Alternatives to antibiotics for preventing bacteria-induced inflammation in early-weaned farm animals are sorely needed. Our previous study showed that Lactiplantibacillus plantarum L47 and inulin could alleviate dextran sulfate sodium (DSS)-induced colitis in mice. To explore the protective effects of L. plantarum L47 and inulin on the ileal inflammatory response in weaned piglets challenged with en-terotoxigenic Escherichia coli (ETEC), 28 weaned piglets were assigned into four groups, namely, CON groupdorally given 10 mL/d phosphate buffer saline (PBS), LI47 group—orally given a mixture of 10 mL/d L. plantarum L47 and inulin, ECON group—orally given 10 mL/d PBS and challenged by ETEC, and ELI47 groupdorally given 10 mL/d L. plantarum L47 and inulin mixture and challenged by ETEC. The results demonstrated that the combination of L. plantarum L47 and inulin reduced inflammatory responses and relieved the inflammatory damage caused by ETEC, including ileal morphological damage, reduced protein expression of ileal tight junction, decreased antioxidant capacity, and decreased anti-inflammatory factors. Transcriptome analysis revealed that L. plantarum L47 and inulin up-regulated the gene expression of phospholipase A2 group IIA (PLA2G2A) (P < 0.05) as well as affected alpha-linolenic acid (ALA) metabolism and linoleic acid metabolism. Moreover, L. plantarum L47 and inulin increased the levels of ALA (P< 0.05), lipoteichoic acid (LTA) (P< 0.05), and 12,13-epoxyoctadecenoic acid (12,13-EpOME) (P< 0.05) and the protein expression of Toll-like receptor 2 (TLR2) (P = 0.05) in the ileal mucosa. In conclusion, L. plantarum L47 and inulin together alleviated ETEC-induced ileal inflammation in piglets by up-regulating the levels of ALA and 12,13-EpOME via the LTA/TLR2/PLA2G2A pathway.

Intensive production can cause immunological stress in commercial broilers. Chlorogenic acid (CGA) regulates the intestinal microbiota, barrier function, and immune function in chickens. As complex interrelations regulate the dynamic interplay between gut microbiota, the host, and diverse health outcomes, the aim of this study was to elucidate the immunoregulatory mechanisms of CGA using multi-omics approaches. A total of 240 one-day-old male broilers were assigned to a 2 × 2 factorial design with 2 CGA levels (0 or 500 mg/kg) either with or without dexamethasone (DEX) injection for a 21-day experimental period. Therefore, there were 4 dietary treatments: control, DEX, CGA, and DEX + CGA, with 6 replicates per treatment. CGA supplementation improved (P< 0.05) growth performance, jejunal morphology, jejunal barrier function, and immune function in DEX-treated broilers. Moreover, in DEX + CGA-treated broilers, the increase in gut microbiome diversity (P< 0.05) was consistent with a change in taxonomic composition, especially in the Clostridiales vadin BB60_group. Additionally, the levels of short-chain fatty acids increased remarkably (P< 0.01) after CGA supplementation. This was consistent with the Kyoto Encyclopedia of Genes and Genomes analysis results that the "pyruvate fermentation to butanoate" pathway was more enriched (P< 0.01) in the DEX + CGA group than in the DEX group. Proteomics revealed that CGA treatment increased the expression of several health-promoting proteins, thymosin beta (TMSB4X) and legumain (LGMN), which were verified by multiple reaction monitoring. Metabolomics revealed that CGA treatment increased the expression of health-promoting metabolites (2,4-dihydroxy benzoic acid and homogentisic acid). Proteomic and metabolic analyses showed that CGA treatment regulated the peroxisome proliferator-activated receptor (PPAR) and mitogen-activated protein kinase (MAPK) pathways. Western blotting results support these findings. Pearson’s correlation analyses showed correlations (P< 0.01) between altered immune function, jejunal barrier function, different microbiota, proteins, and metabolites parameters. Overall, our data indicate that CGA treatment increased growth performance and improved the immunological functions of DEX-treated broilers by regulating gut microbiota and the PPAR and MAPK pathways. The results offer novel insights into a CGA-mediated improvement in immune function and intestinal health.

