Dietary berberine supplementation improves growth performance and alleviates gut injury in weaned piglets by modulating ileal microbiota and metabolites.

This study investigated the effects of dietary berberine (BBR) supplementation on the growth performance, intestinal health, and ileal microbiome and metabolomic profile in weaned piglets challenged with enterotoxigenic Escherichia coli (ETEC). Dietary BBR supplementation significantly attenuated the reduced average daily gain (ADG) and attenuated the increased feed to gain ratio (F/G) and the incidence of diarrhea induced by ETEC K88 (P < 0.05). Dietary BBR supplementation significantly increased the villus height and the villus height to crypt depth ratio in the ileum (P < 0.05). Moreover, the mRNA expression of ZO-1 and occludin as well as aquaporins (AQP1, AQP3, AQP4, AQP7, and AQP10) and Na+/H+ exchanger 3 (NHE3) in ileal mucosa was significantly upregulated by BBR treatment (P < 0.05). Additionally, BBR treatment significantly inhibited the increase of interleukin-1ß (IL-1ß) in jejunal mucosa caused by ETEC and reduced the levels of tumor necrosis factor-α (TNF-α) and IL-1ß and increased interleukin-10 (IL-10) in colonic mucosa (P < 0.05). Dietary BBR treatment significantly increased the Observed_species, Chao 1, abundance based coverage estimators (ACE), and PD_whole tree in the ileal digesta of weaned piglets challenged with ETEC. At the genus level, the relative abundance of unidentified Clostridiales was decreased, while Weissella, Alloprevotella, unidentified Prevotellaceae, and Catenibacterium were increased in the BBR + ETEC group when compared to the ETEC group (P < 0.05). Spearman correlation analysis showed that the relative abundance of unidentified Clostridiales (genus) was negatively correlated with the ileal villus height but negatively correlated with diarrhea and intestinal IL-1ß and TNF-α concentrations (P < 0.05). The ileal metabolome analysis showed that the metabolic pathways including primary and secondary bile acid biosynthesis and bile secretion were significantly enriched by BBR treatment. Collectively, dietary BBR supplementation effectively improved the growth performance and alleviated the diarrhea and intestinal injury induced by ETEC K88 in weaned piglets, which might closely involve the modulation of ileal microbiota and metabolites.

Dietary supplementation with anthocyanin attenuates lipopolysaccharide-induced intestinal damage through antioxidant effects in yellow-feathered broiler chicks.

This study investigated the protective effects of anthocyanin (AC) supplementation on lipopolysaccharide (LPS)-challenged yellow-feathered broiler chicks. A total of 480 1-d female broiler chicks were randomly assigned to 4 treatment groups: basal diet (CON), basal diet + LPS-challenge (LPS), supplementation with 100 or 400 mg/kg AC + LPS-challenge (AC100, AC400). On d 17 and d 19, birds in LPS, AC100 and AC400 received an intramuscular dose of LPS, while birds in CON received saline. The result showed that (1) LPS injection significantly decreased (P < 0.05) body weight on d 21 and average daily gain of broiler chicks from 1 to 21 days of age, and supplementation with 100 mg/kg AC increased (P < 0.05) those of LPS-challenged broilers. (2) There were no differences among the treatments (P > 0.05) in relative weights of immune organs. (3) Supplementation with AC (AC100 and AC400) increased (P < 0.05) the jejunal villus height and villus height/crypt depth ratio (AC100) of LPS-challenged birds. Challenge with LPS decreased the relative expression of OCLN (Occludin), ZO-1, JAM2, and MUC2 in jejunal mucosa of broilers, and supplementation with AC offset the relative expression of ZO-1, JAM2 (AC100 and AC400), and OCLN (AC400) in LPS-injected broilers. (4) LPS-induced increase in the malondialdehyde (MDA) concentration and decreases in activity of total superoxide dismutase (T-SOD), and expression of SOD1, CAT and GPX in jejunal mucosa, were attenuated by dietary AC supplementation. In conclusion, in yellow-feathered broiler chicks, dietary supplementation with AC alleviated LPS-induced declined growth performance and mucosal damage of the intestine through antioxidant effects.

Arginine Promotes the Expression of Aquaporin-3 and Water Transport in Porcine Trophectoderm Cells Through NO- and cAMP-Dependent Mechanisms.

