Colon Of Piglets Research Articles

Lactation-related dynamics of bacterial and fungal microbiomes in feces of sows and gut colonization in suckling and newly weaned piglets.

Changes in the gut microbial composition of the sow during lactation may influence the gut microbial colonization in their offspring, for which less information was available in the literature. This study aimed to assess: 1) the changes that occur in the bacterial and fungal communities in sow feces during the 28-day lactation period as well as in gastric and cecal digesta of piglets until one week after weaning, and 2) bacterial and fungal taxa in cecal digesta of the piglets postweaning that associate with fecal consistency. Aside from sow milk, piglets had access to creep feed from day of life (DoL) 3. Fecal samples from sows for microbial analysis were collected (n=20) on days postpartum (DPP) 1, 6, 13, 20 and 27, as well as from weaned piglets for fecal scoring on DoL 30 and 34. Gastric and cecal digesta of piglets was collected on DoL3, 7, 14, 21, 28, 31 and 35 (n=5/sex/DoL). Progressing lactation affected bacterial and fungal communities in sow feces, including 10.3- and 3.0-fold increases in the relative abundances of Lactobacillus from DPP1 to 6 and Kazachstania from DPP1 to 13, respectively (P < 0.001). Although time- and gut site-related differences existed, bacterial and fungal taxa found in sow feces were also present in gastric and cecal digesta of piglets, which supports their role for gut colonization in neonatal piglets. In piglets, bacterial and fungal alpha-diversities showed certain fluctuations during the suckling period, whereby weaning affected more the fungal than bacterial diversity at both gut sites (P < 0.05). At both gut sites, Lactobacillus largely increased from DoL3 to 7 and remained a dominating taxon until DoL35 (P < 0.05). Postweaning, plant-glycan fermenters (e.g., Prevotella-9) seemed to replace milk-glycan fermenting Fusobacterium and Bacteroides (P < 0.05). In gastric and cecal digesta, Kazachstania, Tausonia, Candida and Blumeria were dominating fungi from DoL3 to 35, with Kazachstania becoming even more dominant postweaning (P < 0.001). Fecal consistency was softer on DoL34 than 30 (P < 0.05). Correlation analysis identified that softer feces were linked to the relative abundances of plant-glycan and proteolytic bacterial taxa including pathobionts (e.g., Clostridium sensu stricto) in the cecum on DoL34. However, the potential association between cecal mold and plant-pathogenic fungi Talaromyces, Mrakia, and Blumeria and softer feces are worth to investigate in the future in relation to (gut) health of piglets.

Effects of Microencapsulated Essential Oils on Growth and Intestinal Health in Weaned Piglets.

The study investigated the effects of microencapsulated essential oils (MEO) on the growth performance, diarrhea, and intestinal microenvironment of weaned piglets. The 120 thirty-day-old weaned piglets (Duroc × Landrace × Yorkshire, 8.15 ± 0.07 kg) were randomly divided into four groups and were fed with a basal diet (CON) or CON diet containing 300 (L-MEO), 500 (M-MEO), and 700 (H-MEO) mg/kg MEO, respectively, and data related to performance were measured. The results revealed that MEO supplementation increased the ADG and ADFI in weaned piglets (p < 0.05) compared with CON, and reduced diarrhea rates in nursery pigs (p < 0.05). MEO supplementation significantly increased the duodenum's V:C ratio and the jejunal villi height of weaned piglets (p < 0.05). The addition of MEO significantly increased the T-AOC activity in the jejunum of piglets (p < 0.05), but only L-MEO decreased the MDA concentration (p < 0.01). H-MEO group significantly increases the content of isobutyric acid (p < 0.05) in the piglet colon, but it does not affect the content of other acids. In addition, MEO supplementation improved appetite in the nursery and increased the diversity and abundance of beneficial bacteria in the intestinal microbiome. In conclusion, these findings indicated that MEO supplementation improves growth and intestinal health in weaned piglets.

