25. Supplementation of Nucleotides and Branched Chain Amino Acids in the Diets of Weaned Pigs Fed Low Protein Diets and Its Effects on Intestinal Morphology, Inflammatory Markers, and Colonic Microbiota

Journal of Animal Science · 2026-04-01

Abstract Feeding low protein (LP) diets to weanling pigs without supplementation of limiting amino acids reduces growth performance and alters fecal microbiota composition. This study evaluated tissue expression of metabolic and inflammatory genes, gut histomorphometry and microbial composition of pigs fed LP diets after weaning. One hundred and sixty piglets with an initial body weight of 5.8 ± 0.92 kg were randomly divided into 5 treatment groups (n = 8/group) in a randomized complete block design for a 42-day nursery trial. The diets included a normal crude protein (CP) positive control (PC) diet, a low CP negative control (NC) diet, NC supplemented with branched chain amino acids (BCAA; NC+BCAA), NC supplemented with 9 g/kg of dietary nucleotides (NC09), NC supplemented with both BCAA and 9 g/kg of dietary nucleotides (NC+BAA+09). On d 42, intestinal tissues were collected from one pig per pen to evaluate morphology and mRNA expression, and colonic digesta were collected to assess microbiome composition. Pigs fed the NC diet had decreased (P < 0.05) villus height (VH), villus height-to-crypt depth ratio (VH:CD), villus perimeter (VP), and area (VA) relative to PC. The NC+BCAA+09 group showed intermediate values for these parameters, which were not different from either PC or NC. The NC+BCAA group had intermediate VH, VP and VA, whereas NC09 had VH that did not differ from PC or NC. Low protein diets without BCAA supplementation (NC and NC09) increased (P < 0.05) jejunal mRNA expression of phosphoglycerate dehydrogenase (PHGDH), and NC increased (P < 0.05) phosphoserine aminotransferase 1 (PSAT1) expression. In the liver, LP diets without BCAA increased (P < 0.0001) tumor necrosis factor-alpha (TNFA) expression, while NC09 increased (P < 0.0001) IL-1β expression. Additionally, NC, NC+BCAA, and NC09 increased (P < 0.0001) hepatic IL-10 expression relative to PC and NC+BCAA+09. β-diversity analysis of microbial community structures showed distinct differences (P < 0.05) between PC and NC diets, and between NC09 and NC+BCAA. At the phylum level, the relative abundance of Spirochaetes and Verrucomicrobia was higher (P < 0.05) in the NC+BCAA treatment group relative to others. Also, genus Bifidobacterium was more abundant in the NC+BCAA and NC09 groups compared to others (P < 0.05). There was greater abundance of Escherichia across all diets relative to NC, whereas Akkermansia was enriched in NC+BCAA relative to NC (P < 0.05). These results suggest that supplementing LP diets with nucleotides and BCAA partially ameliorated the negative effects of protein restriction on growth by improving jejunal morphology, modulating metabolic and inflammatory responses, and altering gut microbiota composition.

Effect of Pre-Exposure to Deoxynivalenol on the Response of Porcine Intestinal Epithelial Cells to F18 E. coli Infection

