A fruitful alliance: the synergy between Atopobium vaginae and Gardnerella vaginalis in bacterial vaginosis-associated biofilm

Liselotte Hardy, Vicky Jespers, Said Abdellati, Irith De Baetselier, Lambert Mwambarangwe, Viateur Musengamana, Janneke van de Wijgert, Mario Vaneechoutte, Tania Crucitti, Liselotte Hardy, Vicky Jespers, Said Abdellati, Irith De Baetselier, Lambert Mwambarangwe, Viateur Musengamana, Janneke van de Wijgert, Mario Vaneechoutte, Tania Crucitti

Abstract

Objectives: Bacterial vaginosis (BV) is characterised by a change in the microbial composition of the vagina. The BV-associated organisms outnumber the health-associated Lactobacillus species and form a polymicrobial biofilm on the vaginal epithelium, possibly explaining the difficulties with antibiotic treatment. A better understanding of vaginal biofilm with emphasis on Atopobium vaginae and Gardnerella vaginalis may contribute to a better diagnosis and treatment of BV.

Methods: To this purpose, we evaluated the association between the presence of both bacteria by fluorescence in situ hybridisation (FISH) and BV by Nugent scoring in 463 vaginal slides of 120 participants participating in a clinical trial in Rwanda.

Results: A bacterial biofilm was detected in half of the samples using a universal bacterial probe. The biofilm contained A. vaginae in 54.1% and G. vaginalis in 82.0% of the samples. A. vaginae was accompanied by G. vaginalis in 99.5% of samples. The odds of having a Nugent score above 4 were increased for samples with dispersed G. vaginalis and/or A. vaginae present (OR 4.5; CI 2 to 10.3). The probability of having a high Nugent score was even higher when a combination of adherent G. vaginalis and dispersed A. vaginae was visualised (OR 75.6; CI 13.3 to 429.5) and highest when both bacteria were part of the biofilm (OR 119; CI 39.9 to 360.8).

Conclusions: Our study, although not comprehensive at studying the polymicrobial biofilm in BV, provided a strong indication towards the importance of A. vaginae and the symbiosis of A. vaginae and G. vaginalis in this biofilm.

Trial registration number: NCT01796613.

Keywords: ANTIBIOTIC SENSITIVITY; BACTERIAL VAGINOSIS; DIAGNOSIS; GENITAL TRACT INFECT; MICROBIOLOGY.

Conflict of interest statement

Competing interests: None declared.

Published by the BMJ Publishing Group Limited. For permission to use (where not already granted under a licence) please go to http://www.bmj.com/company/products-services/rights-and-licensing/.

Figures

Figure 1
Figure 1
Superimposed confocal laser scanning images with 400× magnification of Atopobium vaginae+Gardnerella vaginalis biofilm in six vaginal samples (A–F): vaginal epithelial cells DAPI in blue, A. vaginae-specific peptide nucleic acid (PNA)-probe AtoITM1 with Alexa Fluor 488 in green and G. vaginalis-specific PNA-probe Gard162 with Alexa Fluor 647 in red. For clarity, we omitted the BacUni-1 plane, such that the bacteria that did not hybridise with Gard162 and AtoITM1 are visible in DAPI blue only.

