Subtilis PB6 Research Articles

Effects of replacing zinc oxide with a combination of β-glucan, Bacillus subtilis PB6 and formic acid on the performance of weaner pigs

Effects of replacing zinc oxide with a combination of β-glucan, Bacillus subtilis PB6 and formic acid on the performance of weaner pigs

Effect of probiotics and humate substances on blood parameters, intestinal development and immune organs of growing quail

This study aimed to evaluate impacts of dietary probiotics (Bacillus subtilis PB6) and humate substances (HS) supplementation on histomorphometry of small intestine and immune organs, blood parameters of growing quail. A total of 216 unsexed quails (seven days old) were randomly distributed to six groups. The 1st group did not receive any supplements (control), 2nd group received B. subtilis (CloSTAT: 0.5 g/kg diet), 3rd and 4th groups received HS (4 and 8 g/kg diet, respectively), 5th and 6th groups received CloSTAT + 4g HS and CloSTAT + 8g HS, respectively. Results showed that the inclusion of B. subtilis alone in quail diets significantly improved histomorphometry indices of intestine and immune organs compared to the control. Dietary supplementation of HS alone led to deteriorating histomorphometry indices of intestinal segments and immune organs compared to the control. CloSTAT, HS or both improved lipid profile and antioxidant parameters. Serum mineral levels did not differ significantly among groups except for Ca levels. In conclusion, dietary probiotics supplementation enhanced histomorphometry of intestine and immune organs and improved serum Ca, lipid profile and antioxidant indices. Moreover, the addition of HS (4 or 8 g/kg diet) improved lipid profile and antioxidant indices, but led to undesirable results in intestinal development and immune organs.

Effects of feeding CLOSTAT (Bacillus subtilis PB6) on the clinical health, performance, and carcass characteristics of feedlot steers1.

The objectives of this experiment were to evaluate the effects of feeding Bacillus subtilis PB6 on clinical health, performance, and carcass characteristics of feedlot steers. Bos indicus crossbred steer calves (n = 397; 342 kg initial body weight [BW]) were randomly assigned to pens by initial BW; pens (n = 24) were randomly assigned to one of two of the following experimental treatments: 1) no supplemental dietary direct-fed microbial, control (CON; n = 12 pens) or 2) 13 g/steer daily B. subtilis PB6 (CLO; CLOSTAT, Kemin Industries, Des Moines, IA; n = 12 pens). Steers were housed in 12.2 × 30.5 m soil-surfaced pens; pen served as the experimental unit. The percentage of cattle treated once or twice for bovine respiratory disease (BRD) did not differ among treatments (P ≥ 0.27); BRD mortality also did not differ between CON and CLO (P = 0.34). During the receiving period, final BW (P = 0.97), average daily gain (ADG; P = 0.91), dry matter intake (DMI; P = 0.77), and gain:feed (P = 0.79) were not different among treatments. There was a tendency (P = 0.09) for CLO-supplemented steers to be 14% more efficient from days 0 to 14 of the receiving period. Final BW, overall finishing phase ADG, and DMI did not differ by treatment (P ≥ 0.14); ADG was 0.14 kg greater for CLO than CON (P = 0.03) from days 29 to 56 of the finishing period. Gain: feed tended (P = 0.07) to be 7% greater (0.144 vs. 0.141) for CLO than CON throughout the duration of the finishing period, and 6.7% greater (P = 0.08; 0.152 for CLO vs. 0.150 for CON) for the entirety of the experiment. Carcass traits did not differ among treatments (P ≥ 0.31). The results of this experiment suggest that supplementing 13 g/steer daily B. subtilis PB6 may improve feed efficiency in feedlot cattle.

54 The Effect of Supplementing Clostat 500 (Bacillus Subtilis PB6) to Yearling Steers in a Commercial Feedyard on Health, Salmonella spp. Prevalence, Feedlot Performance and Carcass Characteristics

