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Influence of Dietary Supplementation with an Amino Acid Mixture on Inflammatory Markers, Immune Status and Serum Proteome in LPS-Challenged Weaned Piglets.

ABSTRACT: In order to investigate the effect of a dietary amino acid mixture supplementation in lipopolysaccharide (LPS)-challenged weaned piglets, twenty-seven 28-day-old (8.2 ± 1.0 kg) newly weaned piglets were randomly allocated to one of three experimental treatments for five weeks. Diet 1: a CTRL treatment. Diet 2: an LPS treatment, where piglets were intraperitoneally administered LPS (25 μg/kg) on day 7. Diet 3: an LPS+MIX treatment, where piglets were intraperitoneally administered LPS on day 7 and fed a diet supplemented with a mixture of 0.3% of arginine, branched-chain amino acids (leucine, valine, and isoleucine), and cystine (MIX). Blood samples were drawn on day 10 and day 35, and serum was analysed for selected chemical parameters and proteomics. The LPS and LPS+MIX groups exhibited an increase in haptoglobin concentrations on day 10. The LPS group showed an increased cortisol concentration, while this concentration was reduced in the LPS+MIX group compared to the control group. Similarly, the LPS+MIX group showed a decreased haptoglobin concentration on day 35 compared to the two other groups. Immunoglobulin concentrations were affected by treatments. Indeed, on day 10, the concentrations of IgG and IgM were decreased by the LPS challenge, as illustrated by the lower concentrations of these two immunoglobulins in the LPS group compared to the control group. In addition, the supplementation with the amino acid mixture in the LPS+MIX further decreased IgG and increased IgM concentrations compared to the LPS group. Although a proteomics approach did not reveal important alterations in the protein profile in response to treatments, LPS-challenged piglets had an increase in proteins linked to the immune response, when compared to piglets supplemented with the amino acid mixture. Overall, data indicate that LPS-challenged piglets supplemented with this amino acid mixture are more protected against the detrimental effects of LPS.

SUBMITTER: Prates JAM

PROVIDER: S-EPMC8073091 | biostudies-literature |

REPOSITORIES: biostudies-literature

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Project description:BackgroundThe benefits of combining benzoic acid and essential oils (BAO) to mitigate intestinal impairment during the weaning process have been well established, while the detailed underlying mechanism has not been fully elucidated. Previous research has primarily focused on the reparative effects of BAO on intestinal injury, while neglecting its potential in enhancing intestinal stress resistance.MethodsIn this study, we investigated the pre-protective effect of BAO against LPS-induced stress using a modified experimental procedure. Piglets were pre-supplemented with BAO for 14 d, followed by a challenge with LPS or saline to collect blood and intestinal samples.ResultsOur findings demonstrated that BAO supplementation led to significant improvements in piglets' final weight, average daily gain, and feed intake/body gain ratio. Additionally, BAO supplementation positively influenced the composition of intestinal microbiota, increasing beneficial Actinobacteriota and Alloprevotella while reducing harmful Desulfobacterota, Prevotella and Oscillospira. Furthermore, BAO supplementation effectively mitigated oxidative disturbances and inflammatory responses induced by acute LPS challenge. This was evidenced by elevated levels of T-AOC, SOD, and GSH, as well as decreased levels of MDA, TNF-α, and IL-6 in the plasma. Moreover, piglets subjected to LPS challenge and pre-supplemented with BAO exhibited significant improvements in intestinal morphological structure and enhanced integrity, as indicated by restored expression levels of Occludin and Claudin-1 compared to the non-supplemented counterparts. Further analysis revealed that BAO supplementation enhanced the jejunal antioxidative capacity by increasing GSH-Px levels and decreasing MDA levels under the LPS challenge and stimulated the activation of the Nrf2 signaling pathway. Additionally, the reduction of TLR4/NF-κB/MAPK signaling pathways activation and proinflammatory factor were also observed in the jejunal of those piglets fed with BAO.ConclusionsIn summary, our study demonstrates that pre-supplementation of BAO enhances the anti-stress capacity of weaned piglets by improving intestinal microbiota composition, reinforcing the intestinal barrier, and enhancing antioxidative and anti-inflammatory capabilities. These effects are closely associated with the activation of Nrf2 and TLR4/NF-κB/MAPK signaling pathways.