References

    1. Eschenbach DA. History and review of bacterial vaginosis. Am J Obstet Gynecol 1993;169:441–5. 10.1016/0002-9378(93)90337-I
    1. Hillier SL, Marrazzo JM, Holmes KK. Bacterial vaginosis. In: Holmes KK, Sparling PF, Mardh PA, et al., eds Sexually transmitted diseases. New York: McGraw-Hill, 2008;737–68.
    1. Martin HL, Richardson BA, Nyange PM, et al. . Vaginal lactobacilli, microbial flora, and risk of human immunodeficiency virus type 1 and sexually transmitted disease acquisition. J Infect Dis 1999;180:1863–8. 10.1086/315127
    1. van de Wijgert JH, Borgdorff H, Verhelst R, et al. . The vaginal microbiota: what have we learned after a decade of molecular characterization? PLoS ONE 2014;9:e105998 10.1371/journal.pone.0105998
    1. Srinivasan S, Hoffman NG, Morgan MT, et al. . Bacterial communities in women with bacterial vaginosis: high resolution phylogenetic analyses reveal relationships of microbiota to clinical criteria. PLoS ONE 2012;7:e37818 10.1371/journal.pone.0037818
    1. Patterson JL, Stull-Lane A, Girerd PH, et al. . Analysis of adherence, biofilm formation and cytotoxicity suggests a greater virulence potential of Gardnerella vaginalis relative to other bacterial-vaginosis-associated anaerobes. Microbiology 2010;156:392–9. 10.1099/mic.0.034280-0
    1. Jespers V, van de Wijgert J, Cools P, et al. . The significance of Lactobacillus crispatus and L. vaginalis for vaginal health and the negative effect of recent sex : a cross-sectional descriptive study across groups of African women. BMC Infect Dis 2015;15:1–14. 10.1186/s12879-014-0722-x
    1. Jespers V, Crucitti T, van de Wijgert JH, et al. . A DNA tool for early detection of vaginal dysbiosis in African women. Res Microbiol 2016;167:133–41. 10.1016/j.resmic.2015.10.006
    1. Swidsinski A, Mendling W, Loening-Baucke V, et al. . Adherent biofilms in bacterial vaginosis. Obstet Gynecol 2005;106:1013–23. 10.1097/01.AOG.0000183594.45524.d2
    1. Swidsinski A, Loening-Baucke V, Mendling W, et al. . Infection through structured polymicrobial Gardnerella biofilms (StPM-GB). Histol Histopathol 2014;29:567–87.
    1. Hardy L, Jespers V, Dahchour N, et al. . Unravelling the bacterial vaginosis-associated biofilm: a multiplex Gardnerella vaginalis and Atopobium vaginae fluorescence in situ hybridization assay using peptide nucleic acid probes. PLoS ONE 2015;10:e0136658 10.1371/journal.pone.0136658
    1. Machado A, Cerca N. The influence of biofilm formation by Gardnerella vaginalis and other anaerobes on bacterial vaginosis. J Infect Dis 2015;2015;212:1856–61. 10.1093/infdis/jiv338
    1. Ferris MJ, Masztal A, Aldridge KE, et al. . Association of Atopobium vaginae, a recently described metronidazole resistant anaerobe, with bacterial vaginosis. BMC Infect Dis 2004;4:5 10.1186/1471-2334-4-5
    1. Burton JP, Devillard E, Cadieux PA, et al. . Detection of Atopobium vaginae in postmenopausal women by cultivation-independent methods warrants further investigation. J Clin Microbiol 2004;42:1829–31. 10.1128/JCM.42.4.1829-1831.2004
    1. Verstraelen H, Verhelst R, Claeys G, et al. . Culture-independent analysis of vaginal microflora: the unrecognized association of Atopobium vaginae with bacterial vaginosis. Am J Obstet Gynecol 2004;191:1130–2. 10.1016/j.ajog.2004.04.013
    1. Menard JP, Fenollar F, Henry M, et al. . Molecular quantification of Gardnerella vaginalis and Atopobium vaginae loads to predict bacterial vaginosis. Clin Infect Dis 2008;47:33–43. 10.1086/588661
    1. Bradshaw CS, Tabrizi SN, Fairley CK, et al. . The association of Atopobium vaginae and Gardnerella vaginalis with bacterial vaginosis and recurrence after oral metronidazole therapy. J Infect Dis 2006;194:828–36. 10.1086/506621
    1. De Backer E, Verhelst R, Verstraelen H, et al. . Antibiotic susceptibility of Atopobium vaginae. BMC Infect Dis 2006;6:51 10.1186/1471-2334-6-51
    1. Nugent RP, Krohn MA, Hillier SL. Reliability of diagnosing bacterial vaginosis is improved by a standardized method of gram stain interpretation. J Clin Microbiol 1991;29:297–301.
    1. Cook RL, Reid G, Pond DG, et al. . Clue cells in bacterial vaginosis: immunofluorescent identification of the adherent gram-negative bacteria as Gardnerella vaginalis. J Infect Dis 1989;160:490–6. 10.1093/infdis/160.3.490
    1. Machado A, Almeida C, Salgueiro D, et al. . Fluorescence in situ hybridization method using peptide nucleic acid probes for rapid detection of Lactobacillus and Gardnerella spp. BMC Microbiol 2013;13:82 10.1186/1471-2180-13-82
    1. Perry-O'Keefe H, Stender H, Broomer A, et al. . Filter-based PNA in situ hybridization for rapid detection, identification and enumeration of specific micro-organisms. J Appl Microbiol 2001;90:180–9. 10.1046/j.1365-2672.2001.01230.x
    1. Schurmans C, De Baetselier I, Kestelyn E, et al. . The ring plus project: safety and acceptability of vaginal rings that protect women from unintented pregnancy. BMC Public Health 2015;15:348 10.1186/s12889-015-1680-y
    1. Verhelst R, Verstraelen H, Claeys G, et al. . Cloning of 16S rRNA genes amplified from normal and disturbed vaginal microflora suggests a strong association between Atopobium vaginae, Gardnerella vaginalis and bacterial vaginosis. BMC Microbiol 2004;4:16 10.1186/1471-2180-4-16
    1. Gardner HL, Dukes CD. New etiologic agent in nonspecific bacterial vaginitis. Science 1954;120:853 10.1126/science.120.3125.853
    1. Høiby N, Bjarnsholt T, Moser C, et al. . ESCMID guideline for the diagnosis and treatment of biofilm infections 2014. Clin Microbiol Infect 2015;21(Suppl 1):S1–25. 10.1016/j.cmi.2014.10.024

Source: PubMed

3
購読する