Dataset Information

Combinatorial Glycomic Analyses to Direct CAZyme Discovery for the Tailored Degradation of Canola Meal Non-Starch Dietary Polysaccharides.

ABSTRACT: Canola meal (CM), the protein-rich by-product of canola oil extraction, has shown promise as an alternative feedstuff and protein supplement in poultry diets, yet its use has been limited due to the abundance of plant cell wall fibre, specifically non-starch polysaccharides (NSP) and lignin. The addition of exogenous enzymes to promote the digestion of CM NSP in chickens has potential to increase the metabolizable energy of CM. We isolated chicken cecal bacteria from a continuous-flow mini-bioreactor system and selected for those with the ability to metabolize CM NSP. Of 100 isolates identified, Bacteroides spp. and Enterococcus spp. were the most common species with these capabilities. To identify enzymes specifically for the digestion of CM NSP, we used a combination of glycomics techniques, including enzyme-linked immunosorbent assay characterization of the plant cell wall fractions, glycosidic linkage analysis (methylation-GC-MS analysis) of CM NSP and their fractions, bacterial growth profiles using minimal media supplemented with CM NSP, and the sequencing and de novo annotation of bacterial genomes of high-efficiency CM NSP utilizing bacteria. The SACCHARIS pipeline was used to select plant cell wall active enzymes for recombinant production and characterization. This approach represents a multidisciplinary innovation platform to bioprospect endogenous CAZymes from the intestinal microbiota of herbivorous and omnivorous animals which is adaptable to a variety of applications and dietary polysaccharides.

SUBMITTER: Low KE

PROVIDER: S-EPMC7761036 | biostudies-literature | 2020 Nov

REPOSITORIES: biostudies-literature

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Combinatorial Glycomic Analyses to Direct CAZyme Discovery for the Tailored Degradation of Canola Meal Non-Starch Dietary Polysaccharides.

Low Kristin E KE Xing Xiaohui X Moote Paul E PE Inglis G Douglas GD Venketachalam Sivasankari S Hahn Michael G MG King Marissa L ML Tétard-Jones Catherine Y CY Jones Darryl R DR Willats William G T WGT Slominski Bogdan A BA Abbott D Wade DW

Microorganisms 20201129 12

Canola meal (CM), the protein-rich by-product of canola oil extraction, has shown promise as an alternative feedstuff and protein supplement in poultry diets, yet its use has been limited due to the abundance of plant cell wall fibre, specifically non-starch polysaccharides (NSP) and lignin. The addition of exogenous enzymes to promote the digestion of CM NSP in chickens has potential to increase the metabolizable energy of CM. We isolated chicken cecal bacteria from a continuous-flow mini-biore ...[more]

PMID: 33260318

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Project description:Soluble fibres (non-starch polysaccharides, NSP) from edible plants but particularly plantain banana (Musa spp.), have been shown in vitro and ex vivo to prevent various enteric pathogens from adhering to, or translocating across, the human intestinal epithelium, a property that we have termed contrabiotic. Here we report that dietary plantain fibre prevents invasion of the chicken intestinal mucosa by Salmonella. In vivo experiments were performed with chicks fed from hatch on a pellet diet containing soluble plantain NSP (0 to 200 mg/d) and orally infected with S.Typhimurium 4/74 at 8 d of age. Birds were sacrificed 3, 6 and 10 d post-infection. Bacteria were enumerated from liver, spleen and caecal contents. In vitro studies were performed using chicken caecal crypts and porcine intestinal epithelial cells infected with Salmonella enterica serovars following pre-treatment separately with soluble plantain NSP and acidic or neutral polysaccharide fractions of plantain NSP, each compared with saline vehicle. Bacterial adherence and invasion were assessed by gentamicin protection assay. In vivo dietary supplementation with plantain NSP 50 mg/d reduced invasion by S.Typhimurium, as reflected by viable bacterial counts from splenic tissue, by 98.9% (95% CI, 98.1-99.7; P<0.0001). In vitro studies confirmed that plantain NSP (5-10 mg/ml) inhibited adhesion of S.Typhimurium 4/74 to a porcine epithelial cell-line (73% mean inhibition (95% CI, 64-81); P<0.001) and to primary chick caecal crypts (82% mean inhibition (95% CI, 75-90); P<0.001). Adherence inhibition was shown to be mediated via an effect on the epithelial cells and Ussing chamber experiments with ex-vivo human ileal mucosa showed that this effect was associated with increased short circuit current but no change in electrical resistance. The inhibitory activity of plantain NSP lay mainly within the acidic/pectic (homogalacturonan-rich) component. Supplementation of chick feed with plantain NSP was well tolerated and shows promise as a simple approach for reducing invasive salmonellosis.