Project description:Lactobacillus rhamnosus GG (LGG) has been regarded as a safe probiotic strain. The aim of this study was to investigate whether dietary LGG supplementation could alleviate diarrhea via improving jejunal mucosal barrier function in the weaned piglets challenged by RV, and further analyze the potential roles for apoptosis of jejunal mucosal cells and intestinal microbiota. A total of 24 crossbred barrows weaned at 21 d of age were assigned randomly to 1 of 2 diets: the basal diet and LGG supplementing diet. On day 11, all pigs were orally infused RV or the sterile essential medium. RV infusion increased the diarrhea rate, increased the RV-Ab, NSP4 and IL-2 concentrations and the Bax mRNA levels of jejunal mucosa (P<0.05), decreased the villus height, villus height: crypt depth, the sIgA, IL-4 and mucin 1 concentrations and the ZO-1, occludin and Bcl-2 mRNA levels of jejunal mucosa (P<0.05), and affected the microbiota of ileum and cecum (P<0.05) in the weaned pigs. Dietary LGG supplementation increased the villus height and villus height: crypt depth, the sIgA, IL-4, mucin 1 and mucin 2 concentrations, and the ZO-1, occludin and Bcl-2 mRNA levels of the jejunal mucosa (P<0.05) reduced the Bax mRNA levels of the jejunal mucosa (P<0.05) in weaned pigs. Furthermore, dietary LGG supplementation alleviated the increase of diarrhea rate in the weaned pigs challenged by RV (P<0.05), and relieve the effect of RV infection on the villus height, crypt depth and the villus height: crypt depth of the jejunal mucosa (P<0.05), the NSP4, sIgA, IL-2, IL-4, mucin 1 and mucin 2 concentrations of jejunal mucosa (P<0.05), the ZO-1, occludin, Bax and Bcl-2 mRNA levels of the jejunal mucosa (P<0.05), and the microbiota of ileum and cecum (P<0.05) in the weaned pigs challenged by RV. These results suggest that supplementing LGG in diets alleviated the diarrhea of weaned piglets challenged by RV via inhibiting the virus multiplication and improving the jejunal mucosal barrier function, which was possibly due to the decreasing apoptosis of jejunal mucosal cells and the improvement of intestinal microbiota.

Project description:Two experiments were conducted to investigate the effects of exogenous catalase (CAT) in the diet of weaned piglets on growth performance, oxidative capacity, and hepatic apoptosis after challenge with lipopolysaccharide (LPS). In experiment 1, 72 weaned piglets [Duroc × Landrace × Yorkshire, 6.90 ± 0.01 kg body weight (BW), 21 d of age] were randomly assigned to be fed either a basal diet (CON group) or a basal diet supplemented with 2,000 mg/kg CAT (CAT group; dietary CAT activity, 120 U/kg) for 35 d. Blood samples were collected on day 21 and day 35. At the end of this experiment, 12 pigs were selected from each of the CON and CAT groups, and six pigs were injected with LPS (50 μg/kg BW), while the remaining six pigs were injected with an equal amount of sterile saline, resulting in a 2 × 2 factorial arrangement of treatments (experiment 2). Blood samples and rectal temperature data were collected 0 and 4 h after challenge, and liver samples were obtained after evisceration. The gain-to-feed ratio was higher (P < 0.05) in piglets in the CAT group than in those in the CON group from day 1 to 35. Catalase and total superoxide dismutase (T-SOD) activities were higher (P < 0.05), whereas malondialdehyde (MDA) concentrations were lower (P < 0.05), in piglets in the CAT group than in those in the CON group at day 35. During challenge, rectal temperature and liver MDA and H2O2 concentrations increased significantly (P < 0.05), whereas plasma CAT and glutathione peroxidase (GSH-Px) activities and liver CAT activity decreased markedly (P < 0.05), in LPS-challenged piglets 4 h post-challenge. Increased CAT activity and decreased MDA concentration were observed in the plasma and liver of piglets in the CAT group 4 h post-challenge (P < 0.05). Dietary CAT supplementation markedly suppressed the LPS-induced decrease in plasma GSH-Px activity and liver CAT activity to levels observed in the CON group (P < 0.05) as well as significantly decreasing the concentration and mRNA expression of caspase-3 and caspase-9 (P < 0.05). LPS-induced liver injury was also attenuated by dietary CAT supplementation, as demonstrated by a decrease in liver caspase-3 mRNA expression (P < 0.05). Overall, dietary supplementation with 2,000 mg/kg exogenous CAT (dietary CAT activity, 120 U/kg) improves growth performance and has a beneficial effect on antioxidant capacity in weaned piglets; alleviates oxidative stress and reduces liver damage by suppressing hepatic apoptosis in LPS-challenged piglets.

