Andrew. G. Van Kessel

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.

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 performance of broilers. 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 challenged with a coccidiosis vaccine on d 5. Each treatment was assigned to one floor pen in each of nine rooms. In Experiment 1, the soluble β-glucan weight 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. 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 (BWG) 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 BWG after d 11, and lower and higher feed efficiency before and after d 11, respectively, in unmedicated treatments. In conclusion, BGase and medication caused the depolymerization of soluble ileal β-glucan. Beta-glucanase appeared as a partial replacement for diet medication to increase coccidiosis challenged broiler performance.

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 (PP < 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.

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 2 blocks, whereas Study 2, was conducted using feces collected after a period of 3 (Exp. 1) or 4 weeks (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 (VFA) 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.

As an alternative to antibiotic growth promoters, live yeast supplementation has proven useful in reducing weaning stress and improving performance parameters of piglets. Here, we compared the performance and hindgut microbiota of weanling piglets subjected to different pre- and post-weaning yeast supplementation regimens using a live strain of Saccharomyces cerevisiae (Actisaf Sc 47). Average feed intake and average daily weight gain of piglets within Yeast-Control and Yeast-Yeast groups were higher than those in the Control-Control group. Yeast supplementation resulted in development of microbial communities that were phylogenetically more homogenous and less dispersed compared to the microbiota of control piglets. Key bacterial taxa overrepresented in the microbiota of yeast supplemented piglets included phylum Actinobacteria, specifically family Coriobacteriaceae, as well as Firmicutes families Ruminococcaceae, Clostridiaceae, Peptostreptococcaceae, and Peptococcaceae. Correlation network analysis revealed that yeast supplementation was associated with enrichment of positive correlations among proportions of different bacterial genera within the hindgut ecosystem. In particular, within the cecal microbiota of supplemented piglets, higher numbers of positive correlations were observed among potentially beneficial genera of the phyla Actinobacteria and Firmicutes, suggesting a mechanism by which yeast supplementation may contribute to regulation of intestinal homeostasis and improved performance of piglets.

Salmonella enterica Typhimurium (ST) is of concern in the swine industry with relevance for animal health and consumer safety. Nutritional strategies might help to reduce ST infection and transmission. This study examined the potential of wheat (Triticum aestivum) distillers dried grains with solubles (DDGS) and sugar beet (Beta vulgaris) pulp (SBP) to alter intestinal microbial communities and ST shedding using a Trojan model. Weaned pigs (n = 105; 28.5 ± 3.5 d of age) were separated into 3 treatment groups (7 pigs/pen) and fed a wheat-based control diet or the control diet formulated with 15% wheat DDGS or 6% SBP inclusion. Following 12 d of diet adaptation, 2 pigs/pen were inoculated with 2 x 10⁹ cfu ST, resistant to novobiocin and nalidixic acid. Fecal swabs were taken from infected pigs and pen-mates (contact pigs) for 9 d following challenge, enriched in nutrient broth for 24 h, and plated on selective media to determine prevalence of ST. The ranges of prevalence of ST in feces were from 90 to 100% in challenged pigs and 74 to 78% in contact pigs. No influence of treatment on rectal temperature and prevalence of ST in contact pigs were observed. Fifteen contact pigs were euthanized per treatment group on 9 and 10 d postchallenge to enumerate in intestinal contents (ileum, cecum, and proximal colon), Lactobacillus spp., Enterobacteriaceae, and Clostridium clusters I, VI, and XVIa by quantitative PCR (qPCR) and to determine ST prevalence by selective culture. No significant effects of diet were observed with respect to ST prevalence in feces, ileum, cecum, colon, and lymph nodes of contact pigs. Compared with the control diet, DGGS and SBP diets showed a trend towards increased (P < 0.1) number of Lactobacillus species in the cecum and colon. Although both wheat DGGS and SBP tended to increase the Lactobacillus spp. neither of the feed ingredients affected ST prevalence. Copyright © 2013. American Society of Animal Science .

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 β-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.

Summary: To study microbial influence on intestinal development pertaining to nutrient digestion, two separate gnotobiotic experiments were performed, each with 16 piglets allocated to four treatment groups: germfree (GF), monoassociation with Escherichia coli, monoassociation with Lactobacillus fermentum or conventionalization with faecal bacteria (CV). Enzyme activity and gene expression of lactase phlorizin hydrolase (LPH) and aminopeptidase N (APN) were measured in isolated enterocytes, harvested on day 14, using specific substrates and quantitative PCR respectively. Enterocytes of CV pigs had reduced APN activity, but had increased gene expression relative to GF, making the specific activity:mRNA (A:G) ratio dramatically lower (p < 0.05). Similarly, LPH A:G ratio was significantly reduced (p < 0.05) in enterocytes of CV pigs as compared with GF. The results of co‐incubation of L. fermentum, E. coli and faecal bacteria with APN indicate a direct relationship between enzyme inactivation and specific A:G ratio in enterocytes. We conclude that enterocyte up‐regulation of APN expression occurs as either a direct response to microbial colonization or as a feedback mechanism in response to reduced enzyme activity through microbial degradation. This mechanism may play a role in ensuring effective competition of the host with the intestinal microbiota for available nutrients.

We previously reported marked differences in small intestinal morphology, including changes in crypt depth and villous height, after inoculation of germ-free pigs with different bacterial species. In an attempt to identify the mechanisms governing changes in villous morphology associated with bacterial colonization, 2 gnotobiotic experiments were performed. In each experiment, 16 piglets were allocated to 4 treatment groups including germ-free (GF), monoassociation with Lactobacillus fermentum (LF) or Escherichia coli (EC), or conventionalized with sow feces (SF). Piglets were reared under gnotobiotic conditions until 14 d of age, at which time whole intestinal tissue and enterocytes were collected for histological, gene expression, and protein analysis. Proliferating cell nuclear antigen, tumor necrosis factor α (TNFα), Fas ligand (FasL), CD3ε, caspase 3 (casp3), and toll-like receptors (TLR)2, 4, and 9 expression were measured by quantitative PCR. Activated casp3 was measured by Western blot. Increased abundance of activated casp3 and transcripts encoding proliferating cell nuclear antigen, TNFα, CD3ε, and FasL was observed in SF and EC treatment groups compared with GF and LF. Expression of TLR2 was increased (P < 0.05) in the SF treatment and tended to be greater (P < 0.08) in EC relative to LF and GF. Results indicate that conventional bacteria and E. coli but not L. fermentum increase overall cell turnover by stimulating increased apoptosis through the expression of FasL and TNFα and by increasing cell proliferation. The differential regulation of TLR expression indicates that microbially induced changes may be mediated in part by these receptors. Induction of inflammatory responses and activation of apoptosis through death receptors appears to play a significant role in enterocyte turnover mediated by commensal bacteria. Copyright © 2007. . Copyright 2007 Journal of Animal Science