Toxins · 2026-03-14

The mycotoxin deoxynivalenol (DON) is a common contaminant found in swine diets, causing decreased growth performance and poor health. Additionally, F18 enterotoxigenic E. coli is a leading cause of post-weaning diarrhea. Nursery pigs are often exposed to each of them after weaning; however, it is unknown what impact the combination of these stressors has on gastrointestinal health. Therefore, the objective of this study was to investigate the effect of pre-exposure to DON on the response of intestinal porcine epithelial cells (IPEC-J2) to challenge with enterotoxigenic F18 E. coli. Four groups were compared: Control (untreated cells), DON (cells treated with 0.5 μM DON for 24 h), F18 E. coli (multiplicity of infection 5:1, varied duration) and DON + E. coli (DON treatment with subsequent E. coli infection). Gene expression of IL-8, IL-6 and TNFα was significantly increased in cells infected with E. coli for 3 h vs. uninfected cells (p < 0.0001, p < 0.0001 and p < 0.0001, respectively). There was an interactive effect between DON and E. coli on IL-8 gene expression; cells pretreated with DON before E. coli infection had a higher expression of IL-8 than those not pretreated (p < 0.05). The concentration of IL-8 protein was significantly increased by E. coli (p < 0.0001). Claudin 1 and Occludin protein abundance were reduced by E. coli as measured by Western blot. Cytotoxicity was increased by E. coli vs. Control (p < 0.05). Pretreatment with DON increased the amount of E. coli that adhered to IPEC-J2 cells (p < 0.01) 30 min post-infection. FITC-dextran passage was increased in the DON + E. coli treatment vs. E. coli alone (p < 0.0001). Transepithelial electrical resistance (TEER) was decreased by DON when compared to untreated cells at 0 h (p < 0.0001). Similarly, DON + E. coli exhibited lower TEER vs. E. coli alone at 2 h post-infection (p < 0.0001). Taken together, these results indicate that DON pre-exposure increased the severity of E. coli infection on endpoints such as barrier permeability and E. coli adhesion.

4-Methylcatechol prevents deoxynivalenol-induced claudin-4 internalization and restores barrier integrity via Nrf2 activation

Toxicon · 2026-03-13

163 Evaluating the interaction of branched-chain amino acids and nucleotide supplementation in nursery pigs fed low protein diets

Journal of Animal Science · 2025-05-01

Abstract In a six-week study, a total of 160 piglets (initial body weight, 5.79 ± 0.01 kg) were used to investigate the effects of dietary nucleotides and branched-chain amino acids (BCAA) supplementation of low protein diets on growth performance, blood metabolites, short-chain fatty acids (SCFA), apparent ileal and total tract digestibility of nutrients (AID and ATTD). Following a 3-d adaptation period, pigs were randomly assigned to one of five dietary treatments using a randomized complete block design. Each treatment consisted of 8 replicate pens with 4 piglets per pen. The experimental diets included a high-crude protein (CP) Positive Control (PC) diet with CP at 22%, 20.5%, 19.4% for phases 1, 2 and 3, respectively; a BCAA-deficient low-CP Negative Control (NC) diet with CP at 16%, 14.5%, 13.4% for phases 1, 2 and 3, respectively; NC diet supplemented with crystalline isoleucine and valine (NC+BCAA); NC diet supplemented with 9 g/kg of dietary nucleotides (NC09); and NC diet supplemented with both BCAA and nucleotides (NC+BCAA+09). Body weights and feed intake were determined to calculate average daily feed intake (ADFI), average daily gain (ADG), and gain-feed ratio (G:F). Fecal samples were collected on d 39, 40 and 41 to determine the ATTD of nutrients. Blood samples were collected on day 42 to measure blood metabolites and pigs were euthanized to determine the AID of nutrients and colonic SCFA. Low CP (LP) diets decreased the ADG, G:F and final BW in pigs (P <, 0.05). Pigs fed the PC diet had the highest overall ADG, G:F and final BW (397 g/d, 561 g/kg and 22.9 kg), with intermediate values in NC+BCAA+09 (337 g/d, 489 g/kg and 20.2 kg) and NC+BCAA (303 g/d, 470 g/kg and 18.6 kg), and the lowest values in the NC09 (246 g/d, 421 g/kg and 16.1 kg) and NC (227 g/d, 406 g/kg and 15.4 kg). Blood urea nitrogen (BUN) concentrations decreased (P < 0.05) with reduction in CP, while serum glucose, insulin and glutathione concentrations were not affected by treatments. The NC diet had decreased (P < 0.05) serum insulin-like growth factor-1 levels, with partial recovery in the NC+BCAA diet and complete recovery in the NC+BCAA+09 diet, compared to PC. Additionally, the NC+BCAA+09 diet showed a tendency (P = 0.09) for increased serum uric acid concentration but had the highest ATTD of gross energy (GE), dry matter (DM) and phosphorus (P) as well as the highest AID of N and P. The NC diets had decreased the colonic SCFA concentration. In conclusion, supplementation with nucleotides and BCAA has beneficial additive effects on growth performance and nutrient digestibility in pigs fed BCAA-deficient low-CP diets.

