Figure 1.
Oral cavity and sites of sampling.
A. Anatomic chart of the oral cavity with arrows indicating the sites of sampling (A to F) and a red circle labelling the tumour region of patient FA_T1. Samples C and D were collected from the ventral portion of the tongue (indicated by arrows with hatched centres). B. Photography of the tumour on the FA_T1 patient's left side of the tongue encroaching the centre line. The red circle indicates the sampling site of the tumour region as described in Figure 1, A.
Table 1.
Total number of reads generated per sample.
Figure 2.
Microbiota detected by NGS on the surface of distinct oral sites of the analysed cohort.
DNA of samples, which derived from site B of five healthy probands (H1-H5) and sites A, B, E and F of two FA patients (FA_L1 with a leukoplakia at site F and FA_T1 with a tumour at site B), was subjected to V1-V2 16S rDNA based NGS analysis. A., phyla and families detected in the 13 samples. Families are indicated in shades of the same colour as the respective phylum; B., alpha diversity of all samples (H1-H5 in blue, FA_L1 in green and FA_T1 in red); C., beta diversity plot of all samples (H1-H5 as blue circles, FA_L1 as green squares and FA_T1 as red triangles).
Figure 3.
Dominant families detected by NGS after OTU clustering in the microbiotas of patients and healthy individuals.
Frequency of families (in %) present in at least one of the 13 analysed samples analysed at a frequency >4%; A., site B of healthy probands H1-H5; B., sites A, B, E and F of FA patient FA_L1; and C., sites A, B, E and F of FA patient FA_T1.
Figure 4.
Real time PCRs for the detection of M. salivarium, P. aeruginosa, and Candida.
DNA samples, which were derived from healthy probands (H1-H5, site B), FA-patients with leukoplakia (FA_L1 with a leukoplakia at site F and FA_ L2 with leukoplakia at site B) and FA patients with tumour (FA_T1 with a tumour at site B and FA_T2 with tumour at site D) were subjected to real time PCR in duplicates. A. TaqMan qPCR-derived copy numbers of M. salivarium (red), P. aeruginosa (hatched) and Candida (grey). Due to targeting a single copy gene, copy numbers correspond to genome equivalents for M. salivarium and P. aeruginosa. B., scatter plot of M. salivarium load in the different proband groups: Healthy (H1-H5, blue circles), samples of FA-patients with leukoplakia (FA_L1 and FA_L2, green squares) and with tumour (FA_T1 and FA_T2, red triangles). C., scatter plot of M. salivarium load of FA patients FA_T1 (dark red) and FA_T2 (red) at the tumour surface (Tumour) in relation to the contralateral non-tumour surface (Non-T.) and of the FA patients FA_L1 (green) and FA_L2 (dark green) at the site of leukoplakia (Leukopl.) in relation to the contralateral non-leukoplakia site (Non-L.).