BACKGROUND: Dietary supplementation with L-arginine (Arg) has been shown to increase the volume of fetal fluids in gestating swine. Aquaporins (AQPs), known as water channel proteins, are essential for embryonic growth and development. It was not known if Arg mediates water transport through AQPs in porcine conceptus trophectoderm (pTr2) cells. METHODS: pTr2 cells derived from pregnant gilts on day 12 of gestation were cultured in customized Arg-free Dulbecco's modified Eagle's Ham medium (DMEM) supplemented with either 0.00, 0.25, or 0.50 mM Arg. RESULTS: Arg treatment increased water transport and the expression of AQP3, which was abundantly expressed in pTr2 cells at both the mRNA and protein levels. Arg also increased the expression of iNOS and the synthesis of nitric oxide (NO) in pTr2 cells. The presence of Nω-nitro-L-arginine methyl ester hydrochloride (L-NAME; an inhibitor of NO synthase) significantly attenuated the Arg-induced expression of AQP3. Furthermore, 0.50 mM Arg increased the concentrations of cAMP and the abundances of phosphorylated cAMP-dependent protein kinase A (PKA), phosphorylated PKA α/ß/γ, and phosphorylated CREB. These effects of Arg were mimicked by Forskolin (a cell-permeable activator of adenylyl cyclase), but inhibited by H-89 (an inhibitor of cAMP-dependent protein kinase). CONCLUSIONS: The results of this study demonstrate that Arg regulates AQP3 expression and promotes water transport in pTr2 cells through NO- and cAMP-dependent signaling pathways.

Optimal Level of Supplemental Manganese for Yellow-Feathered Broilers during the Growth Phase.

This experiment investigated the effect of an optimized supplemental dietary manganese (Mn) on growth performance, tibial characteristics, immune function and meat quality, of yellow-feathered broilers. In three rearing periods, birds were fed for 21-d periods, from d 1 (starter), d 22 (grower) and d 43 (finisher), respectively, with basal diets (containing 16, 17, and 14 mg/kg analyzed Mn, respectively) supplemented with 0, 20, 40, 60, 80, 100, 120 and 140 mg/kg Mn. For starter phase broilers, supplemental manganese affected feed to gain ratio (F/G), and the minimum value was observed with 120 mg/kg manganese. During the grower phase, ADG increased quadratically (p < 0.05) with supplemental Mn and was maximal with 54 mg/kg additional manganese estimated using the regression equation. There was no influence of supplemental manganese on growth performance of broilers during the finisher phase (p > 0.05). The thymic relative weight of broilers were linearly (p < 0.05) and quadratically (p < 0.05) increased with supplemental Mn and maxima were obtained with 95 and 110 mg/kg additional Mn at 42 d and 63 d. The bone density of the tibia in broilers at d 21, 42 and 63 were increased quadratically (p < 0.05) by supplemental Mn, and optimal supplementation for the three phases was 52, 60 and 68 mg/kg, respectively. The weight, diameter, breaking strength and bone density of the tibia of 63-d broilers were influenced (p < 0.05) by supplemental manganese. The lightness (L*) value (linear, p < 0.05) and yellowness (b*) value (p < 0.05) of the breast muscle were decreased by dietary manganese supplementation, and the optimal supplementation, based on L*, was 86 mg/kg. In conclusion, supplemental Mn affected the growth performance, thymic relative weight, tibial characteristics, and the meat color of yellow-feathered broilers. From the quadratic regressions, the optimal supplementation of yellow-feathered broilers at the starter, grower and finisher phases to achieve the best performance was 52, 60, and 68 mg/kg, respectively.

High dietary copper induces oxidative stress and leads to decreased egg quality and reproductive performance of Chinese Yellow broiler breeder hens.