148 Dietary gestational and lactational live yeast supplementation to sows impacts milk and colostrum proteome profiles and piglet gut development.

Journal of Animal Science · 2025-10-01

Abstract Sow milk serves as the sole source of nutrients, immune-stimulating and growth-promoting factors for suckling piglets. Milk composition and production are largely affected by sow nutrition during gestation and lactation. Due to the dependence of piglets on milk, the suckling period presents a critical opportunity for maternal nutritional intervention to support piglet intestinal development. This study investigated the effects of live yeast (LY, Saccharomyces cerevisiae) supplementation to sows during late gestation and lactation on intestinal development of piglets, and the proteome profiles of sow colostrum and milk. On d 77 of gestation, 40 sows were allotted to each of two dietary treatments: without (CON) or with (LY) live yeast supplementation at 0.05% of diet during gestation and 0.1% during lactation. Colostrum and milk samples were collected on d 0, 10, and 18 of the lactation (n = 6). Intestinal tissue samples were collected from one piglet per litter on postnatal days (PND) 0, 10, 18, and postweaning days (PWD) 7 and 14 (n = 10). Results showed that maternal LY supplementation increased mRNA expression of interleukin (IL)-6 on PND18 and IL-1β on PWD14 in the ileal mucosa (P < 0.05), with a trend towards higher IL-10 expression on PND18 in the ileal mucosa (P = 0.08). Additionally, LY piglets had increased mRNA expression of superoxide dismutase 1 (SOD1) in the ileal mucosa on PND10 and PWD14 (P < 0.05). Compared to CON piglets, LY piglets had increased protein abundance of E-cadherin on PWD7 and PWD14 in the jejunal mucosa (P < 0.05), with a tendency for greater E-cadherin on PND0 (P = 0.09). The LY piglets had a higher protein abundance of occludin and claudin-4 in the jejunal mucosa on PWD14 (P < 0.05). Shotgun proteomic analysis revealed a higher abundance of immune-associated proteins, including Ig-like domain-containing protein and complement proteins (Complement C8 alpha chain and C1q domain-containing proteins) in the colostrum, and IgG heavy chain in d 10 milk of LY sows than CON sows, whereas the abundance of prostaglandin synthetase was greater in d 10 milk of CON sows than LY sows (P < 0.05). Additionally, milk fat globule EGF and factor V/VIII domain containing protein and Niemann-Pick C2 were found to be more abundant in both d 10 and d 18 milk of LY sows (P < 0.05). In conclusion, dietary live yeast supplementation to sows during gestation and lactation enhanced piglet intestinal development by increasing immune-related proteins in colostrum and proteins involved in lipid uptake and processing in mature milk, thereby supporting neonatal immune and tissue development at different growth stages.

188. Assessment of dietary Spirulina supplementation on growth, jejunal morphology, nutrient digestibility, and intestinal health-related genes in LPS-challenged weanling pigs

Animal - science proceedings · 2025-08-01

Impact of partially defatted black soldier fly larvae meal on coccidia-infected chickens: effects on growth performance, intestinal health, and cecal short-chain fatty acid concentrations

Journal of Animal Science and Biotechnology/Journal of animal science and biotechnology · 2025-02-26 · 5 citations

