The Amino Acids Sensing and Utilization in Response to Dietary Aromatic Amino Acid Supplementation in LPS-Induced Inflammation Piglet Model

Dietary supplementation with aromatic amino acids (AAAs) has been demonstrated to alleviate intestinal inflammation induced by lipopolysaccharide (LPS) in the piglets. But the mechanism of AAA sensing and utilization under inflammatory conditions is not well-understood. The study was conducted with 32 weanling piglets using a 2 × 2 factorial arrangement (diet and LPS challenge) in a randomized complete block design. Piglets were fed as basal diet or the basal diet supplemented with 0.16% tryptophan (Trp), 0.41% phenylalanine (Phe), and 0.22% tyrosine (Tyr) for 21 days. The results showed that LPS treatment significantly reduced the concentrations of cholecystokinin (CCK) and total protein but increased leptin concentration, the activities of alanine transaminase, and aspartate aminotransferase in serum. Dietary supplementation with AAAs significantly increased the serum concentrations of CCK, peptide YY (PYY), and total protein but decreased the blood urea nitrogen. LPS challenge reduced the ileal threonine (Thr) digestibility, as well as serum isoleucine (Ile) and Trp concentrations, but increased the serum concentrations of Phe, Thr, histidine (His), alanine (Ala), cysteine (Cys), and serine (Ser) (P < 0.05). The serum-free amino acid concentrations of His, lysine (Lys), arginine (Arg), Trp, Tyr, Cys, and the digestibilities of His, Lys, Arg, and Cys were significantly increased by feeding AAA diets (P < 0.05). Dietary AAA supplementation significantly increased the serum concentrations of Trp in LPS-challenged piglets (P < 0.05). In the jejunal mucosa, LPS increased the contents of Ala and Cys, and the mRNA expressions of solute carrier (SLC) transporters (i.e., SLC7A11, SLC16A10, SLC38A2, and SLC3A2), but decreased Lys and glutamine (Gln) contents, and SLC1A1 mRNA expression (P < 0.05). In the ileal mucosa, LPS challenge induced increasing in SLC7A11 and SLC38A2 and decreasing in SLC38A9 and SLC36A1 mRNA expressions, AAAs supplementation significantly decreased mucosal amino acid (AA) concentrations of methionine (Met), Arg, Ala, and Tyr, etc. (P < 0.05). And the interaction between AAAs supplementation and LPS challenge significantly altered the expressions of SLC36A1 and SLC38A9 mRNA (P < 0.05). Together, these findings indicated that AAAs supplementation promoted the AAs absorption and utilization in the small intestine of piglets and increased the mRNA expressions of SLC transports to meet the high demands for specific AAs in response to inflammation and immune response.

Supplementation of oligosaccharide-based polymer enhanced growth and disease resistance of weaned pigs by modulating intestinal integrity and systemic immunity

Background There is a great demand for antibiotic alternatives to maintain animal health and productivity. The objective of this experiment was to determine the efficacy of dietary supplementation of a blood group A6 type 1 antigen oligosaccharides-based polymer (Coligo) on growth performance, diarrhea severity, intestinal health, and systemic immunity of weaned pigs experimentally infected with an enterotoxigenic Escherichia coli (ETEC), when compared with antibiotics. Results Pigs in antibiotic carbadox or Coligo treatment groups had greater (P < 0.05) body weight on d 5 or d 11 post-inoculation (PI) than pigs in the control group, respectively. Supplementation of antibiotics or Coligo enhanced (P < 0.05) feed efficiency from d 0 to 5 PI and reduced (P < 0.05) frequency of diarrhea throughout the experiment, compared with pigs in the control group. Supplementation of antibiotics reduced (P < 0.05) fecal β-hemolytic coliforms on d 2, 5, and 8 PI. Pigs in antibiotics or Coligo groups had reduced (P < 0.05) neutrophil counts and serum haptoglobin concentration compared to pigs in the control group on d 2 and 5 PI. Pigs in Coligo had reduced (P < 0.05) total coliforms in mesenteric lymph nodes on d 5 and 11 PI, whereas pigs in antibiotics or Coligo groups had reduced (P < 0.05) total coliforms in spleen on d 11 PI compared with pigs in the control group. On d 5 PI, pigs in the Coligo group had greater (P < 0.05) gene expression of ZO1 in jejunal mucosa, but less (P < 0.05) mRNA expression of IL1B, IL6, and TNF in ileal mucosa, in comparison with pigs in the control group. Supplementation of antibiotics enhanced (P < 0.05) the gene expression of OCLN in jejunal mucosa but decreased (P < 0.05) IL1B and IL6 gene expression in ileal mucosa, compared with the control. On d 11 PI, supplementation of antibiotics or Coligo up-regulated (P < 0.05) gene expression of CLDN1 in jejunal mucosa, but Coligo reduced (P < 0.05) IL6 gene expression in ileal mucosa compared to pigs in the control group. Conclusions Supplementation of Coligo improved growth performance, alleviated diarrhea severity, and enhanced gut health in weaned pigs infected with ETEC F18 in a manner similar to in-feed antibiotics.

