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Effect of Bacillus species-fermented products and essential oils on growth performance, gut morphology, cecal short-chain fatty acid levels, and microbiota community in broilers.

ABSTRACT: In this study, the effects of Bacillus species-fermented products (synbiotics [SYNs]) and essential oils (EOs) on the growth performance, gut morphology, cecal short-chain fatty acid (SCFA) levels, and microbiota of broilers were investigated. A total of 180 one-day-old unsexed broiler chicks (Ross 308) were randomly assigned to 5 dietary treatments as follows: basal diet (control group), basal diet plus enramycin (10 mg/kg; A group), basal diet plus SYNs (3 × 10¹¹ CFU spore/kg of feed; SYN group), basal diet plus EOs (100 mg/kg; EO group), and basal diet plus SYNs and EOs (SYN + EO group), with 6 replicate cages per treatment group and 6 birds per cage. The SYN+EO treatment resulted in a higher (P = 0.003) average daily gain at 1 to 14 d of age than did the control and EO treatments. SYNs had a significant effect on the average daily gain at 1 to 14 d (P < 0.001) and 1 to 35 d (P = 0.045) of age. EOs had a significant effect on the villus height of the duodenum (P = 0.015) and jejunum (P = 0.027). Superoxide dismutase (SOD) and mucin 2 (MUC2) mRNA expression in the duodenum, jejunum, and ileum in the SYN + EO group was higher (P < 0.001) than that in any of the other groups. The SYN+EO treatment resulted in higher (P < 0.001) 2-methylbutyric acid and 3-methylbutyric acid levels in the cecal digesta of the broilers than did the control treatment. Cecal species evenness in the SYN + EO group was higher (P < 0.001) than that in the control group. The abundance of the phylum Firmicutes in the cecal digesta of the broilers was higher (P < 0.001) in the SYN+EO group than in the control group. SYNs had a significant effect (P < 0.001) on the abundance of the genus Lactobacillus in the cecal digesta of the broilers. The abundance of the genus Lactobacillus was positively associated with 2-methylbutyric acid and 3-methylbutyric acid levels. The 2-methylbutyric acid and 3-methylbutyric acid levels were positively correlated with the villus height of the duodenum and ileum. These results suggest that simultaneous supplementation with SYNs and EOs can increase the average daily gain, improve gut health-associated gene expression, increase SCFA levels, and modulate the gut microbiota composition of broilers.

SUBMITTER: Chang WY

PROVIDER: S-EPMC9241036 | biostudies-literature |

REPOSITORIES: biostudies-literature

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Project description:As the poultry industry recedes from the use of antibiotic growth promoters, the need to evaluate the efficacy of possible alternatives and the delivery method that maximizes their effectiveness arises. This study aimed at expounding knowledge on the effect of the delivery method of a probiotic product (Bacillus subtilis fermentation extract) on performance and gut parameters in broiler chickens. A total of 450 fertile eggs sourced from Cobb 500 broiler breeders were randomly allotted to 3 groups: in ovo probiotic (n = 66), in ovo saline (n = 66), and noninjection (n = 200) and incubated for 21 d. On day 18.5 of incubation, 200 μL of either probiotic (10 × 106 cfu) or saline was injected into the amnion. At hatch, chicks were reallotted to 6 new treatment groups: in ovo probiotic, in ovo saline, in-feed antibiotics, in-water probiotic, in-feed probiotics, and control (corn-wheat-soybean diet) in 6 replicate cages and raised for 28 d. Of all hatch parameters evaluated, only percentage pipped eggs was found significant (P < 0.05) with the noninjection group having higher percentage pipped eggs than the other groups. Treatments did not affect the incidence of necrotic enteritis on day 28 (P > 0.05). Irrespective of the delivery method, the probiotic treatments had no significant effect on growth performance. The ileum villus width of the in ovo probiotic treatment was 18% higher than the in ovo saline group (P = 0.05) but not statistically higher than other groups. The jejunum villus height was 23% higher (P = 0.000) in the in ovo probiotic group than in the control group. There was no effect of treatment on total cecal short-chain fatty acid concentration and cecal gut microbiota composition and diversity (P > 0.05), although few unique bacteria differential abundance were recorded per treatment. Conclusively, although probiotic treatments (irrespective of the delivery route) did not affect growth performance, in ovo delivery of the probiotic product enhanced intestinal morphology, without compromising hatch performance and gut homeostasis.

