1887

Abstract

Members of the genus , mainly , can cause severe disease in man, especially after intestinal perforation in the course of abdominal surgery. Treatment is based on a small number of antibiotics, including metronidazole, which has proved to be highly reliable throughout the last 40 to 50 years. Nevertheless, metronidazole resistance does occur in and has been mainly attributed to Nim proteins, a class of proteins with a suggested nitroreductase function. Despite the potentially high importance of Nim proteins for human health, information on the expression of genes in is still lacking. It was the aim of this study to demonstrate expression of genes in at the protein level and, furthermore, to correlate Nim levels with the magnitude of metronidazole resistance. By the application of 2D gel electrophoresis, Nim proteins could be readily identified in -positive strains, but their levels were not elevated to a relevant extent after induction of resistance with high doses of metronidazole. Thus, the data herein do not provide evidence for Nim proteins acting as nitroreductases using metronidazole as a substrate, because no correlation between Nim levels and levels of metronidazole resistance could be observed. Furthermore, no evidence was found that Nim proteins protect . from metronidazole by sequestering the activated antibiotic.

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2014-03-01
2019-10-15
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References

  1. Aldridge K. E., Sanders C. V.. ( 2002;). Susceptibility trending of blood isolates of the Bacteroides fragilis group over a 12-year period to clindamycin, ampicillin-sulbactam, cefoxitin, imipenem, and metronidazole. . Anaerobe 8:, 301–305. [CrossRef][PubMed]
    [Google Scholar]
  2. Bradford M. M.. ( 1976;). A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. . Anal Biochem 72:, 248–254. [CrossRef][PubMed]
    [Google Scholar]
  3. Brazier J. S., Stubbs S. L. J., Duerden B. I.. ( 1999;). Metronidazole resistance among clinical isolates belonging to the Bacteroides fragilis group: time to be concerned. ? J Antimicrob Chemother 44:, 580–581. [CrossRef][PubMed]
    [Google Scholar]
  4. Breuil J., Dublanchet A., Truffaut N., Sebald M.. ( 1989;). Transferable 5-nitroimidazole resistance in the Bacteroides fragilis group. . Plasmid 21:, 151–154. [CrossRef][PubMed]
    [Google Scholar]
  5. Carlier J. P., Sellier N., Rager M. N., Reysset G.. ( 1997;). Metabolism of a 5-nitroimidazole in susceptible and resistant isogenic strains of Bacteroides fragilis. . Antimicrob Agents Chemother 41:, 1495–1499.[PubMed]
    [Google Scholar]
  6. Gal M., Brazier J. S.. ( 2004;). Metronidazole resistance in Bacteroides spp. carrying nim genes and the selection of slow-growing metronidazole-resistant mutants. . J Antimicrob Chemother 54:, 109–116. [CrossRef][PubMed]
    [Google Scholar]
  7. Haggoud A., Reysset G., Azeddoug H., Sebald M.. ( 1994;). Nucleotide sequence analysis of two 5-nitroimidazole resistance determinants from Bacteroides strains and of a new insertion sequence upstream of the two genes. . Antimicrob Agents Chemother 38:, 1047–1051. [CrossRef][PubMed]
    [Google Scholar]
  8. Hedberg M., Nord C. E..on behalf of the ESCMID Study Group on Antimicrobial Resistance in Anaerobic Bacteria ( 2003;). Antimicrobial susceptibility of Bacteroides fragilis group isolates in Europe. . Clin Microbiol Infect 9:, 475–488. [CrossRef][PubMed]
    [Google Scholar]
  9. Husain F., Veeranagouda Y., Hsi J., Meggersee R., Abratt V., Wexler H. M.. ( 2013;). Two multidrug-resistant clinical isolates of Bacteroides fragilis carry a novel metronidazole resistance nim gene (nimJ). . Antimicrob Agents Chemother 57:, 3767–3774. [CrossRef][PubMed]
    [Google Scholar]
  10. Jousimies-Somer H. R., Summanen P., Citron D. M., Baron E. J., Wexler H. M., Finegold S. M.. ( 2002;). Wadsworth - KTL Anaerobic Bacteriology Manual, , 6th edn.. Belmont, CA:: Star Publishing;.
    [Google Scholar]
  11. Kolarich D., Jensen P. H., Altmann F., Packer N. H.. ( 2012;). Determination of site-specific glycan heterogeneity on glycoproteins. . Nat Protoc 7:, 1285–1298. [CrossRef][PubMed]