Abstract Crossbred beef steers, n = 2,100; 313 ± 38 kg initial body weight (BW) were used to evaluate Bacillus Subtilis PB6 supplementation to yearling steers. Cattle were blocked by arrival date and assigned randomly to pen within block; pens were randomly assigned to treatment within block. Treatments, replicated in 15 pens/treatment with 70 steers/pen, included: 1) control (CON), diets containing no supplemental direct fed microbials; 2) CLOSTAT (CLO), diets supplemented with 0.5 g/animal/d Bacillus subtilis PB6 (CLOSTAT 500, Kemin Industries, Des Moines, IA). Supplementing CLO decreased morbidity (P = 0.03), 10.38% (CLO) vs. 13.43% (CON), decreased the percentage of cattle treated once for bovine respiratory disease (BRD; P < 0.01), 9.14% (CST) vs. 12.76% (CON), and decreased BRD re-treatment rate (P = 0.03). Mortality did not differ among treatments (P = 0.23). Cattle removed from the study tended to be less for CLO than CON (53 vs. 73 animals, respectively, P = 0.06). The prevalence of fecal Salmonella was not different among treatments, (P ≥ 0.35). Overall fecal Salmonella counts tended to be less for CLO (1.59 log CFU/g) than for CON (2.04 log CFU/g; P = 0.07). Concentration of Salmonella in subiliac lymph nodes did not differ by treatment (P = 0.62); however, mean prevalence of lymph node Salmonella decreased 46% by CLO (28.66% vs. 15.48%, CON vs. CLO, respectively, P = 0.46). With dead animals and removals included, final BW was heavier for CLO steers than CON, (P = 0.05), and average daily gain (ADG; P = 0.08), and gain efficiency (G:F, P = 0.06) tended to be greater for CLO than CON. With dead animals and removals excluded, final BW, ADG, and G:F did not differ among treatments (P ≥ 0.30). Carcass traits were not different between treatments (P ≥ 0.15). Supplementing CLO improved health outcomes of yearling steers.

The effect of supplementing CLOSTAT 500 (Bacillus subtilis PB6) to yearling steers in a commercial feedyard on health, Salmonella spp. prevalence, feedlot growth performance and carcass characteristics.

British and British × Continental crossbred beef steers, n = 2,100; 313 ± 38 kg of initial body weight (BW) were used to evaluate the effects of Bacillus subtilis PB6 supplementation to yearling steers in a commercial feedyard on health, prevalence of Salmonella spp., growth performance, and carcass characteristics. Steers were blocked by arrival date and assigned randomly to pens within the block; pens were randomly assigned to 1 of 2 dietary treatments within block. Treatments, replicated in 15 pens/treatment with 70 steers/pen, included: 1) control (CON), diets containing no supplemental direct-fed microbials; 2) CLOSTAT (CLO), diets supplemented with 0.5 g/steer/d Bacillus subtilis PB6 (CLOSTAT 500, Kemin Industries, Des Moines, IA) to provide 6.6 × 109 CFU/g of the active ingredient. Supplementing CLO decreased the overall incidence of morbidity (P = 0.03), 10.38% (CLO) vs. 13.43% (CON), decreased the percentage of steers treated once for bovine respiratory disease (BRD; P < 0.01), 9.14% (CLO) vs. 12.76% (CON), and decreased the incidence of BRD retreatment (P = 0.03) compared with CON. Mortality did not differ among treatments (P = 0.23); however, overall deads and removals tended to be less for CLO than CON (53 heads vs. 73 heads respectively, P = 0.06). Prevalence of fecal Salmonella did not differ among treatments, (P ≥ 0.35); overall fecal Salmonella counts tended to be less for CLO (1.59 log (10) CFU/g) than CON (2.04 log (10) CFU/g; P = 0.07). Salmonella concentration in subiliac lymph nodes (n =150/treatment) was not different (P = 0.62) between CON (0.22 log (10) CFU/g) or CLO (0.19 log (10) CFU/g); however, there was a 46% reduction in the overall mean prevalence of lymph node Salmonella (P = 0.46; 15.48% vs. 28.66%) for CLO and CON, respectively. With deads and removals included, final BW was heavier for CLO steers than CON, (654 kg vs. 641 kg, respectively, P = 0.05), and average daily gain (ADG; P = 0.08) and gain efficiency (G:F; P = 0.06) tended to be greater for CLO than CON. With deads and removals excluded, final BW, ADG, and G:F did not differ among treatments (P ≥ 0.30). Carcass traits were not different between treatments (P ≥ 0.15). Supplementing CLO throughout the feeding period in a commercial feedyard improved the health outcomes of yearling steers by decreasing BRD and overall treatment rates, reducing the overall abundance of Salmonella, and resulting in fewer steers removed from the study compared with CON.

Safety and efficacy of a feed additive consisting of Bacillus velezensis ATCC PTA-6737 (Bacillus velezensis PB6) for turkeys for fattening, turkeys reared for breeding, laying hens, minor poultry species for laying, piglets (weaned), weaned minor porcine species and sows (Kemin Europe N.V.).

Following a request from the European Commission, EFSA was asked to deliver a scientific opinion on the safety and efficacy of Bacillus subtilis ATCC PTA‐6737 (Bacillus subtilis PB6) when used as a feed additive for turkeys for fattening, turkeys reared for breeding, laying hens, minor poultry species for laying, piglets (weaned), weaned minor porcine species and sows. With this application, the company requested the modification of the current authorisations as regards the strain taxonomy from B. subtilis ATCC PTA‐6737 to Bacillus velezensis ATCC PTA‐6737. The FEEDAP Panel concluded that the active agent of B. subtilis PB6 should be taxonomically designated as B. velezensis ATCC PTA‐6737. The Panel also concluded that Bacillus velezensis PB6 is presumed safe for the target species, consumers and the environment and is not a dermal/eye irritant or a skin sensitiser but should be considered a respiratory sensitiser. The Panel reiterated the previous conclusions reached on the efficacy of the additive when used with the target species.

