Knowledge about the modulation of gut microbiota improves our understanding of the underlying mechanism by which probiotic treatment benefits the chickens. This study examined the effects of Bacillus subtilis DSM 32315 on intestinal structure and microbial composition in broilers. Broiler chicks were fed basal diets without or with B. subtilis supplementation (1.0 × 10⁹ spores/kg of diet). Supplemental B. subtilis increased average body weight and average daily gain, as well as elevated villus height and villus height to crypt depth ratio of ileum in broilers. Multi-dimension analysis showed a certain degree of separation between the cecal microbiota from treatment and control groups. Increased Firmicutes abundance and reduced Bacteroidetes abundance in cecum were observed responded to B. subtilis addition, which also increased the abundances of Christensenellaceae and Caulobacteraceae, and simultaneously decreased the abundances of potentially harmful bacteria such as Vampirovibrio, Escherichia/Shigella and Parabacteroides. Network analysis signified that B. subtilis addition improved the interaction pattern within cecal microbiota of broilers, however, it exerted little influence on the metabolic pathways of cecal microbiota by comparison of the functional prediction of metagenomes. In conclusion, supplemental B. subtilis DSM 32315 improved growth performance and intestinal structure of broilers, which could be at least partially responsible by the manipulation of cecal microbial composition.

The effects of live yeast (LY) and mannan-oligosaccharide (MOS) supplementation on intestinal disruption induced by Escherichia coli in broilers were investigated. The experimental design was a 3×2 factorial arrangement with three dietary treatments (control, 0·5 g/kg LY (Saccharomyces cerevisiae, 1·0×10¹⁰ colony-forming units/g), 0·5 g/kg MOS) and two immune treatments (with or without E. coli challenge from 7 to 11 d of age). Samples were collected at 14 d of age. The results showed that E. coli challenge impaired (P<0·05) growth performance during the grower period (1–21 d) and the overall period (1–35 d) of broilers, increased (P<0·05) serum endotoxin and diamine oxidase levels coupled with ileal myeloperoxidase and lysozyme activities, whereas reduced (P<0·05) maltase activity, and compromised the morphological structure of the ileum. Besides, it increased (P<0·05) the mRNA expressions of several inflammatory genes and reduced occludin expression in the ileum. Dietary treatment with both LY and MOS reduced (P<0·05) serum diamine oxidase and ileal myeloperoxidase levels, but elevated villus height (P<0·10) and the ratio of villus height:crypt depth (P<0·05) of the ileum. It also alleviated (P<0·05) E. coli-induced increases (P<0·05) in ileal Toll-like receptor 4, NF-κB and IL-1β expressions. Moreover, LY supplementation reduced (P<0·05) feed conversion ratio of birds during the grower period and enhanced (P<0·05) the community diversity (Shannon and Simpson indices) of ileal microbiota, whereas MOS addition counteracted (P<0·05) the decreased ileal IL-10 and occludin expressions in challenged birds. In conclusion, both LY and MOS supplementation could attenuate E. coli-induced intestinal disruption by alleviating intestinal inflammation and barrier dysfunction in broilers. Moreover, LY addition could improve intestinal microbial community structure and feed efficiency of broilers.

Dietary inclusions of baicalin and chlorogenic acid were beneficial for intestinal health in pigs. Nevertheless, it is unknown whether these plant-derived products had protection for intestine against bacterial challenge in chickens. This study was aimed at evaluating the potential mitigating effects of plant extracts (PE) from Flos lonicerae combined with Baikal skullcap (the active components are chlorogenic acid and baicalin) on intestinal disruption and dysbacteriosis induced by Salmonella pullorum in laying hens. A total of 216 41-week-old layers were randomly divided into 3 groups (6 replicates per group): negative control (NC), S. pullorum-infected positive control (PC), and the S. pullorum-infected group with supplementation of PE at 1000 mg/kg. All birds except those in NC were challenged with S. pullorum at the end of 4 weeks of the experiment. S. pullorum challenge impaired (P < 0.05) the production performance (egg production, feed intake, and feed efficiency) of laying hens, increased (P < 0.05) serum endotoxin content and frequency of Salmonella-positive organs, as well as up-regulated (P < 0.05) ileal expression of pro-inflammatory cytokines including IFNG, TNFA, IL8, and IL1B, whereas PE addition reversed (P < 0.05) these changes and increased (P < 0.05) ileal IL10 expression. Supplemental PE moderated ileal microbiota dysbiosis in challenged birds, characterized by a reduced abundance of Firmicutes along with increased abundances of Bacteroidetes (Bacteroides), Deferribacteres and several butyrate-producers such as Prevotellaceae, Faecalibacterium, Blautia, Butyricicoccus, Lachnoclostridium, and Olsenella, which may assist with energy harvesting and boost anti-inflammatory capacity of host. The decreased abundance of Firmicutes with the increased abundance of Bacteroidetes caused by PE addition had positive correlations with the decreased expression of ileal pro-inflammatory cytokines. The increased abundances of Bacteroidetes (Bacteroides) and Prevotellaceae following PE addition were also positively correlated with the improvement of performance (egg production and feed intake) of laying hens. Collectively, supplemental PE from Flos lonicerae in combination with Baikal skullcap alleviated S. pullorum-induced intestinal disruption and performance impairment in laying hens, which could be at least partially responsible by the modulation of gut microbial composition.