Endogenous protein leaving the ileum largely consists of accrued mucins from the upper gastrointestinal tract (GIT) that had resisted digestion. The amounts released rely on their mucosal generation during enteral feeding which vary with age as well as diet. These digestion resistant proteins of endogenous origin continue to be unavailable in the large intestine, whereas those of dietary origin provide amino acids that largely support the existing microbial population while denying limited amounts for absorption. Other mucins pre-exist within the large intestine as two layers at the lumen surface. A loose layer harboring a diverse microbial population is superimposed on the unstirred water layer (USWL) which simultaneously acts as an obstacle to microbes at the loose layer while performing as a molecular sieve for nutrients. The USWL is formed through interplay between enterocyte and goblet cells; however, the basis for presence of the loose layer is elusive. Large intestinal fermentation predominates within the colon of swine, whereas fowl employ their ceca. Motility within the colon of swine segregates fine materials into haustrae out-pocketings that parallel their placement within the ceca of fowl. Viscous mucins from small intestinal endogenous losses may envelop microbes within the large intestinal lumen to present successive adherents on the USWL that assemble its loose layer. The loose layer continually functions as a microbial reservoir in support of lumen fermentation. Microbial catabolism of mucin within the loose layer is known to be slow, but its proximity to the enterocyte is of advantage to enterocyte absorption with by-product amino acids fostering the USWL.

This study aimed to investigate the effects of different levels of black soldier fly (BSF) replacing soybean meal (SBM) in diets on the performance and health condition of piglets. A total of 180 weaned piglets were allocated into 5 treatments: BSF0 (corn-soybean meal basal diet), BSF25 (BSF replacing 25% SBM), BSF50 (BSF replacing 50% SBM), BSF75 (BSF replacing 75% SBM) and BSF100 (BSF replacing 100% SBM). During the whole period, in comparison with BSF0, average daily gain (ADG) and average daily feed intake increased in the BSF25 and BSF50 groups, whereas ADG decreased in the BSF75 and BSF100 groups (P< 0.05). The result of quadratic fitting curve showed that piglets exhibited the highest ADG when BSF replaced around 20% SBM. Compared with BSF0, organic matter and dry matter digestibility improved in the BSF25 group, whereas ether extract digestibility decreased in the BSF100 group (P< 0.05). In comparison with BSF0, piglets from the BSF25 group showed a higher duodenal ratio of villus height to crypt depth, increased jejunal sucrase activity, serum neuropeptide Y and ghrelin levels, elevated ileal immunoglobulin (Ig) A, IgG and IgM contents and a lower leptin level, and piglets from the BSF100 group exhibited an increased relative weight of kidney (P< 0.05). However, no significant differences were observed in the expression level of tight junction proteins and chitin-degrading enzyme. Additionally, compared with BSF0, the abundance of short chain fatty acid producing bacteria such as Ruminococcaceae, Faecalibacterium and Butyricicoccus increased, and potential pathogenic bacteria decreased in piglets from the BSF25 group, whereas piglets from the BSF100 group had a greater abundance of harmful bacteria. In conclusion, BSF replacing 25% SBM in diets could improve digestive parameters, immune function and intestinal microbiota, and thus improved growth performance of piglets. However, BSF replacing 100% SBM showed an adverse effect on piglet performance, and the reason might be related to the limited amount of chitin-degrading enzyme.

