1887

Abstract

is a major bacterial foodborne-pathogen. Ciprofloxacin is an important antibiotic for the treatment of , albeit high rates of fluoroquinolone resistance have limited its usefulness. Persister-cells are transiently antibiotic-tolerant fractions of bacterial populations and their occurrence has been associated with recalcitrant and persistent bacterial infections. Here, time-kill assays with ciprofloxacin (200×MIC, 25 µg ml) were performed in strains 81–176 and RM1221 and persister-cells were found. The frequency of survivors after 8 h of ciprofloxacin exposure was approx. 10 for both strains, while after 22 h the frequency was between 10–10, depending on the strain and growth-phase. Interestingly, the stationary-phase cultures did not display more persister-cells compared to exponential-phase cultures, in contrast to what has been observed in other bacterial species. Persister-cells after ampicillin exposure (100×MIC, 200 µg ml) were not detected, implying that persister-cell formation in is antibiotic-specific. In attempts to identify the mechanism of ciprofloxacin persister-cell formation, stringent or SOS responses were not found to play major roles. Overall, this study reports ciprofloxacin persister-cells in and challenges the notion of persister-cells as plainly dormant non-growing cells.

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/content/journal/micro/10.1099/mic.0.000953
2020-07-22
2020-10-20
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References

  1. Carron M, Chang Y-M, Momanyi K, Akoko J, Kiiru J et al. Campylobacter factors in the fast-evolving chicken meat system of Nairobi, Kenya. PLoS Negl Trop Dis 2018; 12:1–18
    [Google Scholar]
  2. Wieczorek K, Osek J. Antimicrobial resistance mechanisms among Campylobacter . Biomed Res Int 2013; 2013:1–12
    [Google Scholar]
  3. Kim Y, Shin JA, Han SB, Cho B, Jeong DC et al. Recurrent Campylobacter jejuni bacteremia in a patient with hypogammaglobulinemia: A case report. Medicine 2017; 96:1–3
    [Google Scholar]
  4. Sproston EL, Wimalarathna HML, Sheppard SK. Trends in fluoroquinolone resistance in Campylobacter . Microb Genomics. 2018; 4:
    [Google Scholar]
  5. Tang Y, Sahin O, Pavlovic N, Lejeune J, Carlson J et al. Rising fluoroquinolone resistance in Campylobacter isolated from feedlot cattle in the United States. Sci Rep 2017; 7:1–8
    [Google Scholar]
  6. Fauvart M, de Groote VN, Michiels J. Role of persister cells in chronic infections: clinical relevance and perspectives on anti-persister therapies. J Med Microbiol 2011; 60:699–709
    [Google Scholar]
  7. Fisher RA, Gollan B, Helaine S. Persistent bacterial infections and persister cells. Nat Rev Microbiol 2017; 15:453–464
    [Google Scholar]
  8. Levin BR, Rozen DE. Non-inherited antibiotic resistance. Nat Rev Microbiol 2006; 4:556–562
    [Google Scholar]
  9. Dörr T, Lewis K, Vulić M. SOS response induces persistence to fluoroquinolones in Escherichia coli . PLoS Genet 2009; 5:1–9
    [Google Scholar]
  10. Gefen O, Gabay C, Mumcuoglu M, Engel G, Balaban NQ. Single-Cell protein induction dynamics reveals a period of vulnerability to antibiotics in persister bacteria. Proc Natl Acad Sci U S A 2008; 105:6145–6149
    [Google Scholar]
  11. Maisonneuve E, Gerdes K. Molecular mechanisms underlying bacterial persisters. Cell 2014; 157:539–548
    [Google Scholar]
  12. Maisonneuve E, Castro-Camargo M, Gerdes K. p)ppGpp controls bacterial persistence by stochastic induction of toxin-antitoxin activity. Cell 2013; 154:1140–1150
    [Google Scholar]
  13. Hauryliuk V, Atkinson GC, Murakami KS, Tenson T, Gerdes K. Recent functional insights into the role of (p)ppGpp in bacterial physiology. Nat Rev Microbiol 2015; 13:298–309
