The aim of this study was to clarify the effect of the spore-forming and lactic acidproducing probiotic strain, Bacillus coagulans SANK 70258 on human colonic microbiota of healthy subjects and ulcerative colitis patients. A model culture system was employed to construct the in vitro human colonic microbiota, to retain the bacterial species richness and simulate the patient's disordered composition, from the fecal inoculum. Bacterial 16S rRNA gene sequencing confirmed that administration of B. coagulans SANK 70258 (at an initial concentration of 4×10 7 -total cells/mL) suppressed bacteria related to the family Enterobacteriaceae in the microbiota models for both healthy subjects (P = 0.016) and ulcerative colitis patients (P = 0.023). In addition, administration of B. coagulans SANK 70258 increased bacteria related to the family Lachnospiraceae (P = 0.031), thereby enhancing butyrate production (P = 0.031) in the microbiota models of healthy subjects. However, these changes were not observed in the microbiota models of ulcerative colitis patients, likely owing to the low abundance of Lachnospiraceae species. This study demonstrates the potential of B. coagulans SANK 70258 to exhibit antimicrobial activity against harmful organisms in patients with ulcerative colitis; while improving the intestinal microenvironment by increasing butyrogenesis in healthy persons.

To determine whether it could also improve the production performance of Eimeria-infected broilers, Weizmannia coagulans strain SANK70258 (WC) supplementation was compared with coccidiostat lasalocid-A sodium (AM) administration. First, to determine the optimum WC dose, newly hatched broiler chick groups (n = 10) were untreated or consecutively given WC (0.005%, 0.01%, 0.03%, and 0.1%) and AM until slaughter (31 days of age). At day 21, all chicks were infected with coccidia. From the economical and practical viewpoints, 0.03% WC supplementation was the best dose. Second, newly hatched broiler chick groups (n = 10) were untreated or given 0.03% WC and AM. Each group was run in triplicate. At day 21, two chicks/pen with the farthest body weights as per the group’s mean body weight were spared, and the remaining inoculated with coccidia. At days 42 and 49, the WC and AM groups had significantly greater body weights and daily weight gains. Intestinal lesion scores were lower in 29-day-old AM and WC. Oocyst numbers were lower in 29- and 49-day-old AM and WC, but only 29- and 49-day-old AM had higher Escherichia coli levels. To conclude, although WC and AM induced similar growth performance in coccidium-infected chicks, unlike AM, the E. coli levels did not increase with WC.

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To determine the mechanisms by which Weizmannia coagulans SANK70258 (WC) supplementation improved growth performance and coccidial symptoms, we assessed the gene expressions and the microbiota compositions in the small intestinal tissues and digestas of coccidium-infected broilers previously given WC or lasalocid-A sodium (AM). WC supplementation significantly upregulated the gene expressions related to intestinal immunity and barrier functions, such as IL17A, IL17F, IL10, cathelicidin-2 and pIgR. Body weights, and Claudin-1 and IL10 expressions were positively correlated (r = 0.41, p < 0.05 and r = 0.37, p = 0.06, respectively), whereas lesion scores of the small intestine and IL17A expression were negatively correlated (r = −0.33, p = 0.09). The microbiota analysis detected that genus Alistipes was more abundant in WC-supplemented broilers than in control, and positively correlated with body weights and Claudin-1 expression (r = 0.61, p < 0.05 and r = 0.51, p < 0.05, respectively). Intriguingly, genus Enterococcus was most abundant in WC-supplemented broilers and positively correlated with IL17A expression (r = 0.49, p < 0.05). Interestingly, Escherichia-Shigella was significantly more abundant in the small intestinal digestas of AM-administered broilers than in those of control. To summarize, WC supplementation modulated and immunostimulated the microbiotas of broilers, specifically genera Alistipes and Enterococcus, which led to the improvement of weight gain and coccidial symptoms, without disrupting the intestinal microbiota compositions, as AM did.

Specific intestinal bacteria modulate immunoresponses through various pathways, and several probiotic bacteria have been identified as immunostimulants by screening. In the present study, we evaluated the immunomodulating effects ofBacillus coagulansSANK70258 (B. coagulansSANK70258), a spore-forming and lactic acid-producing bacterium usable as food supplement for human and animals. We found that treatment of mouse splenocytes with γ-ray irradiatedB. coagulansSANK70258 induced high amount of IFN-γ in comparison with 7 kinds of typical lactic acid bacteria. Further analyses using splenocytes revealed that NK cell is a major source of IFN-γ, andB. coagulansSANK70258-induced IFN-γ production was inhibited by neutralization of IL-12 or IL-23, depletion of CD11c+cells, and inhibition of NFκB.B. coagulansSANK70258 also induced release of IFN-γ from activated CD8+T cells, and increased expression of chemokine receptors in CD8+T cells.B. coagulansSANK70258-treatment induced production of cytokines from bone marrow-derived dendritic cells, which is reduced by knockdown ofTlr2andNod2.B. coagulansSANK70258-treatment also induced IgA production from Peyer′s patch cells with high level among tested lactic bacteria. The oral intake of γ-ray irradiatedB. coagulansSANK70258 significantly increased intestinal IgA levels and IgA-expressing B cells in the Peyer′s patch of mice. Taken together, we conclude thatB. coagulansSANK70258 possesses high activity as immunostimulant inducing production of IFN-γ and IgA.