Laying hen responses to multi-strain Bacillus-based probiotic supplementation from 25 to 37 weeks of age.

Objective: Investigate the efficacy of Bacillus-based probiotics supplemented at two different levels to modulate the productive performance, egg quality, tibia traits, and specific cecal bacteria counts of Hy-Line Brown layers from 25 to 37 weeks of age. Methods: A total of 216 twenty-five-week-old hens were randomly distributed into 3 experimental diets with 12 replicates of 6 birds per cage. Diets included basal diet supplemented with 0 (CON), 3 × 108 (PRO1), or 3 × 109 (PRO2) CFU of the test probiotic containing Bacillus subtilis PB6, Bacillus subtilis FXA, and Bacillus licheniformis G3 per kilogram of feed. Results: Improved egg weights and mass at 29 weeks; and feed intake at 31 weeks (p < 0.10) were noticed with the probiotic-supplemented PRO1 and PRO2 diets. Considering egg quality, the shell thickness, Haugh units, and yolk color were improved; but yolk cholesterol was lowered (p < 0.05) with PRO1 and PRO2 diets at 29 weeks. At both 33 and 37 weeks, the egg-breaking strength, shell color and thickness, albumen height, Haugh units, and yolk color were improved; but yolk cholesterol was similarly lowered (p < 0.05) with the PRO1 and PRO2 diets. Improved tibia Ca, ash, weights, and density; and raised cecal counts of Bifidobacteria and Lactobacilli (p < 0.05) were noticed with PRO1 and PRO2 diets. Improved tibia P but reduced Clostridia counts (p < 0.10) were also observed with the PRO1 and PRO2 diets. Conclusion: Probiotic supplementation of Bacillus subtilis PB6, Bacillus subtilis FXA, and Bacillus licheniformis G3 at 3 ×108 CFU/kg of feed is adequate to significantly improve egg quality, lower yolk cholesterol, enhance several tibia traits, and raise the populations of beneficial cecal bacteria. Modest improvements in several productive parameters and tibia P but reduced Clostridia were also observed; and could warrant further investigation of probiotic effects beyond the current test period.

Dietary Bacillus spp. supplementation to both sow and progenies improved post-weaning growth rate, gut function, and reduce the pro-inflammatory cytokine production in weaners challenged with Escherichia coli K88.

BACKGROUND: The use of probiotics (PRO) in late gestation sow and their impact on progenies' performance during the post-weaning stage has received more attention from the researchers recently. This study aimed to analyze the effect of probiotic mixture (Bacillus subtilis and Bacillus licheniformis) on both sow and offspring's performance. METHODS: First experiment (Exp.1) was conducted from the 100th day of gestation through to post-weaning. A total of twenty sows and their litters were assigned to one of two dietary treatments, Control (CON) based diet and PRO- CON+ 0.05% probiotic mixture. Dietary treatments were arranged in a split-plot pattern with sow and weaner treatment (CON and PRO diet) as the main and sub plot. Exp.2. E. coli challenge study was carried out two weeks after weaning with 40 piglets. Dietary treatments remained same while all pigs were orally administered with a 1.5 ml suspension of 1010 CFU of K88 strain of E. coli per ml. RESULT: PRO group sow showed significantly decreased backfat thickness difference and body weight difference after farrowing and at the end of weaning d21. The nutrient digestibility of PRO group sows was significantly higher at the end of weaning. Moreover, piglets born from PRO group sow showed higher weaning weight and tend to increase average daily gain at the end of d21. The addition of mixed probiotic in sow and weaner diet had suppressed the production of TNF-α and interleukin-6 in E. coli challenged pigs. The phyla Firmicutes and Bacteroidetes in E. coli -challenged pigs were highly abundant while, the relative abundance of clostridium_sensu_stricto_1 at genus level was significantly reduced by the inclusion of probiotic in both the sow and weaner diet. Also, taxonomic distribution analysis showed significantly lower prevalence of Clostridium and Brachyspira and higher prevalence of Lactobacilli in E. coli-challenged pigs that were born from PRO group sow and fed CON and PRO weaner diet. CONCLUSION: This study reveals that the inclusion of 0.05% mixed probiotics (Bacillus spp.) to both sow and their progenies diet would be more beneficial to enhance the post-weaning growth rate, gut health, and immune status of E. coli challenged pigs.

Effect of dietary inclusion of Bacillus-based probiotics on performance, egg quality, and the faecal microbiota of laying hen.