Alterations of Suckling Piglet Jejunal Microbiota Due to Infection With Porcine Epidemic Diarrhea Virus and Protection Against Infection by Lactobacillus salivarius

The high mortality of neonatal piglets due to porcine epidemic diarrhea virus (PEDV) infection has caused huge economic losses to the pig industry. The intestinal microbiota is an important barrier against invaders entering the gastrointestinal route. In this study, we examined the differences between intestinal microbiota of PEDV-infected and healthy piglets. According to the viral copy numbers, 16 crossbred (Landrace-Yorkshire) piglets were divided into three groups: uninfected, low virus load, and high virus load groups. Next, 16S rRNA sequencing was performed to determine the microbiota composition in jejunal content and jejunal mucosal samples from the three groups. PEDV infection induced an imbalance in the microbiota of both jejunal content and jejunal mucosa. The abundance of phylum Firmicutes was higher in uninfected piglets than in infected piglets, whereas the abundance of Proteobacteria was lower in uninfected piglets. Principal coordinate analysis showed significant separation of jejunal microbiota between different groups. Linear discriminant analysis (LDA) effect size (LEfSe) identified Lactobacillus salivarius as a potential biomarker among three groups at the level of species. Then, in vitro, L. salivarius was able to suppress the infection of PEDV to IPEC-J2 cells and decreased the expression of GRP78 (Glucose-regulating protein 78). In addition, we detected the mRNA expression of genes involved in the FAK/PI3K/Akt signaling pathway. When IPEC-J2 cells were treated with L. salivarius before PEDV infection, the mRNA expression levels of ITGA1, ITGA5, ITGB5, FAK, PIK3R1, PIK3CA and AKT1 were significantly higher than those in the control cells (without treatment) at different times post-infection, indicating that L. salivarius may upregulate the FAK/PI3K/Akt signaling pathway in IPEC-J2 cells to resist PEDV infection. In summary, PEDV infection altered microbial communities in both jejunal content and jejunal mucosa. L. salivarius has a protective effect against PEDV infection in IPEC-J2 cells. This study provides a potentially effective strategy to prevent the occurrence and control the spread of PED in the pig production.

Nutritional and functional values of lysed Corynebacterium glutamicum cell mass for intestinal health and growth of nursery pigs

The objective was to determine the nutritional and functional values of lysed Corynebacterium glutamicum cell mass (CGCM) as a protein supplement and a source of cell wall fragments supporting the growth and intestinal health of nursery pigs. Thirty-two pigs (21 d of age) were allotted to four treatments (n = 8) based on the randomized block design with sex and initial body weight (BW) as blocks. The main effect was the dietary supplementation of lysed CGCM (0, 0.7, 1.4, and 2.1%) replacing blood plasma and fed in two phases (10 and 11 d, respectively). Feed intake and BW were measured at the end of each phase. Pigs were euthanized on day 21 to collect jejunal tissue and mucosa to evaluate intestinal health. Ileal digesta were collected to measure the apparent ileal digestibility of nutrients in diets. Data were analyzed using Proc Mixed and Reg of SAS. Increasing daily intake of CGCM increased (linear; P &lt; 0.05) ADG of pigs. Increasing CGCM supplementation affected (quadratic; P &lt; 0.05) the relative abundance of Lactobacillaceae (minimum: 26.4% at 1.2% CGCM), Helicobacteraceae (maximum: 29.3% at 1.2% CGCM), and Campylobacteraceae (maximum: 9.0% at 1.0% CGCM). Increasing CGCM supplementation affected (quadratic; P &lt; 0.05) the concentrations of immunoglobulin G (maximum: 4.94 µg/mg of protein at 1.0% CGCM) and protein carbonyl (PC; maximum: 6.12 nmol/mg of protein at 1.1% CGCM), whereas linearly decreased (P &lt; 0.05) malondialdehyde (MDA) in the proximal jejunal mucosa. Increasing CGCM supplemention affected (quadratic; P &lt; 0.05) intestinal enterocyte proliferation rate (maximum: 13.3% at 1.0% CGCM), whereas it did not affect intestinal morphology and the nutrient digestibility. In conclusion, supplementing 1.0% to 1.2%, reducing blood plasma supplementation by 0.7% to 0.9%, respectively, increased potential pathogenic microbiota associated in the jejunal mucosa resulting in increased immune response, enterocyte proliferation, and PC concentration. However, supplementing diets with 2.1% CGCM, replacing 1.5% blood plasma, improved growth performance, and reduced MDA without affecting nutrient digestibility, intestinal morphology, and microbiota in the jejunal mucosa. In this study, based on the polynomial contrast, supplementing 1.0% to 1.2% CGCM suppressed the benefits from blood plasma, whereas supplementing 2.1% CGCM showed functional benefits of CGCM with similar effects from blood plasma supplementation.

