Frequent carriage of resistance mechanisms to β-lactams and biofilm formation in Haemophilus influenzae causing treatment failure and recurrent otitis media in young children

J Antimicrob Chemother. 2014 Sep;69(9):2394-9. doi: 10.1093/jac/dku158. Epub 2014 Jun 2.

Abstract

Objectives: Non-typeable Haemophilus influenzae are a major cause of acute otitis media (AOM), including chronic and recurrent otitis in young children. The objective of this study was to determine whether non-typeable H. influenzae isolates causing these infections produce biofilms and carry resistance mechanisms to β-lactams.

Methods: A collection of 48 H. influenzae isolates was obtained by tympanocentesis or from otorrhoea samples from individual patients <3 years of age and diagnosed with recurrent or treatment failure AOM. Each isolate was surveyed for the presence of blaTEM genes, amino acid substitutions in the transpeptidase domain of penicillin-binding protein 3 (PBP3) and biofilm formation in microtitre plates.

Results: In 43 of the 48 isolates (89.6%), at least one of the three tested conditions was identified: biofilm formation (83.3%) and resistance mechanisms to β-lactams (33.3%), modifications in the transpeptidase domain of PBP3 being the most prevalent (22.9%), followed by β-lactamase production (10.4%). Additionally, 13 (27.1%) isolates had two or more of these three traits. In relation to biofilm formation, those isolates with an amoxicillin MIC ≤ 0.5 mg/L had higher optical density values than isolates with an amoxicillin MIC ≥ 1 mg/L (Mann-Whitney U-test, P=0.048).

Conclusions: These findings suggest that the successful treatment of non-typeable H. influenzae causing chronic and recurrent AOM in young children may be compromised by the high biofilm-forming capacity of the isolates and the presence of β-lactam resistance mechanisms, particularly PBP3 mutations.

Keywords: AOM; H. influenzae; gBLNAR.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Biofilms / growth & development*
  • Child, Preschool
  • Genes, Bacterial*
  • Haemophilus Infections / epidemiology
  • Haemophilus Infections / microbiology*
  • Haemophilus influenzae / drug effects*
  • Haemophilus influenzae / genetics
  • Haemophilus influenzae / isolation & purification
  • Haemophilus influenzae / physiology*
  • Humans
  • Infant
  • Otitis Media / epidemiology
  • Otitis Media / microbiology*
  • Recurrence
  • Treatment Failure
  • beta-Lactam Resistance*