Publications by authors named "Andrew G Van Kessel"

33 Publications

A longer adaptation period to a functional amino acid-supplemented diet improves growth performance and immune status of Salmonella Typhimurium-challenged pigs.

J Anim Sci 2021 May;99(5)

Prairie Swine Centre, Inc., S7H 5N9, Saskatoon,Canada.

We recently showed that dietary supplementation with key functional amino acids (FAA) improves growth performance and immune status of Salmonella Typhimurium (ST)-challenged pigs. It is not known if ST-challenged pigs will benefit from a longer adaptation period to FAA. The objective of this study was to evaluate the effects of different adaptation periods to diets containing FAA above requirements for growth on performance and immune response of weaned pigs subsequently challenged with ST. A total of 32 mixed-sex weanling pigs (11.6 ± 0.3 kg) were randomly assigned to 1 of 4 dietary treatments, being a basal amino acid (AA) profile fed throughout the experimental period (FAA-) or a functional AA profile (FAA+; Thr, Met, and Trp at 120% of requirements) fed only in the postinoculation (FAA+0), for 1 wk pre- and postinoculation (FAA+1), or throughout the experimental period (FAA+2). After a 14-d adaptation period, pigs were inoculated with ST (2.15 × 109 CFU/mL). Growth performance, body temperature, fecal score, acute-phase proteins, oxidant/antioxidant balance, score for ST shedding in feces and intestinal colonization, and fecal and digesta myeloperoxidase (MPO) were measured pre- and postinoculation. Postinoculation body temperature and fecal score, serum haptoglobin, plasma superoxide dismutase (SOD), malondialdehyde (MDA), and fecal MPO were increased while serum albumin and plasma reduced glutathione (GSH):oxidized glutathione (GSSG) were reduced compared to pre-inoculation (P < 0.05). Average daily gain and G:F were greater in FAA+2 pigs compared to FAA- pigs (P < 0.05). Serum albumin was higher in FAA+2 and FAA+1 compared to FAA+0 and FAA- pigs (P < 0.05) while FAA+2 pigs had lower haptoglobin compared to FAA- (P < 0.05). Plasma SOD was increased and GSH:GSSG was decreased in FAA- pigs compared to the other treatments (P < 0.05). Score for ST shedding in feces was progressively lower from d 1 to 6 regardless of treatment (P < 0.05) and was lower in FAA+2 pigs compared to FAA- and FAA+0 (P < 0.05). Counts of ST in colon digesta were higher in FAA- and FAA+0 pigs compared to FAA+2 (P < 0.05). Fecal and colonic digesta MPO were lower in FAA+2 and FAA+1 pigs compared to FAA- (P < 0.05). These results demonstrate a positive effect of a longer adaptation period to FAA-supplemented diets on performance and immune status of weaned pigs challenged with Salmonella.
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http://dx.doi.org/10.1093/jas/skab146DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8153703PMC
May 2021

Effects of exogenous β-glucanase on ileal digesta soluble β-glucan molecular weight, digestive tract characteristics, and performance of coccidiosis vaccinated broiler chickens fed hulless barley-based diets with and without medication.

PLoS One 2021 3;16(5):e0236231. Epub 2021 May 3.

Department of Animal and Poultry Science, University of Saskatchewan, Saskatoon, Saskatchewan, Canada.

Introduction: Limited use of medication in poultry feed led to the investigation of exogenous enzymes as antibiotic alternatives for controlling enteric disease. The objective of this study was to evaluate the effects of diet β-glucanase (BGase) and medication on β-glucan depolymerization, digestive tract characteristics, and growth performance of broilers.

Materials And Methods: Broilers were fed hulless barley (HB) based diets with BGase (Econase GT 200P from AB Vista; 0 and 0.1%) and medication (Bacitracin and Salinomycin Na; with and without) arranged as a 2 × 2 factorial. In Experiment 1, 160 broilers were housed in cages from d 0 to 28. Each treatment was assigned to 10 cages. In Experiment 2, broilers (2376) were housed in floor pens and vaccinated for coccidiosis on d 5. Each treatment was assigned to one floor pen in each of nine rooms.

Results: In Experiment 1, the soluble β-glucan weighted average molecular weight (Mw) in the ileal digesta was lower with medication in the 0% BGase treatments. Peak molecular weight (Mp) and Mw were lower with BGase regardless of medication. The maximum molecular weight for the smallest 10% β-glucan (MW-10%) was lower with BGase addition. In Experiment 2, Mp was lower with medication in 0% BGase treatments. Beta-glucanase resulted in lower Mp regardless of medication, and the degree of response was lower with medication. The MW-10% was lower with BGase despite antibiotic addition. Body weight gain and feed efficiency were higher with medication regardless of BGase use through-out the trial (except d 11-22 feed efficiency). Beta-glucanase resulted in higher body weight gain after d 11 and worsened and improved feed efficiency before and after d 11, respectively, in unmedicated treatments.

Conclusion: BGase and medication caused the depolymerization of soluble ileal β-glucan. Beta-glucanase acted as a partial replacement for diet medication by increasing growth performance in coccidiosis vaccinated broilers.
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http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0236231PLOS
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8092798PMC
May 2021

Functional amino acid supplementation, regardless of dietary protein content, improves growth performance and immune status of weaned pigs challenged with Salmonella Typhimurium.

J Anim Sci 2021 Feb;99(2)

Prairie Swine Centre, Inc., Saskatoon, Canada.

High dietary protein may increase susceptibility of weaned pigs to enteric pathogens. Dietary supplementation with functional amino acids (FAA) may improve growth performance of pigs during disease challenge. The objective of this study was to evaluate the interactive effects of dietary protein content and FAA supplementation above requirements for growth on performance and immune response of weaned pigs challenged with Salmonella. Sixty-four mixed-sex weanling pigs (13.9 ± 0.82 kg) were randomly assigned to dietary treatments in a 2 × 2 factorial arrangement with low (LP) or high protein (HP) content and basal (AA-) or FAA profile (AA+; Thr, Met, and Trp at 120% of requirements) as factors. After a 7-d adaptation period, pigs were inoculated with either a sterile saline solution (CT) or saline solution containing Salmonella Typhimurium (ST; 3.3 × 109 CFU/mL). Growth performance, body temperature, fecal score, acute-phase proteins, oxidant/antioxidant balance, ST shedding score in feces and intestinal colonization, fecal and digesta myeloperoxidase (MPO), and plasma urea nitrogen (PUN) were measured pre- and postinoculation. There were no dietary effects on any measures pre-inoculation or post-CT inoculation (P > 0.05). Inoculation with ST increased body temperature and fecal score (P < 0.05), serum haptoglobin, plasma superoxide dismutase (SOD), malondialdehyde (MDA), PUN, and fecal MPO, and decreased serum albumin and plasma reduced glutathione (GSH):oxidized glutathione (GSSG) compared with CT pigs (P < 0.05). ST-inoculation reduced average daily gain (ADG) and feed intake (ADFI) vs. CT pigs (P < 0.05) but was increased by AA+ vs. AA- in ST pigs (P < 0.05). Serum albumin and GSH:GSSG were increased while haptoglobin and SOD were decreased in ST-inoculated pigs fed AA+ vs. AA- (P < 0.05). PUN was higher in HP vs. LP-fed pigs postinoculation (P < 0.05). Fecal ST score was increased in ST-inoculated pigs on days 1 and 2 postinoculation and declined by day 6 (P < 0.05) in all pigs while the overall score was reduced in AA+ vs. AA- pigs (P < 0.05). Cecal digesta ST score was higher in HP vs. LP-fed pigs and were lower in AA+ compared with AA- fed pigs in the colon (P < 0.05). Fecal and digesta MPO were reduced in ST pigs fed AA+ vs. AA- (P < 0.05). These results demonstrate a positive effect of FAA supplementation, with minimal effects of dietary protein, on performance and immune status in weaned pigs challenged with Salmonella.
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http://dx.doi.org/10.1093/jas/skaa365DOI Listing
February 2021

Hulless barley and beta-glucanase levels in the diet affect the performance of coccidiosis-challenged broiler chickens in an age-dependent manner.

