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Dietary Bacillus subtilis supplementation alleviates alcohol-induced liver injury by maintaining intestinal integrity and gut microbiota homeostasis in mice.

ABSTRACT: Alcoholic liver disease (ALD) is a worldwide health problem with limited therapeutic options, which is associated with gut-derived endotoxins, particularly lipopolysaccharide (LPS) and intestinal microbiota dysbiosis. Recently, probiotics, synbiotics and other food additive interventions have been shown to be effective in decreasing or preventing the progression of ALD. Bacillus subtilis (B. subtilis) and its metabolic products are widely used as food additives to maintain intestinal health, but the protective effects of B. subtilis against alcohol-induced liver injury are poorly understood. In the present study a chronic alcohol-induced liver injury model was constructed based on the Lieber-DeCarli diet and it aimed to determine whether dietary B. subtilis supplementation may alleviate alcohol-induced liver injury. Results revealed that prophylactic B. subtilis supplementation partially restored gut microbiota homeostasis and relieved alcohol-induced intestinal barrier injury, which significantly decreased the translocation of bacterial endotoxins to the blood. In addition, the decreased serum LPS alleviated hepatic inflammation via the toll-like receptor 4 pathway, resulting in improved hepatic structure and function. These results demonstrated that dietary B. subtilis supplementation imparts novel hepatoprotective functions by improving intestinal permeability and homeostasis.

SUBMITTER: Zhao M

PROVIDER: S-EPMC8461600 | biostudies-literature |

REPOSITORIES: biostudies-literature

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Project description:Bacillus licheniformis (B. Licheniformis) has been considered to be an effective probiotic to maintain gut health and boost productivity in the pig industry, but there is no complete understanding of its mechanisms. We determined whether weaned piglets exposed to BL-S6 (probiotic) had altered intestinal barrier function or microbiota composition. In our study, 108 weaned piglets (54 barrows and 54 gilts) were divided equally into three groups, each with six pens and six piglets/pen, and fed a basal diet supplemented without or with antibiotic (40 g/t of Virginiamycin and 500 g/t of Chlortetracycline) or probiotic (1000 g/t of B. Licheniformis) for a 14-day trial. On day 14, one piglet was chosen from each pen to collect blood and intestinal samples. Compared with the control group, dietary supplementation with a probiotic promoted body weight (BW) gain and average daily gains (ADG) while reducing diarrhea incidence (p < 0.05). Probiotics enhanced superoxidase dismutase (SOD) activity and decreased malondialdehyde (MDA) levels in serum (p < 0.05), and increased the level of mRNA expression of SOD1, Nrf2, and HO-1 (p < 0.05) in the jejunum mucosa. Moreover, supplementation with probiotics improved intestinal mucosal integrity as evidenced by higher villus heights and a higher ratio of villus heights to crypt depths (duodenum and jejunum) and higher mRNA and protein levels of occludin and ZO-1 in jejunum mucosa (p < 0.05). The intestinal sIgA levels (p < 0.05) were elevated in the probiotic group, and that of serum immunoglobulin A (IgA) tended to be higher (p = 0.09). Furthermore, weaning piglets who were given probiotics had a better balance of the cecum microbiota, with lactobacillus abundance increased and clostridium_sensu_stricto_1 abundance decreased. In conclusion, dietary supplementation with the probiotic BL-S6 promoted intestinal integrity, which was associated, in part, with modulating intestinal barrier function and microbial diversity in weaned piglets; it may offer a promising alternative to antibiotics to prevent diarrhea.