The objective of this study was to investigate the effects of dietary copper (Cu) on production, egg quality, and hatchability of Chinese Yellow broiler breeder hens and growth performance of their offspring. A total of 576 30-week-old hens were randomly allotted into 6 groups, each with 6 replicates (8 cages for each replicate with 2 birds per cage). The basal diet contained 3.50 mg/kg Cu, and the other 5 treatment diets contained 8.5, 13.5, 23.5 43.5, and 83.5 mg/kg Cu, respectively, additionally supplemented with Cu on the basal diet. The trial lasted for 15 wk. Qualified egg rate of birds fed 23.5 or 83.5 mg/kg Cu was significantly decreased (P < 0.05) compared with those given 3.5, 8.5, or 13.5 mg/kg Cu. Plasma malondialdehyde concentration showed quadratic effect (P = 0.002) which that decreased first then increased with dietary Cu increased. Highest values of Cu content and hepatic activity of Cu-ATPase occurred in hens fed 83.5 mg/kg dietary Cu with linear (P = 0.001) and quadratic (P = 0.001) effects. Shell strength and proportion on 18th day of live embryos of hens fed 13.5 mg/kg Cu were the greatest compared with other groups respectively (P < 0.05); rate of qualified eggs for hatch and hatchability of fertilized eggs of hens fed 83.5 mg/kg Cu were the least (P < 0.05). In conclusion, both inadequate (3.5 mg/kg diet) and excess (83.5 mg/kg) of dietary Cu can induce oxidative stress in hens and lead to decreased egg quality. Hatchability and growth performance of offspring were decreased when breeder hens were fed excess Cu in spite of greater hatching weight. The appropriate dietary Cu level for Chinese Yellow broiler breeder hens during the egg-laying period is 15.7 to 21.2 mg/kg (1.81-2.44 mg Cu fed per day) when based on Cu level and Cu-ATPase activity in the liver. This dietary Cu requirement is approximately doubled (∼40 mg/kg, ∼4.60 mg Cu per bird per day) for maximal response of eggshell thickness.

Potential Effects of Acidifier and Amylase as Substitutes for Antibiotic on the Growth Performance, Nutrient Digestion and Gut Microbiota in Yellow-Feathered Broilers.

This study was conducted to evaluate the effects of acidifier (benzoic acid, BA), amylase (AL) and their combination as substitutes for antibiotics on growth performance, antioxidation, nutrient digestion and gut microbiota of yellow-feathered broilers. A total of 1440 twenty-one-day-old broilers were randomly allocated to six treatments. Broilers in the control group (CON) were fed a basal diet, whereas birds in the other five groups were fed the basal diet supplemented with antibiotic (zinc bacitracin, AT, 40 mg/kg), BA (2000 mg/kg), low level AL (AL-L, 300 mg/kg), high level AL (AL-H, 500 mg/kg) and the combination of AL-H and BA (BA+AL-H). The experimental animals were killed at the end of the trial (21 day-63 day) then blood samples were collected from two birds per pen. Bird weight, feed intake and survival rate were recorded on pen basis. Growth performance was not significantly influenced by AT, BA, AL-L, AL-H or BA+AL-H. Plasma uric acid (UA) was decreased from CON by all treatments; the activity of AKP in plasma was also lowered by AT, BA, AL-H and BA+AL-H. Plasma activity of LDH was reduced by BA. In the jejunal mucosa, Na+K+-ATP activity was increased by BA, AL-L, AL-H and BA+AL-H. Mucosal activities of T-AOC and CAT were increased with AL-L and AT supplementation, respectively. Additionally, the relative abundance of Escherichia coli (E. coli) in the cecal contents was reduced by BA+AL-H and, with the exception of AL-H, all treatments increased the relative abundance of Lactobacillus. In conclusion, dietary AT, BA, AL-L, AL-H or BA+AL were effective in improving the antioxidant capacity, nutrient digestion and gut microbiota composition. No significant differences were observed in the tested variables between AT and other treatments, indicating that BA, AL and their combination may be alternatives to dietary inclusion of zinc bacitracin. Dietary addition of 500 mg/kg AL and 2000 mg/kg BA was an optimum supplementation dose.

Effects of maternal and dietary vitamin A on growth performance, meat quality, antioxidant status, and immune function of offspring broilers.