Project description:This study investigated the differential effects of Bacillus subtilis–fermented products (SFPs) and Bacillus licheniformis–fermented products (LFPs) on the growth performance, intestinal morphology, intestinal gene expression, cecal microbiota community, and microbial carbohydrate-active enzyme composition of broilers. In total, 160 one-day-old unsexed Arbor Acres broiler chicks were randomly allocated to 4 treatment groups (with 8 replicates per group and 5 chicks per replicate): control (CON), enramycin (ENM), SFP, and LFP groups, which were fed a basal diet, basal diet supplemented with 10 mg/kg ENM, basal diet supplemented with 108 colony-forming units (CFU) of B. subtilis spores per gram of feed, and basal diet supplemented with 108 CFU of B. licheniformis spores per gram of feed, respectively. LFP treatment resulted in a higher (P < 0.01) body weight at the age of 35 d and higher average daily gain over 15 to 35 (P < 0.05) and 1 to 35 (P < 0.05) d of age than did the CON and SFP treatments. The average villus heights in the jejunum were longer (P < 0.05) in the LFP group than those in the SFP group. Moreover, the LFP group exhibited a higher jejunal barrier function gene expression (mucin 2, zonula occludens-1, and occludin) and cecal antioxidant gene expression (superoxide dismutase) than did the CON group (P < 0.05). A principal coordinate analysis of cecal microbiota and carbohydrate-active enzyme composition demonstrated distinct clustering among the groups. Lactobacillus crispatus abundance in the cecal digesta was higher (P < 0.01) in the SFP group than in the CON and LFP groups. Finally, microbial glycosyltransferase 2 gene expression in the cecal digesta was higher in the LFP group than in the CON and ENM groups. In conclusion, LFPs can improve the growth performance, increase intestinal barrier function and antioxidant gene expression, and modulate cecal microflora composition and carbohydrate-active enzyme composition of broilers. The overall effect of LFPs on growth promotion in broilers was superior to that of SFPs.

Project description:BackgroundThis study investigated a novel in ovo feeding strategy to determine the prebiotic effects of xylo- and mannan- oligosaccharides (XOS and MOS) differing in the degree of polymerization. A total of 192 fertilized eggs were divided into 6 treatment groups: i) normal saline control (NSC), ii) xylotriose (XOS3), iii) xylotetraose (XOS4), iv) mannotriose (MOS3), v) mannotetraose (MOS4), and vi) no injection control (NIC), each containing 4 replicate trays with 8 eggs per replicate. On d 17 of incubation, 3 mg of oligosaccharides (except for controls) dissolved in 0.5 mL of 0.85% normal saline were injected into the amnion of Cobb 500 broilers eggs. After hatch, the chicks were raised for 28 d under standard husbandry practices and were fed a commercial broilers diet ad libitum, and samples were collected periodically.ResultsThe hatchability, growth performance, and relative weights of breast, drumstick, liver, and proventriculus were not different among the treatments (P > 0.05). The XOS3 injection increased the total short-chain fatty acid production at d 28 compared with both control groups (P < 0.05). The villus height to crypt depth ratio was significantly higher in the XOS4 group than both controls on the hatch day (P < 0.01) but were not different among any treatments on d 7 and 28 (P > 0.05). On the hatch day, the expression level of the CD3 gene (a T cell marker) was increased by XOS3, while the IL-10 gene (a marker of anti-inflammatory cytokine) was reduced by MOS4 (P < 0.05) compared with both controls. Compared with both controls, XOS3 exhibited a trend of reduction for IL-10 (P = 0.074). No cytokines or lymphocyte markers were affected by the treatments on d 7 (P > 0.05), except XOS4 increased IL-4 compared with NSC (P < 0.05). The broilers in the MOS4 group had higher operational taxonomic units (OTUs) and had more differentially abundant taxa, including order Lactobacillales and family Leuconostocaceae (P < 0.05) than both controls on d 28. The predictive functional profiling indicated that the linoleic acid metabolism pathway was enriched in the cecal microbiota of the XOS3 group compared with both controls (P < 0.05).ConclusionThe effects of these XOS and MOS on ileal mucosa and immunity are transient, but the effects on fermentation and cecal microbiota are prolonged, and further research is warranted to determine their use as a gut health promoter in poultry.

Project description:The gut microbiota play an important role in the growth and intestinal health of broilers. The present study was to investigate the gut microbiota, short-chain fatty acids, and intestinal morphology of broilers at different ages. A total of 320 one-day-old male broilers were raised in 8 replicates and fed the same corn-soybean diets for 42 D. The duodenal, jejunal, and ileal segments and their and cecal microbiota were collected on day 1, 7, 14, 21, and 42, respectively. The villous height (VH), crypt depth (CD), and their ratio of VH:CD in the duodenum, jejunum, and ileum all increased (P < 0.05) with age. Caecal acetate, propionate, butyrate, valerate, and isovalerate increased (P < 0.01), but isobutyrate decreased (P < 0.001) with age. The cecum had the greatest (P < 0.001) alpha diversity of bacterial community in broilers at different ages. Beta diversities showed distinct differences in gut microbial compositions among different ages (R = 0.55, P < 0.002) and different intestinal segments (R = 0.53, P < 0.002). Lactobacillus was the most abundant genus in the duodenum (36∼97%), jejunum (39∼72%), and ileum (24∼96%) at all ages, and in the ileum, it was positively correlated with VH (R = 0.559, P < 0.03), VH:CD (R = 0.55, P < 0.03), and acetate contents (R = 0.541, P < 0.04) but negatively correlated (R = -0.50, P < 0.05) with isobutyrate contents. Escherichia-Shigella and Salmonella dominated in the cecum of newly hatched broilers, and then the Bacteroides dominated in the cecum on day 42. In the cecum, Escherichia-Shigella was positively correlated (R = 0.577∼0.662, P < 0.05) with isobutyrate contents and Salmonella negatively correlated (R = -0.539∼-0.843, P < 0.05) with isovalerate, butyrate, and acetate contents. These aforementioned results indicated that the most abundant Lactobacillus from the small intestine and the most diversity of microflora community and short-chain fatty acids in the cecum might contribute to the development of intestinal structure in the whole growing period of broilers.