The Impact of Bacillus subtilis PB6 and Chromium Propionate on the Performance, Egg Quality and Nutrient Metabolizability of Layer Breeders.

Simple SummaryEnvironmental conditions can generate heat stress in poultry production. Feed additives have been supplemented in diets for layers and broilers to improve their health status, immune system and nutrition, as well as productive performance. This paper shows the effects of probiotic Bacillus subtilis PB6 and chromium propionate diet supplementation on productive parameters, egg and eggshell quality, excreta moisture, cortisol and serum biochemistry of laying breeder hens. Diets supplemented with Bacillus subtilis PB6, chromium propionate or a combination of the two resulted in improved egg production, feed conversion ratio, eggshell quality and nutrient metabolizability, without modifying the main serum biochemical parameters of hens from 55 to 70 weeks of age. Highlighted improvements in hen performance and eggshell quality were observed when hens were fed diets supplemented with a combination of the probiotic and chromium. This study expands our understanding concerning the combined supplementation of probiotics and organic chromium for poultry.The objective of this study was to evaluate the effects of Bacillus subtillis PB6, chromium propionate or a combination of the two on the performance, egg and eggshell quality, nutrient metabolizability and serum biochemistry of layer breeders. White Plymouth Rock and Red Rhodes Island breeder hens at 55 weeks of age were allocated in individual cages using a completely randomized block design with 16 replicates. Hens were fed control, control + probiotic (500 g/ton of Bacillus subtilis PB6), control + CrProp (50 g/ton of chromium propionate) and control + probiotic + CrProp diets from 55 to 70 weeks of age. Productive parameters and eggshell quality as well as cortisol and blood biochemistry were grouped each 28 d as well as for the overall period. The metabolizability of nutrients and energy was determined at 70 weeks of age. In the overall period, hens fed the control + probiotic or control + probiotic + CrProp diets had significantly higher egg production, egg mass, shell percentage, thickness and shell strength. The metabolizability of dry matter, nitrogen and energy increased in hens that were fed the control + probiotic + CrProp diet. In conclusion, diets supplemented with Bacillus subtillis PB6 and chromium propionate resulted in improved productive performance, eggshell quality and nutrient metabolizability of layer breeders, without modifying serum cortisol, albumin and triglycerides.

Supplemental Bacillus subtilis PB6 Improves Growth Performance and Gut Health in Broilers Challenged with Clostridium perfringens.

Clostridium perfringens (CP) is the principal pathogenic bacterium of chicken necrotic enteritis (NE), which causes substantial economic losses in poultry worldwide. Although probiotics are known to provide multiple benefits, little is known about the potential effects of Bacillus subtilis (B. subtilis) application in preventing CP-induced necrotic enteritis. In this study, 450 male Arbor Acres broilers were divided into 5 experimental treatments: A: basal diet (control group); B: basal diet and CP challenge (model group); C: CP challenge+10 mg/kg enramycin (positive control group); D: CP challenge+4 × 107 CFU/kg of feed B. subtilis PB6 (PB6 low-dosage group); and E: CP challenge+6 × 107 CFU/kg of feed B. subtilis PB6 (PB6 high-dosage group). There were 6 replicate pens per treatment with 15 broilers per pen. The present research examined the effect of Bacillus subtilis PB6 (B. subtilis PB6) on growth performance, mRNA expression of intestinal cytokines and tight junctions, and gut flora composition in broilers challenged with CP. The entire experiment was divided into two phases: the non-CP challenge phase (d0–18) and the CP challenge phase (d18–26). PB6 did not increase the growth performance during the first stage, but the PB6 high-dosage group was found to have larger body weight gain and ADFI during the CP challenge stage. Feed supplementation with PB6 reduced the lesion score of challenged chicks, with increased tight junction-related gene expression (occludin and ZO-1) and decreased TNF-α expression compared with CP-infected birds. A decrease in the abundance of Clostridium XI, Streptococcus, and Staphylococcus was observed after CP infection (P < 0.05), while supplementation with PB6 restored the ileal microbial composition. In conclusion, administration of B. subtilis PB6 improved growth performance, enhanced intestinal barrier function, and mitigated intestinal inflammation/lesions, which might be due to its restoring effects on the ileal microbial composition in CP-challenged broilers.

