The clone library analysis showed consistent decrease in the Firmicutes and consistent increase in Bacteroidetes in both the families with an increase in age (Figure 2). The family level variation in
microflora in individuals is shown in Additional file 1: Table S1. The genera which were dominant in the individual samples are represented in Figure 3. The heat map represented in Figure 3 shows that the individuals within a same family cluster together when genus level distribution of gut flora is considered. Within family T, Fecalibacterium and Roseburia dominated in subject T1 (age 14) Dialister, Prevotella dominated in subject T2 (age 42) and Prevotella in subject T3 (age 62). Within family S the genus BTSA1 chemical structure Streptococcus and Weissella dominated in the Napabucasin purchase infant and Fecalibacterium and Roseburia dominated in adult subjects (age 26 and 62 years respectively). The phylogenetic tree of the OTU’s obtained from all the subjects are represented in Additional files 2: Figures S1, Additional file 3: Figures S2, Additional file 4: Figure S3, Additional file
5: Figure S4, Additional file 6: Figure S5, Additional file 7: Figure S6. The phylogenetic trees consist of clades representing the presence of potential novel bacterial species in the gut flora of the subjects. Figure 2 Phylum level comparison of gut flora of the subjects . The learn more stacked bars describe the percent distribution of each phylum across the subjects. Figure many 3 Genus level comparison of gut flora . The heat map represents clustering of bacterial communities across the subjects at the genus level. Family S: S1 (26 years), S2 (8 months), S3 (56 years) and Family T: T1 (14 years), T2 (42 years), T3 (62 years). Real time PCR The slopes for the standards for all the genus specific primers were in the range of −3.1019 to −3.460 with the R2 value >0.99. The PCR efficiency ranged from 96% to 106%. The qPCR quantification
confirmed that the Firmicutes number is decreasing and Bacteroidetes number is increasing with increasing age. The pattern of change in Firmicutes/Bacteroidetes ratio with age within a Family is represented in Figure 4. The copy numbers of different genera are represented in Table 3. The copy number of Roseburia was more than Clostridium and Lactobacillus group, suggesting dominance of Roseburia in the gut flora, which is consistent with the report by Arumugam et al. showing that Fecalibacterium and Roseburia are the dominant genera in the gut flora [35]. Figure 4 Firmicutes to Bacteroidetes ratio by qPCR, A- The pattern of change in Firmicutes/ Bacteroidetes in family S and B- The pattern of change in Firmicutes/ Bacteroidetes in family T. Table 3 Copy numbers of different genera in the gut flora of individual samples Subjects S2 (8 months) S1 (26 yrs) S3 (56 yrs) T1 (14 yrs) T2 (42 yrs) T3 (62 yrs) ClEub 2.17 ± 0.9 E + 07 1.91 ± 0.01E + 08 7.85 ± 0.06E + 03 1.08 ± 0.01E + 09 2.19 ± 0.1E + 08 1.17 ± 0.01E + 08 Prev 7.83 ± 0.9 E + 07 3.55 ± 0.4E + 07 1.