A 90-day feeding trial was conducted to assess the effects of black soldier fly larvae meal (BSFLM) as a replacement for soybean meal (SM) on growth performance and flesh quality of grass carp. A total of 420 grass carp (299.93 ± 0.85g) were randomly divided into 7 groups (triplicate) and fed 7 diets with SM substitution of 0% (SM, control), 15% (BSFLM15), 30% (BSFLM30), 45% (BSFLM45), 60% (BSFLM60), 75%(BSFLM75) and 100% (BSFLM100) by BSFLM. The growth performance of grass carp in the BSFLM75 and BSFLM100 groups were significantly lower compared to other groups (P < 0.05). The mid-gut villus height was the lowest in the BSFLM100 group (P < 0.05). Muscle nutritional value was improved due to increased DHA (docosahexaenoic acid), EPA (eicosapentaenoic acid), total HUFA (highly unsaturated fatty acids) and glycine levels, and reached the optimum in the BSFLM100 group (P < 0.05). According to the results of principal component analysis and weight analysis of muscle texture and body color, all the BSFLM diets except BSFLM15 could improve muscle texture and body color and reached the optimum level in the BSFLM100 group. Muscle drip loss and hypoxanthine content were the lowest and muscle antioxidant capacity was the highest in the BSFLM75 group, and water- and salt-soluble protein contents reached the optimum level in the BSFLM60 group (P < 0.05). Dietary BSFLM significantly reduced muscle fiber area and diameter, and increased muscle fiber density and the proportion of small fiber (diameter <20μm) (P < 0.05). Additionally, sarcomere lengths in the BSFLM75 and BSFLM100 groups were significantly higher than that in the SM group (P < 0.05). The mRNA relative expression levels of MyoD, Myf5, MyHC and FGF6b were remarkably up-regulated at an appropriate dietary BSFLM level (P < 0.05). In conclusion, BSFLM could replace up to 60% SM without an adverse effect on growth performance and improve the flesh quality of grass carp. The optimum levels of dietary BSFLM were 71.0 and 69.1g/kg diet based on the final body weight and feed conversion ratio. The flesh quality was optimal when dietary SM was completely replaced with BSFLM (227g/kg diet).

As a foodborne pathogen of global importance, Salmonella enterica serovar Enteritidis (S. Enteritidis) is a threat to public health that is mainly spread by poultry products. Intestinal Enterobacteriaceae can inhibit the colonization of S. Enteritidis and are regarded as a potential antibiotic substitute. We investigated, in chicks, the anti-S. Enteritidis effects of Escherichia coli (E. coli) Nissle 1917, the most well-known probiotic member of Enterobacteriaceae. Eighty 1-d-old healthy female AA broilers were randomly divided into 4 groups, with 20 in each group, namely the negative control (group P), the E. coli Nissle 1917-treated group (group N), the S. Enteritidis-infected group (group S) and the E. coli Nissle 1917-treated and S. Enteritidis-infected group (group NS). From d 5 to 7, chicks in groups N and NS were orally gavaged once a day with E. coli Nissle 1917 and in groups P and S were administered the same volume of sterile PBS. At d 8, the chicks in groups S and NS were orally gavaged with S. Enteritidis and in groups P and N were administered the same volume of sterile PBS. Sampling was conducted 24 h after challenge. Results showed that gavage of E. coli Nissle 1917 reduced the spleen index, Salmonella loads, and inflammation (P < 0.05). It improved intestinal morphology and intestinal barrier function (P < 0.05). S. Enteritidis infection significantly reduced mRNA expression of angiotensin-converting enzyme 2 (ACE2) and solute carrier family 6-member 19 (SLC6A19) in the cecum and the content of Gly, Ser, Gln, and Trp in the serum (P < 0.05). Pretreatment with E. coli Nissle 1917 yielded mRNA expression of ACE2 and SLC6A19 in the cecum and levels of Gly, Ser, Gln, and Trp in the serum similar to that of uninfected chicks (P < 0.05). Additionally, E. coli Nissle 1917 altered cecum microbiota composition and enriched the abundance of E. coli, Lactobacillales, and Lachnospiraceae. These findings reveal that the probiotic E. coli Nissle 1917 reduced S. Enteritidis infection and shows enormous potential as an alternative to antibiotics.