Probiotic Clostridium butyricum could affect lipid metabolism in broilers. However, it is not clear whether C. butyricum could improve lipid metabolism through shaping gut microbiota and bile acid (BA) profile of laying hens. We aimed to evaluate the contributions of gut microbiota and BA profile to the potential effect of C. butyricum on lipid metabolism of aged laying hens. A total of 192 60-week-old Hy-Line Brown laying hens were divided into two groups (eight replicates per group). Birds were fed a basal diet supplemented with 0 or 2.7 g/kg C. butyricum (1.0 × 10⁹ CFU/g). Samples were collected at the end of week 8 of the experiment. The results showed elevated (P < 0.05) concentrations of glucagon-like peptide 1, insulin and thyroid hormones in serum responded to C. butyricum addition, which also decreased (P < 0.05) hepatic free fatty acids contents, as well as increased (P < 0.05) the expression of hepatic acyl-CoA oxidase, farnesoid X receptor (FXR) and PPARα. C. butyricum addition increased (P < 0.05) Bacteroidetes abundance but tended to decrease (P < 0.10) Firmicutes abundance in the ileum. Besides, C. butyricum addition resulted in higher (P < 0.05) abundances of Clostridia (Clostridiales) and Prevotellaceae, concurrent with an increasing trend (P < 0.10) of Bifidobacteriaceae abundance and decreased the abundances of several harmful bacteria such as Klebsiella (P < 0.05). Regarding ileal BA profile, there was a reduced (P < 0.05) content of tauro-α-muricholic acid, increased (P < 0.05) contents of tauroursodeoxycholic acid and lithocholic acid, along with increasing trends (P < 0.10) of glycochenodeoxycholic acid and hyodeoxycholic acid contents due to C. butyricum addition, which also increased (P < 0.05) ileal FXR expression. Collectively, supplemental C. butyricum accelerated hepatic fatty acid oxidation, and shaped gut microbiota and BA profile, thus reducing fat deposition in the liver of aged laying hens.

Catalase (CAT) can eliminate oxygen radicals, but it is unclear whether exogenous CAT can protect chickens against deoxynivalenol (DON)-induced oxidative stress. This study aimed to investigate the effects of supplemental CAT on antioxidant property and gut microbiota in DON-exposed broilers. A total of 144 one-day-old Lingnan yellow-feathered male broilers were randomly divided into three groups (six replicates/group): control, DON group, and DON + CAT (DONC) group. The control and DON group received a diet without and with DON contamination, respectively, while the DONC group received a DON-contaminated diet with 200 U/kg CAT added. Parameter analysis was performed on d 21. The results showed that DON-induced liver enlargement (p < 0.05) was blocked by CAT addition, which also normalized the increases (p < 0.05) in hepatic oxidative metabolites contents and caspase-9 expression. Additionally, CAT addition increased (p < 0.05) the jejunal CAT and GSH-Px activities coupled with T-AOC in DON-exposed broilers, as well as the normalized DON-induced reductions (p < 0.05) of jejunal villus height (VH) and its ratio for crypt depth. There was a difference (p < 0.05) in gut microbiota among groups. The DON group was enriched (p < 0.05) with some harmful bacteria (e.g., Proteobacteria, Gammaproteobacteria, Enterobacteriales, Enterobacteriaceae, and Escherichia/Shigella) that elicited negative correlations (p < 0.05) with jejunal CAT activity, and VH. DONC group was differentially enriched (p < 0.05) with certain beneficial bacteria (e.g., Acidobacteriota, Anaerofustis, and Anaerotruncus) that could benefit intestinal antioxidation and morphology. In conclusion, supplemental CAT alleviates DON-induced oxidative stress and intestinal damage in broilers, which can be associated with its ability to improve gut microbiota, aside from its direct oxygen radical-scavenging activity.

CLIC4/mtCLIC (referred to here as CLIC4) is one of the seven-member family of chloride intracellular channels (CLIC). CLIC4 localizes to the mitochondria, nucleus, cytoplasm and other organellular compartments and participates in the apoptotic response to stress. However, the role of CLIC4 in oxidative stress and apoptosis is not well understood. In this study, we showed the important role of CLIC4 in apoptosis of C6 glioma cells induced by hydrogen peroxide (H2O2). Our results showed that CLIC4 protein expression was upregulated following H2O2-induced C6 cell apoptosis. The upregulation of CLIC4 protein expression was paralleled with an increased Bax/Bcl-2 ratio, cytochrome c and cleaved caspase-3 protein expression upon H2O2-induced C6 cell apoptosis. Suppression of CLIC4 expression by RNA interference enhanced cell apoptosis, but the ratio of Bax/Bcl-2 was not involved in this process. Dissipation of mitochondrial membrane potential and nuclear translocation of CLIC4 were involved in the activation of apoptosis induced by H2O2. Our data indicate that CLIC4 protein may be a key element in the apoptotic response to oxidative stress.