Project description:Background/aimsThe gut microbiota regulates intestinal immune homeostasis through host-microbiota interactions. Multiple factors affect the gut microbiota, including age, sex, diet, and use of drugs. In addition, information on gut microbiota differs depending on the samples. The aim of this study is to investigate whether changes in cecal microbiota depend on aging.MethodsGut microbiota in cecal contents of 6-, 31-, and 74-week-old and 2-year-old male Fischer-344 rats (corresponding to 5-, 30-, 60-, and 80-year-old humans in terms of age) were analyzed using 16S ribosomal RNA metagenome sequencing and phylogenetic investigation of communities by reconstruction of unobserved states (PICRUSt) based on the Kyoto Encyclopedia of Genes and Genomes orthology. Moreover, short-chain fatty acid (SCFA) level in cecum and inflammation related factors were measured using real-time quantitative polymerase chain reaction and enzyme linked immunosorbent assay.ResultsAlpha and beta diversity did not change significantly with age. At the family level, Lachnospiraceae and Ruminococcaceae, which produce SCFAs, showed significant change in 31-week-old rats: Lachnospiraceae significantly increased at 31 weeks of age, compared to other age groups, while Ruminococcaceae decreased. Butyrate levels in cecum were significantly increased in 31-week-old rats, and the expression of inflammation related genes was increased followed aging. Especially, EU622775_s and EU622773_s, which were highly abundance species in 31-week-old rats, showed significant relationship with butyrate concentration. Enzymes required for producing butyrate-acetyl-CoA transferase, butyryl-CoA dehydrogenase, and butyrate kinase-were not predicted by PICRUSt.ConclusionsMajor bacterial taxa in the cecal lumen, such as Lachnospiraceae, well-known SCFAs-producing family, changed in 31-week-old rats. Moreover, unknown species EU622775_s and EU622773_s showed strong association with cecal butyrate level at 31 weeks of age.

Project description:BackgroundFermented Aloe leaf juice is a commonly used food supplement in Japan. In a previous study, fermentation of A. arborescence juice was performed and the presence of short-chain fatty acids (SCFAs) was confirmed and quantified. Samples were collected before and after the fermentation process to be subjected, in the present study, to DNA extraction, 16S rRNA gene (V3-V4 regions) amplification, and sequencing by the next-generation Illumina MiSeq sequencer. Our work aims to analyze the sequences to assess the bacterial diversity in the juice before and after fermentation, identify the beneficial microbes responsible for the production of SCFAs, and evaluate some of the biological activities of the fermented juice.ResultsData revealed the richness and diversity of the bacterial community in the fermented juice compared to the unfermented control. Relative abundance of bacterial phyla showed that the majority of the microbial community in the test samples corresponded to Pseudomonadota (unfermented; 10.4%, fermented; 76.36%), followed by Bacillota (unfermented; 4.71%, fermented; 17.13%) and then Bacteroidota (unfermented; 0.57%, fermented; 1.64%). For the fermented sample, 84% of Bacillota were lactobacilli. A hierarchically clustered heatmap revealed that Lactobacillus was the most abundant genus in both samples suggesting its involvement in the production of SCFAs. To assess potential health benefits, the anticancer efficacy of the fermented product of A. arborescens was investigated against colorectal cancer (IC50 = 3.5 µg/ml) and liver cancer (IC50 = 6.367 µg/ml) compared to the normal peripheral blood mononuclear cells (PBMCs). Flow cytometric analysis of the cell cycle pattern revealed remarkable population arrest in G0 and G1, however, the highest percentages were mainly in the G1 phase for Hep-G2 (40.1%) and HCT-116 (53.2%) cell lines. This effect was accompanied by early apoptotic profiles of HCT-116 (36.9%) and late apoptosis for Hep-G2 (17.3%). Furthermore, immunomodulatory properties demonstrated a significantly (p < 0.001) reduced percentage of induced TNF-α while enhancing IFN-γ dramatically. For antimicrobial activities, marked broad-spectrum activities were recorded against some bacterial and fungal pathogens (17-37 mm inhibition zone diameter range).ConclusionTherefore, this study affords the basis of bacterial community composition in fermented A. arborescens juice as well as its potential biological benefits.

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Effect of Bacillus species-fermented products and essential oils on growth performance, gut morphology, cecal short-chain fatty acid levels, and microbiota community in broilers.

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