159 Evaluation of Bacillus Subtilis PB6 Probiotic (CLOSTAT® 500) on Feedlot Phase Growth Performance, Efficiency of Dietary Net Energy Utilization, and Fecal and Subiliac Lymph Node Salmonella Prevalence

Abstract Yearling beef steers (n = 238; initial BW=402 ± 31.2 kg) were used to evaluate a Bacillus subtilis probiotic on growth performance, dietary net energy (NE) utilization, carcass characteristics, and fecal and subiliac lymph node Salmonella prevalence during a 140-d finishing period. Steers were allotted to 24 pens (n = 9 to 10 steers/pen) and assigned to one of two treatments (12 pens/treatment): no probiotic (CON) or 0.50 g·steer-1·d-1 of a Bacillus subtilis PB6 probiotic (CLOSTAT® 500, Kemin Industries, Des Moines, IA; CLO). Steers were transitioned to a 90% concentrate diet (DM basis) over 14-d. Steers were fed once daily at 0700 h; bunks were managed according to a slick bunk management. Fecal samples were collected on d 1, 28, 56, 112, and 140 from each pen (n = 5 steers/pen) via rectal palpation and composited by pen for determination of Salmonella prevalence. Upon harvest, subiliac lymph nodes were obtained from 60 steers in CON and 57 steers in CLO. Data were analyzed as a randomized complete block design; pen was the experimental unit; α of 0.05 determined significance. No differences were detected (P ≥ 0.25) between treatments for live or carcass-adjusted average daily gain, dry matter intake, gain efficiency, dietary NE utilization, nor calculated dietary NE content based upon performance. No differences were detected between treatments for any carcass traits (P ≥ 0.15). Salmonella was not recovered in any fecal samples except on d 112, where steers from CLO had a numerically lower (P = 0.17; 8.3 vs. 25.0%) incidence of fecal Salmonella compared to CON and on d 140 fecal, where Salmonella incidence did not differ (P = 0.34; 0.0 vs. 8.3%) for CON and CLO, respectively. Salmonella was not recovered in any subiliac lymph nodes. These data indicate that CLO did not influence growth performance or Salmonella prevalence.

Impact of humate substances and Bacillus subtilis PB6 on thyroid activity and histomorphometry, iron profile and blood haematology of quail

The purpose of this study was to assess the effects of dietary humate substances (HS) and CloSTAT (Bacillus subtilis PB6) on the thyroid activity and histology, iron profile, blood haematology and performance of growing Japanese quail. A total of 216 unsexed 7-day-old quail chicks were randomly assigned to six groups. The first group was fed a basal diet (BD) without any additives (control); the 2nd group received BD plus 0.05% CloSTAT, the 3rd and 4th groups were given BD plus 0.4% and 0.8% HS, respectively; and the 5th and 6th groups were administered BD plus CloSTAT+0.4% HS and BD plus CloSTAT+0.8% HS, respectively. The results showed that the growth performance was improved with the addition of CloSTAT alone or in combination with 0.4% HS compared with the control. Haematological parameters, iron level and transferrin saturation % were significantly (p<0.001) increased by feeding HS compared with the control group. Serum thyroxin and triiodothyronine levels were significantly (p=0.001) increased by adding CloSTAT relative to the control. Supplementation of 0.8% HS caused deterioration in histomorphometry parameters of the thyroid gland, but these parameters were improved in response to CloSTAT compared with the control. In conclusion, dietary B.subtilis PB6 as CloSTAT or CloSTAT+0.4% HS supplementation may be efficacious in enhancing the growth performance and boosting the thyroid activity of growing Japanese quail. Moreover, the addition of 0.4% or 0.8% HS to quail diets boosted their iron profile and haematological parameters.

Quantifying the impact of eight unit operations on the survival of eight Bacillus strains with claimed probiotic properties

This study assessed the impact of eight unit operations [slow pasteurization, high-temperature short time (HTST) pasteurization, cooking, baking, drying, fermentation, supercritical carbon dioxide (CO2), irradiation and extrusion] in different food matrices (milk, orange juice, meatballs, bread, crystallized pineapple, yogurt, orange juice, ground black pepper, snacks, and spaghetti) on the resistance of eight (Bacillus flexus Hk1 Bacillus subtilis Bn1, Bacillus licheniformis Me1, Bacillus mojavensis KJS3, Bacillus subtilis PXN21, Bacillus subtilis PB6, Bacillus coagulans MTCC 5856 and Bacillus coagulans GBI-30, 6086) Bacillus strains with claimed probiotic properties (PB). The number of decimal reductions (γ) caused by the unit operations varied (p < 0.05) amongst the PB. Most of the unit operations caused ≤ 2 γ of PB in the food matrices evaluated. Irradiation caused up to 4.9 γ (p < 0.05) amongst the PB tested. B. subtilis Bn1, B. mojavensis KJS3, B. licheniformis Me1, and B. coagulans GBI-30 showed higher resistance to most of the tested unit operations. These results indicate that the choice of PB for application in foods should also be based on their resistance to unit operations employed during processing. Finally, the high resistance of PB to the unit operations tested comprise valuable data for the development and diversification of probiotic foods with sporeforming strains with claimed probiotic properties.