The aim of this study was to evaluate the effects of maternal and dietary vitamin A (VA) level on growth performance, meat quality, antioxidant status, and immune function of offspring broilers. Chinese yellow-feathered breeder hens were fed a basal diet supplemented with 0, 5,400, 10,800, and 21,600 IU/kg VA for 8 wk, with 6 replicates of 22 hens per replicate. Then the offspring hatched from each of the 4 maternal groups were fed a basal diet supplemented with 0 or 5,000 IU/kg VA for 63 D. Overall, there were 8 treatment combinations, each with 6 replicate pens of 20 birds. Results showed that (1) providing VA in offspring diets increased final body weight (FW), average daily gain, and average daily feed intake but reduced feed-to-gain ratio and mortality of offspring broilers (P < 0.05), whereas maternal provision of VA did not significantly affect the growth performance and mortality of offspring broilers. Maternal or offspring VA did not affect proportion of breast or thigh muscle (P > 0.05). (2) Maternal feeding with 21,600 IU/kg VA increased (P < 0.05) pH 24 h postmortem of breast muscle, compared with those without maternal supplication of VA. Dietary provision of 5,000 IU/kg VA in the posthatching diet decreased (P < 0.05) drip loss, yellowness (b∗) value and lightness (L∗) value, and increased shear force and pH of breast muscle compared with those without dietary VA supplication. (3) Maternal or offspring VA did not affect the activities of total superoxide dismutase and glutathione peroxidase (GSH-Px) or the content of malondialdehyde; however, there was a significant interaction (P < 0.05) between maternal and offspring VA on the activity of GSH-Px in serum. (4) Dietary provision of 5,000 IU/kg VA increased (P < 0.05) the weight proportion of liver and bursa of fabricius, whereas maternal feeding with 21,600 IU/kg VA increased the hatchling BW. Maternal feeding with 5,400 and 21,600 IU/kg VA decreased (P < 0.05) splenic interferon-γ (IFN-γ) transcripts and increased (P < 0.05) those of interleukin-2 (IL-2) in the progeny. There were interactions (P < 0.05) between maternal and offspring VA on splenic IL-2, IL-1ß, and IFN-γ expression. In summary, maternal and offspring provision of VA both had influence on meat quality and immune function in progeny broilers. Dietary VA increased growth performance, whereas the maternal VA affected the initial body weight of progeny when hatched, but the difference in performance caused by maternal VA level was able to be eliminated by dietary VA supplementation. Therefore, offspring provision had greater importance than maternal VA in the production; however, both should be considered in broiler nutrition to achieve good meat quality and immune status of broilers.

Influence of Mushroom Polysaccharide, Nano-Copper, Copper Loaded Chitosan, and Lysozyme on Intestinal Barrier and Immunity of LPS-mediated Yellow-Feathered Chickens.

This study investigated the influence of dietary supplementation with some antibiotic alternatives on growth performance, intestinal barrier, and immunity of lipopolysaccharide (LPS) challenged chicks. Wenshi females, aged 4 days, were allocated randomly into eight groups, each with six replicates of 20 birds (n = 120/treatment), which received a basal diet supplemented with 0 (control), 0 (LPS), 200 mg/kg aureomycin, 50 mg/kg mushroom polysaccharide, 100 mg/kg mushroom polysaccharide, 500 mg/kg nano-copper, 300 mg/kg copper loaded chitosan, and 500 mg/kg lysozyme for 21 days. On day 18 and 20, the control birds were injected with 0.5 mL saline solution, the other treatments were injected with 0.5 mL saline containing 500 µg LPS/kg body weight (BW). The results indicated that LPS treatment reduced the BW, average daily gain (ADG), and daily feed intake (ADFI) than the controls (p < 0.05), and the antibiotic and the tested alternatives could not retrieve the normal BW, ADG, and ADFI. The tested additives reduced several negative effects of LPS; they reduced diamine oxidase activity and inflammatory mediators in plasma, jejunal mucosa, spleen and thymus, increased content of immunoglobulin in plasma and jejunal mucosa, and decreased gene expression of inducible nitric oxide synthase and Cyclooxygenase 2 in jejunal mucosa.

mTORC1 Mediates Lysine-Induced Satellite Cell Activation to Promote Skeletal Muscle Growth.

As the first limiting amino acid, lysine (Lys) has been thought to promote muscle fiber hypertrophy by increasing protein synthesis. However, the functions of Lys seem far more complex than that. Despite the fact that satellite cells (SCs) play an important role in skeletal muscle growth, the communication between Lys and SCs remains unclear. In this study, we investigated whether SCs participate directly in Lys-induced skeletal muscle growth and whether the mammalian target of rapamycin complex 1 (mTORC1) pathway was activated both in vivo and in vitro to mediate SC functions in response to Lys supplementation. Subsequently, the skeletal muscle growth of piglets was controlled by dietary Lys supplementation. Isobaric tag for relative and absolute quantitation (iTRAQ) analysis showed activated SCs were required for longissimus dorsi muscle growth, and this effect was accompanied by mTORC1 pathway upregulation. Furthermore, SC proliferation was governed by medium Lys concentrations, and the mTORC1 pathway was significantly enhanced in vitro. After verifying that rapamycin inhibits the mTORC1 pathway and suppresses SC proliferation, we conclude that Lys is not only a molecular building block for protein synthesis but also a signal that activates SCs to manipulate muscle growth via the mTORC1 pathway.