OBJECTIVE: Our study examined the impact of propriety blends of Bacillus strain probiotics on the performance, egg quality, and faecal microflora of laying hens. METHODS: A total of 183 Institut de selection Animale (ISA) brown laying hens aged 23 weeks with an average body weight of 1,894±72 g were randomly allocated into 3 groups as control (corn-soybean meal based diet, CON), 0.5 g/kg Enterosure probiotics (ET1, 3×108 colony-forming unit [CFU]/kg feed), and 5 g/kg Enterosure probiotics (ET2, 3×109 CFU/kg feed) administered in mashed form. At the completion of each phase hen day egg production (HDEP), average egg weight (AEW), feed intake, and faecal microbiota were evaluated. RESULTS: HDEP and AEW were higher (p<0.05) in the ET2-supplemented diet in phase 3 (week 9 to 12) compared with CON. Egg mass (EM) was higher (p<0.05) in phase 2 at ET2, and also higher (p<0.05) in phase 3 at the ET1 and ET2-supplemented diets compared with CON. Feed conversion ratio was lower (p<0.05) in phase 3 at the ET1 and ET2-supplemented diets, with ET2 being the lowest compared with ET1 and CON. Yolk colour was higher (p<0.05) in the ET-supplemented diets at phase 3 compared with CON. Bifidobacterium spp. was higher (p<0.05) in the ET2- supplemented diet compared with CON in phase 2, while in In phase 3, Lactobacillus spp. and Bifidobacterium spp. were higher (p<0.05) in the ET-supplemented diets compared with CON. Coliforms were lower (p<0.05) in the ETsupplemented diets compared with CON in phase 3. CONCLUSION: The propriety blends of Bacillus strain probiotics supplements at 0.5 g/kg and 5 g/kg could improve the production and quality of eggs with more significance at 5 g/kg for HDEP, AEW and EM, which was achieved via the increase of beneficial microbiomes such as Lactobacillus spp., Bifidobacterium spp., and the decrease of pathogenic microbiomes like Escherichia coli and Coliforms which was speculated to improve gut barrier function and the reproductive hormone.

Increasing doses of phytase from Citrobacter braakii in diets with reduced inorganic phosphorus and calcium improve growth performance and lean meat of growing and finishing pigs.

The objective of this study was to evaluate the effect of increasing doses of bacterial phytase (RONOZYME HiPhos) on performance and carcass characteristics of growing and finishing pigs. The study included 120 castrated males with initial weight of 23.21 ± 1.91 kg and 68 days of age, distributed in a randomized block design with five treatments and eight replicates with three animals each. The pigs were fed five corn-soybean meal-based diets: positive control (PC), supplemented with inorganic phosphorus and calcium; negative control (NC), with 0.13% reduction in available phosphorus and 0.11% in calcium; and three NC diets supplemented with 1,000, 2,000, and 3,000 phytase units (FYT)/kg in the feed. Compared with the NC diets without phytase, diets with 1,000, 2,000, and 3,000 FYT/kg inclusion increased the daily weight gain by +12% (quadratic, p<0.05) during the growing I period; +2.9, +2.9, and +10.5% (linear, p<0.01), respectively, during the growing II period; and +4.1, +5.1, and +8.2% (linear, p<0.001), respectively, over the entire experimental period. The daily feed intake increased by 0, +2.8, and +4.3% (linear, p<0.05), respectively, considering the entire experimental period; and the final live weight increased by +3.2, +4.2, and +6.1% (linear, p<0.001), respectively. The phytase treatments did not influence feed conversion ratio, carcass weight and yield, backfat thickness, loin depth and carcass lean meat. According to the European Carcass Classification (SEUROP), however, the animals fed the PC diet and the three phytase levels had more carcasses classified as E (between 55-60% lean meat) when compared to carcasses of pigs fed the NC. Supplementing increasing levels of phytase to a corn- and soybean meal-based diet with inorganic P and Ca reduction improved daily weight gain and feed intake of growing pigs, and such effects were maintained until slaughter age.

Effect of exogenous amylase on lactation performance of dairy cows fed a high-starch diet.