Impact of dietary Laminaria digitata with alginate lyase or carbohydrase mixture on nutrient digestibility and gut health of weaned piglets

Laminaria digitata is a brown seaweed rich in prebiotic polysaccharides, mainly laminarin, but its alginate-rich cell wall could compromise nutrient access. Carbohydrase supplementation, such as individual alginate lyase and carbohydrases mixture (Rovabio® Excel AP), could enhance nutrient digestibility and prebiotic potential. This study aimed to evaluate the effect of these enzymes on nutrient digestibility and gut health of weaned piglets fed with 10% L. digitata. Diets did not affect growth performance (P > 0.05). The majority of the feed fractions had similar digestibility across all diets, but the supplementation of alginate lyase increased hemicellulose digestibility by 3.3% compared to the control group (P = 0.047). Additionally, we observed that algal zinc was more readily available compared to the control group, even without enzymatic supplementation (P < 0.001). However, the increased digestibility of some minerals, such as potassium, raises concerns about potential mineral imbalance. Seaweed groups had a higher abundance of beneficial bacteria in colon contents, such as Prevotella, Oscillospira and Catenisphaera. Furthermore, the addition of alginate lyase led to a lower pH in the colon (P < 0.001) and caecum (P < 0.001) of piglets, which is possibly a result of released fermentable laminarin, and is consistent with the higher proportion of butyric acid found in these intestinal compartments. L. digitata is a putative supplement to enhance piglet gut health due to its prebiotic polysaccharides. Alginate lyase supplementation further improves nutrient digestibility and prebiotic potential. These results suggest the potential use of L. digitata and these enzymatic supplements in commercial piglet-feeding practices.

Cold stress changes the composition and function of microbiota in the content and mucosa of the ileum and colon in piglets

Context Cold stress is a significant factor that contributes to the imbalance of energy in piglets during their early life. Recent studies have shown that gut microbiota plays a crucial role in maintaining energy homeostasis under cold-stress conditions. Aims This study aims to investigate the effects of cold stress on the microbiota and expression pathways in the colon and ileum of weaned piglets, providing new insights and methods for helping piglets resist cold stress. Methods In total, 10 piglets in the cold-stress group (n = 5) and room-temperature group (n = 5) were sacrificed. Intestinal contents and mucosa samples were collected for 16S rRNA analysis. Key results The results showed that cold stress increased the observed features and chao1 index in the colonic mucosa. The beta diversity of ileum, colon and ileum mucosa was significantly changed. Under cold stress, the relative abundance of Acholeplasma, Proteiniphilum, and Olsenella increased in the contents of the ileum and colon. The relative abundance of Ruminococcaceae sp., Butyricicoccus, and Lachnospiraceae FCS020 increased in the mucosa of the colon. Sphingomonas, Helicobacter, Cutibacterium, and Bradyrhizobium were significantly increased in the mucosa of the ileum. In predicted functions, after cold stress, the purine metabolism and degradation increased in the content and mucosa of the ileum and mucosa of the colon. The fat biosynthesis pathway increased in the content of the colon. Complex carbohydrate degradation increased in the mucosa of both. Conclusions These findings suggest that cold stress has a significant impact on the species richness, composition, and predicted functions of the microbiota in the ileum and colon of piglets, with these effects varying depending on the location within the gut. Implications Therefore, we can help piglets resist cold stress by modifying the structure of gut microbiota through the addition of probiotics or adjusting the composition of their diet.

Improvements of weaned pigs barn hygiene to reduce the spread of antimicrobial resistance.

The spread of antimicrobial resistance (AMR) in animal husbandry is usually attributed to the use of antibiotics and poor hygiene and biosecurity. We therefore conducted experimental trials to improve hygiene management in weaned pig houses and assessed the impact on the spread. For each of the two groups examined, the experimental group (EG) and the control group (CG), three replicate batches of piglets from the same pig breeder, kept in pre-cleaned flat decks, were analyzed. In the flat decks of the experimental groups, the hygiene conditions (cleaning, disinfection, dust removal and fly control) were improved, while regular hygiene measures were carried out in the control groups. The occurrence and spread of AMR were determined in Escherichia coli (E. coli; resistance indicator) using cultivation-dependent (CFU) and -independent (qPCR) methods as well as whole genome sequencing of isolates in samples of various origins, including feces, flies, feed, dust and swabs. Surprisingly, there were no significant differences (p > 0.05) in the prevalence of resistant E. coli between the flat decks managed with conventional techniques and those managed with improved techniques. Selective cultivation delivered ampicillin- and sulfonamide-resistant E. coli proportions of up to 100% and 1.2%, respectively. While 0.5% E. coli resistant to cefotaxime and no ciprofloxacin resistance were detected. There was a significant difference (p < 0.01) in the abundance of the blaTEM-1 gene in fecal samples between EG and CG groups. The colonization of piglets with resistant pathogens before arrival, the movement of flies in the barn and the treatment of bacterial infections with antibiotics obscured the effects of hygiene improvement. Biocide tolerance tests showed no development of resistance to the farm regular disinfectant. Managing hygiene alone was insufficient for reducing antimicrobial resistances in piglet rearing. We conclude that the complex factors contributing to the presence and distribution of AMR in piglet barns underscore the necessity for a comprehensive management strategy.