Since starch digestion in the small intestine provides more energy than digestion in the rumen of ruminants, reducing dietary rumen degradable starch (RDS) content is beneficial for improving energy utilization of starch in ruminants. The present study tested whether the reduction of rumen degradable starch by restricting dietary corn processing for growing goats could improve growth performance, and further investigated the possible underlying mechanism. In this study, twenty-four 12-wk-old goats were selected and randomly allocated to receive either a high RDS diet (HRDS, crushed corn-based concentrate, the mean of particle sizes of corn grain = 1.64 mm, n = 12) or a low RDS diet (LRDS, non-processed corn-based concentrate, the mean of particle sizes of corn grain >8 mm, n = 12). Growth performance, carcass traits, plasma biochemical indices, gene expression of glucose and amino acid transporters, and protein expression of the AMPK-mTOR pathway were measured. Compared to the HRDS, LRDS tended to increase the average daily gain (ADG, P = 0.054) and decreased the feed-to-gain ratio (F/G, P < 0.05). Furthermore, LRDS increased the net lean tissue rate (P < 0.01), protein content (P < 0.05) and total free amino acids (P < 0.05) in the biceps femoris (BF) muscle of goats. LRDS increased the glucose concentration (P < 0.01), but reduced total amino acid concentration (P < 0.05) and tended to reduce blood urea nitrogen (BUN) concentration (P = 0.062) in plasma of goats. The mRNA expression of insulin receptors (INSR), glucose transporter 4 (GLUT4), L-type amino acid transporter 1 (LAT1) and 4F2 heavy chain (4F2hc) in BF muscle, and sodium-glucose cotransporters 1 (SGLT1) and glucose transporter 2 (GLUT2) in the small intestine were significantly increased (P < 0.05) in LRDS goats. LRDS also led to marked activation of p70-S6 kinase (S6K) (P < 0.05), but lower activation of AMP-activated protein kinase (AMPK) (P < 0.05) and eukaryotic initiation factor 2α (P < 0.01). Our findings suggested that reducing the content of dietary RDS enhanced postruminal starch digestion and increased plasma glucose, thereby improving amino acid utilization and promoting protein synthesis in the skeletal muscle of goats via the AMPK-mTOR pathway. These changes may contribute to improvement in growth performance and carcass traits in LRDS goats.

This paper was to determine the effects of dietary Litsea cubeba essential oil (LEO) supplementation on growth performance, immune function, antioxidant level, intestinal morphology and microbial composition in weaned piglets. One hundred and ninety-two piglets (Duroc × [Large White × Landrace]) with 6.85 ± 0.22 kg mean body weight weaned at 21 d of age were randomly assigned to 4 treatment groups with 8 replicates and were fed with a basal diet (CON) or CON diet containing 100 (LLEO), 200 (MLEO) and 400 (HLEO) mg/kg LEO. The results revealed that HLEO supplementation (P < 0.05) increased the average daily gain on d 28 compared with CON. MLEO and HLEO supplementation decreased (P < 0.05) feed conversion ratio. LEO-containing diets had a lower (P < 0.05) diarrhea rate. Supplementation with HLEO increased (P < 0.05) total antioxidant capacity (T-AOC) both in the serum and liver. Meanwhile, the supplementation of MLEO and HLEO resulted in higher (P < 0.05) glutathione peroxidase (GPx) activities both in serum and liver. Supplementation of HLEO increased (P < 0.05) serum immunoglobulin A, immunoglobulin G and interleukin-10, whereas supplementation with MLEO and HLEO decreased (P < 0.05) tumor necrosis factor-α. Villus height in the duodenum or jejunum was increased (P < 0.05) in the HLEO group, and the villus height to crypt depth ratio in the jejunum was also improved (P < 0.05) in the MLEO group. The addition of LEO increased (P < 0.05) the richness and diversity of the microbial community in the cecum, which mainly increased the relative abundance of Oscillospiraceae_UCG-005, Faecalibacterium, Blautia and Coprococcus. Piglets supplemented with HLEO increased (P < 0.05) the concentration of short chain fatty acids (SCFA), including acetic acid in the cecum and propionic acid in the colon. In conclusion, these findings indicated that LEO supplementation improved growth performance and intestinal health in weaned piglets.