PSVII-9 Influence of lactose and milk oligosaccharides in whey permeate on jejunal mucosa-associated microbiota in nursery pigs during 7 to 11 kg BW

This study aimed to evaluate supplemental effects of milk carbohydrates in whey permeate on jejunal mucosa-associated microbiota in nursery pigs during 7 to 11 kg BW. A total of 720 pigs at 7.5 kg BW were allotted to 6 treatments (6 pens/treatment and 20 pigs/pen). Treatments were 6 levels of whey permeate supplementation (0, 3.75, 7.50, 11.25, 15.00, and 18.75%) and fed to pigs for 11 d. On d 11, 36 pigs representing median BW of each pen were euthanized to collect the jejunal mucosa to evaluate microbiota in the jejunum by 16S rDNA sequencing. Data were analyzed using contrasts in MIXED procedure of SAS. Whey permeate contained 76.3% lactose and 0.4% milk oligosaccharides. Increasing whey permeate supplementation from 0 to 18.75% did not affect the alpha-diversity estimates of microbiota. Whey permeate supplementation tended to decrease (P = 0.073, 1.59 to 1.22) Firmicutes:Bacteroidetes compared with no addition of whey permeate. Increasing whey permeate supplementation tended to linearly increase Bifidobacteriaceae (P = 0.089, 0.73 to 1.11), decrease Enterobacteriaceae (P = 0.091, 1.04 to 0.52), decrease Stretococcaceae (P = 0.094, 1.50 to 0.71), and caused quadratic changes (P &lt; 0.05) on Lactobacillaceae (maximum: 9.14% at 12.91% whey permeate). Increasing whey permeate supplementation caused a quadratic change (P &lt; 0.05) on Lactobacillus_Salivarius (maximum: 0.92% at 7.35% whey permeate) and tended to cause quadratic changes on Lactobacillus_Rogosae (P = 0.083; maximum: 0.53% at 8.45% whey permeate) and Lactobacillus_Mucosae (P = 0.092; maximum: 0.70% at 6.98% whey permeate). In conclusion, supplementation of whey permeate as sources of lactose and milk oligosaccharides at a range from 7 to 13% seems to be beneficial to nursery pigs by increasing the abundance of lactic acid-producing bacteria in the jejunal mucosa.

PSVII-2 Effects of xylanase and its optimal supplementation level on viscosity of jejunal digesta, nutrient digestibility, intestinal health, and growth performance of nursery pigs

This study aimed to determine supplemental effects of xylanase (endo-β-1,4-xylanase, CJ BIO, Korea) and its optimal supplementation level on viscosity of jejunal digesta, nutrient digestibility, intestinal health, and growth performance of pigs. Sixty weaned pigs (6.9 ± 0.8 kg BW) were randomly allotted to 5 treatments based on a RCBD with initial BW and sex as block and fed in 3 phases (P1/2/3 for 10/14/14 d, respectively). Dietary treatment were the supplementation levels of xylanase providing (0, 220, 440, 880, and 1,760 XU/kg feed). Titanium dioxide (0.4%) was added to P3 diets as an indigestible marker to measure AID. On d 38, all pigs were euthanized to collect ileal and jejunal digesta to measure AID and viscosity, respectively; jejunal mucosa and tissue to measure intestinal health parameters. Data were analyzed using SAS 9.4. Xylanase supplementation from 0 to 350 XU/kg increased (P &lt; 0.05) ADG (596 to 746 g/d) during the last week, whereas there was no effect by xylanase on the overall growth performance. Increasing xylanase supplementation reduced (P &lt; 0.05) digesta viscosity (1.91 to 1.48 mPa.s); increased (P &lt; 0.05) the AID of EE (83.9 to 89.5%), NDF (52.9 to 56.9%) and ADF (35.3 to 39.3%); tended to reduce Cupriavidus (P = 0.073; 1.33 to 0.63%) and Megasphaera (P = 0.063; 1.26 to 0.23%); and tended to increase Succinivibrio (P = 0.076; 1.10 to 2.71%) and Pseudomonas (P = 0.060; 4.89 to 13.29%). Xylanase supplementation from 0 to 520 XU/kg reduced (P &lt; 0.05) jejunal MDA (0.99 to 0.58 µmol/mg protein). In conclusion, xylanase supplementation showed benefits on intestinal health by reducing digesta viscosity, oxidative stress status, and harmful bacteria in the jejunal mucosa and by increasing the AID of nutrients. Xylanase supplementation at a range of 350 to 520 XU/kg feed provided the most benefits.