Effect of Low Protein Diets Supplemented with Sodium Butyrate, Medium-Chain Fatty Acids, or n-3 Polyunsaturated Fatty Acids on the Growth Performance, Immune Function, and Microbiome of Weaned Piglets.

This study aimed to investigate the effects of low-protein (LP) diets supplemented with sodium butyrate (SB), medium-chain fatty acids (MCT), or n-3 polyunsaturated fatty acids (n-3 PUFA) on the growth performance, immune function, and the microbiome of weaned piglets. A total of 120 healthy weaned piglets ((Landrace × Large White × Duroc); 7.93 ± 0.7 kg initial body weight), were randomly divided into five groups. Each group consisted of six replications with four piglets per replication. Dietary treatments included control diet (CON); LP diet (LP); LP + 0.2% SB diet (LP + SB); LP + 0.2% MCT diet (LP + MCT); and LP + PUFA diet (LP + PUFA). The experimental period lasted for 4 weeks. Compared with the CON diet, LP, LP + SB, LP + MCT, and LP + PUFA diets decreased the final weight and average daily gain (ADG) of piglets (p < 0.05). There were lower (p < 0.05) concentrations of IL-8 and higher (p < 0.05) Glutathione peroxidase (GSH-Px) activity in the plasma of piglets fed with LP + SB, LP + MCT, and LP + PUFA diets than those fed with the LP diet. The piglets in the LP + SB and LP + PUFA groups had lower IKK-alpha (IKKa) mRNA expression in the colonic mucosa compared with those in the CON and LP groups (p < 0.05). The mRNA expression of TLR4 in the colonic mucosa of piglets in the LP + SB, LP + MCT, and LP + PUFA groups was decreased when compared with the CON and LP groups (p < 0.05). The LP + MCT diets increased the gene expression of nuclear factor erythroid 2-related factor 2 (Nrf2) in the colonic mucosa of piglets compared with CON, LP, and LP + SB diets (p < 0.05). The abundance of Erysipelotrichaceae in the colonic microbiome of piglets in the LP group was higher than that in the other four groups (p < 0.05). Collectively, this study showed that LP diets supplemented with SB, MCT, or n-3 PUFA reduced plasma inflammatory factor levels, increased plasma GSH-Px activity, and declined mRNA expression of TLR4 and IKKa in the colonic epithelium, whereas it reduced the abundance of Erysipelotrichaceae in the colon of piglets.

Transcriptomics yields valuable information regarding the response mechanisms of Chinese Min pigs infected with PEDV.