It has been demonstrated that the fiber content of oilseed meals and total amino acids (AA) in distillers' grains products affects standardized ileal digestibility (SID) values from swine assays and that total protein concentration affects the AA digestibility in assays using chickens. This analysis was conducted to test the hypothesis that the total AA content of test samples affects the SID assay results. Databases containing total AA profiles and SID values of 20 feedstuffs commonly fed to pigs from 2 sources, AMINODat 5.0 (Evonik Industries, 2015) and the Nutrient Requirements of Swine, 12th edition (NRC, 2012) were used to compare AA concentration effects on standardized ileal digestibility coefficients. Databases were compared with AA and SID available in both data sets. The total AA values were similar for the 2 databases with an R² of 0.979 (P < 0.001). The linear relationship between digestibility coefficients from the 2 databases was highly significant, an R² of 0.810 (P < 0.001). Both databases had increased SID values with increasing AA contents within and across feed ingredients. The SID = f(AA concentration) relationship was confirmed with an individual paper. Since SID, as typically measured, is a function of both digestion and absorption, both processes following Michaelis-Menten kinetics, SID = f(AA concentration) may simply be a natural phenomenon. Other reasons for the relationship were explored. Methods of estimating endogenous AA losses and misapplication statistical procedures may contribute to variation in results and at least partially explain why SID = f(AA concentration).

The goal of this review article, based on a systematic literature search, is to critically assess the state of knowledge and experimental methodologies used to delineate the conversion and metabolism of the 2 methionine (Met) sources DL-methionine (DL-Met) and DL-2-hydroxy-4-(methylthio) butanoic acid (HMTBa). The difference in the chemical structures of HMTBa and DL-Met indicates that these molecules are absorbed and metabolized differently in animals. This review explores the methodologies used to describe the 2-step enzymatic conversion of the 3 enantiomers (D-HMTBa, L-HMTBa and D-Met) to L-Met, as well as the site of conversion at the organ and tissue levels. Extensive work was published documenting the conversion of HMTBa and D-Met into L-Met and, consequently, the incorporation into protein using a variety of in vitro techniques, such as tissue homogenates, cell lines, primary cell lines, and everted gut sacs of individual tissues. These studies illustrated the role of the liver, kidney, and intestine in the conversion of Met precursors into L-Met. A combination of in vivo studies using stable isotopes and infusions provided evidence of the wide conversion of HMTBa to L-Met by all tissues and how some tissues are net users of HMTBa, whereas others are net secreters of L-Met derived from HMTBa. Conversion of D-Met to L-Met in organs other than the liver and kidney is poorly documented. The methodology cited in the literature to determine conversion efficiency ranged from measurements of urinary, fecal, and respiratory excretion to plasma concentration and tissue incorporation of isotopes after intraperitoneal and oral infusions. Differences observed between these methodologies reflect differences in the metabolism of Met sources rather than differences in conversion efficiency. The factors affecting conversion efficiency are explored in this paper and are mostly associated with extreme dietary conditions, such as noncommercial crystalline diets that are very deficient in total sulfur amino acids with respect to requirements. Implications in the diversion of the 2 Met sources toward transsulfuration over transmethylation pathways are discussed. The strengths and weaknesses of some methodologies used are discussed in this review. From this review, it can be concluded that due to the inherent differences in conversion and metabolism of the 2 Met sources, the experimental methodologies (e.g., selecting different organs at different time points or using diets severely deficient in Met and cysteine) can impact the conclusions of the study and may explain the apparent divergences of conclusion found in the literature. It is recommended when conducting studies or reviewing the literature to properly select the experimental models that allow for differences in how the 2 Met precursors are converted to L-Met and metabolized by the animal to enable a proper comparison of their bioefficacy.