PSXVI-3 Supplemental effects of Bacillus sp. in prevention of post-weaning diarrhea caused by enterotoxigenic F18+ Escherichia coli in nursery pigs

This study investigated the effects of Bacillus sp. on intestinal health and prevention of diarrhea by F18+ Escherichia coli (ETEC) in nursery pigs. Forty-eight weaned pigs (7.9 ± 0.5 kg BW) were allotted to 4 treatments with 12 replicates (NC: no-challenge; PC: challenge/no-treat; BMD: challenge/bacitracin; BAC: challenge/Bacillus sp. at 109 CFU/kg feed). All pigs were fed diets for 28 d. At d 7, challenged groups were orally inoculated with ETEC (1.8 × 1010 CFU) and NC group received sterile solution. Growth performance was analyzed weekly and pigs were euthanized on d 28 to measure intestinal health. Data were analyzed using SAS 9.4. There were no difference on growth performance among treatments during pre-challenge period. From d 7 to 14, PC tended to reduce (P = 0.067) ADG (352 to 440 g/d), whereas no effect was observed during the overall period. The PC increased (P &lt; 0.05) fecal score at d 7 (post-challenge) (4.1 to 3.3) and d 8 (4.1 to 3.3), whereas BMD decreased (P &lt; 0.05) it at d 9 (4.3 to 3.5) and d 11 (3.8 to 3.3). The PC increased (P &lt; 0.05) MDA (0.56 to 0.30 μmol/mg protein) and tended to increase TNFα (P = 0.084; 0.89 to 1.04 pg/mg protein), whereas BMD and BAC reduced (P &lt; 0.05) MDA (0.56 to 0.31, 0.29 μmol/mg protein) and TNFα (1.04 to 0.81, 0.77 pg/mg protein) in jejunal mucosa. The PC reduced (P &lt; 0.05) α-diversity (Chao1: 32.3 to 19.4), whereas BAC increased it (Chao1: 19.4 to 30.7) whereas BMD increased (P &lt; 0.05) it (Shannon: 1.85 to 2.44; Simpson: 0.55 to 0.70) of jejunal mucosa-associated microbiota. In conclusion, ETEC challenge caused diarrhea, increased immune response, and oxidative stress by disrupting mucosa-associated microbiota. The BMD and BAC reduced jejunal immune response and oxidative stress by restoring mucosa-associated microbiota.

PSXVI-6 Supplemental effects of phytase on bone parameters, intestinal health, nutrient digestibility and growth performance on broiler chickens