Project description:Our previous research reported that supplementation of Bacillus subtilis DSM 25841 promoted growth and disease resistance of weaned pigs under enterotoxigenic Escherichia coli (ETEC) challenge and its efficacy is comparable to carbadox. This follow-up study aimed to characterize the effects of ETEC infection, supplementing B. subtilis DSM 25841 or carbadox on intestinal microbiota of pigs. Forty-eight weaned pigs (6.17 ± 0.36 kg BW) were randomly allotted to one of four treatments: negative control (NC), positive control (PC), antibiotics (AGP, 50 mg/kg of carbadox), and direct fed microbials (DFM, 2.56 × 109 CFU/kg of B. subtilis). The experiment lasted 28 days with 7 days before and 21 days after first E. coli inoculation (day 0). Pigs in the PC, AGP, and DFM groups were orally inoculated with F18 ETEC for 3 consecutive days with 1010 CFU per dose per day. Fecal samples were collected on day -7, and day 7 and day 21 post inoculation, digesta samples were collected from jejunum, ileum, and distal colon on day 21 post inoculation to perform 16S rRNA sequencing. Sampling days and locations influenced (p < 0.05) Chao1 index and beta-diversity. Age increased (p < 0.05) the relative abundance of Firmicutes but decreased (p < 0.05) the relative abundance of Bacteroidetes in feces. ETEC infection increased (p < 0.05) the relative abundance of Proteobacteria in feces on day 7 post inoculation. AGP reduced (p < 0.05) relative abundance of Firmicutes and Lactobacillaceae in feces compared with PC and DFM. AGP reduced (p < 0.05) relative abundance of Bifidobacteriaceae in jejunum and ileum, while DFM reduced (p < 0.05) relative abundance of Actinomycetaceae in jejunum and Lachnospiraceae in ileum, compared with PC. Pigs fed with DFM had greater (p < 0.05) relative abundance of Ruminococcaceae, Veillonellaceae, Bifidobacteriaceae in jejunum, Lactobacillaceae in ileum and colon, and Bifidobacteriaceae in colon than pigs in AGP. Current results indicate that carbadox or B. subtilis had stronger influences on microbial diversity and composition in ileum than other intestinal segments and feces. Supplementation of B. subtilis could increase or maintain the relative abundance of beneficial bacteria in ileum compared with carbadox.

Project description:This study was conducted to investigate the effects of dietary supplementation with different types of Bacillus subtilis (B. subtilis) on the growth and gut health of weaned piglets. A total of 160 piglets were randomly assigned into four groups: control group (a basal diet), BS-A group (a basal diet supplemented with B. subtilis A at 1 × 106 CFU/g feed), BS-B group (a basal diet supplemented with B. subtilis B at 1 × 106 CFU/g feed), and BS-C group (a basal diet supplemented with B. subtilis C at 1 × 106 CFU/g feed). All groups had five replicates with eight piglets per replicate. On days 7, 21, and 42 of the trial, blood plasma and intestinal tissues and digesta samples were collected to determine plasma cytokine concentrations, intestinal morphology, gut microbiota community and metabolic activity, and the expression of genes related to gut physiology and metabolism. The results showed that dietary B. subtilis supplementation improved (P < 0.05) the body weight and average daily gain (in BS-B and BS-C groups) of weaned piglets and decreased (P < 0.05) the diarrhea rates (in BS-A, BS-B, and BS-C groups). In the intestinal morphology analysis, B. subtilis supplementation improved (P < 0.05) the size of villus height and villus height to crypt depth ratio in the ileum of weaned piglets. Firmicutes, Bacteroidetes, and Tenericutes were the most dominant microflora in piglets' colon whatever the trial group and time of analysis. Dietary BS-C supplementation increased (P < 0.05) the relative abundances of Anaerovibrio and Bulleidia and decreased (P < 0.05) the relative abundances of Clostridium and Coprococcus compared with the control group. In addition, dietary B. subtilis supplementation increased (P < 0.05) the indicators of intestinal health, including plasma levels of interleukin (IL)-2 and IL-10, as well as the colonic levels of short-chain fatty acids. Furthermore, dietary B. subtilis supplementation also up-regulated (P < 0.05) the expression of genes involved in metabolic pathways related to intestinal microbiota maturation. In conclusion, these findings suggest that a diet containing BS-B or BS-C can efficiently promote growth performance, decrease diarrhea incidence, and ameliorate several indicators of intestinal health through the modulation of gut microbiota composition and metabolic activity in weaned piglets.