Evaluation of Bacillus subtilis PB6 on feedlot phase growth performance, efficiency of dietary net energy utilization, and fecal and subiliac lymph node Salmonella prevalence in spring placement yearling beef steers fed in southeastern South Dakota,.

Yearling crossbred beef steers [N = 238; initial shrunk body weight (BW) = 402 ± 31.2 kg] were used to investigate the influence of a Bacillus subtilis probiotic on animal growth performance, efficiency of dietary net energy (NE) utilization, carcass characteristics, and fecal and subiliac lymph node Salmonella prevalence during a 140-d finishing period at the Southeast Research Farm in Beresford, SD. Steers were allotted to 1 of 24 pens (N = 9–10 steers/pen) and assigned to 1 of 2 dietary treatments (12 pens/treatment): no probiotic (CON) or 0.5 g/steer/d of a B. subtilis PB6 probiotic (CLOSTAT500, Kemin Industries, Des Moines, IA; CLO). Bunks were managed according to a slick bunk management approach. Fecal samples were collected on study days 1, 28, 56, 112, and 140 from a subsample of steers from each pen (N = 5 steers/pen) via rectal palpation and composited by pen for the determination of Salmonella prevalence using selective enrichment and culture media. Upon harvest, subiliac lymph nodes were obtained from an equal number of steers from each treatment (collected from every other steer) following evisceration and hide removal. Data were analyzed as a randomized complete block design and pen served as the experimental unit; an α of 0.05 determined significance. Live-basis final BW and average daily gain tended (P ≤ 0.06) to be reduced for CLO. No differences were detected (P ≥ 0.11) between treatments for dry matter intake or gain efficiency. Treatment neither altered the efficiency of dietary NE utilization nor calculated dietary NE content based upon observed performance (P ≥ 0.46). No differences were detected between treatments for any carcass traits (P ≥ 0.15). No Salmonella was recovered in any fecal samples collected on study days 1, 28, or 56. On day 112, steers from CLO had a numerically lower (P = 0.17; 25.0 vs. 8.3%) incidence of fecal Salmonella compared to CON. On study day 140, fecal Salmonella incidence did not differ between treatments (P = 0.34; 0.0 vs. 8.3%) for CON and CLO, respectively. Upon harvest, no Salmonella was recovered in any subiliac lymph nodes. These data indicate that B. subtilis PB6 did not influence feedlot phase growth performance or fecal Salmonella prevalence. Additionally, Salmonella was not observed in the subiliac lymph nodes of any steers upon harvest.

Effects of Bacillus subtilis PB6 supplementation on productive performance, egg quality and hatchability in broiler breeder hens under commercial farm condition

ABSTRACT The aim of the present study was to determine the effects of Bacillus subtilis PB6 supplementation on productive performance, egg quality and hatchability of broiler breeder hens. A total of 20,000 broiler breeder hens (ROSS 308), 57 weeks old, were assigned to the two equal groups as follows: The first group was fed a basal diet and the second group was fed the basal diet that supplemented with the Bacillus subtilis PB6 (2 × 107 cfu/g) at the rate of 1 g/kg of diet. Results indicated that adding Bacillus subtilis significantly improves egg production (P < 0.05). The body weights and egg weight were not affected by the dietary treatments (P > 0.05) but Bacillus subtilis improved the egg haugh unit (P < 0.05). Adding Bacillus subtilis significantly increased hatching egg, eggshell thickness and egg hatchability (P < 0.05). A highly significant improvement was noticed in the faecal score of the group fed with Bacillus subtilis (P < 0.01). Adding Bacillus subtilis increased the level of total cholesterol (P < 0.01) and decreased the blood heterophil to lymphocyte ratio (P < 0.01) in breeders’ serum. It is concluded that Bacillus subtilis PB6 supplementation at the level of 1 g/kg of broiler breeder diet, significantly improve the egg production, egg quality, hatchability, faecal score and return on investment.

211 Evaluation of CLOSTAT-500® on growth performance and fecal Salmonella prevalence in yearling beef steers during the feedlot receiving phase