Poult Sci 2021 Feb 2;100(2):776-787. Epub 2020 Nov 2.

Department of Animal and Poultry Science, University of Saskatchewan, Saskatoon, SK S7N5A8, Canada.

Diet β-glucanase (BGase) depolymerizes viscous β-glucan into lower molecular weight carbohydrates, which might act as a prebiotic in chickens exposed to enteric disease. Coccidiosis-challenged broiler chickens were fed graded levels of hulless barley (HB) and BGase to determine their effects on growth performance. Broilers were fed high β-glucan HB (CDC Fibar; 0, 30, and 60% replacing wheat) and BGase (Econase GT 200P; 0, 0.01, and 0.1%) in a 3 × 3 factorial arrangement. A total of 5,346 broilers were raised in litter floor pens and vaccinated for coccidiosis in feed and water on day 5. Each treatment was assigned to 1 pen (66 birds) in each of 9 rooms. Statistical significance was set at P ≤ 0.05. Overall, HB decreased body weight gain (BWG) and increased feed: gain ratio (F:G) of broilers. From day 0 to 11, BGase did not affect BWG and F:G, at the 0 and 30% HB. However, at 60% HB, the 0.01% BGase improved them, and the 0.1% BGase had no effect on BWG and increased F:G. For the day 22 to 32 and 0 to 32 periods, BGase did not affect BWG for 0 and 30% HB levels, but for the 60% HB, both BGase levels increased gain. The 0.1% level of BGase resulted in the lowest F:G for all HB levels, with the degree of response increasing with HB. No interaction was found for ileal digesta viscosity at day 11; the level of HB did not affect viscosity, but both levels of BGase decreased viscosity. At day 33, BGase did not affect viscosity at 0 and 30% HB levels, but viscosity was lowered for the 0.1% BGase treatment at the 60% HB level. In conclusion, HB reduced broiler performance, and BGase alleviated most but not all the effects. In young birds fed 60% HB, 0.1% BGase did not impact BWG and increased F:G.
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http://dx.doi.org/10.1016/j.psj.2020.10.036DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7858129PMC
February 2021

Intestinal Health and Threonine Requirement of Growing Pigs Fed Diets Containing High Dietary Fibre and Fermentable Protein.

Animals (Basel) 2020 Nov 6;10(11). Epub 2020 Nov 6.

Prairie Swine Centre, Inc., Saskatoon, SK S7H5N9, Canada.

Dietary fibre (DF) and fermentable crude protein (fCP) are dietary factors which affect nutrient utilization and intestinal health in pigs. A nitrogen (N)-balance study was conducted to determine the impact of DF and fCP on threonine (Thr) requirement for protein deposition (PD) and indicators of intestinal health. A total of 160 growing pigs (25 kg) were randomly assigned to 1 of 20 dietary treatments in a 2 × 2 × 5 factorial arrangement in a randomized complete block design with dietary fibre (low (LF) or high fibre (HF)], fCP [low (LfCP) or high fCP (HfCP)) and Thr (0.52, 0.60, 0.68, 0.76, or 0.82% standardized ileal digestible) as factors. Then, 4-day total urine and fecal collection was conducted, and pigs were euthanized for intestinal tissue and digesta sampling. Feeding high DF, regardless of fCP content, increased Thr requirement for PD ( < 0.05). High fCP, regardless of DF content, reduced Thr requirement for PD. Serum antioxidant capacity increased as dietary Thr level increased ( < 0.05). Cecal digesta short-chain fatty acids (SCFA) increased ( < 0.05) with HF and branched-chain fatty acids (BCFA) increased with HfCP and reduced with HF ( < 0.05). HfCP reduced ( < 0.05) mucin-2 (MUC2) expression in the colon of the HF but not the LF fed pigs and HF increased MUC2 in the LfCP but not the HfCP fed pigs. Feeding HF diet increased ( < 0.05) expression of zonula occludens-1 in the LfCP with no effect on HfCP fed pigs. Ammonia concentration in both cecum and colon increased ( < 0.05) in the HfCP fed pigs. Overall, high DF reduced the negative impact of HfCP on intestinal health, as indicated by alterations in SCFA and BCFA production and gut barrier gene expression. While increased dietary Thr content is required for PD in pigs fed high DF, feeding high fCP reduced Thr requirements.
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http://dx.doi.org/10.3390/ani10112055DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7694666PMC
November 2020

Effect of dietary fiber and threonine content on intestinal barrier function in pigs challenged with either systemic lipopolysaccharide or enteric Typhimurium.

J Anim Sci Biotechnol 2020 15;11:38. Epub 2020 Apr 15.

1Prairie Swine Centre, Inc., Saskatoon, SK S7H 5N9 Canada.

Background: The independent and interactive effects of dietary fiber (DF) and threonine (Thr) were investigated in growing pigs challenged with either systemic lipopolysaccharide (LPS) or enteric Typhimurium (ST) to characterise their effect on intestinal barrier function.

Results: In experiment 1, intestinal barrier function was assessed via oral lactulose and mannitol (L:M) gavage and fecal mucin analysis in pigs challenged with LPS and fed low fiber (LF) or high fiber (HF) diets with graded dietary Thr. Urinary lactulose recovery and L:M ratio increased ( < 0.05) during the LPS inoculation period in LF fed pigs but not in HF fed pigs. Fecal mucin output was increased ( < 0.05) in pigs fed HF compared to LF fed pigs. In experiment 2, RT-qPCR, ileal morphology, digesta volatile fatty acid (VFA) content, and fecal mucin output were measured in Typhimurium challenged pigs, fed LF or HF diets with standard or supplemented dietary Thr. inoculation increased ( < 0.05) fecal mucin output compared to the unchallenged period. Supplemental Thr increased fecal mucin output in the HF-fed pigs (Fib × Thr;  < 0.05). Feeding HF increased ( < 0.05) VFA concentration in cecum and colon. No effect of either Thr or fiber on expression of gene markers was observed except a tendency ( = 0.06) for increased MUC2 expression with the HF diet. Feeding HF increased goblet cell numbers ( < 0.05).

Conclusion: Dietary fiber appears to improve barrier function through increased mucin production capacity (i.e., goblet cell numbers, MUC2 gene expression) and secretion (i.e., fecal mucin output). The lack of effect of dietary Thr in -challenged pigs provides further evidence that mucin secretion in the gut is conserved and, therefore, Thr may be limiting for growth under conditions of increased mucin production.
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http://dx.doi.org/10.1186/s40104-020-00444-3DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7158091PMC
April 2020

The influence of indigestible protein on broiler digestive tract morphology and caecal protein fermentation metabolites.

J Anim Physiol Anim Nutr (Berl) 2020 May 18;104(3):847-866. Epub 2019 Dec 18.

Department of Animal and Poultry Science, University of Saskatchewan, Saskatoon, SK, Canada.

Indigestible dietary protein fermentation products have been suggested to negatively influence broiler performance due to their impact on health and digestive tract morphology. This study evaluated the digestive tract morphology and caecal protein fermentation metabolites of broiler fed 3 dietary protein levels (24%, 26% and 28%) with low or high indigestible protein (LIP, HIP). Two completely randomized 3 × 2 factorial trials were conducted with protein level (PL) and indigestible protein (IDP) as the main factors. In both trials, birds received six diets (24-LIP, 24-HIP, 26-LIP, 26-HIP, 28-LIP and 28 HIP) formulated with no medication. On day 5, trial 1 birds were vaccinated with Coccivac-B52, while trial 2 received no vaccine. Tissue and caecal samples were collected and caecal contents analysed for fermentation metabolites. Differences were considered significant when p ≤ .05. The LIP treatment caecal content in trial 1 at 14 days had greater histamine, agmatine and cadaverine levels, while HIP diets resulted in increased serotonin, tryptamine and spermidine. Histamine, serotonin and tryptamine at day 28 were not affected by IDP, and ammonia was not affected by treatments at day 14 or day 28. At day 14, HIP birds had lower total short-chain fatty acids, higher caecal pH and heavier pancreas, proventriculus, gizzard, jejunum and ileum weights. The same effects of IDP found in trial 1 were observed for histamine, agmatine, cadaverine, serotonin, tryptamine and spermidine at day 21 in trial 2. Trial 2 had a PL-by-IDP interaction influencing tyramine, spermidine (28-LIP > 24-LIP) and spermine with values increasing with PL for LIP diets and remaining constant for HIP diets. An interaction between PL and IDP was found for ammonia level and was similar to interactions for biogenic amines. In conclusion, dietary PL and IDP influence broiler caecal protein fermentation metabolites and those effects varied with coccidiosis vaccination and rearing environment.
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http://dx.doi.org/10.1111/jpn.13256DOI Listing
May 2020

Effect of live yeast Saccharomyces cerevisiae supplementation on the performance and cecum microbial profile of suckling piglets.

PLoS One 2019 22;14(7):e0219557. Epub 2019 Jul 22.

University of Saskatchewan, Department of Animal and Poultry Science, Saskatoon, Saskatchewan, Canada.

One mechanism through which S. cerevisiae may improve the performance of pigs is by altering the composition of the gut microbiota, a response that may be enhanced by early postnatal supplementation of probiotics. To test this hypothesis, newborn piglets (16 piglets/group) were treated with either S. cerevisiae yeast (5 x 109 cfu/pig: Low) or (2.5 x 1010 cfu/piglet: High) or equivalent volume of sterile water (Control) by oral gavage every other day starting from day 1 of age until weaning (28±1 days of age). Piglet body weight was recorded on days 1, 3, 7, 10, 17, 24 and 28 and average daily gain (ADG) calculated for the total period. At weaning, piglets were euthanized to collect cecum content for microbial profiling by sequencing of the 16S rRNA gene. ADG was higher in both Low and High yeast groups than in Control group (P<0.05). Alpha diversity analyses indicated a more diverse microbiota in the Control group compared with Low yeast group; the High yeast being intermediate (P < 0.01). Similarly, Beta diversity analyses indicated differences among treatments (P = 0.03), mainly between Low yeast and Control groups (P = 0.02). The sparse Partial Least Squares Discriminant Analysis (sPLS-DA) indicated that Control group was discriminated by a higher abundance of Veillonella, Dorea, Oscillospira and Clostridium; Low yeast treated pigs by higher Blautia, Collinsella and Eubacterium; and High yeast treated pigs by higher Eubacterium, Anaerostipes, Parabacteroides, Mogibacterium and Phascolarctobacterium. Partial Least Squares (PLS) analysis showed that piglet ADG was positively correlated with genus Prevotella in High yeast group. Yeast supplementation significantly affected microbial diversity in cecal contents of suckling piglets associated with an improvement of short chain fatty acid producing bacteria in a dose-dependent manner. In conclusion, yeast treatment improved piglet performance and shaped the piglet cecum microbiota composition in a dose dependent way.
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http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0219557PLOS
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6645501PMC
February 2020

Effect of supplemental threonine above requirement on growth performance of Salmonella typhimurium challenged pigs fed high-fiber diets1.

J Anim Sci 2019 Sep;97(9):3636-3647

Prairie Swine Centre, Inc. Saskatoon, SK, Canada.

It was shown previously that high dietary fiber (DF) and immune system stimulation (ISS) with systemic Escherichia coli lipopolysaccharide independently increased the threonine (Thr) requirement to maximize growth performance and protein deposition (PD). However, no additive effects on the Thr requirement were observed when both DF and ISS were present. The objective of the present study was to investigate whether supplementing Thr to meet previously estimated requirements for high DF and systemic immune challenge would maintain performance of pigs exposed to an enteric immune challenge when fed high DF. A total of 128 pigs (22.6 ± SD = 1.6 kg initial BW) were assigned to 1 of 4 dietary treatments in a 2 × 2 factorial arrangement in a randomized complete block design (n = 8 pens/treatment and 4 pigs/pen) for 28 d. Treatments were a low-fiber (LF; 13% total DF) or high-fiber (HF; 20% total DF) diet with either a standard (STD; 0.65% SID) or supplemental (SUP; 0.78% SID) Thr level. After a 7-d adaptation, pigs were orally inoculated with 2 mL (2.3 × 109 CFU/mL) of Salmonella typhimurium (ST). Blood samples and rectal swabs were obtained and rectal temperature recorded to determine clinical responses and ST shedding. On day 7 postinoculation, 1 pig/pen was euthanized and mesenteric lymph nodes, spleen, and digesta (ileum, cecum, and colon) were sampled to assess ST colonization and translocation. Body weight and feed intake were recorded on day 0, 7, and 21 postinoculation to calculate ADG, ADFI, and G:F. Rectal temperature increased (P < 0.05) 24 h postinoculation and remained elevated at day 6. Serum albumin concentration decreased (P < 0.05), whereas haptoglobin concentration increased (P < 0.05) postinoculation. There was no fiber or Thr effect (P > 0.05) on ST counts in the ileum and cecum, but a fiber × Thr interaction (P < 0.05) was observed in the colon. Supplemental Thr improved (P < 0.05) growth performance in LF- and HF-fed challenged pigs. However, performance of supplemented HF challenged pigs was less than (P < 0.05) supplemented LF challenged pigs. These results suggest that Thr supplemented to meet requirements for high DF and systemic immune challenge was not sufficient to maintain growth performance of pigs fed HF diets and challenged with an enteric pathogen.
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http://dx.doi.org/10.1093/jas/skz225DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6735783PMC
September 2019

Phytate degradation in gnotobiotic broiler chickens and effects of dietary supplements of phosphorus, calcium, and phytase.

Poult Sci 2019 Nov;98(11):5562-5570

Institute of Animal Science, University of Hohenheim, 70599 Stuttgart, Germany.

Gnotobiotic broiler chickens were used to study interactive effects of supplemented phosphorus, calcium (PCa), and phytase (Phy) on myo-inositol 1,2,3,4,5,6-hexakis (dihydrogen phosphate) (InsP6) degradation and release of myo-inositol in the digestive tract. In 2 subsequent runs, the chickens were subjected to 1 of 4 dietary treatments with and without PCa and Phy supplementation. Sanitized eggs were hatched in 8 germfree isolators, and a minimum of 9 male Ross 308 chickens were placed in each pen (total 16 pens). Treatments implemented on day 10 included gamma-irradiated diets without (PCa-; 4.1 g P and 6.2 g Ca/kg DM) or with (PCa+; 6.9 g P and 10.4 g Ca/kg DM) monosodium phosphate and limestone supplementation and without (Phy-) or with (Phy+) 1,500 FTU Phy/kg feed in a factorial arrangement. On day 15, digesta was collected from different sections of the intestinal tract and analyzed for InsP isomers and myo-inositol. The isolators did not remain germfree, but analysis of contaminants and results of InsP degradation indicated no or minor effects of the identified contaminants. Prececal InsP6 disappearance was 42% with the PCa-Phy- treatment and 17% with PCa+Phy-. No InsP3-4 isomers were found in the digesta of the terminal ileum in PCa-Phy-. The concentration of myo-inositol in the ileal digesta from PCa-Phy- (6.1 μmol/g DM) was significantly higher than that from PCa+Phy- (1.7 μmol/g DM), suggesting rapid degradation of the lower InsP isomers by mucosal phosphatases and their inhibition by PCa. Phytase supplementation increased InsP6 disappearance and prevented inhibitory effects of PCa supplements (72% in PCa-Phy+ and 67% in PCa+Phy+). However, PCa supplementation reduced the degradation of lower InsP isomers mainly in the posterior intestinal sections in the presence of Phy, resulting in significantly lower myo-inositol concentrations. It is concluded that mucosa-derived phosphatases might significantly contribute to InsP6 degradation in broiler chickens. The potential of mucosa-derived phosphatases to degrade InsP6 and lower InsP is markedly reduced by dietary PCa supplementation.
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http://dx.doi.org/10.3382/ps/pez309DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6771544PMC
November 2019

Supplementation with live yeast increases rate and extent of in vitro fermentation of nondigested feed ingredients by fecal microbiota.

J Anim Sci 2019 Apr;97(4):1806-1818

Department of Animal and Poultry Science, University of Saskatchewan, Saskatoon, SK, Canada.

Two studies were conducted to investigate the effect of live yeast (LY) on the in vitro fermentation characteristics of wheat, barley, corn, soybean meal (SBM), canola meal, and distillers dried grains with solubles (DDGS). In Study 1, LY yeast was added directly to in vitro fermentations inoculated with feces from lactating sows, whereas as in study 2, feces collected from lactating sows fed LY as a daily supplement was used. Selected feedstuffs were digested and the residue added to separate replicated (n = 3) fermentation reactions. Study 1 was conducted in two blocks, whereas study 2 was conducted using feces collected after a period of 3 (Exp. 1) or 4 wk (Exp. 2) of LY supplementation. Accumulated gas produced over 72 h was modeled for each substrate and the kinetics parameters compared between LY and control groups. The molar ratio of the volatile fatty acids (VFAs) produced in vitro were also compared at 12 and 72 h of incubation. In study 1, in vitro addition of yeast increased (P < 0.001) the rate of gas production (Rmax). However, a yeast × substrate effect (P < 0.05) observed for total gas accumulated (A), time to half asymptote (B), and time required to reach maximum rate of fermentation (Tmax) suggested that yeast-mediated increases in extent and rate of fermentation varied by substrate. Greater total gas production was observed only for corn and SBM, associated with greater B and Tmax. Supplementation with LY appeared to increase A and Rmax although with variation between experiments and substrates. In Exp. 1, LY decreased (P < 0.05) B and Tmax. However, a yeast × substrate effect (P < 0.05) was observed for only A (for wheat, barley, corn, and corn DDGS) and Rmax (wheat, barley, corn, and wheat DDGS). In Exp. 2, LY increased (P < 0.0001) A and decreased B. However, an interaction (P < 0.05) with substrates was observed for Rmax (except SBM) and Tmax. With exception of the DDGS samples, LY supplementation increased (P < 0.05) VFA production at 12 and 72 h of incubation. Yeast increased (P < 0.05) the molar ratios of acetic acid and branch-chain fatty acids at 12 h of incubation; however, this response was more variable by substrate at 72 h. In conclusion, LY supplementation increased the rate and extent of in vitro fermentation of a variety of substrates prepared from common feedstuffs. Greater effects were observed when LY was fed to sows than added directly in vitro, suggesting effects on fermentation were not mediated directly.
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http://dx.doi.org/10.1093/jas/skz073DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6447280PMC
April 2019

Impact of dietary fiber and immune system stimulation on threonine requirement for protein deposition in growing pigs.

J Anim Sci 2018 Dec;96(12):5222-5232

Department of Animal and Poultry Science, University of Saskatchewan, Saskatoon, SK, Canada.

High dietary fiber (DF) and immune system stimulation (ISS) are thought to limit amino acid availability for protein deposition (PD) in growing pigs. A nitrogen-balance study was conducted to determine Thr requirement for optimal PD when DF and ISS were present alone and in combination. A total of 90 barrows (20.5 kg initial BW; SD = 0.75 kg) were randomly assigned to one of 10 dietary treatments (n = 9) in nine blocks. Diets consisted of a low fiber (LF; 12.5% total DF) or high fiber (HF; 18.5% total DF by adding 10% sugar beet pulp and 5% wheat bran to the LF diet) with graded levels of Thr (0.49%, 0.57%, 0.65%, 0.73%, and 0.81% standardized ileal digestible [SID]) fed at 2.2 × maintenance ME requirements. After an 8-d adaptation, two 4-d nitrogen balance collection periods (pre-ISS and ISS) were conducted. ISS was induced by repeated injections (i.m.) of increasing doses of Escherichia coli lipopolysaccharide. Blood samples were taken during both periods to assess acute phase proteins and complete blood cell count. Data were analyzed by PROC MIXED with fixed effects of period, Thr, fiber, and their interactions, with block as a random effect. Nitrogen balance was analyzed separately for each period. Threonine requirement was estimated using PROC NLIN quadratic break-point model. Serum concentration of albumin, haptoglobin, fibrinogen, whole blood white blood cell, and platelet count were affected by ISS (P < 0.001) confirming successful ISS. During pre-ISS, PD increased linearly (P < 0.01) as Thr concentration in the diet increased, with a significant interaction (P < 0.05) between fiber and Thr. During ISS, PD increased linearly (P < 0.05) as Thr concentration in the diet increased. Quadratic break-point model estimated SID Thr required to maximize PD of pigs fed LF and HF diets during pre-ISS period was 0.68% (R2 = 0.88) and 0.78% (R2 = 0.99), respectively. During ISS, the SID Thr requirement was estimated at 0.76% (R2 = 0.76) for LF diet and 0.72% (R2 = 0.95) for HF fed pigs. HF and ISS independently increased Thr requirement for maximum PD, but these effects were not additive. HF may therefore mask the effects of ISS on Thr requirement for immune response and PD.
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http://dx.doi.org/10.1093/jas/sky381DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6276565PMC
December 2018

Effect of ruminal acidosis and short-term low feed intake on indicators of gastrointestinal barrier function in Holstein steers.

J Anim Sci 2018 Feb;96(1):108-125

Department of Animal and Poultry Science, University of Saskatchewan, Saskatoon, SK, Canada.

The objective of this study was to determine effect of ruminal acidosis (RA) and low feed intake [LFI] on the regional barrier function of the gastrointestinal tract. Twenty-one Holstein steers were fed for ad libitum intake for 5 d (control [CON]), fed at 25% of ad libitum intake for 5 d (LFI), or provided 2 d of ad libitum intake followed by 1-d of feed restriction (25% of ad libitum intake), 1 d where 30% of ad libitum dry matter intake (DMI) was provided as pelleted barley followed by the full allocation (RA) and fed for ad libitum intake the following day. Tissues and digesta from the rumen, omasum, duodenum, jejunum, ileum, cecum, proximal, and distal colon were collected. Permeability was assessed using the mucosal-to-serosal flux of inulin (JMS-inulin) and mannitol (JMS-mannitol). Digesta pH was 0.81, 0.63, and 0.42 pH units less for RA than CON in the rumen, cecum, and proximal colon; while, LFI had pH that was 0.47 and 0.36 pH units greater in the rumen and proximal colon compared to CON. Total ruminal short-chain fatty acid (SCFA) concentration were less for LFI (92 mM; P = 0.010) and RA (87 mM; P = 0.007) than CON (172 mM) steers. In the proximal colon, the proportion of butyrate (P = 0.025 and P = 0.022) and isobutyrate (P = 0.019 and P = 0.019) were greater, and acetate (P = 0.028 and P = 0.028) was less for LFI and RA, respectively, when compared to CON steers. Ruminal papillae length, width, perimeter, and surface area were 1.21 mm, 0.78 mm, 3.84 mm, and 11.15 mm2 less for LFI than CON; while, RA decreased papillae width by 0.52 mm relative to CON. The JMS-mannitol was less for LFI steers than CON in the proximal colon (P = 0.041) and in the distal colon (P = 0.015). Increased gene expression for claudin 1, occludin, tight-cell junction protein 1 and 2, and toll-like receptor 4 were detected for LFI relative to CON in the rumen, jejunum, and proximal colon. For RA steers, expression of toll-like receptor 4 in the rumen, and occludin and tight-cell junction protein 1 were greater in the jejunum than CON. An acute RA challenge decreased pH in the rumen and large intestine but did not increase tissue permeability due to increases in the expression of genes related to barrier function within 1 d of the challenge. Steers exposed to LFI for 5 d had reduced ruminal SCFA concentrations, smaller ruminal papillae dimensions, and increased tissue permeability in the proximal and distal colon despite increases for genes related to barrier function and immune function.
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http://dx.doi.org/10.1093/jas/skx049DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6140865PMC
February 2018

Claudin-4 undergoes age-dependent change in cellular localization on pig jejunal villous epithelial cells, independent of bacterial colonization.

Mediators Inflamm 2015 8;2015:263629. Epub 2015 Apr 8.

Vaccine and Infectious Disease Organization (VIDO), Home of the International Vaccine Centre (InterVac), 120 Veterinary Road, University of Saskatchewan, Saskatoon, SK, Canada S7N 5E3.

Newborn piglets are immunologically naïve and must receive passive immunity via colostrum within 24 hours to survive. Mechanisms by which the newborn piglet gut facilitates uptake of colostral cells, antibodies, and proteins may include FcRn and pIgR receptor-mediated endocytosis and paracellular transport between tight junctions (TJs). In the present study, FcRn gene (FCGRT) was minimally expressed in 6-week-old gut and newborn jejunum but it was expressed at significantly higher levels in the ileum of newborn piglets. pIgR was highly expressed in the jejunum and ileum of 6-week-old animals but only minimally in neonatal gut. Immunohistochemical analysis showed that Claudin-5 localized to blood vessel endothelial cells. Claudin-4 was strongly localized to the apical aspect of jejunal epithelial cells for the first 2 days of life after which it was redistributed to the lateral surface between adjacent enterocytes. Claudin-4 was localized to ileal lateral surfaces within 24 hours after birth indicating regional and temporal differences. Tissue from gnotobiotic piglets showed that commensal microbiota did not influence Claudin-4 surface localization on jejunal or ileal enterocytes. Regulation of TJs by Claudin-4 surface localization requires further investigation. Understanding the factors that regulate gut barrier maturation may yield protective strategies against infectious diseases.
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http://dx.doi.org/10.1155/2015/263629DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4407623PMC
February 2016

Diets high in heat-treated soybean meal reduce the histamine-induced epithelial response in the colon of weaned piglets and increase epithelial catabolism of histamine.

PLoS One 2013 19;8(11):e80612. Epub 2013 Nov 19.

Institute of Animal Nutrition, Department of Veterinary Medicine, Freie Universität Berlin, Berlin, Germany.

We examined the influence of dietary fermentable protein (fCP) and fermentable carbohydrates (fCHO) on the colonic epithelial response to histamine in pigs. Thirty-two weaned piglets were fed 4 diets in a 2 × 2 factorial design with low fCP/low fCHO, low fCP/high fCHO, high fCP/low fCHO and high fCP/high fCHO. After 21-23 days, the pigs were killed and tissue from the proximal colon was stimulated with carbachol, histamine, PGE2 or sodium hydrogen sulphide in Ussing chambers. Changes in short-circuit current and tissue conductance were measured. Diamine oxidase, histamine N-methyltransferase, stem cell growth factor receptor, Fc-epsilon receptor I and cystic fibrosis transmembrane conductance regulator gene expression was determined. Activities of diamine oxidase and histamine N-methyltransferase and numbers of colonic mast cells were measured. The change in the short-circuit current in response to histamine was lower (P = 0.002) and tended to be lower for PGE2 (P = 0.053) in high fCP groups compared to low fCP groups, irrespective of fCHO. Additionally, the change in tissue conductance after the application of histamine was lower (P = 0.005) in the high fCP groups. The expression of histamine N-methyltransferase mRNA (P = 0.033) and the activities of diamine oxidase (P = 0.001) and histamine N-methyltransferase (P = 0.006) were higher with high fCP in comparison with low fCP. The expression of mast cell markers, stem cell growth factor receptor (P = 0.005) and Fc-epsilon receptor I (P = 0.049) was higher with high fCP diets compared to diets low in fCP, whereas the mast cell count did not differ between groups. The expression of the cystic fibrosis transmembrane conductance regulator was reduced (P = 0.001) with high fCP diets compared to low fCP diets. The lower epithelial response to histamine and PGE2 and elevated epithelial histamine inactivation suggests an adaptation to high fCP diets.
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http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0080612PLOS
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3833947PMC
August 2014

Diets high in fermentable protein and fibre alter tight junction protein composition with minor effects on barrier function in piglet colon.

Br J Nutr 2014 Mar 11;111(6):1040-9. Epub 2013 Nov 11.

Department of Animal and Poultry Science, University of Saskatchewan, 51 Campus Drive, Saskatoon, Saskatchewan, Canada S7N 5A8.

Protein fermentation end products may damage the colonic mucosa, which could be counteracted by dietary inclusion of fermentable carbohydrates (fCHO). Although fermentable crude protein (fCP) and fCHO are known to affect microbial ecology, their interactive effects on epithelial barrier function are unknown. In the present study, in a 2 × 2 factorial experiment, thirty-two weaned piglets were fed low-fCP/low-fCHO (14·5 % crude protein (CP)/14·5 % total dietary fibre (TDF)), low-fCP/high-fCHO (14·8 % CP/16·6 % TDF), high-fCP/low-fCHO (19·8 % CP/14·5 % TDF) and high-fCP/high-fCHO (20·1 % CP/18·0 % TDF) diets. After 21-23 d, samples of proximal and distal colonic mucosae were investigated in Ussing chambers with respect to the paracellular and transcytotic passages of macromolecules and epithelial ion transport. The high-fCHO diets were found to reduce the permeability of the distal colon to the transcytotic marker horseradish peroxidase (HRP, 44 kDa; P <0·05) and also reduce the paracellular permeation of N-hydroxysuccinimide-biotin into the submucosa (443 Da; P <0·05), whereas that of HRP was decreased by the high-fCP diets (P <0·01). Short-circuit current (active ion transport), transepithelial resistance (barrier function) and charge selectivity were largely unaffected in both the segments. However, the high-fCP diets were found to suppress the aldosterone-induced epithelial Na channel activity (P <0·01) irrespective of fCHO inclusion. The high-fCP diets generally reduced the expression of colonic claudin-1, claudin-2 and claudin-3 (P <0·01), while that of claudin-4 was increased by the high-fCHO diets (P <0·01). The high-fCHO diets also altered the ratio between occludin forms (P <0·05) and increased the expression of tricellulin in the proximal colon, which was not observed with high-fCP diets. In conclusion, dietary fCHO and fCP exerted few and largely independent effects on functional measurements, but altered tight junction protein composition in a compensatory way, so that colonic transport and barrier properties were only marginally affected.
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http://dx.doi.org/10.1017/S0007114513003498DOI Listing
March 2014

Influence of different carbohydrate composition in barley varieties on Salmonella Typhimurium var. Copenhagen colonisation in a "Trojan" challenge model in pigs.

Arch Anim Nutr 2012 Jun;66(3):163-79

Department of Animal and Poultry Science, College of Agriculture and Bioressources, University ofSaskatchewan, Saskatoon, Canada.

Based on previously performed in vitro studies, which showed that hulless barley varieties could reduce large intestinal Salmonella Typhimurium var. Copenhagen proliferation in pigs, two in vivo experiments were conducted to prove these observations. In Experiment (Exp.) 1, 126 weaning piglets were randomly allocated into pens of seven animals each and fed one of six experimental diets. Three diets contained (75% as-fed) one of three hulless barley varieties with beta-glucan (BG) contents ranging from 5 to 11% and amylose from 5 to 40%, and two diets contained a low BG and amylose hulless barley supplemented with isolated barley BG or raw potato starch. A hulled barley diet served as a control. Two piglets per pen ("Trojan" pigs) were orally infected with Salmonella Typhimurium var. Copenhagen (ST). The remaining five pigs per pen were designated "Contact" pigs. The ST shedding was determined over one week after infection. On day 6, the two Trojans and two random Contacts from each pen were euthanised and intestinal contents and mesenteric lymph nodes cultured for ST. Intestinal volatile fatty acids and microbial composition were determined. In Exp. 2, 126 piglets were assigned to one of three diets based on hulled or hulless barleys. The timeline, infection, sampling and analyses were similar as in Exp. 1 except samples were taken from four Contact pigs. Hulless barley varieties with high BG and amylose tended to decrease ST persistence in Exp. 1. Clostridia from cluster I in the colon were reduced with high amylose hulless barley or diets supplemented with potato starch (p < 0.05), whereas other microbial groups were not. Propionate increased (p < 0.05) and acetate decreased (p < 0.05) with hulless barley inclusion. Exp. 2 revealed a reduced ST shedding and reduced number of clostridia for high BG hulless barley as compared to common hulled barley and a low BG variety (p < 0.05). In conclusion, high BG hulless barley do not prevent ST colonisation but might help to reduce transmission in pigs, likely by supporting an intestinal environment limiting growth of this zoopathogen.
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http://dx.doi.org/10.1080/1745039x.2012.676814DOI Listing
June 2012

Fermentable fiber ameliorates fermentable protein-induced changes in microbial ecology, but not the mucosal response, in the colon of piglets.

J Nutr 2012 Apr 22;142(4):661-7. Epub 2012 Feb 22.

Institute of Animal Nutrition, Freie Universität Berlin, Berlin, Germany.

Dietary inclusion of fermentable carbohydrates (fCHO) is reported to reduce large intestinal formation of putatively toxic metabolites derived from fermentable proteins (fCP). However, the influence of diets high in fCP concentration on epithelial response and interaction with fCHO is still unclear. Thirty-two weaned piglets were fed 4 diets in a 2 × 2 factorial design with low fCP/low fCHO [14.5% crude protein (CP)/14.5% total dietary fiber (TDF)]; low fCP/high fCHO (14.8% CP/16.6% TDF); high fCP low fCHO (19.8% CP/14.5% TDF); and high fCP/high fCHO (20.1% CP/18.0% TDF) as dietary treatments. After 21-23 d, pigs were killed and colon digesta and tissue samples analyzed for indices of microbial ecology, tissue expression of genes for cell turnover, cytokines, mucus genes (MUC), and oxidative stress indices. Pig performance was unaffected by diet. fCP increased (P < 0.05) cell counts of clostridia in the Clostridium leptum group and total short and branched chain fatty acids, ammonia, putrescine, histamine, and spermidine concentrations, whereas high fCHO increased (P < 0.05) cell counts of clostridia in the C. leptum and C. coccoides groups, shifted the acetate to propionate ratio toward acetate (P < 0.05), and reduced ammonia and putrescine (P < 0.05). High dietary fCP increased (P < 0.05) expression of PCNA, IL1β, IL10, TGFβ, MUC1, MUC2, and MUC20, irrespective of fCHO concentration. The ratio of glutathione:glutathione disulfide was reduced (P < 0.05) by fCP and the expression of glutathione transferase was reduced by fCHO (P < 0.05). In conclusion, fermentable fiber ameliorates fermentable protein-induced changes in most measures of luminal microbial ecology but not the mucosal response in the large intestine of pigs.
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http://dx.doi.org/10.3945/jn.111.156190DOI Listing
April 2012

Nonstarch polysaccharide-degrading enzymes alter the microbial community and the fermentation patterns of barley cultivars and wheat products in an in vitro model of the porcine gastrointestinal tract.

FEMS Microbiol Ecol 2011 Jun 22;76(3):553-63. Epub 2011 Mar 22.

Prairie Swine Centre Inc., Saskatoon, SK, Canada.

An in vitro experiment was carried out to assess how nonstarch polysaccharide (NSP)-degrading enzymes influence the fermentation of dietary fiber in the pig large intestine. Seven wheat and barley products and cultivars with differing carbohydrate fractions were hydrolyzed using pepsin and pancreatin in the presence or not of NSP-degrading enzymes (xylanase and β-glucanase) and the filter retentate was subsequently fermented with sow fecal bacteria. Dry matter, starch, crude protein and β-glucan digestibilities during hydrolysis were measured. Fermentation kinetics of the hydrolyzed ingredients were modelled. Short-chain fatty acids (SCFA) production and molar ratio were compared after 12, 24 and 72 h. Microbial communities were analyzed after 72 h of fermentation using terminal restriction fragment length polymorphism. The results showed an increase of nutrient digestibility (P<0.001), whereas fermentability and SCFA production decreased (P<0.001) with addition of the enzyme. SCFA and bacterial community profiles also indicated a shift from propionate to acetate and an increase in cellulolytic Ruminococcus- and xylanolytic Clostridium-like bacteria. This is explained by the increase in slowly fermentable insoluble carbohydrate and the lower proportion of rapidly fermentable β-glucan and starch in the retentate when grains were incubated with NSP-degrading enzymes. Shifts were also different for the four barley varieties investigated, showing that the efficiency of the enzymes depends on the structure of the carbohydrate fractions in cereal products and cultivars.
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http://dx.doi.org/10.1111/j.1574-6941.2011.01074.xDOI Listing
June 2011

Improvement of the representation of bifidobacteria in fecal microbiota metagenomic libraries by application of the cpn60 universal primer cocktail.

Appl Environ Microbiol 2010 Jul 30;76(13):4550-2. Epub 2010 Apr 30.

University of Saskatchewan, 52 Campus Drive, Saskatoon, SK, Canada S7N 5B4.

Actinobacteria, particularly bifidobacteria, are widely observed to be underrepresented in metagenomic studies of microbial communities. We have compared human fecal microbiota clone libraries based on 16S rRNA and cpn60 PCR products. Taxonomic profiles were similar except that the cpn60 libraries contained large numbers of bifidobacterial sequences.
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http://dx.doi.org/10.1128/AEM.01510-09DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2897446PMC
July 2010

Nonstarch polysaccharides modulate bacterial microbiota, pathways for butyrate production, and abundance of pathogenic Escherichia coli in the pig gastrointestinal tract.

Appl Environ Microbiol 2010 Jun 9;76(11):3692-701. Epub 2010 Apr 9.

Department of Agricultural, Food and Nutritional Science, University of Alberta, 4-10 Agriculture/Forestry Centre, Edmonton, AB, Canada.

The impact of nonstarch polysaccharides (NSP) differing in their functional properties on intestinal bacterial community composition, prevalence of butyrate production pathway genes, and occurrence of Escherichia coli virulence factors was studied for eight ileum-cannulated growing pigs by use of terminal restriction fragment length polymorphism (TRFLP) and quantitative PCR. A cornstarch- and casein-based diet was supplemented with low-viscosity, low-fermentability cellulose (CEL), with high-viscosity, low-fermentability carboxymethylcellulose (CMC), with low-viscosity, high-fermentability oat beta-glucan (LG), and with high-viscosity, high-fermentability oat beta-glucan (HG). Only minor effects of NSP fractions on the ileal bacterial community were observed, but NSP clearly changed the digestion in the small intestine. Compared to what was observed for CMC, more fermentable substrate was transferred into the large intestine with CEL, LG, and HG, resulting in higher levels of postileal dry-matter disappearance. Linear discriminant analysis of NSP and TRFLP profiles and 16S rRNA gene copy numbers for major bacterial groups revealed that CMC resulted in a distinctive bacterial community in comparison to the other NSP, which was characterized by higher gene copy numbers for total bacteria, Bacteroides-Prevotella-Porphyromonas, Clostridium cluster XIVa, and Enterobacteriaceae and increased prevalences of E. coli virulence factors in feces. The numbers of butyryl-coenzyme A (CoA) CoA transferase gene copies were higher than those of butyrate kinase gene copies in feces, and these quantities were affected by NSP. The present results suggest that the NSP fractions clearly and distinctly affected the taxonomic composition and metabolic features of the fecal microbiota. However, the effects were more linked to the individual NSP and to their effect on nutrient flow into the large intestine than to their shared functional properties.
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http://dx.doi.org/10.1128/AEM.00257-10DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2876444PMC
June 2010

Effect of barley and oat cultivars with different carbohydrate compositions on the intestinal bacterial communities in weaned piglets.

FEMS Microbiol Ecol 2008 Dec 16;66(3):556-66. Epub 2008 Jun 16.

Research Unit for Nutritional Physiology 'Oskar Kellner', Research Institute for the Biology of Farm Animals, Dummerstorf, Germany.

This experiment was aimed at comparing the intestinal microbial community composition in pigs fed hulled common barley supplemented with isolated barley mixed-linked beta-glucan, four hulless barley varieties and breeding lines with mixed-linked beta-glucan contents ranging from 41 to 84 g kg(-1) and different amylose/amylopectin ratios as well as two oat varieties. Seventy-two weaned piglets were allocated to one of nine diets composed of 81.5% cereal, 6% whey, 9% soy protein isolate and 3.5% minerals. After 15 days, pigs were sacrificed and ileum and colon contents were collected for quantitative real-time PCR (qPCR) and denaturing gradient gel electrophoresis to evaluate microbial communities. Shifts in intestinal microbial communities were observed with the hulless barley cultivars with a normal to high beta-glucan content and from normal starch toward either high-amylopectin or high-amylose starch. These hulless barleys had the lowest (P<0.05) microbial diversity, whereas oats had intermediate diversity compared with low-beta-glucan hulless cultivars and hulled varieties. Furthermore, hulless varieties favoured xylan- and beta-glucan-degrading bacteria whereas mixed-linked beta-glucan-supplemented hulled barley favoured lactobacilli. Numbers of lactobacilli decreased in the ileum of pigs fed hulless/high mixed-linked beta-glucan barley-based diets. Thus, cultivar differences in both the form and the quantity of carbohydrates affect gut microbiota in pigs, which provides information for future feeding strategies.
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http://dx.doi.org/10.1111/j.1574-6941.2008.00605.xDOI Listing
December 2008

Commensal bacteria and expression of two major intestinal chemokines, TECK/CCL25 and MEC/CCL28, and their receptors.

PLoS One 2007 Jul 25;2(7):e677. Epub 2007 Jul 25.

Lymphocyte et Immunité des Muqueuses, UR 1282, Infectiologie Animale et Santé Publique, Institut National de la Recherche Agronomique, Nouzilly, France.

Background: CCL25/TECK and CCL28/MEC are CC chemokines primarily expressed in thymic dendritic cells and mucosal epithelial cells. Their receptors, CCR9 and CCR10, are mainly expressed on T and B lymphocytes. In human, mouse, pig and sheep CCL25 and CCL28 play an important role in the segregation and the compartmentalization of the mucosal immune system. As evidenced by early comparisons of germ-free and conventional animals, the intestinal bacterial microflora has a marked effect on host intestinal immune functions. However, little is known about the impact of bacterial colonization on constitutive and induced chemokine expressions as well as on the generation of anti-inflammatory mechanisms.

Methodology/principal Findings: Therefore, we decided to focus by qPCR on the mRNA expression of two main gut chemokines, CCL25 and CCL28, their receptors CCR9 and CCR10, the Tregs marker Foxp3 and anti-inflammatory cytokines TGF-beta and IL-10 following colonization with different bacterial species within the small intestine. To accomplish this we used an original germ-free neonatal pig model and monoassociated pigs with a representative Gram-negative (Escherichia coli) or Gram-positive (Lactobacillus fermentum) commensal bacteria commonly isolated from the neonatal pig intestine. Our results show a consistent and marked effect of microbial colonization on the mRNA expression of intestinal chemokines, chemokine receptors, Foxp3 and TGF-beta. Moreover, as evidenced by in vitro experiments using two different cell lines, the pattern of regulation of CCL25 and CCL28 expression in the gut appears complex and suggests an additional role for in vivo factors.

Conclusions/significance: Taken together, the results highlight the key role of bacterial microflora in the development of a functional intestinal immune system in an elegant and relevant model for human immune system development.
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http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0000677PLOS
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC1919421PMC
July 2007

Transcriptome profiling of the small intestinal epithelium in germfree versus conventional piglets.

BMC Genomics 2007 Jul 5;8:215. Epub 2007 Jul 5.

Department of Animal Sciences, University of Illinois, Urbana, Illinois 61801, USA.

Background: To gain insight into host-microbe interactions in a piglet model, a functional genomics approach was used to address the working hypothesis that transcriptionally regulated genes associated with promoting epithelial barrier function are activated as a defensive response to the intestinal microbiota. Cesarean-derived germfree (GF) newborn piglets were colonized with adult swine feces, and villus and crypt epithelial cell transcriptomes from colonized and GF neonatal piglets were compared using laser-capture microdissection and high-density porcine oligonucleotide microarray technology.

Results: Consistent with our hypothesis, resident microbiota induced the expression of genes contributing to intestinal epithelial cell turnover, mucus biosynthesis, and priming of the immune system. Furthermore, differential expression of genes associated with antigen presentation (pan SLA class I, B2M, TAP1 and TAPBP) demonstrated that microbiota induced immune responses using a distinct regulatory mechanism common for these genes. Specifically, gene network analysis revealed that microbial colonization activated both type I (IFNAR) and type II (IFNGR) interferon receptor mediated signaling cascades leading to enhanced expression of signal transducer and activator of transcription 1 (STAT1), STAT2 and IFN regulatory factor 7 (IRF7) transcription factors and the induction of IFN-inducible genes as a reflection of intestinal epithelial inflammation. In addition, activated RNA expression of NF-kappa-B inhibitor alpha (NFkappaBIA; a.k.a I-kappa-B-alpha, IKBalpha) and toll interacting protein (TOLLIP), both inhibitors of inflammation, along with downregulated expression of the immunoregulatory transcription factor GATA binding protein-1 (GATA1) is consistent with the maintenance of intestinal homeostasis.

Conclusion: This study supports the concept that the intestinal epithelium has evolved to maintain a physiological state of inflammation with respect to continuous microbial exposure, which serves to sustain a tight intestinal barrier while preventing overt inflammatory responses that would compromise barrier function.
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http://dx.doi.org/10.1186/1471-2164-8-215DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC1949829PMC
July 2007

Effects of bacterial colonization on the porcine intestinal proteome.

J Proteome Res 2007 Jul 2;6(7):2596-604. Epub 2007 Jun 2.

Department of Animal Health, Welfare and Nutrition, Faculty of Agricultural Sciences, University of Aarhus, 8830 Tjele, Denmark.

The gastrointestinal tract harbors a complex community of bacteria, of which many may be beneficial. Studies of germ-free animal models have shown that the gastrointestinal microbiota not only assists in making nutrients available for the host but also contributes to intestinal health and development. We studied small intestinal protein expression patterns in gnotobiotic pigs maintained germ-free, or monoassociated with either Lactobacillus fermentum or non-pathogenic Escherichia coli. A common reference design in combination with labeling with stable isobaric tags allowed the individual comparison of 12 animals. Our results showed that bacterial colonization differentially affected mechanisms such as proteolysis, epithelial proliferation, and lipid metabolism, which is in good agreement with previous studies of other germ-free animal models. We have also found that E. coli has a profound effect on actin remodeling and intestinal proliferation, which may be related to stimulated migration and turnover of enterocytes. Regulations related to L. fermentum colonization involved individual markers for immunoregulatory mechanisms.
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http://dx.doi.org/10.1021/pr070038bDOI Listing
July 2007

Dietary encapsulated glycine influences Clostridium perfringens and Lactobacilli growth in the gastrointestinal tract of broiler chickens.

J Nutr 2007 Jun;137(6):1408-14

Department of Animal and Poultry Science, University of Saskatchewan, Saskatoon, Saskatchewan, Canada.

Three experiments were conducted to determine whether there is a causative relation between dietary glycine concentration and intestinal Clostridium perfringens growth in broiler chickens. Expt. 1 showed that glycine concentrations were higher (P < 0.05) in jejunum and ileum of birds fed fat-encapsulated glycine compared with crystalline glycine. In Expt. 2, 2 cages of 6 birds were assigned to 1 of 6 experimental diets formulated to contain 7.6 and 10.6, 17.8 and 40.6, 27.8 and 30.6, 37.8 and 20.6, 47.7 and 10.6, and 7.8 and 50.6 g/kg total glycine and proline, respectively, provided primarily by supplementation with encapsulated glycine or proline as required. In Expt. 3, 12 groups of 6 birds were fed 4 different diets supplemented with encapsulated glycine to achieve 7.6, 21.0, 34.3, or 47.7 g/kg total glycine. The birds were orally challenged with C. perfringens type A on d 1 and d 14-21 and killed on d 28. In Expt. 2, C. perfringens populations were higher (P < 0.05) in ileum and cecum of birds, which received either 37.8 or 47.7 g/kg total glycine compared with those fed 7.6 g/kg glycine. In Expt. 3, C. perfringens numbers were higher (P < 0.05) in ileum of birds fed either 34.3 or 47.7 g/kg dietary glycine than those given either 7.6 or 21.0 g/kg glycine. Conversely, lactobacilli counts in ileum and cecum were significantly lower in birds fed the higher levels of glycine in both experiments. High C. perfringens colonization and high intestinal lesion scores were associated with reduced performance (P < 0.05). We conclude that glycine is an important determinant of C. perfringens growth in the intestinal tract of broiler chickens.
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http://dx.doi.org/10.1093/jn/137.6.1408DOI Listing
June 2007

Molecular characterization of microbial communities in Canadian pulp and paper activated sludge and quantification of a novel Thiothrix eikelboomii-like bulking filament.

Can J Microbiol 2006 May;52(5):494-500

Department of Animal and Poultry Science, University of Saskatchewan, Canada.

We examined the microbial community structure and quantified the levels of the filamentous bulking organism Thiothrix eikelboomii in samples of activated sludge mixed liquor suspended solids (MLSS) from Canadian pulp and paper mills. Libraries of chaperonin 60 (cpn60) gene sequences were prepared from MLSS total microbial community DNA and each was compared with cpnDB, a reference database of cpn60 sequences (http://cpndb.cbr.nrc.ca) for assignment of taxonomic identities. Sequences similar to but distinct from the type strain of T. eikelboomii AP3 (ATCC 49788T) (approximately 89% identity over 555 bp) were recovered at high frequency from a mill sample that was experiencing bulking problems at the time of sample collection, which corresponded to microscopic observations using fluorescent in situ hybridization with commercially available 16S rDNA-based probes. We enumerated this strain in five mill-derived MLSS samples using real-time quantitative PCR (qPCR) and found that two samples had high levels of the bulking strain (>1012 genomes/g MLSS) and two contained lower but detectable levels of this organism. None of the mill samples contained cpn60 sequences that were identical to the type strain of T. eikelboomii. This technique shows promise for monitoring pulp and paper mill wastewater treatment systems by detecting and enumerating this strain of T. eikelboomii, which may be specific to pulp and paper mill wastewater treatment systems.
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http://dx.doi.org/10.1139/w05-160DOI Listing
May 2006

Characterization of intestinal microbiota and response to dietary virginiamycin supplementation in the broiler chicken.

Appl Environ Microbiol 2006 Apr;72(4):2815-23

Department of Animal and Poultry Science, University of Saskatchewan, 51 Campus Drive, Saskatoon, SK S7N 5A8, Canada.

The inclusion of antibiotic growth promoters, such as virginiamycin, at subtherapeutic levels in poultry feeds has a positive effect on health and growth characteristics, possibly due to beneficial effects on the host gastrointestinal microbiota. To improve our understanding of the chicken gastrointestinal microbiota and the effect of virginiamycin on its composition, we characterized the bacteria found in five different gastrointestinal tract locations (duodenal loop, mid-jejunum, proximal ileum, ileocecal junction, and cecum) in 47-day-old chickens that were fed diets excluding or including virginiamycin throughout the production cycle. Ten libraries (five gastrointestinal tract locations from two groups of birds) of approximately 555-bp chaperonin 60 PCR products were prepared, and 10,932 cloned sequences were analyzed. A total of 370 distinct cpn60 sequences were identified, which ranged in frequency of recovery from 1 to 2,872. The small intestinal libraries were dominated by sequences from the Lactobacillales (90% of sequences), while the cecum libraries were more diverse and included members of the Clostridiales (68%), Lactobacillales (25%), and Bacteroidetes (6%). To assess the effects of virginiamycin on the gastrointestinal microbiota, 15 bacterial targets were enumerated using quantitative, real-time PCR. Virginiamycin was associated with increased abundance of many of the targets in the proximal gastrointestinal tract (duodenal loop to proximal ileum), with fewer targets affected in the distal regions (ileocecal junction and cecum). These findings provide improved profiling of the composition of the chicken intestinal microbiota and indicate that microbial responses to virginiamycin are most significant in the proximal small intestine.
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http://dx.doi.org/10.1128/AEM.72.4.2815-2823.2006DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC1448984PMC
April 2006

Enumeration of specific bacterial populations in complex intestinal communities using quantitative PCR based on the chaperonin-60 target.

J Microbiol Methods 2006 Jan 19;64(1):46-62. Epub 2005 Aug 19.

Department of Animal and Poultry Science, University of Saskatchewan, 51 Campus Drive, Saskatoon SK, Canada, S7N 5A8.

We used qPCR and the target gene chaperonin-60 (cpn60) to enumerate Clostridium perfringens genomes in DNA extracts from contents of the chicken gastrointestinal tract with the aim of optimizing this methodology to enumerate any bacterium of interest. To determine the most accurate protocols for determining target species abundance, we compared various DNA extraction methods in combination with four methods for producing standard curves. Factors affecting accuracy included the co-purification of PCR inhibitors and/or fluorescence quenchers and the yield of target DNA in the extract. Anion exchange chromatography of the spiked test samples enabled accurate enumeration of C. perfringens using a standard curve comprised of a plasmid containing a fragment of C. perfringens cpn60. We used qPCR to enumerate C. perfringens and other intestinal bacteria in ileum and cecum samples from chickens that had been challenged with C. perfringens and compared the results with viable counts on corresponding selective agars. We conclude that qPCR-based molecular enumeration of target species in the gastrointestinal tract is feasible, but care must be taken in order to mitigate the effects of confounding factors that can affect the apparent cell count.
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http://dx.doi.org/10.1016/j.mimet.2005.04.006DOI Listing
January 2006

Comparison of ileum microflora of pigs fed corn-, wheat-, or barley-based diets by chaperonin-60 sequencing and quantitative PCR.

Appl Environ Microbiol 2005 Feb;71(2):867-75

National Research Council Plant Biotechnology Institute, Saskatoon, Saskatchewan S7N 5A8, Canada.

We have combined the culture-independent methods of high-throughput sequencing of chaperonin-60 PCR product libraries and quantitative PCR to profile and quantify the small-intestinal microflora of pigs fed diets based on corn, wheat, or barley. A total of 2,751 chaperonin-60 PCR product clones produced from samples of ileum digesta were examined. The majority (81%) of these clones contained sequences independently recovered from all three libraries; 372 different nucleotide sequences were identified, but only 14% of the 372 different sequences were recovered from all three libraries. Taxonomic assignments of the library sequences were made by comparison to a reference database of chaperonin-60 sequences combined with phylogenetic analysis. The taxa identified are consistent with previous reports of pig ileum microflora. Frequencies of each sequence in each library were calculated to identify taxa that varied in frequency between the corn, barley, and wheat libraries. The chaperonin-60 sequence inventory was used as a basis for designing PCR primer sets for taxon-specific quantitative PCR. Results of quantitative PCR analysis of ileum digesta confirmed the relative abundances of targeted taxa identified with the library sequencing approach. The results of this study indicate that chaperonin-60 clone libraries can be valid profiles of complex microbial communities and can be used as the basis for producing quantitative PCR assays to measure the abundance of taxa of interest during experimentally induced or natural changes in a community.
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http://dx.doi.org/10.1128/AEM.71.2.867-875.2005DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC546709PMC
February 2005
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