Skip to main content
PMC home page
Antimicrobial Agents and Chemotherapy logoLink to Antimicrobial Agents and Chemotherapy
. 1995 Sep;39(9):2019–2022. doi: 10.1128/aac.39.9.2019

Evidence for an efflux pump in multidrug-resistant Campylobacter jejuni.

E Charvalos 1, Y Tselentis 1, M M Hamzehpour 1, T Köhler 1, J C Pechere 1
PMCID: PMC162874  PMID: 8540709

Abstract

Mechanisms of drug resistance in Campylobacter jejuni were investigated. Mutant strains 34PEFr, which was resistant to pefloxacin (128-fold increase in the MIC), and 34CTXr, which was resistant to cefotaxime (32-fold increase in the MIC) and which was derived from the susceptible parent 34s, were obtained by serial passages on pefloxacin and cefotaxime gradient plates, respectively. Both mutants showed cross-resistance to erythromycin, chloramphenicol, tetracycline, beta-lactams, and quinolones. While the quinolone resistance of strain PEFr could be explained by a mutation at codon 86 of the gyrA gene, the multidrug resistance phenotype of both strains was further investigated. Accumulation of pefloxacin, ciprofloxacin, and minocycline was measured by fluorometry and was found to be lower in the mutant strains than in the parent strain. Preincubation of the cells with carbonyl cyanide m-chlorophenylhydrazone, however, completely abolished this difference. Analysis by sodium dodecyl sulfate-polyacrylamide gel electrophoresis of outer membrane preparations from both mutant strains showed overexpression of two proteins of 55 and 39 kDa which were absent from the outer membranes of the wild-type strain. These results indicate that in C. jejuni 34PEFr and 34CTXr, multidrug resistance is associated with an efflux system with a broad specificity.