Project description:The purpose of this study was to investigate the effects of Bacillus subtilis on growth performance, intestinal morphology, and cecal microbial composition of broilers. A total of 270 healthy one-day-old Arbor Acres male broiler chicks were randomly divided into 3 dietary treatment groups, with 6 replicates per group and 15 chickens per replicate. The dietary treatment groups were as follows: 1) basal diet, negative control group; 2) basal diet +250 g/t of zinc bacitracin, positive control group; and 3) basal diet +750 g/t of B. subtilis, B. subtilis group. Results of this experiment showed that compared with the negative control group, body weight at 42 d, average daily gain and European Production Efficiency Factor over the 42 d phase in the B. subtilis group and positive control group were significantly increased (P < 0.05); feed conversion rates in the B. subtilis group and positive control group were significantly decreased (P < 0.05); and average daily feed intake and mortality were not significantly different (P > 0.05). The villus height to crypt depth ratio in the ileum of the B. subtilis group was significantly higher (P < 0.05) than that of the negative control group. The results of cecal microflora at genus level were as follows. As compared with the negative control group, the abundance of Blautia, Faecalibacterium, Flavonifractor, and Hydrogenoanaerobacterium of the B. subtilis group and positive control group was significantly higher (P < 0.05), whereas the abundance of Odoribacter was significantly lower (P < 0.05). Moreover, abundance of the genera Romboutsia in the B. subtilis group was higher (P < 0.05) than that in the positive control group. The abundance of Flavonifractor, Erysipelatoclostridium, and Hydrogenoanaerobacterium were positively correlated with body weight and average daily gain by Spearman correlation analysis. In conclusion, dietary supplementation with B. subtilis improved growth performance of broilers which may be related to the increased abundance of Blautia, Faecalibacterium, Flavonifractor, Hydrogenoanaerobacterium, and Romboutsia, along with the decreased abundance of Odoribacter. In addition, the effect of B. subtilis was superior to zinc bacitracin in improving intestinal microbial composition of broilers. Therefore, B. subtilis may act as an effective antibiotic substitute in broilers.

Project description:This study aimed to investigate the effects of chitooligosaccharide (COS) on intestinal barrier, antioxidant capacity, and immunity of lipopolysaccharide (LPS)-challenged laying hens. A total of 360 Hy-line Brown laying hens (80-wk-old) were randomly divided into 5 groups with 6 replicates of 12 birds. Hens were fed a corn-soybean meal basal diet supplemented with different COS levels (0; 5; 10; 15; 20 mg/kg) for 8 wk. The results showed that 15 mg/kg COS administration elevated albumen height and Haugh unit (P < 0.05), and numerically optimized productive performance (P > 0.05), therefore, the dosage of 15 mg/kg was chosen for the subsequent experiment. Thereafter, 12 birds from non-supplemented group were randomly selected and assigned into 2 groups, and birds in each group were administered (1.5 mg/kg BW, i.p.) with saline (control group) or LPS (challenge group). Another 6 hens from 15 mg/kg COS-supplemented group were selected and injected with LPS in the same way. Compared with the control group, LPS-challenged birds exhibited elevated circulating diamine oxidase activity, and reduced jejunal villus height and ratio of villus height to crypt depth, and these indices were reversed to control levels by COS (P < 0.05). Also, LPS increased malondialdehyde accumulation and reduced several antioxidant enzyme activities in the intestinal mucosa (P < 0.05). Additionally, LPS increased jejunal secretory IgA and interferon-γ (IFN-γ), and ileal secretory IgA, IgM, and interleukin-1β (IL-1β) concentrations, whereas COS reduced jejunal IFN-γ and IL-1β, and ileal IgM levels (P < 0.05). Moreover, LPS down-regulated mRNA abundance of jejunal occludin and claudin 2, and upregulated expression of jejunal nuclear factor erythroid-2 related factor 2, superoxide dismutase 1, and IFN-γ as well as ileal IL-1β (P < 0.05). Besides, COS increased jejunal occludin and ileal claudin 2, nuclear factor erythroid-2 related factor 2, and heme oxygenase-1 expression, and decreased jejunal IFN-γ and IL-1β abundance (P < 0.05). These results suggested that COS could alleviate LPS-induced intestinal barrier impairment, and oxidative and immunological stress in laying hens.

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Dietary Bacillus subtilis supplementation alleviates alcohol-induced liver injury by maintaining intestinal integrity and gut microbiota homeostasis in mice.

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