    [Google Scholar]
  14. Keren I, Minami S, Rubin E, Lewis K. Characterization and transcriptome analysis of Mycobacterium tuberculosis persisters. MBio 2011; 2:1–10
    [Google Scholar]
  15. Shan Y, Gandt AB, Rowe SE, Deisinger JP, Conlon BP et al. ATP-dependent persister formation in Escherichia coli . MBio 2017; 8:1–14
    [Google Scholar]
  16. Conlon BP, Rowe SE, Gandt AB, Nuxoll AS, Donegan NP et al. Persister formation in Staphylococcus aureus is associated with ATP depletion. Nat Microbiol 2016; 1:1–7
    [Google Scholar]
  17. Ma C, Sim S, Shi W, Du L, Xing D et al. Energy production genes sucB and ubiF are involved in persister survival and tolerance to multiple antibiotics and stresses in Escherichia coli . FEMS Microbiol Lett 2010; 303:33–40
    [Google Scholar]
  18. Spoering LA, Vulić M, Lewis K. GlpD and PlsB participate in persister cell formation in Escherichia coli . J Bacteriol 2006; 188:5136–5144
    [Google Scholar]
  19. Leung V, Lévesque CM. A stress-inducible quorum-sensing peptide mediates the formation of persister cells with noninherited multidrug tolerance. J Bacteriol 2012; 194:2265–2274
    [Google Scholar]
  20. Möker N, Dean CR, Tao J. Pseudomonas aeruginosa increases formation of multidrug-tolerant persister cells in response to quorum-sensing signaling molecules. J Bacteriol 2010; 192:1946–1955
    [Google Scholar]
  21. Levin BR, Concepción-Acevedo J, Udekwu KI. Persistence: a copacetic and parsimonious hypothesis for the existence of non-inherited resistance to antibiotics. Curr Opin Microbiol 2014; 21:18–21
    [Google Scholar]
  22. Kaldalu N, Hauryliuk V, Tenson T. Persisters—as elusive as ever. Appl Microbiol Biotechnol 2016; 100:6545–6553
    [Google Scholar]
  23. Kim J-S, Wood TK. Persistent persister misperceptions. Front Microbiol. 2016; 7:1–7
    [Google Scholar]
  24. Wood TK, Song S, Yamasaki R. Ribosome dependence of persister cell formation and resuscitation. J Microbiol 2019; 57:213–219
    [Google Scholar]
  25. CLSI Performance standards for antimicrobial susceptibility testing; 20th informational supplement. CLSI document M45-A2. CLSI, Wayne, PA 2010; 30: Vol.
    [Google Scholar]
  26. Amato SM, Brynildsen MP. Persister heterogeneity arising from a single metabolic stress. Curr Biol 2015; 25:2090–2098
    [Google Scholar]
  27. Luidalepp H, Jõers A, Kaldalu N, Tenson T. Age of inoculum strongly influences persister frequency and can mask effects of mutations implicated in altered persistence. J Bacteriol 2011; 193:3598–3605
    [Google Scholar]
  28. Goneau LW, Yeoh NS, MacDonald KW, Cadieux PA, Burton JP et al. Selective target inactivation rather than global metabolic dormancy causes antibiotic tolerance in uropathogens. Antimicrob Agents Chemother 2014; 58:2089–2097
    [Google Scholar]
  29. Keren I, Kaldalu N, Spoering A, Wang Y, Lewis K. Persister cells and tolerance to antimicrobials. FEMS Microbiol Lett 2004; 230:13–18
    [Google Scholar]
  30. Mina EG, Marques CNH. Interaction of Staphylococcus aureus persister cells with the host when in a persister state and following awakening. Sci Rep 2016; 6:1–10
    [Google Scholar]
  31. Spoering A, Lewis K. Biofilms and planktonic cells of Pseudomonas aeruginosa have similar resistance to killing by antimicrobials. Society 2001; 183:6746–6751
    [Google Scholar]
  32. Nguyen D, Joshi-Datar A, Lepine F, Bauerle E, Olakanmi O et al. Active starvation responses mediate antibiotic tolerance in biofilms and nutrient-limited bacteria. Science 2011; 334:982–986
    [Google Scholar]
  33. Goneau LW, Yeoh NS, MacDonald KW, Cadieux PA, Burton JP et al. Selective target inactivation rather than global metabolic dormancy causes antibiotic tolerance in uropathogens. Antimicrob Agents Chemother 2014; 58:2089–2097
