Screening and characterization of Bacillus velezensis LB-Y-1 toward selection as a potential probiotic for poultry with multi-enzyme production property.

Bacillus spp. have gained increasing recognition as an option to use as antimicrobial growth promoters, which are characterized by producing various enzymes and antimicrobial compounds. The present study was undertaken to screen and evaluate a Bacillus strain with the multi-enzyme production property for poultry production. LB-Y-1, screened from the intestines of healthy animals, was revealed to be a Bacillus velezensis by the morphological, biochemical, and molecular characterization. The strain was screened out by a specific screening program, possessed excellent multi-enzyme production potential, including protease, cellulase, and phytase. Moreover, the strain also exhibited amylolytic and lipolytic activity in vitro. The dietary LB-Y-1 supplementation improved growth performance and tibia mineralization in chicken broilers, and increased serum albumin and serum total protein at 21 days of age (p < 0.05). Besides, LB-Y-1 enhanced the activity of serum alkaline phosphatase and digestive enzyme in broilers at 21 and 42 days of age (p < 0.05). Analysis of intestinal microbiota showed that a higher community richness (Chao1 index) and diversity (Shannon index) in the LB-Y-1 supplemented compared with the CON group. PCoA analysis showed that the community composition and structure were distinctly different between the CON and LB-Y-1 group. The beneficial genera such as Parasutterella and Rikenellaceae were abundant, while the opportunistic pathogen such as Escherichia-Shigella were reduced in the LB-Y-1 supplemented group (p < 0.05). Collectively, LB-Y-1 can be considered as a potential strain for further utilization in direct-fed microbial or starter culture for fermentation.

Escherichia coli O88 induces intestinal damage and inflammatory response through the oxidative phosphorylation and ribosome pathway in Pekin ducks.

Colibacillosis is one of the major health threats in the poultry industry worldwide. Understanding the pathogenic mechanisms involved in Escherichia coli-induced inflammatory response may lead to the development of new therapies to combat the disease. To address this, a total of 96 1-day-old male lean Pekin ducklings were employed and randomly allocated to two treatments, each with six replicates of eight ducks. Ducks in the experiment group (EG) and the control group (CG) were separately orally administered with 0.2 ml of pathogenic E. coli O88 (3 × 109 CFU/ml) or equivalent volumes of 0.9% sterile saline solution on day 7, two times with an 8-h interval. Serum and intestinal samples were collected on days 9, 14, and 28. Results showed that ducks challenged with E. coli had lower average daily gain and higher feed intake/weight gain during days 9-14 and overall (P < 0.05). Histopathological examination showed that E. coli decreased the villus height and the ratio of villus height/crypt depth in the jejunum (P < 0.05) on days 9 and 14. The intestinal barrier was disrupted, presenting in E. coli ducks having higher serum DAO and D-LA on days 9 and 14 (P < 0.05) and greater content of serum LPS on day 9 (P < 0.05). Escherichia coli infection also triggered a systemic inflammatory response including the decrease of the serum IgA, IgM, and jejunal sIgA on day 14 (P < 0.05). In addition to these, 1,062 differentially expressed genes were detected in the jejunum tissues of ducks by RNA-seq, consisting of 491 upregulated and 571 downregulated genes. Based on the KEGG database, oxidative phosphorylation and the ribosome pathway were the most enriched. These findings reveal the candidate pathways and genes that may be involved in E. coli infection, allow a better understanding of the molecular mechanisms of inflammation progression and may facilitate the genetic improvement of ducks, and provide further insights to tackle the drug sensitivity and animal welfare issues.

Comparing the potential of Bacillus amyloliquefaciens CGMCC18230 with antimicrobial growth promoters for growth performance, bone development, expression of phosphorus transporters, and excreta microbiome in broiler chickens.

Bone health of broiler chickens is essential for welfare and production. In this study, the probiotic Bacillus amyloliquefaciens (BA) CGMCC18230 was compared with antimicrobial growth promoters (AGPs) for its ability to promote growth and bone health. To address this, a total of 180 Arbor Acres (AA) 1-day-old, male, broiler chicks were randomly allocated into 3 treatment groups, with 6 replicates, containing 10 chicks in each replicate. The treatment groups were: control group (CON) fed a corn-soybean based diet; BA treatment group fed the basal diet supplemented with 2.5 × 1010 CFU/kg BA CGMCC18230; AGPs treatment group was fed the basal diet containing the antibiotics aureomycin (75 mg/kg), flavomycin (5 mg/kg) and kitasamycin (20 mg/kg). Over the 42 d experiment, broilers fed BA and AGPs diets both had higher BW, and the ADG was significantly (P < 0.05) higher than that of the CON group both in the grower phase (22-42 d) and overall. Moreover, with BA birds had higher (P < 0.05) serum concentrations of phosphorus (P, day 42) and alkaline phosphatase (ALP, days 21 and 42). Conversely, the content of P in excreta decreased significantly (P < 0.05) on days 21 and 42. Tibia bone mineralization was improved in BA, and the mRNA of P transport related genes PiT-1,2 in the duodenum and jejunum were significantly up-regulated in the BA group than in the CON group (P < 0.05). 16S rRNA gene sequencing revealed that dietary BA supplementation increased the relative abundance of butyrate-producing bacteria (Ruminococcaceae) and polyamine-producing bacteria (Akkermansia and Alistipes), which had a positive effect on bone development. These data show that dietary supplementation of BA CGMCC18320 improves broiler growth performance and bone health similar to supplementation with AGPs through up-regulation of intestinal P transporters, microbial modulation and increase P retention. However, no significant influence of BA CGMCC18320 supplementation on the retention of Ca was found.

Enterococcus faecium Modulates the Gut Microbiota of Broilers and Enhances Phosphorus Absorption and Utilization.

Modern broiler chickens have ongoing bone health problems. Phosphorus (P) plays an important role in bone development and increased understanding of P metabolism should improve the skeletal health of broilers. Enterococcus faecium has been widely used as a probiotic in broiler production and is shown to improve skeletal health of rats, but its effect on the bones of broilers remains unclear. This study investigated the effect of E. faecium on P absorption and utilization in broilers and the associated changes in the gut microbiota using 16S rDNA sequencing. Dietary supplementation with E. faecium improved P absorption through upregulation of the expression of intestinal NaP-IIb mRNA and increased the concentration of serum alkaline phosphatase. These actions increased P retention and bone mineralization in E. faecium-treated broilers. The positive effects of E. faecium on P metabolism were associated with changes in the populations of the intestinal microbiota. There was increased relative abundance of the following genera, Alistipes, Eubacterium, Rikenella and Ruminococcaceae and a decrease in the relative abundance of Faecalibacterium and Escherichia-Shigella. Dietary supplementation with E. faecium changed gut microbiota populations of broilers, increased the relative abundance of SCFA (short-chain fatty acid)-producing bacteria, improved intestinal P absorption and bone forming metabolic activities, and decreased P excretion. E. faecium facilitates increased utilisation of P in broilers.