Effect of biofilm phenotype on resistance of Gardnerella vaginalis to hydrogen peroxide and lactic acid

doi:10.1016/j.ajog.2007.02.027

American Journal of Obstetrics and Gynecology

Volume 197, Issue 2, August 2007, Pages 170.e1-170.e7

https://doi.org/10.1016/j.ajog.2007.02.027 Get rights and content

Objective

Bacterial vaginosis is the most common vaginal disorder worldwide. Certain lactobacilli produce H₂0₂ and lactic acid, which normally suppress growth of anaerobes; however, in bacterial vaginosis, Gardnerella vaginalis and other anaerobes proliferate, and the number of lactobacilli decreases. G vaginalis colonizes the vaginal epithelium as a biofilm, which likely plays a role in colonization and relapsing infection.

Study Design

We developed an in vitro model for G vaginalis biofilm formation and compared susceptibilities of biofilms vs planktonic cultures to H₂0₂ and lactic acid. The structure and composition of the biofilm matrix were studied in order to design a method for biofilm dissolution.

Results

Biofilms tolerated 5-fold and 4-8 fold higher concentrations of H₂0₂ and lactic acid (respectively) than planktonic cultures. Proteolytic dissolution of biofilms reduced sensitivity to H₂0₂ and lactic acid.

Conclusion

Increased tolerance to H₂0₂ and lactic acid suggests that biofilm formation contributes to the survival of G vaginalis in the presence of lactobacilli.

Section snippets

Strains and culture conditions

G vaginalis strain ATCC 49145 (American Type Culture Collection, Manassas, VA) and strains 5-1 and 465-5 (provided by Dr Robin Ross, Boston, MA) were grown under anaerobic conditions, in approximately 10% CO₂ with the use of the AnaeroPack system (Mitsubishi Gas Chemical Co, Tokyo, Japan). The 96-well tissue culture plate assays were performed essentially as previously described to determine optimal growth conditions for biofilm formation.¹⁴ G vaginalis was grown in Corning Cell-Bind Plates

Biofilm formation by G vaginalis in vitro

To determine optimal conditions for in vitro biofilm formation, G vaginalis was cultured anaerobically in 96-well plates in Luria-Bertani broth, tryptic soy broth, BHI, Mueller-Hinton broth, de Man Rogosa Sharpe broth, or a CDM that resembled vaginal secretions.¹⁷ Because glucose increases biofilm formation in other bacterial species,¹⁸ G vaginalis was also cultured in the presence of 1% glucose (Luria-Bertani broth with glucose, tryptic soy broth with glucose, BHIG, Mueller-Hinton with

Comment

The cause and pathogenesis of BV have remained enigmatic ever since the initial reports of the condition in 1954.²⁰ Confounding issues include the polymicrobial nature of BV and the role of host factors, both genetic and behavioral. The recent demonstration that BV is associated with the formation of a biofilm in which the predominant species is G vaginalis¹² is an important advance towards understanding its pathogenesis. We therefore sought to develop a simple model for studying G vaginalis

Acknowledgment

We thank Dr Robin Ross for providing us with clinical isolates of G vaginalis.

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: Supported by National Institutes of Health grant R21AI61590-02 from the National Institute of Allergy and Infectious Diseases.

: Cite this article as: Patterson JL, Girerd PH, Karjane NW, Jefferson KK. Effect of biofilm phenotype on resistance of Gardnerella vaginalis to hydrogen peroxide and lactic acid. Am J Obstet Gynecol 2007;197:170.e1-170.e7.

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ResearchBasic science: GynecologyEffect of biofilm phenotype on resistance of Gardnerella vaginalis to hydrogen peroxide and lactic acid

Objective

Study Design

Results

Conclusion

Section snippets

Strains and culture conditions

Biofilm formation by G vaginalis in vitro

Comment

Acknowledgment

Infect Dis Clin North Am

Am J Obstet Gynecol

Microbes Infect

Am J Obstet Gynecol

Curr Opin Biotechnol

Managing recurrent bacterial vaginosis

Sex Transm Infect

Association between bacterial vaginosis and preterm delivery of a low-birth-weight infant: the Vaginal Infections and Prematurity Study group

N Engl J Med

Bacterial vaginosis and susceptibility to HIV infection in South African women: a nested case-control study

J Infect Dis

Bacterial vaginosis and disturbances of vaginal flora: association with increased acquisition of HIV

Aids

Growth inhibition of metronidazole-susceptible and metronidazole-resistant strains of Gardnerella vaginalis by lactobacilli in vitro

Appl Environ Microbiol

Molecular identification of bacteria associated with bacterial vaginosis

N Engl J Med

Haemophilus vaginalis: vaginitis by inoculation from culture

Obstet Gynecol

Adherent biofilms in bacterial vaginosis

Obstet Gynecol

Research
Basic science: Gynecology
Effect of biofilm phenotype on resistance of Gardnerella vaginalis to hydrogen peroxide and lactic acid