Methylene blue active substances in plaque of Bacillus subtilis subsp. subtilis and enrichment by supplemental calcium in culture media.

A series of experiments was conducted to identify the molecular species responsible for surface active emulsification (surfactant) bioactivity in Bacillus subtilis subsp. subtilis strain ATCC PTA-125135, and to describe culture conditions to support the enriched production of said bioactivity in cultured plaque of the strain. The assay for methylene blue active substances (MBAS) was found to be suitable for describing surfactant activity, where a solvent-extracted molecular fraction from the biofilm was found to retain surfactant activity and positively quantified as MBAS. Furthermore, an HPLC-refined protein fraction was found to quantify as MBAS with approximately 1·36-fold or greater surfactant activity per mol than sodium dodecyl sulphate, and a proteomic analysis of solvent extracted residues confirmed that biofilm surface layer protein BslA was a primary constituent of extracted residues. Surfactant bioactivity, quantified as MBAS, was enriched in cultured plaque by the supplementation of culture media with calcium chloride or calcium nitrate.

Titration of supplemental Bacillus subtilis subsp. subtilis American Type Culture Collection PTA-125135 to broiler chickens fed diets of 2 different metabolizable energy concentrations.

Bacillus subtilis subsp. subtilis American Type Culture Collection deposit number PTA-125135 has recently been studied by our laboratory as a potential probiotic strain for avian species. The objective of the present study was to evaluate growth performance and feed efficiency in broiler chickens in response to a dose titration of the Bacillus strain in feed. In addition to a nonsupplemented control, Bacillus spores were supplemented into broiler chicken diets at 4 levels, which were 8.1 × 104, 1.6 × 105, 2.4 × 105, and 3.2 × 105 CFU per g of feed. The titration was applied to two different dietary regimes of standard or low metabolizable energy (ME), which differed in ME by 22, 56, and 110 kcal/kg in starter, grower, and finisher dietary phases, respectively. All diets contained 249 g per metric ton of a previously patented synbiotic feed additive. Performance data were collected at day 14, 26, and 40 of age, and the effects of Bacillus and ME treatments were evaluated by factorial ANOVA. Treatment group means were further examined for significant (P < 0.05) pairwise differences among treatments and for significant (P < 0.05) linear and quadratic effects. At day 14 of age, significant linear effects for decreased feed conversion ratio (FCR) with higher CFU of Bacillus supplementation were observed within the standard ME diet. At day 26, a linear trend was observed for increased mortality with increased dose within the standard ME diet only. Bacillus supplementation at day 26 also significantly affected FCR and mortality-adjusted FCR, where supplementation with 3.2 × 105 CFU per g feed produced lower FCR and mortality-adjusted FCR than supplementation with 1.6 × 105 CFU per g feed. We conclude from linear effects related to feed efficiency observed at day 14 and from the significant separation of Bacillus treatment means within the titrated range of supplementation at day 26 that further evaluation for effects on performance should be made of doses at 2.4 × 105, 3.2 × 105, and greater CFU per g in feed.

Avi-Lution® supplemented at 1.0 or 2.0 g/kg in feed improves the growth performance of broiler chickens during challenge with bacitracin-resistant Clostridium perfringens.

Avi-Lution® is a defined, patented, synbiotic product containing Saccharomyces cerevisiae, Enterococcus faecium, and Bacillus spp. Broiler chickens (n = 1,250) were experimentally treated as uninoculated controls (uCon), inoculated controls (iCon) with Clostridium perfringens, or inoculated and treated with bacitracin methylene disalicylate (BMD) at 55 mg/kg as an infected/treated control or Avi-Lution® at 1.0 (AvL1) or 2.0 (AvL2) g/kg in feed for 42 d. Each treatment was applied to 10 replicate pens of 25 straight-run, newly hatched chicks. Pens treated with AvL1, AvL2, or BMD showed improved growth, feed efficiency, or mortality from necrotic enteritis compared with iCon pens at d 14, 28, and 42. No differences in these measurements, however, were observed between pens treated with AvL1 and AvL2, which suggests that Avi-Lution® was effective at 1.0 g/kg in feed. Despite improved performance, BMD, AvL1, and AvL2 treatments did not decrease the severity of intestinal lesion scores through 42 d of age compared with the infected control. These results demonstrate that Avi-Lution® improved growth performance and feed conversion rates in broilers challenged with Clostridium perfringens despite no difference in severity of intestinal lesion scores.

Effects of dietary supplementation of an enzyme blend on the ileal and fecal digestibility of nutrients in growing pigs.

The objective of this experiment was to determine the effect of a beta-glucanase-protease enzyme blend product (EBP) on fecal digestibility (FD), apparent ileal digestibility (AID), standardized ileal digestibility, and digestibility in the hindgut of growing pigs. Twelve ileal-cannulated, growing barrows (38.2 +/- 0.5 kg) were housed in individual metabolism crates, blocked by previous feed intake into 3 groups with 4 pigs each, and randomly assigned to 1 of 4 treatments within a square (group) of 3 replications of 4 x 4 Latin square design. Treatments were basal diet (Basal), Basal + 0.05% of EBP (0.05% EBP), Basal + 0.10% of EBP (0.10% EBP), and hydrolyzed casein for measurement of endogenous amino acids. The Basal diet consisted of corn and soybean meal and was calculated to have 3.36 Mcal of ME/kg and 1.1% of total lysine, as-fed basis. Feed intake of each replicate of the Latin square during the first period was 85% of the minimum feed intake of the 4 pigs during the preliminary period and was equalized within each square. The feeding level was increased by 100 g/d in each subsequent period. Each of the experimental periods was 14 d, including 4 d of dietary adaptation, 5 d of fecal collection, 3 d of transition period, and 2 d of ileal collection. Ileal effluents were collected continuously for the same 12-h interval each day. Pigs fed the EBP demonstrated increased (P < 0.05) FD of DM, OM, energy, CP, nonfiber carbohydrate, total dietary fiber, insoluble dietary fiber, acid-hydrolyzed fat, ash, Ca, and P compared with pigs fed Basal. The AID of NDF and hemicellulose was increased (P < 0.05) by supplying the EBP either at 0.05 or 0.10% in the diets, but AID of DM and energy was not increased. The AID of acid-hydrolyzed fat tended to be greater (P = 0.051) for the pigs fed the EBP than for those fed Basal. Ileal digestibility of most amino acids was not affected by treatment, but the EBP reduced the apparent and standardized digestibility of methionine, alanine, and serine (P < 0.05). The difference between FD and AID of hemicellulose was lower (P < 0.05) for the pigs fed the EBP than for those fed Basal. These results demonstrated that the EBP fed to growing pigs improved the FD of DM, OM, energy, CP, nonfiber carbohydrate, total dietary fiber, acid-hydrolyzed fat, Ca, and P, and the AID of NDF and hemi-cellulose, but the standardized ileal digestibility of amino acids was not improved by supplying the EBP in corn-soybean meal-based diets of growing pigs.