Abstract Two-hundred and thirty eight crossbred beef steers (initial BW = 419 ± 32.4 kg) were used to evaluate the influence of a patented probiotic on growth performance responses and fecal Salmonella prevalence during a 28-d feedlot receiving period at the South East Research Farm (SERF) feedlot located near Beresford, SD. Steers were allotted to 24 pens (9 to 10 steers/pen) and assigned to one of two treatments (12 pens/treatment): no probiotic (CON) or 0.5 g·steer-1·d-1 of a Bacillus subtilis PB6 probiotic (CLOSTAT-500®, Kemin Industries, Des Moines, IA; CLO). Steers were transitioned from a 70% concentrate diet (DM basis) to a 92% concentrate diet (DM basis) over a 14-d period. The final diet contained (DM basis): 12.5% CP, 2.08 Mcal/kg of NEm, 1.40 Mcal/kg of NEg, 30 g/907-kg of monensin sodium, and 0.45 mg/kg of chromium propionate (KemTRACE® Chromium, Kemin Industries, Des Moines, IA). Fecal samples were collected on study d 1 and 28 (6 and 34 d following arrival to the SERF, respectively) from sentinel steers (n = 5/pen) via rectal palpation and composited by pen for determination of Salmonella prevalence using selective enrichment and culture medias. Steers were fed once daily and bunks were managed according to a slick bunk management approach. Data were analyzed as a randomized complete block design; pen served as the experimental unit and an α of 0.05 determined significance. No differences were detected (P ≥ 0.25) between treatments for ADG, DMI, or gain efficiency during the initial 28-d feedlot receiving phase. Additionally, no Salmonella was recovered in any fecal samples collected. These data indicate that Bacillus subtilis PB6 had no influence on receiving phase growth performance, and fecal Salmonella prevalence was not observed in yearling steers placed on feed in March in southeastern South Dakota.

Effects of Spirulina (Arthrospira) platensis and Bacillus subtilis PB6 on growth performance, intestinal microbiota and morphology, and serum parameters in broiler chickens

Context The use of microalgae as prebiotics in poultry diets may improve production efficiency by modifying the gastrointestinal ecosystem. Prebiotic properties of the cyanobacterium Spirulina (Arthrospira) platensis have been confirmed, but effects of its combination with probiotics on broiler chicken performance are unclear. Aims This experiment was designed to study the effects of different levels of a microalga (Spirulina platensis) and a probiotic (Bacillus subtilis PB6, BSPB) on performance, intestinal microbiota and morphology, and blood parameters in broiler chickens. Methods One-day-old Ross 308 broiler chickens (300 in total) were allocated to six treatments in a completely randomised design with 3 × 2 factorial arrangement including three levels of Spirulina (0%, 0.05% or 0.1% of diet) and two levels of BSPB (0% or 0.05% of diet) and were reared for 42 days. Key results Individual use of Spirulina and BSPB improved bodyweight gain and feed conversion ratio of broiler chickens. The relative weight of the breast cut was greater in broilers treated with 0.05% Spirulina than in those receiving Spirulina-free treatments. The count of lactic acid bacteria was higher in the ileum of birds receiving only BSPB than in those receiving diets without BSPB. The use of 0.1% Spirulina in the diet decreased the pH and coliform population in the ileum compared with 0% Spirulina. There was an interaction between Spirulina and BSPB on villus height (VH) and the VH:crypt depth ratio in the small intestine. Co-supplementation with 0.1% Spirulina and BSPB increased VH and the VH:crypt depth ratio in the duodenum compared with diets supplemented, respectively, with 0.1% Spirulina and BSPB alone. Broiler chickens fed diets with BSPB had higher activities of superoxide dismutase and glutathione peroxidase and lower concentrations of cholesterol and triglyceride in serum. Conclusions The results show that supplementation with Spirulina and BSPB had a positive effect on performance and carcass quality of broiler chickens. The synergistic interaction between these supplements leads to enhancement of epithelial morphology in the small intestine. Implications Because of the potential of Spirulina and BSPB to create beneficial changes in the intestinal ecosystem and serum parameters, these dietary additives could be recommended for improving the quantity and quality of meat products and health status in broiler chickens.

Dietary supplementation of Bacillus subtilis PB6 improves sow reproductive performance and reduces piglet birth intervals.

We investigated the effects of dietary supplementation with Bacillus subtilis PB6 (B. subtilis PB6) during late gestation and lactation on sow reproductive performance, antioxidant indices, and gut microbiota. A total of 32 healthy Landrace × Yorkshire sows on d 90 of gestation were randomly assigned to 2 groups, with 16 replicates per group, receiving basal diet (CON) or the basal diet + 0.2% B. subtilis PB6, containing 4.0 × 108 CFU/kg of feed (BS). The litter sizes (total born) and numbers of piglets born alive were larger in the BS group (P < 0.01), whereas the weights of piglets born alive and the piglet birth intervals were lower in the BS group (P < 0.05). Although the litter weights and piglet bodyweights (after cross-fostering) were lower after BS treatment (P < 0.05), the litter sizes, litter weights, lactation survival rate, and litter weight gains at weaning were higher in BS group (P < 0.05). The concentrations of malondialdehyde (MDA) in the sow sera at parturition were lower in the BS group (P < 0.01). The serum total antioxidant capacity (T-AOC) at parturition and the serum catalase (CAT) concentrations on d 21 of lactation were higher in the BS group (P < 0.05). Dietary supplementation with B. subtilis PB6 (P < 0.05) reduced the serum endotoxin concentrations in the sows and the serum cortisol concentrations of the piglets at d 14 of lactation. The α-diversity indices of microbial were higher in the CON group (P < 0.05). At the phylum level, B. subtilis PB6 supplementation increased the relative abundances of Gemmatimonadete and Acidobacteria (both P < 0.01) and reduced those of Proteobacteria, and Actinobacteria (both P < 0.05). At the genus level, B. subtilis PB6 supplementation increased the relative abundance of Ruminococcaceae_UCG-013 cc (P < 0.05) and reduced that of Streptococcus (P < 0.05). This study demonstrated that adding 4.0 × 108 CFU/kg B. subtilis PB6 to sows’ feed during late gestation and lactation could shorten piglet birth intervals, enhance the growth performance of suckling piglets, and improve the gut health of sows during late gestation.