Project description:BackgroundWeanling pigs, with immature immune system and physiological function, usually experience post-weaning diarrhea. This study determined the effects of dietary Clostridium butyricum supplementation on growth performance, diarrhea, and immunity of weaned pigs challenged with lipopolysaccharide (LPS).MethodsIn Experiment (Exp.) 1, 144 weaned piglets were weaned at 21 d and randomly assigned to six groups, with six replicates per group and four pigs per replicate, receiving a control diet (CON) or diet supplemented with antibiotics (AB) or C. butyricum (CB) (0.1%, 0.2%, 0.4%, or 0.8%), respectively. All diets in Exp. 1 were a highly digestible basal diet, with 3,000 mg/kg zinc oxide supplied in the first 2 wk only. In Exp. 2, 180 piglets were weaned at 21 d and randomly assigned to five groups, with six replicates per group and six pigs per replicate, receiving CON, AB, or CB (0.2%, 0.4%, or 0.6%) diets. The digestibility of diets was lower than those in Exp. 1, and did not include zinc oxide. At 36 d of Exp. 2, 12 piglets were selected from each of the CON and 0.4% CB groups, six piglets were intraperitoneally injected with LPS (50 μg/kg body weight) and the other six piglets with normal saline; animals were killed at 4 h after injection to collect blood, intestine, and digesta samples for biochemical analysis.ResultsIn Exp. 1, CB and AB diets had no effect on growth performance of piglets. In Exp. 2, 0.4% CB decreased feed-gain ratio (P < 0.1), diarrhea score (P < 0.05), and increased duodenal, jejunal, and ileal villus height and jejunal villus height/crypt depth (P < 0.05). The 0.4% CB decreased the plasma tumor necrosis factor (TNF) α (P < 0.05) but increased ileal mucosa IL-10 and TLR2 mRNA expression (P < 0.05). Furthermore, 0.4% CB altered the microbial profile, with Bacillus and Ruminococcaceae UGG-003 at genus level and Lactobacillus casei and Parasutterella secunda at species level were higher than CON in colonic content (P < 0.05).ConclusionsDietary C. butyricum supplementation had positive effects on growth of weaned piglets with less digestible diets. There was a tendency to reduce the feed-gain ratio, which could reduce feed costs in pig production. Moreover, C. butyricum decreased post-weaning diarrhea by improving the intestinal morphology, intestinal microflora profile, and immune function.

Project description:This study mainly investigated the effects of berberine (BBR) on the bile acid metabolism in gut-liver axis and the microbial community in large intestine of weaned piglets challenged with enterotoxigenic Escherichia coli (ETEC) by microbiome and metabolome analyses. Sixty-four piglets were randomly assigned to four groups including Control group, BBR group, ETEC group, and BBR + ETEC group. Dietary BBR supplementation upregulated the colonic mRNA expression of Occludin, Claudin-5, trefoil factor 3 (TFF3), and interleukin (IL)-10, and downregulated colonic IL-1β and IL-8 mRNA expression in piglets challenged with ETEC K88 (p < 0.05). The hepatic non-targeted metabolome results showed that dietary BBR supplementation enriched the metabolic pathways of primary bile acid biosynthesis, tricarboxylic acid cycle, and taurine metabolism. The hepatic targeted metabolome analyses showed that BBR treatment increased the hepatic concentrations of taurocholic acid (TCA) and taurochenodeoxycholic acid (TDCA), but decreased the hepatic cholic acid (CA) concentration (p < 0.05). Further intestinal targeted metabolome analyses indicated that the deoxycholic acid (DCA), hyocholic acid (HCA), 7-ketodeoxycholic acid (7-KDCA), and the unconjugated bile acid concentrations in ileal mucosa was decreased by dietary BBR treatment (p < 0.05). Additionally, BBR treatment significantly upregulated the hepatic holesterol 7 α-hydroxylase (CYP7A1) and sterol 27-hydroxylase (CYP27A1) mRNA expression, and upregulated the ileal mRNA expression of farnesoid X receptor (FXR) and apical sodium-dependent bile acid transporter (ASBT) as well as the colonic mRNA expression of FXR, fibroblast growth factor19 (FGF19), takeda G protein-coupled receptor 5 (TGR5) and organic solute transporters beta (OST-β) in piglets (p < 0.05). Moreover, the microbiome analysis showed that BBR significantly altered the composition and diversity of colonic and cecal microbiota community, with the abundances of Firmicutes (phylum), and Lactobacillus and Megasphaera (genus) significantly increased in the large intestine of piglets (p < 0.05). Spearman correlation analysis showed that the relative abundances of Megasphaera (genus) were positively correlated with Claudin-5, Occludin, TFF3, and hepatic TCDCA concentration, but negatively correlated with hepatic CA and glycocholic acid (GCA) concentration (p < 0.05). Moreover, the relative abundances of Firmicute (phylum) and Lactobacillus (genus) were positively correlated with hepatic TCDCA concentration (p < 0.05). Collectively, dietary BBR supplementation could regulate the gut microbiota and bile acid metabolism through modulation of gut-liver axis, and attenuate the decreased intestinal tight junction expression caused by ETEC, which might help maintain intestinal homeostasis in weaned piglets.