Full Text

The Full Text of this article is available as a PDF (227.8 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Adler-Mosca H., Lüthy-Hottenstein J., Martinetti Lucchini G., Burnens A., Altwegg M. Development of resistance to quinolones in five patients with campylobacteriosis treated with norfloxacin or ciprofloxacin. Eur J Clin Microbiol Infect Dis. 1991 Nov;10(11):953–957. doi: 10.1007/BF02005451. [DOI] [PubMed] [Google Scholar]
  2. Blaser M. J., Berkowitz I. D., LaForce F. M., Cravens J., Reller L. B., Wang W. L. Campylobacter enteritis: clinical and epidemiologic features. Ann Intern Med. 1979 Aug;91(2):179–185. doi: 10.7326/0003-4819-91-2-179. [DOI] [PubMed] [Google Scholar]
  3. Blaser M. J., Reller L. B. Campylobacter enteritis. N Engl J Med. 1981 Dec 10;305(24):1444–1452. doi: 10.1056/NEJM198112103052404. [DOI] [PubMed] [Google Scholar]
  4. Bryan J. P., Waters C., Sheffield J., Krieg R. E., Perine P. L., Wagner K. In vitro activities of tosufloxacin, temafloxacin, and A-56620 against pathogens of diarrhea. Antimicrob Agents Chemother. 1990 Feb;34(2):368–370. doi: 10.1128/aac.34.2.368. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Butzler J. P., Skirrow M. B. Campylobacter enteritis. Clin Gastroenterol. 1979 Sep;8(3):737–765. [PubMed] [Google Scholar]
  6. Celesk R. A., Robillard N. J. Factors influencing the accumulation of ciprofloxacin in Pseudomonas aeruginosa. Antimicrob Agents Chemother. 1989 Nov;33(11):1921–1926. doi: 10.1128/aac.33.11.1921. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Chapman J. S., Georgopapadakou N. H. Routes of quinolone permeation in Escherichia coli. Antimicrob Agents Chemother. 1988 Apr;32(4):438–442. doi: 10.1128/aac.32.4.438. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Cohen S. P., Hächler H., Levy S. B. Genetic and functional analysis of the multiple antibiotic resistance (mar) locus in Escherichia coli. J Bacteriol. 1993 Mar;175(5):1484–1492. doi: 10.1128/jb.175.5.1484-1492.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Cohen S. P., McMurry L. M., Hooper D. C., Wolfson J. S., Levy S. B. Cross-resistance to fluoroquinolones in multiple-antibiotic-resistant (Mar) Escherichia coli selected by tetracycline or chloramphenicol: decreased drug accumulation associated with membrane changes in addition to OmpF reduction. Antimicrob Agents Chemother. 1989 Aug;33(8):1318–1325. doi: 10.1128/aac.33.8.1318. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. DuPont H. L., Ericsson C. D., Robinson A., Johnson P. C. Current problems in antimicrobial therapy for bacterial enteric infection. Am J Med. 1987 Apr 27;82(4A):324–328. [PubMed] [Google Scholar]
  11. Endtz H. P., Mouton R. P., van der Reyden T., Ruijs G. J., Biever M., van Klingeren B. Fluoroquinolone resistance in Campylobacter spp isolated from human stools and poultry products. Lancet. 1990 Mar 31;335(8692):787–787. doi: 10.1016/0140-6736(90)90897-e. [DOI] [PubMed] [Google Scholar]
  12. Fukuda H., Hosaka M., Hirai K., Iyobe S. New norfloxacin resistance gene in Pseudomonas aeruginosa PAO. Antimicrob Agents Chemother. 1990 Sep;34(9):1757–1761. doi: 10.1128/aac.34.9.1757. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. George A. M., Levy S. B. Amplifiable resistance to tetracycline, chloramphenicol, and other antibiotics in Escherichia coli: involvement of a non-plasmid-determined efflux of tetracycline. J Bacteriol. 1983 Aug;155(2):531–540. doi: 10.1128/jb.155.2.531-540.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Goodman L. J., Trenholme G. M., Kaplan R. L., Segreti J., Hines D., Petrak R., Nelson J. A., Mayer K. W., Landau W., Parkhurst G. W. Empiric antimicrobial therapy of domestically acquired acute diarrhea in urban adults. Arch Intern Med. 1990 Mar;150(3):541–546. [PubMed] [Google Scholar]
  15. Gootz T. D., Martin B. A. Characterization of high-level quinolone resistance in Campylobacter jejuni. Antimicrob Agents Chemother. 1991 May;35(5):840–845. doi: 10.1128/aac.35.5.840. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Hallett P., Maxwell A. Novel quinolone resistance mutations of the Escherichia coli DNA gyrase A protein: enzymatic analysis of the mutant proteins. Antimicrob Agents Chemother. 1991 Feb;35(2):335–340. doi: 10.1128/aac.35.2.335. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Hooper D. C., Wolfson J. S., Souza K. S., Ng E. Y., McHugh G. L., Swartz M. N. Mechanisms of quinolone resistance in Escherichia coli: characterization of nfxB and cfxB, two mutant resistance loci decreasing norfloxacin accumulation. Antimicrob Agents Chemother. 1989 Mar;33(3):283–290. doi: 10.1128/aac.33.3.283. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  19. Legakis N. J., Tzouvelekis L. S., Makris A., Kotsifaki H. Outer membrane alterations in multiresistant mutants of Pseudomonas aeruginosa selected by ciprofloxacin. Antimicrob Agents Chemother. 