Skip to main content

Minimal biofilm eradication concentration of antimicrobial agents against nontypeable Haemophilus influenzae isolated from middle ear fluids of intractable acute otitis media

Abstract

Nontypeable Haemophilus influenzae (NTHi) makes the clinical course of acute otitis media (AOM) intractable by forming a biofilm that may hamper the clearance of the bacteria from middle ear cavity. In this study, we evaluated the minimum biofilm eradication concentration (MBEC) of antimicrobial agents against biofilm-forming NTHi strains. Twelve NTHi strains isolated from middle ear fluids of Japanese children with intractable AOM before antimicrobial treatment were evaluated for MBEC of fluoroquinolones in comparison with those of β-lactams and macrolides. AMPC and CDTR required much higher concentration, i.e., high MBECs, to suppress the biofilm formation of NTHi. In contrast, fluoroquinolones followed by macrolides showed lower MBECs. MBEC would be a good parameter to infer the efficacies of antimicrobials against NTHi in biofilm.

This is a preview of subscription content, access via your institution.

Fig. 1
Fig. 2
Fig. 3

References

  1. Teele DW, Klein JO, Rosner B. Epidemiology of otitis media during the first seven years of life in children in greater Boston: a prospective, cohort study. J Infect Dis. 1989;160:83–94.

    PubMed  Article  CAS  Google Scholar 

  2. Monasta L, Ronfani L, Marchetti F, Montico M, Vecchi Brumatti L, Bavcar A, et al. Burden of disease caused by otitis media: systematic review and global estimates. PLoS ONE. 2012;7:e36226.

    PubMed  Article  CAS  Google Scholar 

  3. Hotomi M, Yamanaka N, Shimada J, Ikeda Y, Faden H. Factors associated with clinical outcomes in acute otitis media. Ann Otol Rhinol Laryngol. 2004;113:846–52.

    PubMed  Google Scholar 

  4. Murphy TF, Faden H, Bakaletz LO, Kyd JM, Forsgren A, Campos J, et al. Nontypeable Haemophilus influenzae as a pathogen in children. Pediatr Infect Dis J. 2009;28:43–8.

    PubMed  Article  Google Scholar 

  5. Hotomi M, Fujihara K, Billal DS, Suzuki K, Nishimura T, Baba S, et al. Genetic characteristics and clonal dissemination of beta-lactamase-negative ampicillin-resistant Haemophilus influenzae strains isolated from the upper respiratory tract of patients in Japan. Antimicrob Agents Chemother. 2007;51:3969–76.

    PubMed  Article  CAS  Google Scholar 

  6. Moxon ER, Sweetman WA, Deadman ME, Ferguson DJ, Hood DW. Haemophilus influenzae biofilms: hypothesis or fact? Trends Microbiol. 2008;16:95–100.

    PubMed  Article  CAS  Google Scholar 

  7. Post JC. Direct evidence of bacterial biofilms in otitis media. Laryngoscope. 2001;111:2083–94.

    PubMed  Article  CAS  Google Scholar 

  8. Bakaletz LO. Bacterial biofilms in otitis media: evidence and relevance. Pediatr Infect Dis J. 2007;26:S17–9.

    PubMed  Article  Google Scholar 

  9. Costerton JW, Stewart PS, Greenberg EP. Bacterial biofilms: a common cause of persistent infections. Science. 1999;284:1318–22.

    PubMed  Article  CAS  Google Scholar 

  10. Stewart PS, Costerton JW. Antibiotic resistance of bacteria in biofilms. Lancet. 2001;358:135–8.

    PubMed  Article  CAS  Google Scholar 

  11. Anderson GG, O’Toole GA. Innate and induced resistance mechanisms of bacterial biofilms. Curr Top Microbiol Immunol. 2008;322:85–105.

    PubMed  Article  CAS  Google Scholar 

  12. American Academy of Pediatrics Subcommittee on Management of Acute Otitis Media. Diagnosis and management of acute otitis media. Pediatrics. 2004;113:1451–5.

    Article  Google Scholar 

  13. Grubb MS, Spaugh DC. Treatment failure, recurrence, and antibiotic prescription rates for different acute otitis media treatment methods. Clin Pediatr (Phila). 2010;49:970–5.

    Article  Google Scholar 

  14. Moriyama S, Hotomi M, Shimada J, Billal DS, Fujihara K, Yamanaka N. Formation of biofilm by Haemophilus influenzae isolated from pediatric intractable otitis media. Auris Nasus Larynx. 2009;36:525–31.

