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Biofilms of vaginal Lactobacillus reuteri CRL 1324 and Lactobacillus rhamnosus CRL 1332: kinetics of formation and matrix characterization

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Abstract

Adhesion and biofilm formation are strain properties that reportedly contribute to the permanence of lactobacilli in the human vagina. The kinetics of biofilm formation and the chemical nature of the biofilm matrix formed by Lactobacillus reuteri CRL (Centro de Referencia para Lactobacilos Culture Collection) 1324 and Lactobacillus rhamnosus CRL 1332, vaginal beneficial strains, were evaluated in this work. Crystal violet-stained microplate assay and techniques of epifluorescence, electron and confocal microscopy were applied. The highest density and complexity of biofilms of both vaginal lactobacilli were observed at 72 h of incubation. Protease, proteinase K, α-chymotrypsin and trypsin treatments efficiently detached L. reuteri CRL 1324 biofilm that was also partially affected by α-amylase. However, L. rhamnosus CRL 1332 biofilm was slightly affected by protease, proteinase K and α-amylase. Confocal microscopy revealed greater amount of polysaccharides in L. rhamnosus CRL 1332 biofilm matrix than in L. reuteri CRL 1324 biofilm matrix. The results indicate that proteins are one of the main components of the L. reuteri CRL 1324 biofilm, while the biofilm matrix of L. rhamnosus CRL 1332 is composed of carbohydrates and proteins. The results obtained support the knowledge, understanding and characterization of two biofilm-forming vaginal Lactobacillus strains.

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Acknowledgments

This paper was supported by CONICET (Consejo Nacional de Investigaciones Científicas y Técnicas, Argentina) (PIP 632 and PIP 744) and ANPCyT (Agencia Nacional de Promoción Científica y Tecnológica) (PICT 2012-1187), and was partially performed within the framework of a Bilateral Cooperation Project (A12-B01) between Mincyt (Ministerio de Ciencia, Tecnología e Innovación Productiva de Argentina) and ECOS-SUD (France). The strains were licensed through a CONICET-BIOLIFE agreement. We thank Elena Bru for her collaboration in the statistical analyses.

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Authors and Affiliations

  1. Centro de Referencia para Lactobacilos (CERELA)-CONICET, Chacabuco 145, 4000, Tucumán, Argentina

    María Cecilia Leccese Terraf, María Silvina Juárez Tomás & María Elena Fátima Nader-Macías

  2. UMR 1253 STLO, INRA, Rennes, France

    Lucie Rault, Yves Le Loir & Sergine Even

  3. UMR1253 STLO, Agrocampus Ouest, Rennes, France

    Lucie Rault, Yves Le Loir & Sergine Even

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  1. María Cecilia Leccese Terraf
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  2. María Silvina Juárez Tomás
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  3. Lucie Rault
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  4. Yves Le Loir
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  5. Sergine Even
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  6. María Elena Fátima Nader-Macías
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Correspondence to María Elena Fátima Nader-Macías.

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Communicated by Erko Stackebrandt.

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Leccese Terraf, M.C., Juárez Tomás, M.S., Rault, L. et al. Biofilms of vaginal Lactobacillus reuteri CRL 1324 and Lactobacillus rhamnosus CRL 1332: kinetics of formation and matrix characterization. Arch Microbiol 198, 689–700 (2016). https://doi.org/10.1007/s00203-016-1225-5

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  • DOI: https://doi.org/10.1007/s00203-016-1225-5

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