This study aimed to determine supplemental effects of phytase (VTR BIO-TECH, China) on bone parameters, jejunal mucosa-associated microbiota, intestinal morphology, nutrient digestibility, and growth performance of broiler chickens (Ross 308). Newly hatched broiler chickens (360, 44 ± 6 g BW) were randomly allotted in 6 treatments with 10 birds per cage based on CRD and fed for 27 d. Dietary treatments were the supplementation levels of phytase providing (0; 500; 1,000; 2,000; 4,000 FTU/kg feed) and 0.15% less Ca and 0.15% less P than the recommended requirements by ROSS 2019. Titanium dioxide (0.4%) was added to the diets as an indigestible marker to measure AID. On d 27, 3 birds were randomly selected from each cage and euthanized to collect jejunal mucosa and tissue to measure microbiota diversity and morphology, ileal digesta to measure AID, and left tibia to measure bone parameters. Data were analyzed using SAS. Phytase supplementation tended to increase overall ADG (P = 0.078; 39.7 to 42.2 g/d), whereas without affecting the overall ADFI and G:F. Phytase supplementation reduced (P &lt; 0.05) Pelomonas (10.01 to 2.64%) and tended to reduce Helicobacter (P = 0.085; 10.67 to 0.88%); Pseudomonas (P = 0.090; 2.02 to 0.31%); Sphingomonas (P = 0.071; 1.03 to 0.19%). Phytase supplementation increased (P &lt; 0.05) villus height (884 to 989 µm); the AID of CP (65.4 to 70.6%) and P (36.3 to 43.7%). Increasing phytase supplementation from 0 to 1,296 FTU/kg increased (P &lt; 0.05) breaking strength (184 to 213 N) and P content in the tibia (9.5 to 10.0%). In conclusion, phytase supplementation up to 4,000 FTU/kg feed showed benefits on intestinal health by reducing harmful bacteria in the jejunal mucosa, enhancing intestinal morphology and by increasing the AID of nutrients; and on the bone parameters by increasing the breaking strength and P content.

Probiotics and Achyranthes bidentata Polysaccharides Improve Growth Performance via Promoting Intestinal Nutrient Utilization and Enhancing Immune Function of Weaned Pigs

The experiment aimed to investigate the effects of probiotics and Achyranthes bidentata polysaccharides on the growth performance, nutrients digestibility, and immune function of weaned pigs. One hundred and twenty weaned pigs (about 7 kg BW, 23 ± 2 d) were allotted to five dietary treatments (CON: antibiotics-free basal diet; ANT: CON + antibiotics; PRO: CON + probiotics; ABPS: CON + Achyranthes bidentata polysaccharides; P-ABPS: PRO + ABPS) for a 28-day trial. Compared with CON, pigs in ANT, PRO, ABPS, and P-ABPS had greater (p < 0.05) ADG, ATTD of CP and GE, serum ALB, IgA and IL-2, duodenal intraepithelial lymphocyte, ileal VH and jejunal mucosa sIgA, but lower (p < 0.05) fecal scores, serum BUN, and IL-1β. Meanwhile, ANT, PRO, ABPS, and P-ABPS exhibited similar beneficial roles on growth performance, nutrients digestibility, serum parameters, and immune function. Interestingly, P-ABPS effects were similar to those obtained with ANT rather than with PRO or ABPS. In conclusion, Dietary PRO or ABPS used alone or in combination (P-ABPS), the combination augmenting the positive effect more than the independent supplement, could improve piglets’ growth performance via promoting intestinal nutrient digestion and absorption and enhancing immune function, indicating it had the potential to act as an alternative to in-feed antibiotics used in piglet diets.

Enzyme-Treated Soybean Meal Enhanced Performance via Improving Immune Response, Intestinal Morphology and Barrier Function of Nursery Pigs in Antibiotic Free Diets

This study aims to investigate the effects of ESBM on performance, antioxidant status, immune response, and intestinal barrier function of nursery pigs in antibiotic free diets compared with EFS. A total of 32 Duroc × (Landrace × Yorkshire) barrows (initial body weight of 8.05 ± 0.66 kg, weaned on d 28) were selected and allocated to two treatments with 16 replicates per treatment and one pig per replicate using a complete random design. The treatments included an EFS group (basal diet + 24% EFS; EFS) and an ESBM group (basal diet + 15% ESBM; ESBM). Corn was used to balance energy and diets were iso-nitrogenous at about 18% crude protein. The experiment lasted for 14 days and pigs were slaughtered for sampling on d 14. Compared with EFS, pigs fed ESBM showed enhanced (p < 0.05) gain to feed ratio and average daily gain and a reduced (p < 0.05) diarrhea score. These pigs had increased (p < 0.05) contents of glutathione peroxidase, catalase, superoxide dismutase, IgG, interleukin-10, and ferric reducing ability of plasma, as well as decreased (p < 0.05) malondialdehyde, IL-6, IL-1β, tumor necrosis factor (TNF-α), interferon-γ, thiobarbituric acid-reactive substances, and diamine oxidase level in serum and TNF-α level in the jejunal mucosa. Moreover, these pigs also showed enhanced (p < 0.05) villus height/crypt depth in ileum, villus height in duodenum, protein expression of zonula-occludens-1 in jejunal mucosa, and fecal total volatile fatty acids and butyric acid contents. In conclusion, ESBM replacing EFS could enhance performance via improving immune response, antioxidant status, gut morphology, and barrier function of nursery pigs in antibiotic free diets.