Exogenous amylase supplementation can increase starch and fiber digestibility in lactating dairy cows. We evaluated the effect of exogenous amylase supplementation on diets with high starch concentration (32% of dry matter). Twenty-eight Holstein cows (171 ± 80 d in milk, 4 primiparous) received a standard diet for 14 d and then a treatment for 63 d, in a covariate-adjusted randomized block design with repeated measures over time. Treatments were amylase [0.5 g of Ronozyme RumiStar (DSM Nutritional Products, Basel, Switzerland) per kg of total mixed ration dry matter] or control. The diets contained (% of dry matter): 39.4% corn silage, 11.2% rehydrated and ensiled mature corn grain, and 11.7% finely ground mature corn. Amylase increased milk yield (32.3 vs. 33.0 kg/d) and reduced dry matter intake (20.7 vs. 19.7 kg/d), increasing feed efficiency (1.52 vs. 1.63). Amylase also increased milk lactose synthesis (1.49 vs. 1.56 kg/d) and plasma glucose concentration (59.3 vs. 68.6 mg/dL). Secretions of milk fat and protein did not differ. Although milk urea N did not differ, amylase reduced the concentration of urea N in blood, suggesting an increase in ruminal starch degradation. However, the total-tract apparent digestibility of starch (96.3% of intake) and neutral detergent fiber (44.4% of intake), ruminal fermentation profile, and microbial yield estimated by urinary allantoin excretion did not differ. Cows fed amylase sorted in favor of long feed particles and against short particles, had shorter chewing activity (780 vs. 699 min/d), and had fewer meals per day (11.5 vs. 9.7). Amylase improved the feed efficiency of lactating cows fed a high-starch diet; the enzyme increased milk yield and reduced intake.

Screening the ability of natural feed ingredients to interfere with the adherence of enterotoxigenic Escherichia coli (ETEC) K88 to the porcine intestinal mucus.

The inhibition of the attachment of bacteria to the intestine by receptor analogues could be a novel approach to prevent enterotoxigenic Escherichia coli (ETEC) K88-induced diarrhoea in piglets. The objective of the present study was to screen the ability of different feed ingredients (FI) to bind to ETEC K88 (adhesion test, AT) and to block its attachment to the porcine intestinal mucus (blocking test, BT) using in vitro microtitration-based models. In the AT, wheat bran (WB), casein glycomacropeptide (CGMP) and exopolysaccharides exhibited the highest adhesion to ETEC K88 (P< 0·001). In the BT, WB, CGMP and locust bean (LB) reduced the number of ETEC K88 attached to the intestinal mucus (P< 0·001). For WB and LB, fractionation based on their carbohydrate components was subsequently carried out, and each fraction was evaluated individually. None of the WB fractions reduced the adhesion of ETEC K88 to the mucus as did the original extract, suggesting that a protein or glycoprotein could be involved in the recognition process. With regard to the LB fractions, the water-extractable material reduced the adhesion of ETEC K88 (P< 0·001) to the mucus similar to the original extract (P< 0·001), indicating, in this case, that galactomannans or phenolic compounds could be responsible for the recognition process. In conclusion, among the FI screened, the soluble extracts obtained from WB, LB and CGMP exhibited the highest anti-adhesive properties against ETEC K88 in the BT. These results suggest that they may be good candidates to be included in diets of weaned piglets for the prevention of ETEC K88-induced diarrhoea.

Casein glycomacropeptide in the diet may reduce Escherichia coli attachment to the intestinal mucosa and increase the intestinal lactobacilli of early weaned piglets after an enterotoxigenic E. coli K88 challenge.

Casein glycomacropeptide (CGMP), a glycoprotein originating during cheese manufacture, has shown promising effects by promoting the growth of some beneficial bacteria in vitro, although its activity has not been well explored. The present study was designed to evaluate the effects of CGMP against enterotoxigenic Escherichia coli (ETEC) K88 in vitro (Trial 1) and in vivo (Trial 2). In Trial 1, increasing concentrations of CGMP (0, 0.5, 1.5 or 2.5 mg/ml) were tested regarding its ability to block the attachment of ETEC K88 to ileal mucosa tissues obtained from piglets. Increasing the concentration of CGMP resulted in a gradual decrease in ETEC K88 attachment to the epithelial surface. In Trial 2, seventy-two piglets were distributed in a 2 × 2 factorial combination including or omitting CGMP in the diet (control diet v. CGMP) and challenged or not with ETEC K88 (yes v. no). Inclusion of CGMP increased crude protein, ammonia and isoacid concentrations in colon digesta. CGMP also increased lactobacilli numbers in ileum and colon digesta, and reduced enterobacteria counts in mucosa scrapings and the percentage of villi with E. coli adherence measured by fluorescence in situ hybridisation. The inclusion of CGMP in the diets of challenged animals also prevented the increase of enterobacteria in ileal digesta. We can conclude that CGMP may improve gut health by diminishing the adhesion of ETEC K88 to the intestinal mucosa, by increasing the lactobacilli population in the intestine and by reducing the overgrowth of enterobacteria in the digestive tract of piglets after an ETEC K88 challenge.

