The association of Streptococcus mutans ATCC 25175 or Streptococcus oralis ATCC 9811, as pioneer microorganisms, with the secondary colonizers Fusobacterium nucleatum ATCC 25586 or Porphyromonas gingivalis ATCC 33277, during biofilms development on titanium surface, was evaluated by flow cytometry (FCM) using specific polyclonal antibodies. Antibody sensitivity and specificity were determined in ELISA and FCM, revealing a high sensitivity and specificity. Biofilm formation of four dual-species combinations was analyzed by crystal violet staining, while the association between streptococci and periodontal pathogens was assessed in FCM. Dual-species association between S. oralis and P. gingivalis or F. nucleatum showed significantly proportional decrease of S. oralis during biofilm development with concomitant increase of P. gingivalis or F. nucleatum. This trend was not observed in both the dual-species associations of S. mutans with the periodontal pathogens. Our dual-species microbial model by FCM revealed to be useful to study partnerships between bacteria in oral associations, evidencing that the presence of primary colonizers is required for the establishment of secondary colonizers into biofilms. Since the proposed experimental approach is technically simple to prepare and analyse, and proved to be reproducible, it is very suitable for studying the development and dynamics of oral communities.
A dual-species microbial model for studying the dynamics between oral streptococci and periodontal pathogens during biofilm development on titanium surfaces by flow cytometry
MANTI, ANITA;CIANDRINI, ELEONORA;CAMPANA, RAFFAELLA;DOMINICI, SABRINA;CIACCI, CATERINA;SISTI, DAVIDE;ROCCHI, MARCO BRUNO LUIGI;PAPA, STEFANO;BAFFONE, WALLY
2016
Abstract
The association of Streptococcus mutans ATCC 25175 or Streptococcus oralis ATCC 9811, as pioneer microorganisms, with the secondary colonizers Fusobacterium nucleatum ATCC 25586 or Porphyromonas gingivalis ATCC 33277, during biofilms development on titanium surface, was evaluated by flow cytometry (FCM) using specific polyclonal antibodies. Antibody sensitivity and specificity were determined in ELISA and FCM, revealing a high sensitivity and specificity. Biofilm formation of four dual-species combinations was analyzed by crystal violet staining, while the association between streptococci and periodontal pathogens was assessed in FCM. Dual-species association between S. oralis and P. gingivalis or F. nucleatum showed significantly proportional decrease of S. oralis during biofilm development with concomitant increase of P. gingivalis or F. nucleatum. This trend was not observed in both the dual-species associations of S. mutans with the periodontal pathogens. Our dual-species microbial model by FCM revealed to be useful to study partnerships between bacteria in oral associations, evidencing that the presence of primary colonizers is required for the establishment of secondary colonizers into biofilms. Since the proposed experimental approach is technically simple to prepare and analyse, and proved to be reproducible, it is very suitable for studying the development and dynamics of oral communities.I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