    [Google Scholar]
  34. Hofsteenge N, van Nimwegen E, Silander OK. Quantitative analysis of persister fractions suggests different mechanisms of formation among environmental isolates of E. coli . BMC Microbiol 2013; 13:1–11
    [Google Scholar]
  35. Orman MA, Brynildsen MP. Dormancy is not necessary or sufficient for bacterial persistence. Antimicrob Agents Chemother 2013; 57:3230–3239
    [Google Scholar]
  36. Völzing KG, Brynildsen MP. Stationary-phase persisters to ofloxacin sustain DNA damage and require repair systems only during recovery. MBio 2015; 6:1–11
    [Google Scholar]
  37. Claudi B, Spröte P, Chirkova A, Personnic N, Zankl J et al. Phenotypic variation of Salmonella in host tissues delays eradication by antimicrobial chemotherapy. Cell 2014; 158:722–733
    [Google Scholar]
  38. Turonova H, Haddad N, Hernould M, Chevret D, Pazlarova J et al. Profiling of Campylobacter jejuni proteome in exponential and stationary phase of growth. Front Microbiol 2017; 8:1–12
    [Google Scholar]
  39. Parkhill J, Wren BW, Mungall K, Ketley JM, Churcher C et al. The genome sequence of the food-borne pathogen Campylobacter jejuni reveals hypervariable sequences. Nature 2000; 403:665–668
    [Google Scholar]
  40. Jeon B. A tangle of poly-phosphate in Campylobacter . Virulence 2014; 5:449–450
    [Google Scholar]
  41. Fields JA, Li J, Gulbronson CJ, Hendrixson DR, Thompson SA. Campylobacter jejuni CsrA regulates metabolic and virulence associated proteins and is necessary for mouse colonization. PLoS One 2016; 11:1–20
    [Google Scholar]
  42. Kelly AF, Park SF, Bovill R, Mackey M. Survival of Campylobacter jejuni during stationary phase: evidence for the absence of a phenotypic stationary-phase response. Appl Environ Microbiol 2001; 67:2248–2254
    [Google Scholar]
  43. Murphy C, Carroll C, Jordan KN. Induction of an adaptive tolerance response in the foodborne pathogen, Campylobacter jejuni . FEMS Microbiol Lett 2003; 223:89–93
    [Google Scholar]
  44. Ferullo DJ, Lovett ST. The stringent response and cell cycle arrest in Escherichia coli . PLoS Genet 2008; 4:1–15
    [Google Scholar]
  45. Germain E, Guiraud P, Byrne D, Douzi B, Djendli M et al. YtfK activates the stringent response by triggering the alarmone synthetase SpoT in Escherichia coli . Nat Commun 2019; 10:1–12
    [Google Scholar]
  46. Gaynor EC, Wells DH, MacKichan JK, Falkow S. The Campylobacter jejuni stringent response controls specific stress survival and virulence-associated phenotypes. Mol Microbiol 2005; 56:8–27
    [Google Scholar]
  47. Gaasbeek EJ, Wagenaar JA, Guilhabert MR, Van Putten JPM, Parker CT et al. Nucleases encoded by the integrated elements CJIE2 and CJIE4 inhibit natural transformation of Campylobacter jejuni . J Bacteriol 2010; 192:936–941
    [Google Scholar]
  48. Miller WG, Bates AH, Horn ST, Brandl MT, Wachtel MR et al. Detection on surfaces and in Caco-2 cells of Campylobacter jejuni cells transformed with new gfp, yfp, and cfp marker plasmids. Appl Environ Microbiol 2000; 66:5426–5436
    [Google Scholar]
  49. Chowdhury N, Kwan BW, Wood TK. Persistence increases in the absence of the alarmone guanosine tetraphosphate by reducing cell growth. Sci Rep 2016; 6:1–9
    [Google Scholar]
  50. Shah D, Zhang Z, Khodursky A, Kaldalu N, Kurg K et al. Persisters: a distinct physiological state of E. coli . BMC Microbiol 2006; 6:1–9
    [Google Scholar]
  51. Cohen NR, Lobritz MA, Collins JJ. Microbial persistence and the road to drug resistance. Cell Host Microbe 2013; 13:632–642
    [Google Scholar]
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