Open Access Eradication therapy tore (Immediately after) (3 months after) Firmicutes 76.3 718% Actinobacteria 14.6 Archaea Figure 1 Proportion of the dominant five phyla in the faecal samples. The per cent proportions of the five dominant phyla of a N=ggNz3gN→o Firmicutes, Actinobacteria, Proteobacteria, Bacteroidetes and Archaea in all faecal samples at S1, S2 and S3 are depicted in the bar charts. The S1, S2 and S3 samples include 984 296, 1 187 681 and 1 022 232 high-quality reads, respectively the pretherapy level at S3(table 2). The OTUs gener- Bifidobacterium and Ruminococcaceae including Faecali- 5 ted from the 16s reads were then classified into phyla bacterium in the analysis of similarity in the frequency of according to a similarity search(figure 1). At S2, there identification among samples in the clusterin was a marked increase in the abundance of both Bacte- rial families roidetes((0.9% to 4.5%)%)and Archaea but a decrease in that of both Actinobacteria(20.5% to 15.6%)%) Change in the biomarkers regarding obesity/type 2 diabetes and Proteobacteria. The increase in Bacteroidetes and mellitus(T2DM)and their association with gut microbiota decrease in Actinobacteria persisted even at $3(0.9% to during the therapy 4.0%and 20.5% to 14.6%, respectively). The abundance Among the clinical and laboratory data conce of Firmicutes, the predominant phylum, showed little obesity/T2DM examined in the present study(table 3) change between SI and $3(69.9% to 71.8%)%). Thus, statistically significant decreases and increases were the median B: F ratio was significantly greater at S than observed in the values of active ghrelin(P=0.00%)and respectively, between S3 and Sl. We a quite obvious, with 15 of the 20 patients(75%)showing a therefore examined whether the change in the levels of 3 higher ratio at S3 than at SI (table I and figure 2) 6 We then examined the abundance of particular those adipogenesis-regulating hormones was associated acteria considered to be associated with dysbiosis in with the alteration of the ratio of B: F in each patient A obesity(table 2). The abundance of the genus Bifidobac- (figure 2). The rate of decrease in active ghrelin levels s terium,which is included in phylum Actinobacteria, was among patients with an increased B: F ratio was 14 out 8 the abundance of Faecalibacterium prausnitz was greater levels among patients with a decreased B: F ratio in o significantly lower at S3 than at SI(P<0.01). In contrast, of 15, whereas the rate of decrease in active ghrelin o at S3 than at SI(P-0.02). A double hierarchical clus- only 2 out of 5(table 1). The difference in the rates 2 tering analysis demonstrated an opposite but closely between these two groups was statistically significant relate ed change in the abundance after therapy betwe (P=0.03) by Fisher's test No significant association was T Bifidobacterium and Faecalibacterium(figure 3), with a found between the change in the leptin level and the distinct cluster formed by Bifidobacteriaceae including B: F ratio. Yanagi H, et al. BM Open Gastro 2017: 4: 0000182. doi: 10. 1136/bmigast-2017-0001824 Yanagi H, et al. BMJ Open Gastro 2017;4:e000182. doi:10.1136/bmjgast-2017-000182 Open Access the pretherapy level at S3 (table 2). The OTUs gener￾ated from the 16S reads were then classified into phyla according to a similarity search (figure 1). At S2, there was a marked increase in the abundance of both Bacte￾roidetes ((0.9% to 4.5%)%) and Archaea but a decrease in that of both Actinobacteria ((20.5% to 15.6%)%) and Proteobacteria. The increase in Bacteroidetes and decrease in Actinobacteria persisted even at S3 (0.9% to 4.0%and 20.5% to 14.6%, respectively). The abundance of Firmicutes, the predominant phylum, showed little change between S1 and S3 ((69.9% to 71.8%)%). Thus, the median B:F ratio was significantly greater at S3 than at S1 (P<0.01, table 2). This increase in the B:F ratio was quite obvious, with 15 of the 20 patients (75%) showing a higher ratio at S3 than at S1 (table 1 and figure 2). We then examined the abundance of particular bacteria considered to be associated with dysbiosis in obesity (table 2). The abundance of the genus Bifidobac￾terium, which is included in phylum Actinobacteria, was significantly lower at S3 than at S1 (P<0.01). In contrast, the abundance of Faecalibacterium prausnitzii was greater at S3 than at S1 (P=0.02). A double hierarchical clus￾tering analysis demonstrated an opposite but closely related change in the abundance after therapy between Bifidobacterium and Faecalibacterium (figure 3), with a distinct cluster formed by Bifidobacteriaceae including Bifidobacterium and Ruminococcaceae including Faecali￾bacterium in the analysis of similarity in the frequency of identification among samples in the clustering of bacte￾rial families. Change in the biomarkers regarding obesity/type 2 diabetes mellitus (T2DM) and their association with gut microbiota during the therapy Among the clinical and laboratory data concerning obesity/T2DM examined in the present study (table 3), statistically significant decreases and increases were observed in the values of active ghrelin (P=0.003) and leptin (P=0.03), respectively, between S3 and S1. We therefore examined whether the change in the levels of those adipogenesis-regulating hormones was associated with the alteration of the ratio of B:F in each patient (figure 2). The rate of decrease in active ghrelin levels among patients with an increased B:F ratio was 14 out of 15, whereas the rate of decrease in active ghrelin levels among patients with a decreased B:F ratio was only 2 out of 5 (table 1). The difference in the rates between these two groups was statistically significant (P=0.03) by Fisher’s test. No significant association was found between the change in the leptin level and the B:F ratio. Figure 1 Proportion of the dominant five phyla in the faecal samples. The per cent proportions of the five dominant phyla of Firmicutes, Actinobacteria, Proteobacteria, Bacteroidetes and Archaea in all faecal samples at S1, S2 and S3 are depicted in the bar charts. The S1, S2 and S3 samples include 984 296, 1 187 681 and 1 022 232 high-quality reads, respectively. copyright. on 4 July 2018 by guest. Protected by http://bmjopengastro.bmj.com/ BMJ Open Gastroenterol: first published as 10.1136/bmjgast-2017-000182 on 26 November 2017. Downloaded from