Project description:An experiment was conducted to determine the effects of including canola meal (CM) in nursery pig diets on growth performance, immune response, fecal microbial composition, and gut integrity. A total of 200 nursery pigs (initial body weight = 7.00 kg) were obtained in two batches of 100 pigs each. Pigs in each batch were housed in 25 pens (four pigs per pen) and fed five diets in a randomized complete block design. The five diets were corn-soybean meal (SBM)-based basal diets with 0%, 10%, 20%, 30%, or 40% of CM. The diets were fed in three phases: phase 1: day 0 to 7, phase 2: day 7 to 21, and phase 3: day 21 to 42. Diets in each phase were formulated to similar net energy, Ca, and digestible P and amino acid contents. Feed intake and body weight were measured by phase. Immune response and gut integrity parameters were measured at the end of phases 1 and 2. Fecal microbial composition for diets with 0% or 20% CM was determined at the end of phase 2. Overall average daily gain (ADG) responded quadratically (P < 0.05) to increasing dietary level of CM such that ADG was increased by 17% due to an increase in the dietary level of CM from 0% to 20% and was reduced by 16% due to an increase in the dietary level of CM from 20% to 40%. Pigs fed diets with 0% or 40% CM did not differ in overall ADG. Dietary CM tended to quadratically decrease (P = 0.09) serum immunoglobulin A (IgA) level at the end of phase 2 such that serum IgA level tended to reduce with an increase in dietary CM from 0% to 20% and to increase with an increase in dietary CM from 20% to 40%. Dietary CM at 20% decreased (P < 0.05) the relative abundance of Bacteroidetes phylum and tended to increase (P = 0.07) the relative abundance of Firmicutes phylum. Dietary CM linearly increased (P < 0.05) the lactulose to mannitol ratio in the urine by 47% and 49% at the end of phases 1 and 2, respectively, and tended to linearly decrease (P < 0.10) ileal transepithelial electrical resistance at the end of phase 1 by 64%. In conclusion, CM fed in the current study could be included in corn-SBM-based diets for nursery pigs 20% to improve the growth performance and gut microbial composition and reduce immune response. Also, the CM used in the current study could be included in corn-SBM-based diets for nursery pigs at 30% or 40% without compromising growth performance. Dietary CM increased gut permeability, implying that dietary CM at 20% improves the growth performance of weaned pigs through mechanisms other than reducing gut permeability.

Project description:An experiment was conducted to evaluate the effects of including canola meal (CM) in diets for weaning pigs challenged with a F18 strain of Escherichia coli on growth performance and gut health. A total of 36 individually housed weaned pigs (initial body weight [BW] = 6.22 kg) were randomly allotted to one of the three diets (12 pigs/diet). The three diets were corn-soybean meal (SBM)-based basal diet (control diet) and the basal diet with 0.3% zinc oxide, 0.2% chlortetracycline, and 0.2% tiamulin (antibiotic diet) or with 20% CM diet. The diets were fed in two phases: Phase 1: days 0 to 7 and Phase 2: days 7 to 20. All pigs were given an oral dose of 2 × 109 CFU of F18 strain of E. coli on day 7. Fecal score was assessed daily throughout the trial. Dietary antibiotics increased (P < 0.05) overall average daily gain (ADG) and average daily feed intake (ADFI) compared by 48% and 47%, respectively. Dietary CM increased (P < 0.05) overall ADG and ADFI by 22% and 23%, respectively; but the ADG and ADFI values for CM-containing diet did not reach those for the antibiotics-containing diet. Dietary antibiotics reduced (P < 0.05) fecal score; however, dietary CM unaffected fecal score. Dietary antibiotics decreased (P < 0.05) liver weight per unit live BW by 16% at day 20, whereas dietary CM did not affect liver weight per unit live BW (29.2 vs. 28.6). Also, dietary antibiotics increased (P < 0.05) serum triiodothyronine and tetraiodothyronine levels for day 14, whereas dietary CM did not affect the serum level of these hormones. Dietary antibiotics reduced (P < 0.05) the number white blood cells and neutrophils by 38% and 43% at day 20, respectively, whereas dietary CM tended to reduce (P = 0.09) the number white blood cells by 19% at day 20. The number white blood cells for CM diet tended to be greater (P < 0.10) than that for antibiotics diet. The dietary antibiotics decreased (P < 0.05) the concentration of individual volatile fatty acids and hence of total volatile fatty acid in cecum by 61% at day 20, whereas dietary CM decreased (P < 0.05) cecal butyric acid concentration by 61% and tended to reduce (P < 0.10) total volatile fatty acid concentration by 30% at day 20. In conclusion, the dietary inclusion of 20% CM improved ADG and tended to reduce white blood cell counts. Thus, inclusion of CM in antibiotics-free corn-SBM-based diets for weaned pigs that are challenged with F18 strain of E. coli can result in their improved performance partly through a reduction of the inflammatory response.

Dataset Information

Combinatorial Glycomic Analyses to Direct CAZyme Discovery for the Tailored Degradation of Canola Meal Non-Starch Dietary Polysaccharides.

Publications

Combinatorial Glycomic Analyses to Direct CAZyme Discovery for the Tailored Degradation of Canola Meal Non-Starch Dietary Polysaccharides.

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