson T (2016) Growth resumption from stationary phase reveals memory in Escherichia coli cultures. Sci Rep 6:24055CrossRefGoogle Scholar
  3. 3.
    Balaban NQ, Merrin J, Chait R, Kowalik L, Leibler S (2004) Bacterial persistence as a phenotypic switch. Science 305:1622–1625CrossRefGoogle Scholar
  4. 4.
    Balaban NQ et al (2019) Definitions and guidelines for research on antibiotic persistence. Nat Rev Microbiol 17:441–448CrossRefGoogle Scholar
  5. 5.
    Keren I, Kaldalu N, Spoering A, Wang Y, Lewis K (2004) Persister cells and tolerance to antimicrobials. FEMS Microbiol Lett 230:13–18CrossRefGoogle Scholar
  6. 6.
    Fauvart M, De Groote VN, Michiels J (2011) Role of persister cells in chronic infections: clinical relevance and perspectives on anti-persister therapies. J Med Microbiol 60:699–709CrossRefGoogle Scholar
  7. 7.
    Cabral DJ, Wurster JI, Belenky P (2018) Antibiotic persistence as a metabolic adaptation: stress, metabolism, the host, and new directions. Pharmaceuticals 11:14CrossRefGoogle Scholar
  8. 8.
    Chubukov V, Sauer U (2014) Environmental dependence of stationary-phase metabolism in Bacillus subtilis and Escherichia coli. Appl Environ Microbiol 80:2901–2909CrossRefGoogle Scholar
  9. 9.
    Navarro Llorens JM, Tormo A, Martínez-García E (2010) Stationary phase in gram-negative bacteria. FEMS Microbiol Rev 34:476–495CrossRefGoogle Scholar
  10. 10.
    Wang JD, Levin PA (2009) Metabolism, cell growth and the bacterial cell cycle. Nat Rev Microbiol 7:822–827CrossRefGoogle Scholar
  11. 11.
    Nguyen D et al (2011) Active starvation responses mediate antibiotic tolerance in biofilms and nutrient-limited bacteria. Science 334:982–986CrossRefGoogle Scholar
  12. 12.
    Jubair M, Jr JGM, Ali A (2012) Survival of Vibrio cholerae in nutrient-poor environments is associated with a novel ‘persister’ phenotype. PLoS ONE 7:e45187CrossRefGoogle Scholar
  13. 13.
    Fung DKC, Chan EWC, Chin ML, Chan RCY (2010) Delineation of a bacterial starvation stress response network which can mediate antibiotic tolerance development. Antimicrob Agents Chemother 54:1082–1093CrossRefGoogle Scholar
  14. 14.
    Colwell RR, Spira WM (1992) The ecology of Vibrio cholera. In: Barua D, Greenough WB (eds) Cholera. Springer, US, Boston, pp 107–127CrossRefGoogle Scholar
  15. 15.
    Munro PM, Colwell RR (1996) Fate of Vibrio cholerae O1 in seawater microcosms. Water Res 30:47–50CrossRefGoogle Scholar
  16. 16.
    Ackermann M (2015) A functional perspective on phenotypic heterogeneity in microorganisms. Nat Rev Microbiol 13:497CrossRefGoogle Scholar
  17. 17.
    Sukhi SS, Shashidhar R, Kumar SA, Bandekar JR (2009) Radiation resistance of Deinococcus radiodurans R1 with respect to growth phase. FEMS Microbiol Lett 297:49–53CrossRefGoogle Scholar
  18. 18.
  19. 19.
    Ostling J, Holmquist L, Kjelleberg S (1996) Global analysis of the carbon starvation response of a marine Vibrio species with disruptions in genes homologous to relA and spot. J Bacteriol 178:4901–4908CrossRefGoogle Scholar
  20. 20.
    Wai SN, Mizunoe Y, Yoshida S (1999) How Vibrio cholerae survive during starvation. FEMS Microbiol Lett 180:123–131CrossRefGoogle Scholar
  21. 21.
    Oliver JD (2010) Recent findings on the viable but nonculturable state in pathogenic bacteria. FEMS Microbiol Rev 34:415–425CrossRefGoogle Scholar
  22. 22.
    Kim J-S, Chowdhury N, Yamasaki R, Wood TK (2018) Viable but non-culturable and persistence describe the same bacterial stress state. Environ Microbiol 20:2038–2048CrossRefGoogle Scholar
  23. 23.
    Kjelleberg S (ed) (1993) Starvation in bacteria. Springer, New YorkGoogle Scholar
  24. 24.
    Mizunoe Y, Wai SN, Takade A, Yoshida S-I (1999) Isolation and characterization of rugose form of Vibrio cholerae O139 strain MO10. Infect Immun 67:958–963PubMedPubMedCentralGoogle Scholar
  25. 25.
    Nyström T, Albertson NH, Flärdh K, Kjelleberg S (1990) Physiological and molecular adaptation to starvation and recovery from starvation by the marine Vibrio sp S14. FEMS Microbiol Ecol 7:129–140CrossRefGoogle Scholar
  26. 26.
    Ebrahimi A, Csonka LN, Alam MA (2018) Analyzing thermal stability of cell membrane of salmonella using time-multiplexed impedance sensing. Biophys J 114:609–618CrossRefGoogle Scholar
  27. 27.
    Conlon BP et al (2016) Persister formation in Staphylococcus aureus is associated with ATP depletion. Nat Microbiol 1:16051CrossRefGoogle Scholar
  28. 28.
    Häder D-P, Helbling E, Williamson C, Worrest R (2011) Effects of UV radiation on aquatic ecosystems and interactions with climate change. Photochem Photobiol Sci 10:242–260CrossRefGoogle Scholar
  29. 29.
    Nagar V, Bandekar JR, Shashidhar R (2016) Expression of virulence and stress response genes in Aeromonas hydrophila under various stress conditions. J Basic Microbiol 56:1132–1137CrossRefGoogle Scholar
  30. 30.
    Grant SS, Hung DT (2013) Persistent bacterial infections, antibiotic tolerance, and the oxidative stress response. Virulence 4:273–283CrossRefGoogle Scholar
  31. 31.
    Redza-Dutordoir M, Averill-Bates DA (2016) Activation of apoptosis signalling pathways by reactive oxygen species. Biochem Biophys Acta 1863:2977–2992CrossRefGoogle Scholar
  32. 32.
    Wu Y, Vulić M, Keren I, Lewis K (2012) Role of oxidative stress in persister tolerance. Antimicrob Agents Chemother 56:4922–4926CrossRefGoogle Scholar
  33. 33.
    J.-S. Kim and T. K. Wood (2017) Tolerant, growing cells from nutrient shifts are not persister cells. mBio 8:e00354–17.PubMedPubMedCentralGoogle Scholar
  34. 34.
    Altschuler SJ, Wu LF (2010) Cellular heterogeneity: do differences make a difference? Cell 141:559–563CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  • Shridhar S. Paranjape
    • 1
    • 2
  • Ravindranath Shashidhar
    • 1
    • 2
    Email author
  1. 1.Food Technology DivisionBhabha Atomic Research CentreMumbaiIndia
  2. 2.Life SciencesHomi Bhabha National Institute (Deemed To Be University)MumbaiIndia

Personalised recommendations