1989 Jan;33(1):124–127. doi: 10.1128/aac.33.1.124. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Levy S. B. Active efflux mechanisms for antimicrobial resistance. Antimicrob Agents Chemother. 1992 Apr;36(4):695–703. doi: 10.1128/aac.36.4.695. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Li X. Z., Livermore D. M., Nikaido H. Role of efflux pump(s) in intrinsic resistance of Pseudomonas aeruginosa: resistance to tetracycline, chloramphenicol, and norfloxacin. Antimicrob Agents Chemother. 1994 Aug;38(8):1732–1741. doi: 10.1128/aac.38.8.1732. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Li X. Z., Ma D., Livermore D. M., Nikaido H. Role of efflux pump(s) in intrinsic resistance of Pseudomonas aeruginosa: active efflux as a contributing factor to beta-lactam resistance. Antimicrob Agents Chemother. 1994 Aug;38(8):1742–1752. doi: 10.1128/aac.38.8.1742. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Logan S. M., Trust T. J. Outer membrane characteristics of Campylobacter jejuni. Infect Immun. 1982 Dec;38(3):898–906. doi: 10.1128/iai.38.3.898-906.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Ma D., Cook D. N., Alberti M., Pon N. G., Nikaido H., Hearst J. E. Molecular cloning and characterization of acrA and acrE genes of Escherichia coli. J Bacteriol. 1993 Oct;175(19):6299–6313. doi: 10.1128/jb.175.19.6299-6313.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Oram M., Fisher L. M. 4-Quinolone resistance mutations in the DNA gyrase of Escherichia coli clinical isolates identified by using the polymerase chain reaction. Antimicrob Agents Chemother. 1991 Feb;35(2):387–389. doi: 10.1128/aac.35.2.387. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Page W. J., Huyer G., Huyer M., Worobec E. A. Characterization of the porins of Campylobacter jejuni and Campylobacter coli and implications for antibiotic susceptibility. Antimicrob Agents Chemother. 1989 Mar;33(3):297–303. doi: 10.1128/aac.33.3.297. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Page W. J., Taylor D. E. Comparison of methods used to separate the inner and outer membranes of cell envelopes of Campylobacter spp. J Gen Microbiol. 1988 Nov;134(11):2925–2932. doi: 10.1099/00221287-134-11-2925. [DOI] [PubMed] [Google Scholar]
  28. Poole K., Krebes K., McNally C., Neshat S. Multiple antibiotic resistance in Pseudomonas aeruginosa: evidence for involvement of an efflux operon. J Bacteriol. 1993 Nov;175(22):7363–7372. doi: 10.1128/jb.175.22.7363-7372.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Reina J., Alomar P. Fluoroquinolone-resistance in thermophilic Campylobacter spp isolated from stools of Spanish patients. Lancet. 1990 Jul 21;336(8708):186–186. doi: 10.1016/0140-6736(90)91715-m. [DOI] [PubMed] [Google Scholar]
  30. Segreti J., Gootz T. D., Goodman L. J., Parkhurst G. W., Quinn J. P., Martin B. A., Trenholme G. M. High-level quinolone resistance in clinical isolates of Campylobacter jejuni. J Infect Dis. 1992 Apr;165(4):667–670. doi: 10.1093/infdis/165.4.667. [DOI] [PubMed] [Google Scholar]
  31. Seoane A. S., Levy S. B. Identification of new genes regulated by the marRAB operon in Escherichia coli. J Bacteriol. 1995 Feb;177(3):530–535. doi: 10.1128/jb.177.3.530-535.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Shales S. W., Chopra I., Ball P. R. Evidence for more than one mechanism of plasmid-determined tetracycline resistance in Escherichia coli. J Gen Microbiol. 1980 Nov;121(1):221–229. doi: 10.1099/00221287-121-1-221. [DOI] [PubMed] [Google Scholar]
  33. Taylor D. E., Ng L. K., Lior H. Susceptibility of Campylobacter species to nalidixic acid, enoxacin, and other DNA gyrase inhibitors. Antimicrob Agents Chemother. 1985 Nov;28(5):708–710. doi: 10.1128/aac.28.5.708. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Wang Y., Huang W. M., Taylor D. E. Cloning and nucleotide sequence of the Campylobacter jejuni gyrA gene and characterization of quinolone resistance mutations. Antimicrob Agents Chemother. 1993 Mar;37(3):457–463. doi: 10.1128/aac.37.3.457. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Wolfson J. S., Hooper D. C. Fluoroquinolone antimicrobial agents. Clin Microbiol Rev. 1989 Oct;2(4):378–424. doi: 10.1128/cmr.2.4.378. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Yoshida H., Bogaki M., Nakamura M., Nakamura S. Quinolone resistance-determining region in the DNA gyrase gyrA gene of Escherichia coli. Antimicrob Agents Chemother. 1990 Jun;34(6):1271–1272. doi: 10.1128/aac.34.6.1271. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Antimicrobial Agents and Chemotherapy are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES