Diet medication and beta-glucanase affect ileal digesta soluble beta-glucan molecular weight, carbohydrate fermentation, and performance of coccidiosis vaccinated broiler chickens given wheat-based diets.

Exogenous enzymes as alternatives to feed antibiotics in poultry has become an emerging research area with the emergence of antibiotic resistance. The objective was to evaluate the effects of diet medication (antibiotics) and ß-glucanase (BGase) on digesta soluble ß-glucan depolymerization, carbohydrate fermentation, and performance of coccidiosis-vaccinated broiler chickens fed wheat-based diets. A total of 1,782 broilers were raised on litter floor pens, and each treatment was assigned to 1 pen in each of the 9 rooms. The 3 dietary treatments were based on wheat as the sole grain (control, control + medication and control + 0.1% BGase), and the birds were fed the respective treatments ad libitum from 0 to 33 d. Treatments were arranged in a randomized complete block design and analyzed as a one-way ANOVA. Beta-glucanase reduced the peak molecular weight, weight average molecular weight (Mw) and maximum molecular weight for the smallest 10% ß-glucan molecules (MW-10%) in ileal digesta at d 11 and 33, whereas diet medication reduced Mw and MW-10% at d 33 compared to the control (P < 0.01). Beta-glucanase and medication reduced the ileal viscosity at d 11 compared to the control (P = 0.010). Ileal propionic acid concentration at d 11 and caecal total SCFA, acetic, and butyric acid concentrations at d 33 were lower in the BGase-supplemented diet than in the control (P < 0.05). The BGase-added diet had higher duodenal pH compared to the control at d 33 (P = 0.026). The effect of medication on carbohydrate fermentation was minimal. Diet medication increased weight gain after d 11, whereas BGase increased the gain for the total trial period compared to the control (P < 0.001). Feed intake was not affected by the dietary treatment. Medication and BGase improved feed efficiency after d 11 compared to the control (P < 0.001). The response to diet medication was larger than BGase, considering weight gain and feed efficiency after d 11 (P < 0.001). In conclusion, diet medication and BGase depolymerized high molecular weight ileal soluble ß-glucan and increased overall bird performance. Dietary BGase may benefit bird health in broilers fed wheat-based diets without medication.

Tracking the fecal mycobiome through the lifespan of production pigs and a comparison to the feral pig.

IMPORTANCE: This work provides evidence that early-life fungal community composition, or host genetics, influences long-term mycobiome composition. In addition, this work provides the first comparison of the feral pig mycobiome to the mycobiome of intensively raised pigs.

Functional amino acid supplementation attenuates the negative effects of plant-based nursery diets on the response of pigs to a subsequent Salmonella Typhimurium challenge.

Functional amino acids (FAA) attenuate the effects of Salmonella challenge in pigs. However, this may be affected by protein source (PS). The objective of the present study was to determine the effects of nursery dietary PS and FAA supplementation on growth performance and immune status of pigs subsequently challenged with Salmonella Typhimurium (ST). Thirty-two weanling pigs (8.7 ± 0.23 kg) were assigned to a feeding program for 31 d in a 2 × 2 factorial arrangement. Factors were dietary PS (plant-based [PB] vs. animal-based [AB]) and FAA profile (basal [FAA-] or supplemented [FAA+; Thr, Met, and Trp at 120% of requirements]). Pigs were subsequently placed on a common grower diet and, after a 7-d adaptation, were inoculated with ST and monitored for 7 d postinoculation. Growth performance, rectal temperature, fecal score, gut health, ST shedding score, intestinal colonization and translocation, and blood parameters of acute-phase response and antioxidant balance were measured pre- and postinoculation. Data were analyzed with a 2 (AB vs. PB) × 2 (FAA- vs. FAA+) factorial arrangement of treatments and differences between means were considered significant at P ≤ 0.05. Postinoculation fecal score was worse, ST shedding, cecal myeloperoxidase, and cecal and colonic ST colonization were greater in PB compared to AB pigs (P < 0.05). Translocation of ST to spleen was decreased by FAA+ (P < 0.05), regardless of dietary PS. Postinoculation, AB pigs had greater average daily gain compared to PB-FAA- (P < 0.05). Pigs fed AB-FAA- showed increased average daily feed intake compared to PB-FAA- pigs (P < 0.05) and feed efficiency was increased in AB-FAA+ compared to PB-FAA- pigs (P < 0.05). Feeding PB ingredients in nursery diets seems to increase susceptibility of pigs to Salmonella. Moreover, FAA supplementation partially attenuated the negative effects of PB diets on the response of pigs to ST challenge.

While long-term growth performance of weaned pigs is not negatively affected by feeding nursery diets containing only plant-based protein sources, these pigs may be more susceptible to subsequent disease challenges. It has been previously shown that supplementation with key functional amino acids improves growth performance and the immune status of pigs during intestinal pathogen challenge. A study was performed to determine the effect of feeding nursery diets containing only plant-based protein sources or including animal-based protein sources with or without supplementation with a blend of functional amino acids (methionine, threonine, and tryptophan) on growth and immune status during a subsequent Salmonella challenge. Pigs fed diets containing animal-based protein sources had improved growth performance and immune status compared to pigs fed plant-based diets, regardless of the inclusion of functional amino acids. Pigs fed plant-based diets were more susceptible to the disease challenge, however, this was partially mitigated by the inclusion of functional amino acids. The inclusion of animal-based protein sources may be necessary to optimize pig health and performance, however, functional amino acid inclusion may be beneficial when plant-based diets are fed.

Ileal alkaline phosphatase is upregulated following functional amino acid supplementation in Salmonella Typhimurium-challenged pigs.

We recently showed that functional amino acid (FAA) supplementation improves growth performance and immune status of Salmonella Typhimurium (ST)-challenged pigs, which was further improved by a longer adaptation period. It is expected that the effects are associated with increased activity of intestinal alkaline phosphatase (IAP). The objective of this study was to evaluate the effects of FAA supplementation and adaptation period on the ileal, cecal, and colonic activity of IAP in weaned pigs challenged with ST. In experiment 1, a total of 32 mixed-sex weanling pigs were randomly assigned to dietary treatments in a 2 × 2 factorial arrangement with low (LP) or high protein (HP) content and basal (FAA-) or FAA profile (FAA+; Thr, Met, and Trp at 120% of requirements) as factors. In experiment 2, a total of 32 mixed-sex weanling pigs were randomly assigned to one of four dietary treatments, being FAA- fed throughout the experimental period (FAA-) or an FAA profile fed only in the post-inoculation (FAA + 0), for 1 wk pre- and post-inoculation (FAA + 1), or throughout the experimental period (FAA + 2). In experiments 1 and 2, after a 7- and 14-d adaptation period, respectively, pigs were inoculated with saline solution containing ST (3.3 and 2.2 × 109 CFU/mL, respectively). Plasma alkaline phosphatase was measured on days 0 and 7 post-inoculation in experiment 1, and IAP (ileum, cecum, and colon) was measured in experiments 1 and 2. Correlations among ileal IAP and serum albumin and haptoglobin, plasma superoxide dismutase (SOD), malondialdehyde (MDA), and reduced:oxidized glutathione, ileal myeloperoxidase, ST shedding and ileal colonization, and post-inoculation average daily gain, feed intake (ADFI), and gain:feed were also analyzed. In experiment 1, plasma alkaline phosphatase was decreased with ST inoculation and the overall content was increased in LP-FAA+ compared with LP-FAA- (P < 0.05). Moreover, ileal IAP was increased in FAA+ compared with FAA- pigs in both studies (P < 0.05) regardless of adaptation time (P > 0.05). IAP was positively correlated with MDA and ADFI and negatively correlated with SOD and ST shedding in experiment 1 (P < 0.05). These results demonstrate a positive effect of FAA supplementation, but not adaptation period, on ileal alkaline phosphatase activity in Salmonella-challenged pigs, which may be associated with improvements in antioxidant balance.

Functional amino acid (FAA) supplementation has been shown to improve gut health and antioxidant defense in weaned piglets challenged with Salmonella Typhimurium (ST), regardless of dietary protein content. The beneficial effects were further improved when pigs were adapted to FAA for 2 wk prior to the ST challenge. Recent evidence has shown that intestinal alkaline phosphatase (IAP), which may be influenced by nutritional factors, attenuates intestinal inflammation, possibly due to gut microbiota modulation. This study is the first to identify that ileal IAP activity is increased following FAA supplementation in ST-challenged pigs, regardless of adaptation period. Moreover, ileal IAP activity correlated with systemic markers of antioxidant defense, which highlights the enzyme's role in attenuating systemic infection. Overall, the development of feeding strategies with positive effects on IAP activity is of interest, due to the enzyme's central role on the gut and whole-body homeostasis and health.

Hulless barley and ß-glucanase affect ileal digesta soluble beta-glucan molecular weight and digestive tract characteristics of coccidiosis-vaccinated broilers.

Exogenous ß-glucanase (BGase) in barley-based feed has been shown to reduce digesta viscosity in chickens, and thereby improve performance. Less well studied is the potential for BGase to convert barley ß-glucan into low molecular weight carbohydrates, which might influence digestive tract function and enteric disease. Coccidiosis-vaccinated broiler chickens were fed graded levels of hulless barley (HB) and BGase to determine their effects on ß-glucan depolymerization and digestive tract characteristics. Broilers were fed high ß-glucan HB (0%, 30% and 60% replacing wheat) and BGase (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 on d 5. Each treatment was assigned to 1 pen in each of 9 rooms. The significance level was set at P ≤ 0.05. At both 11 and 33 d of broiler ages, peak molecular weight of ß-glucan in ileal digesta decreased with increasing BGase for 30% and 60% HB. The maximum molecular weight for the smallest 10% ß-glucan molecules (MW-10%) decreased with BGase at both ages for 30% and 60% HB; for birds fed 0% HB, only 0.1% BGase decreased MW-10%. The 0.1% BGase increased caecal short chain fatty acids (SCFA) compared to the 0.01% BGase at d 11 only for the 60% HB. Ileal pH increased with increasing HB and BGase at d 11 and 33. Caecal pH was lower for 0.1% BGase than 0% BGase for 60% HB at d 11. Relative mRNA expression of interleukin 6 (IL-6) and IL-8 in the ileum increased with 0.1% BGase at d 11 and 33, respectively, whereas expression of ileal mucin 2 (MUC2) decreased with 0.1% BGase at d 33. In the caeca, interactions between HB and BGase were significant for monocarboxylate transporter 1 (MCT1) and mucin 5AC (MUC5 AC) on d 11, but no treatment effects were found at d 33. In conclusion, BGase depolymerized high molecular weight ß-glucan in HB in a dose-dependent manner. Hulless barley and BGase did not increase SCFA concentrations (except for 60% HB with 0.1% BGase at d 11) and caused minor effects on digestive tract histomorphological measurements and relative mRNA gene expression.

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

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.

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

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 ß-glucanase (BGase) and medication on ß-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 ß-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% ß-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 ß-glucan. Beta-glucanase acted as a partial replacement for diet medication by increasing growth performance in coccidiosis vaccinated broilers.

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

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.

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

Diet ß-glucanase (BGase) depolymerizes viscous ß-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 ß-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.

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

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 (p < 0.05). High fCP, regardless of DF content, reduced Thr requirement for PD. Serum antioxidant capacity increased as dietary Thr level increased (p < 0.05). Cecal digesta short-chain fatty acids (SCFA) increased (p < 0.05) with HF and branched-chain fatty acids (BCFA) increased with HfCP and reduced with HF (p < 0.05). HfCP reduced (p < 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 (p < 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 (p < 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.

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

BACKGROUND: The independent and interactive effects of dietary fiber (DF) and threonine (Thr) were investigated in growing pigs challenged with either systemic E. coli lipopolysaccharide (LPS) or enteric Salmonella 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 E. coli LPS and fed low fiber (LF) or high fiber (HF) diets with graded dietary Thr. Urinary lactulose recovery and L:M ratio increased (P < 0.05) during the LPS inoculation period in LF fed pigs but not in HF fed pigs. Fecal mucin output was increased (P < 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 Salmonella Typhimurium challenged pigs, fed LF or HF diets with standard or supplemented dietary Thr. Salmonella inoculation increased (P < 0.05) fecal mucin output compared to the unchallenged period. Supplemental Thr increased fecal mucin output in the HF-fed pigs (Fib × Thr; P < 0.05). Feeding HF increased (P < 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 (P = 0.06) for increased MUC2 expression with the HF diet. Feeding HF increased goblet cell numbers (P < 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 Salmonella-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.

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

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.

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

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.

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

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.

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

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.

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

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.

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

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.

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

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.

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

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.

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

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.