IADR Abstract Archives
The Effect of Probiotic Treatments on the Composition of In-Vitro Oral Biofilms
Objectives: Lactobacillus strains have been shown to have antagonistic properties to oral disease-associated bacteria and therefore have potential to be used as probiotics. The aim of this study was to test the effect of treatment with potential probiotic strains on the composition of in-vitro oral biofilms.
Methods: Biofilms were generated by means of the Calgary biofilm device seeded with saliva. Biofilms were cultivated in supplemented Brain Heart Infusion changed every 3.5 days. After 7 days of growth, biofilms were treated with live and heat-attenuated preparations of 14 Lactobacillus species twice daily for 7 days. Biofilm samples were then harvested and community profiling targeting 16S rRNA was performed on all samples by means of 454 pyrosequencing, analysed using the mothur pipeline.
Results: Analysis of molecular variance (AMOVA) showed that there were overall significant differences between the groups for both live and heat-attenuated treatments (P<0.001), although there were no significant differences between individual treatments and negative control. Ordination analysis using non-metric multidimensional scaling (NMDS) showed Lactobacillus plantarum SYBIO-24 and Lactobacillus paracasei SYBIO-15 to be separated from the other treatments. Heat map analysis indicated that these differences were due to reductions in the proportions of Bacteroides, Fusobacterium, Prevotella and Pyramidobacter, and increases in Campylobacter, Corynebacterium, Mobiluncus and Neisseria. Linear Discriminant Analysis Effect Size (LEfSe) indicated that 49 species-level OTUs from these genera were significantly differentially abundant between the L. plantarum SYBIO-24 heat-attenuated treatment and the negative control biofilms, whilst 37 were differentially abundant between L. paracasei SYBIO-15 and the negative control.
Conclusions: The majority of probiotic treatments did not affect the composition of in-vitro oral biofilms. However, treatment with heat-attenuated L. plantarum SYBIO-24 and L paracasei SYBIO-15 reduced the proportions of certain anaerobic bacterial genera in the biofilms which suggests that these strains have potential for use as oral probiotics.
Methods: Biofilms were generated by means of the Calgary biofilm device seeded with saliva. Biofilms were cultivated in supplemented Brain Heart Infusion changed every 3.5 days. After 7 days of growth, biofilms were treated with live and heat-attenuated preparations of 14 Lactobacillus species twice daily for 7 days. Biofilm samples were then harvested and community profiling targeting 16S rRNA was performed on all samples by means of 454 pyrosequencing, analysed using the mothur pipeline.
Results: Analysis of molecular variance (AMOVA) showed that there were overall significant differences between the groups for both live and heat-attenuated treatments (P<0.001), although there were no significant differences between individual treatments and negative control. Ordination analysis using non-metric multidimensional scaling (NMDS) showed Lactobacillus plantarum SYBIO-24 and Lactobacillus paracasei SYBIO-15 to be separated from the other treatments. Heat map analysis indicated that these differences were due to reductions in the proportions of Bacteroides, Fusobacterium, Prevotella and Pyramidobacter, and increases in Campylobacter, Corynebacterium, Mobiluncus and Neisseria. Linear Discriminant Analysis Effect Size (LEfSe) indicated that 49 species-level OTUs from these genera were significantly differentially abundant between the L. plantarum SYBIO-24 heat-attenuated treatment and the negative control biofilms, whilst 37 were differentially abundant between L. paracasei SYBIO-15 and the negative control.
Conclusions: The majority of probiotic treatments did not affect the composition of in-vitro oral biofilms. However, treatment with heat-attenuated L. plantarum SYBIO-24 and L paracasei SYBIO-15 reduced the proportions of certain anaerobic bacterial genera in the biofilms which suggests that these strains have potential for use as oral probiotics.