Project description:BackgroundLactulose as an effective prebiotic protects intestinal mucosal injury. Bacillus coagulans is widely used in feed additives because of its ability to promote intestinal health. Our previous study suggests that the combination of lactulose and Bacillus coagulans may be a good candidate as alternative for antibiotic growth promoters. However, the in vivo effects of lactulose and Bacillus coagulans on growth and intestinal health under immune challenge in piglets remains unclear. The objective of this study is to explore the protective effects of synbiotic containing lactulose and Bacillus coagulans on the intestinal mucosal injury and barrier dysfunction under immune challenge in weaned piglets.MethodsTwenty four weaned piglets were assigned to 4 groups. Piglets in the CON-saline and LPS-LPS group were fed the basal diet, while others were fed either with chlortetracycline (CTC) or synbiotic mixture of lactulose and Bacillus coagulans for 32 d before injection of saline or lipopolysaccharide (LPS). Piglets were sacrificed 4 h after LPS injection to collect samples to determine intestinal morphology, integrity and barrier functions as well as relative genes and proteins.ResultsOur data showed that no differences were observed in the growth performance of the four test groups. LPS injection induced higher serum diamine oxidase activities, D-lactic acid levels, and endotoxin status, lower villus height and ratio of villus height to crypt depth, greater mRNA and lower protein expression related tight junction in both jejunum and ileum. In addition, a higher apoptosis index, and protein expression of Bax and caspase-3 were also observed in the LPS challenge group. Interestingly, dietary synbiotic mixture with lactulose and Bacillus coagulans protected against LPS-induced intestinal damage, barrier dysfunction and higher apoptosis as well as CTC.ConclusionsOur data suggest that dietary supplementation of synbiotic mixture with lactulose and Bacillus coagulans showed resilience to LPS-induced intestinal morphological damage, barrier dysfunction and aggressive apoptosis in piglets as well as the protective effects of CTC. These results indicate that synbiotic mixture of lactulose and Bacillus coagulans showed beneficial effects on performance and resilience to acute immune stress in weaned piglets.

Project description:This study was conducted to investigate the effects of dietary supplementation with different types of Bacillus subtilis (B. subtilis) on the growth and gut health of weaned piglets. A total of 160 piglets were randomly assigned into four groups: control group (a basal diet), BS-A group (a basal diet supplemented with B. subtilis A at 1 × 106 CFU/g feed), BS-B group (a basal diet supplemented with B. subtilis B at 1 × 106 CFU/g feed), and BS-C group (a basal diet supplemented with B. subtilis C at 1 × 106 CFU/g feed). All groups had five replicates with eight piglets per replicate. On days 7, 21, and 42 of the trial, blood plasma and intestinal tissues and digesta samples were collected to determine plasma cytokine concentrations, intestinal morphology, gut microbiota community and metabolic activity, and the expression of genes related to gut physiology and metabolism. The results showed that dietary B. subtilis supplementation improved (P < 0.05) the body weight and average daily gain (in BS-B and BS-C groups) of weaned piglets and decreased (P < 0.05) the diarrhea rates (in BS-A, BS-B, and BS-C groups). In the intestinal morphology analysis, B. subtilis supplementation improved (P < 0.05) the size of villus height and villus height to crypt depth ratio in the ileum of weaned piglets. Firmicutes, Bacteroidetes, and Tenericutes were the most dominant microflora in piglets' colon whatever the trial group and time of analysis. Dietary BS-C supplementation increased (P < 0.05) the relative abundances of Anaerovibrio and Bulleidia and decreased (P < 0.05) the relative abundances of Clostridium and Coprococcus compared with the control group. In addition, dietary B. subtilis supplementation increased (P < 0.05) the indicators of intestinal health, including plasma levels of interleukin (IL)-2 and IL-10, as well as the colonic levels of short-chain fatty acids. Furthermore, dietary B. subtilis supplementation also up-regulated (P < 0.05) the expression of genes involved in metabolic pathways related to intestinal microbiota maturation. In conclusion, these findings suggest that a diet containing BS-B or BS-C can efficiently promote growth performance, decrease diarrhea incidence, and ameliorate several indicators of intestinal health through the modulation of gut microbiota composition and metabolic activity in weaned piglets.

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Influence of Dietary Supplementation with an Amino Acid Mixture on Inflammatory Markers, Immune Status and Serum Proteome in LPS-Challenged Weaned Piglets.

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