BACKGROUND: Black soldier fly larvae meal (BSFLM) stands out as a promising nutritional resource due to its rich bioactive substances and favorable protein profile. Nonetheless, its potential to mitigate coccidia infection in broilers remains uncertain. This study aimed to evaluate the impact of partially defatted BSFLM (pBSFLM) on growth performance, nutrient utilization, and intestinal health, focusing on morphology, immunology, and cecal fermentation in coccidia-infected broilers. METHODS: Over the initial 13 d, 480 newly-hatched Cobb 500 male birds were allocated to three diets with increasing pBSFLM concentrations (0, 60, or 120 g/kg). At d 13 post hatching, chicks within each dietary group were further allotted to non-challenge or challenge subsets, generating six treatments in a 3 × 2 factorial arrangement. Challenged birds were orally administered oocysts of E. maxima, E. acervulina, and E. tenella (25,000:125,000:25,000). RESULTS: During the infection phase (d 13 to 19), linear interactions between Eimeria and pBSFLM were observed in gain to feed ratio (G:F) (P < 0.05) and cecal interferon-γ (IFN-γ, P < 0.05), with a tendency in cecal acetate concentration (P = 0.06). A quadratic interaction was observed in crypt depth (CD, P < 0.05). Incremental pBSFLM inclusion negatively affected G:F, CD, IFN-γ, and acetate productions in the ceca under coccidia challenge. Conversely in non-challenged birds, the impact of pBSFLM varied from neutral (e.g. G:F) to potentially advantageous (e.g. acetate). Challenged birds exhibited decreased (P < 0.01) BW, BW gain, feed intake (FI), and the apparent ileal digestibility and total tract nutrient utilization of DM, gross energy, and nitrogen (N). Eimeria challenge reduced (P < 0.01) serum carotenoid concentrations, decreased the villus height to crypt depth ratio (VH:CD, P < 0.01), and increased concentrations of branched-chain fatty acids, specifically isobutyrate (P = 0.059) and isovalerate (P < 0.05) in the cecum. Dietary pBSFLM addition linearly reduced (P < 0.05) BW, FI, and N utilization. Tendencies (P < 0.06) were observed where pBSFLM linearly decreased VH:CD and reduced goblet cell density. CONCLUSIONS: Increasing pBSFLM supplementation, particularly at 12%, adversely affected growth, ileal morphology, cecal acetate production, and downregulated key cytokine expression in response to coccidia infection.

PSI-6 Temporal analysis of jejunal mucosal proteome reveals profile changes in piglets from birth to early postweaning stage

Journal of Animal Science · 2025-05-01

Abstract The intestinal mucosa is a dynamic and interactive structure that supports physiological processes essential for maintaining intestinal architecture, epithelial renewal, cellular metabolism, and mucosal immunity. As piglets age, the intestinal mucosa adapts to dietary changes and evolving physiological demands across different growth stages, partly through changes in proteome profiles. However, little is known about the dynamic changes of mucosal proteomes during the preweaning and postweaning stages in piglets. The aim of this study was to investigate temporal changes in jejunal mucosal proteomes during the suckling and early postweaning periods. On postnatal day (PND) 0 (within 24 h after birth), 10, 18, postweaning day (PWD) 7 and 14, six barrows were selected and euthanized for jejunal mucosa collection. Total proteins were extracted from mucosal samples and subjected to liquid chromatography-tandem mass spectrometry for shotgun proteomic analysis. A total of 5,498 proteins were detected across all time points. Principal component analysis and hierarchical clustering were applied to capture features representing the entire proteome across developmental stages. These analyses revealed age-related effects on the jejunal mucosal proteomes. Specifically, mucosal proteomes on PND 10 were distinct from those on PWD 7 and PWD 14, with PND 18 intermediate to PND 10 and postweaning profiles, while samples on PND 0 showed considerable variation. Analysis of the top 50 differentially expressed proteins revealed that mucosal proteins on PND 0 had the highest abundance of alpha-fetoprotein, Ig-like domain-containing proteins, MAM domain-containing 4, neural cell adhesion molecule 1, branched-chain-amino-acid aminotransferase, and melanotransferrin (FDR &amp;lt; 0.05). Additionally, the abundance of proteins involved in stress and antioxidant defense responses, amino acid metabolism, protein translation, and post-translational modifications or function as metabolic enzymes and proteasome components, increased from PND 0 to PWD 14 (FDR &amp;lt; 0.05). Gene ontology enrichment analysis indicated age-related changes in proteins associated with translation, tricarboxylic acid cycle, glycolytic process, positive regulation of cell proliferation, immune response, and carbohydrate metabolic process (P &amp;lt; 0.05). Taken together, the shift in proteomes likely reflects an age-related upregulation of metabolic and stress responses and protein turnover that are critical for maintaining cellular homeostasis and resilience against age-associated oxidative stress. Overall, the findings may indicate age-associated physiological changes during intestinal maturation in piglets, including increases in antioxidant capacity, energy production, protein repair, and maintenance of tissue growth and integrity.