Interactive effects of dietary vitamin K3 and Bacillus subtilis PB6 on the growth performance and tibia quality of broiler chickens with sex separate rearing

Both vitamin K and probiotics can promote the bone health of poultry and mammals. The present study was conducted to investigate the interactive effects between vitamin K3 (VK3) and Bacillus subtilis PB6 on the growth performance and tibia quality of broiler chickens with sex separate rearing. In a 3 × 2 × 2 factorial arrangement, 720 one-day-old broiler chicks (Arbor Acres) were assigned to 12 groups with three levels of dietary VK3 (0, 0.5 and 4.0 mg/kg), with or without probiotic supplementation (500 g/t) and with sex separation (male and female). Each group included 3 replicates with 20 birds per replicate. During day 1 to 21, 0.5 and 4.0 mg/kg of VK3 increased average daily gain (ADG) of all birds and average daily feed intake of male birds (P < 0.05). During day 22 to 42, probiotic supplementation increased the ADG of birds (P < 0.05). Probiotic addition increased the weight, length, diameter and strength of tibia in all birds, and 0.5 and 4.0 mg/kg of VK3 increased the tibial breaking strength of male birds at day 21 (P < 0.05). Vitamin K3 and probiotic synergistically increased tibial breaking strength at day 42 and ash content at day 21 (P < 0.05). Three factors exhibited interactive effects on the chemical composition of tibia at day 42, and female birds fed 4 mg/kg of VK3 and probiotic had the highest contents of ash, calcium and phosphorus (P < 0.05). Bacillus subtilis PB6 increased the serum phosphorus level of male birds at day 21 and serum calcium level of female ones at day 42 (P < 0.05). At day 21, in the probiotic-supplemented birds, serum osteocalcin (OCN) and bone-specific alkaline phosphatase (BALP) were increased by 0 and 4.0 mg/kg of VK3, respectively (P < 0.05). Probiotic increased serum OCN and cooperated with VK3 to increase the serum BALP at day 42 (P < 0.05). Vitamin K3 and probiotic synergistically down-regulated the mRNA expression of Runt-related transcription factor 2 and OCN at day 21 (P < 0.05). Vitamin K3 down-regulated the alkaline phosphatase (liver/bone/kidney) expression in male birds at day 21 and 42, but probiotic up-regulated the expression of these genes at day 42 (P < 0.05). In conclusion, VK3 and B. subtilis PB6 promoted the growth performance of broilers during starter and grower phases, respectively. They synergistically improved the physical and chemical traits of tibias, especially in grower phase, by modulating calcium and phosphorus metabolism as well as osteogenic gene expression.

Behavior of different Bacillus strains with claimed probiotic properties throughout processed cheese (“requeijão cremoso”) manufacturing and storage

Even though spore-forming bacteria have been mainly linked to spoilage or foodborne pathogens vehiculated through foods, some strains of Bacillus can potentially present probiotic properties. The advantage of incorporating probiotic Bacillus strains in foods relies mainly on the fact that these microorganisms present high resistance to harsh processing conditions. “Requeijão cremoso” is a type of processed cheese highly appreciated in Brazil. During processing, this product is submitted to several harsh conditions (heating at 90 °C, for instance), leading to the inactivation of probiotic bacteria belonging to Lactobacillus and Bifidobacterium genera. That fact has precluded the development of probiotic “requeijão cremoso” products; however, probiotic Bacillus strains may comprise a promising alternative to overcome the low resistance of traditional probiotics to food processing. The objective of this study was to evaluate the behavior of different Bacillus strains with claimed probiotic properties throughout processed cheese (“requeijão cremoso”) manufacturing. A total of five different Bacillus strains with claimed probiotic properties (B. coagulans MTCC 5856, B. coagulans GBI-30 6086, B. subtilis PXN 21, B. subtilis PB6, and B. flexus HK1) were individually inoculated at different stages of manufacture – curd pasteurization, coagulation, and fusion – of “requeijão cremoso” and their survival in each of these stages was determined. The survival of B. coagulans GBI-30 6086 was further assessed throughout “requeijão cremoso” production and shelf life (45 days at 6 °C). Besides, the chemical composition, level of proteolysis, and fatty acid profile of the treatments during shelf life were evaluated. The fusion stage was found as the most appropriate for the addition of B. coagulans GBI-30 6086, which allows the production of probiotic “requeijão cremoso” and facilitates the technological process while preventing the occurrence of final product recontamination.

