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Background  Bacterial vaginosis (BV) is one of the most common vaginal infections worldwide. It is associated with the presence of a dense polymicrobial biofilm on the vaginal epithelium, formed mainly by  Gardnerella  species. The biofilm also contains other anaerobic species, but little is known about their role in BV development.    Aim  To evaluate the influence of different culture media on the planktonic and biofilm growth of six cultivable anaerobes frequently associated with BV, namely  Gardnerella  sp.,  Atopobium vaginae ,  Lactobacillus iners ,  Mobiluncus curtisii ,  Peptostreptococcus anaerobius  and  Prevotella bivia .    Methods  A total of nine different culture media compositions, including commercially available and chemically defined media simulating genital tract secretions, were tested in this study. Planktonic cultures and biofilms were grown under anaerobic conditions (10% carbon dioxide, 10% helium and 80% nitrogen). Planktonic growth was assessed by optical density measurements, and biofilm formation was quantified by crystal violet staining.    Results  Significant planktonic growth was observed for  Gardnerella  sp.,  A. vaginae  and  L. iners  in New York City III broth, with or without ascorbic acid supplementation. Biofilm quantification showed high in vitro biofilm growth for  Gardnerella  sp.,  P. anaerobius  and  P. bivia  in almost all culture media excluding Brucella broth. Contrary, only New York City III broth was able to promote biofilm formation for  A. vaginae ,  L. iners  and  M. curtisii .    Conclusions  Our data demonstrate that New York City III broth relative to the other tested media is the most conducive for future studies addressing polymicrobial biofilms development as this culture medium allowed the formation of significant levels of single-species biofilms.

Evaluation of different culture media to support in vitro growth and biofilm formation of bacterial vaginosis-associated anaerobes