Influence of dietary ingredients on in vitro inflammatory response of intestinal porcine epithelial cells challenged by an enterotoxigenic Escherichia coli (K88).

Enterotoxigenic Escherichia coli (ETEC) K88 is the main bacterial cause of diarrhea in piglets around weaning and the adhesion of ETEC to the intestinal mucosa is a prerequisite step for its colonization. In this study, the adhesion of a fimbriated ETEC and a non-fimbriated E. coli (NFEC) to the intestinal cells and the activation of the innate immune system were evaluated using a porcine intestinal epithelial cell line (IPEC-J2). The impact of several feedstuffs (wheat bran (WB); casein glycomacropeptide (CGMP); mannan-oligosaccharides (MOS); locust bean extract (LB) and Aspergillus oryzae fermentation extract (AO)) on ETEC attachment and the inflammatory response were also studied. The gene expression of TLR-4; TLR-5; IL-1ß; IL-8; IL-10 and TNF-α were quantified using Cyclophilin-A, as a reference gene, and related to a non-challenged treatment. The fimbriated strain was markedly better than the non-fimbriated strain at adherence to intestinal cells and inducing an inflammatory response. All the feedstuffs studied were able to reduce the adhesion of ETEC, with the greatest decrease with CGMP or MOS at highest concentration. Regarding the inflammatory response, the highest dose of WB promoted the lowest relative expression of cytokines and chemokines. All tested feedstuffs were able to reduce the adhesion of ETEC to IPEC-J2 and interfere on the innate inflammatory response; however WB should be further studied according to the beneficial results on the intestinal inflammatory process evidenced in this study.

Effect and interaction between wheat bran and zinc oxide on productive performance and intestinal health in post-weaning piglets.

The inclusion of fibre has been studied as an alternative to antimicrobials in weaning pig diets, while ZnO is used as an effective method to prevent post-weaning diarrhoea. However, it has not been investigated to what extent these two strategies interact with each other. The present study was designed to evaluate the effects of including wheat bran (WB) and ZnO alone or combined in the diet of early-weaning pigs on productive performance and microbial activity in the gastrointestinal tract (trial 1). A total of sixty-four piglets were distributed in a 2 × 2 factorial combination of two levels of WB (0 v. 40 g/kg) and ZnO (0 v. 3 g/kg) in the diet. The inclusion of ZnO in the diet improved the feed intake and growth of the animals and reduced the incidence of diarrhoea. The inclusion of WB increased SCFA concentrations and decreased Escherichia coli counts. However, simultaneous incorporation of WB and ZnO increased E. coli counts. Two in vitro trials were also designed to clarify hypotheses derived from the in vivo test: (1) the ability of WB and other fibre sources to bind E. coli in vitro (trial 2) and (2) the in vitro interactions between WB and ZnO with respect to E. coli growth (trial 3). We can conclude that incorporation of WB in the diet improved gut health by modulating the activity and composition of the microbial population. The negative interaction between WB and ZnO raises the interest of considering the inclusion of phytase enzymes to reduce the therapeutic levels of ZnO in post-weaning diets.

Administration of loperamide and addition of wheat bran to the diets of weaner pigs decrease the incidence of diarrhoea and enhance their gut maturation.

The influence of fibre inclusion and transit time regulation on the performance, health status, microbial activity and population, physico-chemical characteristics of the hindgut digesta and intestinal morphology in early weaned pigs were examined. For these experiments, wheat bran (WB) was used as fibre source and loperamide as a drug (LOP) to increase the digesta transit time. In Expt 1, a total of 128 early weaned pigs were randomly distributed in a 2 x 2 factorial combination of WB inclusion (0 v. 40 g/kg) and LOP administration (0 v. 0.07 mg/kg body weight) during 13 d. For Expt 2, a total of twenty-four piglets were allotted to three dietary treatments for 15 d with the same basal diet (control diet) as Expt 1; a diet with 80 g/kg of WB and the combination of WB and LOP. In Expt 1, LOP improved the average daily feed intake and average daily gain of the animals (P = 0.001 and 0.007, respectively). The same result was obtained when WB was combined with LOP. The WB-LOP group also showed a higher concentration of SCFA (P = 0.013), acetic acid (P = 0.004) and propionic acid (P = 0.093). On the other hand, WB inclusion reduced the organic matter and crude protein digestibility (P = 0.001) and tended to decrease the enterobacteria population (P = 0.089). In Expt 2, WB increased the butyric acid concentration (P = 0.086). We concluded that the inclusion of WB to modify the intestinal microbiota activity combined with LOP may be beneficial to animal health and performance.