Intestinal Morphologic and Microbiota Responses to Dietary Bacillus spp. in a Broiler Chicken Model.

Dietary inclusion of probiotic Bacillus spp. beneficially affect the broiler chickens by balancing the properties of the indigenous microbiota causing better growth performance. The effects of three Bacillus spp. on the growth performance, intestinal morphology and the compositions of jejunal microflora were investigated in broiler chickens. A total of 480 1-day-old male Arbor Acres broilers were randomly divided into four groups. All groups had six replicates and 20 birds were included in each replicate. The control birds were fed with a corn-soybean basal diet, while three treatment diets were supplemented with Bacillus coagulans TBC169, B. subtilis PB6, and B. subtilis DSM32315 with a dosage of 1 × 109 cfu/kg, respectively. The experiment lasted for 42 days. The compositions and diversity of jejunal microflora were analyzed by MiSeq high-throughput sequencing. The B. coagulans TBC169 group showed marked improvements of growth performance, nutrient digestibility and intestinal morphology compared with the other B. subtilis treatments. B. coagulans TBC169 supplementation improved the average body weight (BW), average daily weight gain (ADG), total tract apparent digestibility of crude protein and gross energy (GE), and reduced feed conversion rate (FCR) compared with the control group (P < 0.05). The villus height to crypt depth ratio (VH/CD) of jejunum and duodenum was increased in the birds fed with B. coagulans TBC169 compared with the control group (P < 0.05). However, two B. subtilis treatments presented more positive variation of the jejunum microflora of chickens than that in the B. coagulans TBC169 group. B. subtilis PB6 and B. subtilis DSM32315 treatments improved the diversity of jejunal microbiota on day 21 compared with the control (P < 0.05), while which were decreased on day 42 (P < 0.05). The supplementation with B. coagulans TBC169 significantly improved the proportion of Firmicutes, otherwise two B. subtilis significantly improved the proportion of Proteobacteria, Bacteroidetes, Actinobacteria, and Acidobacteria at the phylum level during starter phase and decreased the proportion of Bacteroidetes during growing phase compared with the control. The supplementation with B.subtilis DSM32315 significantly improved the proportion of Clostridiales during starter phase, whereas two B. subtilis significantly improved the proportion of Pseudomonas, Burkholderia, Prevotella, DA101 during growing phase at the genus level compared with the control. In conclusion, the dietary supplementation with probiotic Bacillus spp. strains improved body weight and intestinal morphology in broiler chickens, which might be associated with the gut microbiota.

Use of Bacillus Subtilis PB6 as a potential antibiotic growth promoter replacement in improving performance of broiler birds

The intestinal gut health is one of the primary determinants of broiler growth and performance. Among the various enteric diseases, necrotic enteritis (NE) is an enterotoxemic disease caused by Clostridium perfringens, which can result in severe economic losses in poultry farming. Antibiotics like bacitracin methylene disalicylate (BMD) and avilamycin (AVL) are commonly used antibiotic growth promoters (AGP) in poultry feed to control necrotic enteritis in birds. Bacillus subtilis PB6 was reported to prevent necrotic enteritis and improve performance in birds. This paper investigated the influence of Bacillus subtilis PB6 in improving the performance of broiler birds in comparison with BMD and avilamycin. A 35 day trial was conducted with 240 day-old commercial broiler chicks (VenCobb 400), which were divided into four treatment groups, where each treatment group was composed of 6 replicates each containing 10 birds, for a total of 60 birds per treatment. The treatment groups included a negative control (no AGP), Bacillus subtilis PB6, BMD, and avilamycin. The parameters analyzed included body weight, feed conversion ratio (FCR), mortality, villus histomorphometry, and European efficiency factor (EEF). Bacillus subtilis PB6 significantly (P < 0.05) improved body weight and FCR (8 points) compared to the control. The group supplemented with B. subtilis PB6 or BMD had higher (P < 0.05) body weight compared to all other treatment groups. The supplementation of B. subtilis PB6 significantly improved the villus height (P < 0.05) compared to control and other AGP groups. The EEF was found to be the highest in the B. subtilis PB6 supplemented group at 35th day as compared to other treatment groups. The combined data from this study indicate that supplementation of B. subtilis PB6 improves overall performance of broilers compared to BMD and avilamycin, and can be used as potential AGP replacement in poultry farming.