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Background  Biofilm production by  Haemophilus influenzae  and  Streptococcus pneumoniae  has been implicated in the pathogenesis of otitis media, mainly in chronic and recurrent cases. We studied the  “ in vitro ”  biofilm production by these 2 species isolated alone or together from the nasopharynx of children with acute otitis media.    Methods  The studied strains were from 3 pneumococcal conjugate vaccine (PCV) periods: pre-PCV7, post-PCV7/pre-PCV13 and post-PCV13. A modified microtiter plate assay with crystal violet stain was used to study the biofilm production of 182  H. influenzae  and 191  S. pneumonia e strains.    Results  Overall, 117/181 (64.6%)  H. influenzae  and 128/191 (66.8%)  S. pneumoniae  strains produced biofilm. The proportion of biofilm-producing  H. influenzae  strains was greater with than without the isolation of  S. pneumoniae  in the same sample (75.5% vs 52.3%,  p  = 0.001). Conversely, the proportion of biofilm-producing  S. pneumoniae  strains was not affected by the presence or not of  H. influenzae  (66.3% vs 67.4%).  S. pneumoniae  serotypes 6B, 15B/C, 19A, 35F and 35B were the better biofilm producers (80%). Serotypes 11A, 14, 15A, 19F and 19A were more associated with  H. influenzae  biofilm-producing strains. Overall, 89/94 (94.6%) of cases with combined isolation showed biofilm production by  S. pneumoniae  or  H. influenzae .    Conclusion  This study emphasizes the high proportion of biofilm production by  H. influenzae  and  S. pneumoniae  strains isolated from the nasopharynx of children with acute otitis media, which reinforces the results of studies suggesting the importance of biofilm in the pathogenesis of acute otitis media.    Electronic supplementary material  The online version of this article (10.1186/s12879-018-3657-9) contains supplementary material, which is available to authorized users.

Biofilm production by Haemophilus influenzae and Streptococcus pneumoniae isolated from the nasopharynx of children with acute otitis media