PSVI-18 Mycotoxin exposure and its impact on inflammatory response and growth in pigs and intestinal epithelial cells

Journal of Animal Science · 2025-05-01

Abstract The objective of these studies was to determine the effect of pre-exposure to the mycotoxin deoxynivalenol (DON) on the response of intestinal porcine epithelial cells (IPEC-J2) and nursery pigs to immune challenge with F18 E. coli and lipopolysaccharide (LPS), respectively. In-vitro experiments involved four experimental treatments: Control, DON (0.5μM, 24hr), F18 E. coli (multiplicity of infection 5:1, 3hr) and DON + E. coli. Results showed that IL-8, IL-6 and TNFα gene expression were upregulated in the E. coli and DON + E. coli treatments (P&amp;lt; 0.05). Relative to the Control, Claudin-4 protein expression tended to be decreased in the E. coli treatment (P=0.054) and was significantly reduced in the DON + E. coli treatment (P=0.015). Furthermore, Occludin protein expression was downregulated in E. coli (P=0.001) and DON + E. coli (P=0.001) treatments relative to Control. In each case, the DON treatment was not different from Control. A lactate-dehydrogenase assay was completed to evaluate cell death. This assay revealed that the E. coli treatment increased cell death relative to Control and DON treatments (P&amp;lt; 0.05). Additionally, the DON + E. coli treatment had higher cell death relative to the Control, DON and E. coli treatments (P&amp;lt; 0.05), indicating that pre-exposure to DON increased the lethality of E. coli in IPEC-J2 cells. To complement in-vitro work, a 28-day nursery study was conducted using 128 piglets allotted to 4 treatments with each treatment having a total of eight replicates in a randomized-complete block design. The treatments were: Control (fed standard diets, no final LPS injection), Mycotoxin (fed diets formulated to contain 2.5 ppm DON, no final LPS injection), Control + LPS (fed control diets, injected with LPS at 25 μg/kg BW on d 28), and Mycotoxin + LPS (fed mycotoxin contaminated diets, injected with LPS at 25 μg/kg BW on d 28). Body weights (kg) were higher (P &amp;lt; 0.05) in the Control than Mycotoxin pigs on d 14 (9.6 vs. 9.3) and d 28 (17.1 vs. 15.8). Similarly, average daily gain was higher (P &amp;lt; 0.05) in the Control than Mycotoxin pigs from d 1-14, 15-28 and overall (1-28). However, Gain:Feed was not affected by treatments (P &amp;gt; 0.05). Rectal temperatures were collected from pigs hourly for four hours following injection. At 1, 3 and 4 hours post-injection, pigs injected with LPS had elevated temperatures compared to those injected with saline (P=0.03, P=0.01, and P=0.00, respectively). At 4 hours post-injection, the Mycotoxin + LPS injection treatment tended (P=0.09) to have a higher rectal temperature relative to control diet + LPS injection treatment. In sum, prior mycotoxin exposure may impact how intestinal cells and nursery pigs respond to inflammatory or immune challenge with a potential consequence on intestinal function and growth of pigs.

Author response for "Impact of Gestational and Lactational Live Yeast Supplementation to Sows on Litter Performance, Colostrum and Milk Proteome Profiles"

2025-06-10