IADR Abstract Archives
Investigating Oral Microcosm Biofilms Shifts Associated with a 15-day Constant Depth Film Fermentor Model
Objectives: To explore longitudinally the microbial shifts under different in vitro environmental modelling parameters in a 15-day Constant Depth Film Fermentor Model (CDFF).
Methods: Biofilms were sampled regularly over a 15-day period. Bacterial DNA was extracted, PCR amplified using 16S rRNA gene V5-V7 primers (barcoded amplicons 785F;1175R), purified, pooled at equimolar concentrations and sequenced (MiSeq, Illumina). The 16S rRNA reads were filtered, assembled and analysed. Confocal Laser Scanning Microscopy (CLSM) and Scanning Electron Microscopy (SEM) imaging analyses were performed on the microbial communities within steady-state microcosm biofilms, derived from a 15-day CDFF model exhibiting the environmental conditions progressing from peri-implant health to disease in vitro.
Results: CLSM analyses showed that early biofilms, under conditions emulating peri-implant health, were mainly composed of cocci bacteria, and displayed many visible channels and voids. Biofilms at day 6 displayed a compact structure with the presence of bacilli and filamentous bacteria. After peri-implant disease conditions were commenced, a translocation from rods in chains and filaments, to numerous filaments and fusiform bacteria was observed. The metagenomic analyses revealed that in conditions emulating peri-implant health, the steady-state biofilm exhibited high proportions of the genera Streptococcus, Granulicatella, Klebsiella, Haemophilus, Gemella, and Neisseria. In contrast, in conditions emulating peri-implant disease, lower proportions of Streptococcus and Granulicatella were observed.
Conclusions: The model presented here serves as an approach to analyse the impact of environmental-induced factors in the biofilms developed over a longitudinal 15-day period under different in vitro conditions. Imaging analyses revealed a number of characteristic bacterial morphologies across different time-points. During peri-implant health conditions, live bacteria were predominant. Microbiome analyses of the steady-state biofilms revealed bacterial shifts associated with peri-implant disease environmental conditions, characterised by an increase in proportions of Veillonella, Peptostreptococcus, Parvimonas, Fusobacterium, Actinomyces, Klebsiella, Leptotrichia, and Enterobacter.
Methods: Biofilms were sampled regularly over a 15-day period. Bacterial DNA was extracted, PCR amplified using 16S rRNA gene V5-V7 primers (barcoded amplicons 785F;1175R), purified, pooled at equimolar concentrations and sequenced (MiSeq, Illumina). The 16S rRNA reads were filtered, assembled and analysed. Confocal Laser Scanning Microscopy (CLSM) and Scanning Electron Microscopy (SEM) imaging analyses were performed on the microbial communities within steady-state microcosm biofilms, derived from a 15-day CDFF model exhibiting the environmental conditions progressing from peri-implant health to disease in vitro.
Results: CLSM analyses showed that early biofilms, under conditions emulating peri-implant health, were mainly composed of cocci bacteria, and displayed many visible channels and voids. Biofilms at day 6 displayed a compact structure with the presence of bacilli and filamentous bacteria. After peri-implant disease conditions were commenced, a translocation from rods in chains and filaments, to numerous filaments and fusiform bacteria was observed. The metagenomic analyses revealed that in conditions emulating peri-implant health, the steady-state biofilm exhibited high proportions of the genera Streptococcus, Granulicatella, Klebsiella, Haemophilus, Gemella, and Neisseria. In contrast, in conditions emulating peri-implant disease, lower proportions of Streptococcus and Granulicatella were observed.
Conclusions: The model presented here serves as an approach to analyse the impact of environmental-induced factors in the biofilms developed over a longitudinal 15-day period under different in vitro conditions. Imaging analyses revealed a number of characteristic bacterial morphologies across different time-points. During peri-implant health conditions, live bacteria were predominant. Microbiome analyses of the steady-state biofilms revealed bacterial shifts associated with peri-implant disease environmental conditions, characterised by an increase in proportions of Veillonella, Peptostreptococcus, Parvimonas, Fusobacterium, Actinomyces, Klebsiella, Leptotrichia, and Enterobacter.