Core supra-gingival microbiome in root caries and health.

<p>Red field represents health-associated core species with significantly higher prevalence and relative abundance in Healthy_controls than in Patient_cases; green field represents root caries-associated core species with significantly higher prevalence and relative abundance in Patient_cases than in Healthy_controls; brown field represents core species in both health and root caries with no significantly different prevalence and relative abundance between the two groups. In each field, the prevalences of species were at least 1/2. Inner circles labeled 1 contain the species with high prevalence (prevalence ≥ 2/3) and high abundance (average relative abundance ≥ 2%); circles labeled 2 contain species with high prevalence (prevalence ≥ 2/3) but low abundance (average relative abundance < 2%); circles labeled 3 contain species with moderate prevalence (1/2 ≤ prevalence < 2/3) but high abundance (average relative abundance ≥ 2%); circles labeled 4 contain species with moderate prevalence (1/2 ≤ prevalence < 2/3) and low abundance (average relative abundance < 2%).</p

Maximum likelihood phylogenetic tree at the species level.

<p>Inner loop displays species with color ranges for the class level (see color key at the bottom left); middle loop indicates mean relative abundance in each group (see color key at the top left); outer loop indicates significantly different species detected between groups (see color key at the top left). An online tool of iTOL was used to construct this tree.</p

Samples clustering by PCoA.

<p>All PCoA plots were first constructed on the basis of unweighted UniFrac distance (see the panel at the top left). Then the points in PCoA plots were weighted by the relative abundances of the top 12 root caries-associated species and top 12 health-associated species of <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0117064#pone.0117064.g005" target="_blank">Fig. 5</a>.</p

Boxplots for comparison of bacterial loads by qPCR and bacterial relative abundances by 454 pyrosequencing.

<p>(a) Boxplot for total bacterial load. (b) Boxplots for the total loads of genera <i>Bifidobacterium</i>, <i>Lactobacillus</i>, and species <i>Streptococcus mutans</i>. (c) Boxplots for the pyrosequencing relative abundances of the same taxa as (b). Gene copy number shown above was transformed by natural logarithm (ln) and relative abundance was transformed by percent (%). * denotes P < 0.1; ** denotes P < 0.05; *** denotes P < 0.01.</p

Alpha diversity and beta diversity.

<p>(a) Boxplots for alpha diversity metrics of Chao1 (species richness) and Shannon Diversity Index (species diversity). (b) Beta diversity metrics of unweighted UniFrac (community membership) and weighted UniFrac (community structure). The color in the heatmaps indicates the UniFrac distance between each pair of samples (see color key at the left). * denotes P < 0.1; ** denotes P < 0.05; *** denotes P < 0.01.</p

Rarefaction curves for evaluation of species coverage in our study.

<p>The <i>x</i> axis indicates the number of sequences per sample. The <i>y</i> axis indicates the average number of OTUs (0.03 distance threshold) per sample in each group. The error bars represent standard deviation.</p

Distributions of abundance-based significant phyla, genera and species between health and root caries.

<p>Pie charts display the counts distribution of significantly different taxa. Line charts sort significantly different genera and species in the order of magnitude of abundance change (≥0.1%) between Healthy_controls and Patient_cases.</p

Distributions of prevalence-based significant phyla, genera and species between health and root caries.

<p>Pie charts display the counts distribution of significantly different taxa. Line charts sort significantly different genera and species in the order of magnitude of prevalence change between Healthy_controls and Patient_cases.</p

Demographic and clinical characteristics of the study subjects.

<p>SD = standard deviation of the mean.</p><p>NRT = number of remaining teeth.</p><p>RCT = root caries teeth.</p><p>RDFS = root decayed filled surfaces.</p><p>^a Student’s t test.</p><p>^b Fisher’s exact test.</p><p>* p<0.05</p><p>** p<0.01</p><p>Demographic and clinical characteristics of the study subjects.</p

Characteristics of the study population according to HLA genotype^a.

<p>^aFour different groups were included to represent homozygosity and heterozygosity for the AH8.1 haplotype (as defined by the presence of HLA-B*0801 and DRB1*0301), absence of this haplotype as well as homozygosity for other DRB1 haplotypes. There were no significant differences (all p>0.23) for any of the parameters when applying Kruskal-Wallis test for continuous and 2x4 chi square test for categorical parameters.</p><p>^bTransport time as defined by time from sampling to the freezer.</p><p>Characteristics of the study population according to HLA genotype<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0133804#t002fn001" target="_blank">^a</a>.</p