    [Google Scholar]
  12. Kuwahara T., Yamashita A., Hirakawa H., Nakayama H., Toh H., Okada N., Kuhara S., Hattori M., Hayashi T., Ohnishi Y.. ( 2004;). Genomic analysis of Bacteroides fragilis reveals extensive DNA inversions regulating cell surface adaptation. . Proc Natl Acad Sci U S A 101:, 14919–14924. [CrossRef][PubMed]
    [Google Scholar]
  13. Leitsch D., Kolarich D., Wilson I. B. H., Altmann F., Duchêne M.. ( 2007;). Nitroimidazole action in Entamoeba histolytica: a central role for thioredoxin reductase. . PLoS Biol 5:, e211. [CrossRef][PubMed]
    [Google Scholar]
  14. Leitsch D., Kolarich D., Binder M., Stadlmann J., Altmann F., Duchêne M.. ( 2009;). Trichomonas vaginalis: metronidazole and other nitroimidazole drugs are reduced by the flavin enzyme thioredoxin reductase and disrupt the cellular redox system. Implications for nitroimidazole toxicity and resistance. . Mol Microbiol 72:, 518–536. [CrossRef][PubMed]
    [Google Scholar]
  15. Leitsch D., Schlosser S., Burgess A., Duchêne M.. ( 2012;). Nitroimidazole drugs vary in their mode of action in the human parasite Giardia lamblia.. Int J Parasitol Drugs Drug Res 2:, 166–170. [CrossRef]
    [Google Scholar]
  16. Löfmark S., Fang H., Hedberg M., Edlund C.. ( 2005;). Inducible metronidazole resistance and nim genes in clinical Bacteroides fragilis group isolates. . Antimicrob Agents Chemother 49:, 1253–1256. [CrossRef][PubMed]
    [Google Scholar]
  17. Samuelson J.. ( 1999;). Why metronidazole is active against both bacteria and parasites. . Antimicrob Agents Chemother 43:, 1533–1541.[PubMed]
    [Google Scholar]
  18. Sebald M., Reysset G., Breuil J.. ( 1990;). What’s new in 5-nitroimidazole resistance in the Bacteroides fragilis group. ? In Clinical and Molecular Aspects of Anaerobes, pp. 217–225. Edited by Borriello S. P... Petersfield:: Wrightson Biomedical Publishing;.
    [Google Scholar]
  19. Sisson G., Jeong J. Y., Goodwin A., Bryden L., Rossler N., Lim-Morrison S., Raudonikiene A., Berg D. E., Hoffman P. S.. ( 2000;). Metronidazole activation is mutagenic and causes DNA fragmentation in Helicobacter pylori and in Escherichia coli containing a cloned H. pylori RdxA+ (nitroreductase) gene. . J Bacteriol 182:, 5091–5096. [CrossRef][PubMed]
    [Google Scholar]
  20. Sóki J., Fodor E., Hecht D. W., Edwards R., Rotimi V. O., Kerekes I., Urbán E., Nagy E.. ( 2004;). Molecular characterization of imipenem-resistant, cfiA-positive Bacteroides fragilis isolates from the USA, Hungary and Kuwait. . J Med Microbiol 53:, 413–419. [CrossRef][PubMed]
    [Google Scholar]
  21. Sóki J., Gal M., Brazier J. S., Rotimi V. O., Urbán E., Nagy E., Duerden B. I.. ( 2006;). Molecular investigation of genetic elements contributing to metronidazole resistance in Bacteroides strains. . J Antimicrob Chemother 57:, 212–220. [CrossRef][PubMed]
    [Google Scholar]
  22. Sóki J., Eitel Z., Urbán E., Nagy E..on behalf of the ESCMID Study Group on Anaerobic Infections ( 2013;). Molecular analysis of the carbapenem and metronidazole resistance mechanisms of Bacteroides strains reported in a Europe-wide antibiotic resistance survey. . Int J Antimicrob Agents 41:, 122–125. [CrossRef][PubMed]
    [Google Scholar]
  23. Stubbs S. L., Brazier J. S., Talbot P. R., Duerden B. I.. ( 2000;). PCR-restriction fragment length polymorphism analysis for identification of Bacteroides spp. and characterization of nitroimidazole resistance genes. . J Clin Microbiol 38:, 3209–3213.[PubMed]
    [Google Scholar]
  24. Wexler H. M.. ( 2007;). Bacteroides: the good, the bad, and the nitty-gritty. . Clin Microbiol Rev 20:, 593–621. [CrossRef][PubMed]
    [Google Scholar]
  25. Williams C. F., Lloyd D., Kolarich D., Alagesan K., Duchêne M., Cable J., Williams D., Leitsch D.. ( 2012;). Disrupted intracellular redox balance of the diplomonad fish parasite Spironucleus vortens by 5-nitroimidazoles and garlic-derived compounds. . Vet Parasitol 190:, 62–73. [CrossRef][PubMed]
    [Google Scholar]
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