    PubMed  Article  Google Scholar 

  15. Subcommittee of Clinical Practice Guideline for Diagnosis and Management of Acute Otitis Media in Children (Japan Otological Society, Japan Society for Pediatric Otorhinolaryngology, Japan Society for Infectious Diseases in Otolaryngology). Clinical practice guidelines for the diagnosis and management of acute otitis media (AOM) in children in Japan. Auris Nasus Larynx 2012;39:1–8.

    Google Scholar 

  16. Murphy TF, Brauer AL, Sethi S, Kilian M, Cai X, Lesse AL. Haemophilus haemolyticus: a human respiratory tract commensal to be distinguished from Haemophilus influenzae. J Infect Dis. 2007;195:81–9.

    PubMed  Article  CAS  Google Scholar 

  17. Kaji C, Watanabe K, Apicella MA, Watanabe H. Antimicrobial effect of fluoroquinolones for the eradication of nontypeable Haemophilus influenzae isolates within biofilms. Tohoku J Exp Med. 2008;214:121–8.

    PubMed  Article  CAS  Google Scholar 

  18. National Committee for Clinical Laboratory Standards (2004) Performance standards for antimicrobial susceptibility testing. Fourteenth informational supplement. Approved standard M100-S14. Wayne, PA: NCCLS.

  19. Billal DS, Fedorko DP, Yan SS, Hotomi M, Fujihara K, Nelson N, et al. In vitro induction and selection of fluoroquinolone-resistant mutants of Streptococcus pyogenes strains with multiple emm types. J Antimicrob Chemother. 2007;59:28–34.

    PubMed  Article  CAS  Google Scholar 

  20. Ceri H, Olson ME, Stremick C, Read RR, Morck D, Buret A. The Calgary Biofilm Device: new technology for rapid determination of antibiotic susceptibilities of bacterial biofilms. J Clin Microbiol. 1999;37:1771–6.

    PubMed  CAS  Google Scholar 

  21. Craig WA. Pharmacokinetic/pharmacodynamic parameters: rationale for antibacterial dosing of mice and men. Clin Infect Dis. 1998;26:1–10.

    PubMed  Article  CAS  Google Scholar 

  22. Anderl JN, Franklin MJ, Stewart PS. Role of antibiotic penetration limitation in Klebsiella pneumoniae biofilm resistance to ampicillin and ciprofloxacin. Antimicrob Agents Chemother. 2000;44:1818–24.

    PubMed  Article  CAS  Google Scholar 

  23. Fux CA, Costerton JW, Stewart PS, Stoodley P. Survival strategies of infectious biofilms. Trends Microbiol. 2005;13:34–40.

    PubMed  Article  CAS  Google Scholar 

  24. Donlan RM, Costerton JW. Biofilms: survival mechanisms of clinically relevant microorganisms. Clin Microbiol Rev. 2002;15:167–93.

    PubMed  Article  CAS  Google Scholar 

  25. Yamanaka N, Sugita R, Uno Y, Matsubara S, Hayashi Y, Sawada S. Clinical efficacy of tosufloxacin tosilate hydrate for the treatment of acute otitis media in children (in Japanese). Practica Oto-Rhino-Laryngologica 2012;105:381–392

    Google Scholar 

  26. Dagan R, Arguedas A, Schaad UB. Potential role of fluoroquinolone therapy in childhood otitis media. Pediatr Infect Dis J. 2004;23:390–8.

    PubMed  Article  Google Scholar 

  27. Reid G, Sharma S, Advikolanu K, Tieszer C, Martin RA, Bruce AW. Effects of ciprofloxacin, norfloxacin, and ofloxacin on in vitro adhesion and survival of Pseudomonas aeruginosa AK1 on urinary catheters. Antimicrob Agents Chemother. 1994;38:1490–5.

    PubMed  Article  CAS  Google Scholar 

Download references

Acknowledgments

This work was supported by national grants from the Japanese Ministry of Education, Culture, Sports, Science and Technology (grant 24791800).

Conflict of interest

We declare no conflict of interest for this study.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Muneki Hotomi.

Additional information

S. Takei and M. Hotomi contributed equally to this work.

About this article

Cite this article

Takei, S., Hotomi, M. & Yamanaka, N. Minimal biofilm eradication concentration of antimicrobial agents against nontypeable Haemophilus influenzae isolated from middle ear fluids of intractable acute otitis media. J Infect Chemother 19, 504–509 (2013). https://doi.org/10.1007/s10156-013-0592-y

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10156-013-0592-y

Keywords

  • Haemophilus influenzae
  • Biofilm
  • Minimal biofilm eradication concentration
  • Antimicrobial agents