Porcine epidemic diarrhea virus (PEDV) causes porcine epidemic diarrhea (PED), a highly infectious disease, which has resulted in huge economic losses for the pig industry. To date, the pathogenic and immune response mechanism was not particularly clear. The purpose of this study was to investigate the pathogenic and immune responses of pigs infected with PEDV.In this study, 12 Min pigs were randomly selected without taking colostrum. At 3 days old, eight piglets were infected with 1 mL of PEDV solution (10 TCID50/ml), and the remaining four piglets were handled by 1 mL of 0.9% normal saline. Within the age of 7 days old, four piglets died and were considered as the death group. Correspondingly, four alive individuals were classified into the resistance group. Tissues of the duodenum, jejunum, ileum, colon, cecum, and rectum of piglets in the three groups were collected to measure the PEDV content. Additionally, the jejunum was used for the measurements and analyses of Hematoxylin-eosinstaining (HE), immunohistochemical sections, and transcriptomics. The phenotypes of Min piglets infected with PEDV showed that the viral copy numbers and jejunal damage had significant differences between the death and resistance groups. We also observed the transcriptome of the jejunum, and the differentially expressed (DE) analysis observed 6,585 DE protein-coding genes (PCGs), 3,188 DE long non-coding RNAs (lncRNAs), and 350 DE microRNAs (miRNAs), which were mainly involved in immune response and metabolic pathways. Furthermore, the specific expressed molecules for each group were identified, and 97 PCGs,108 lncRNAs, and 51 miRNAs were included in the ceRNA-regulated networks. By weighted gene co-expression network analysis (WGCNA) and transcription factor (TF) prediction, 27 significant modules and 32 significant motifs (E-value < 0.05) annotated with 519 TFs were detected. Of these TFs, 53 were DE PCGs. In summary, the promising key PCGs, lncRNAs, and miRNAs related to the pathogenic and immunological response of pigs infected with PEDV were detected and provided new insights into the pathogenesis of PEDV.

Piglet growth performance improved by dietary supplementation of porous or nano particles of zinc oxide may be related to the gut microbiota

Previous studies on porous or nano particles zinc oxide (ZnO) in the piglets have mainly focused on growth performance and intestinal inflammation, but have scarcely explored the efficacy on gut microbiota. In addition, the efficacy of nano particles ZnO, which is related to its product quality, remains undefined. This study aimed to determine the efficacy of dietary 500 mg/kg porous or nano particles ZnO on the growth performance and gut microbiota of the weaned piglets. A total of 128 weaned piglets were randomly assigned to the dietary groups: NC (basal diet), PC (basal diet + 3,000 mg/kg conventional ZnO), 500HiZ (basal diet + 500 mg/kg porous particles ZnO), and 500ZNP (basal diet + 500 mg/kg nano particles ZnO). Compared with the NC diet group, both 500HiZ and 500ZNP increased (P < 0.05) average daily feed intake (1 to 28 d) and average daily gain (1 to 28 d), and the 500ZNP tended to decrease feed to gain ratio (F:G ratio, 1 to 28 d) (P = 0.09). Both 500HiZ and 500ZNP decreased crypt depth of the ileum and increased claudin-2 in the duodenum and zonula occludens-1 in the ileum (P < 0.05). Moreover, both 500HiZ and 500ZNP decreased IL-1β and tumor necrosis factor-α (TNF-α) in the jejunum and decreased TNF-α and IL-6 in the ileum (P < 0.05). Both 500HiZ and 500ZNP increased microbial β-diversity index in the ileum and microbial α-diversity indices in the colon of piglets (P < 0.05). The probiotic genera Coprococcus (500ZNP) and Blautia (500HiZ) were positively correlated with the F:G ratio (1 to 28 d) in colon of piglets (P < 0.05). In addition, 500HiZ promoted mitochondrial fusion protein 1 (MFN1) and zinc transporter-1 (ZnT-1) in the jejunum (P < 0.05), whilst 500ZNP decreased MFN1 in the jejunum and ZnT-1 in the ileum (P < 0.05). In summary, both 500HiZ and 500ZNP improved the growth performance of piglets, which is likely via the genera Blautia and Coprococcus, respectively. Both 500HiZ and 500ZNP improved barrier function and inflammation of the intestine, and 500HiZ achieved better efficacy than 500ZNP on intestine mitochondrial functions.

Disruption of the primocolonizing microbiota alters epithelial homeostasis and imprints stem cells in the colon of neonatal piglets.

The gut microbiota plays a key role in the postnatal development of the intestinal epithelium. However, the bacterial members of the primocolonizing microbiota driving these effects are not fully identified and the mechanisms underlying their long-term influence on epithelial homeostasis remain poorly described. Here, we used a model of newborn piglets treated during the first week of life with the antibiotic colistin in order to deplete specific gram-negative bacteria that are transiently dominant in the neonatal gut microbiota. Colistin depleted Proteobacteria and Fusobacteriota from the neonatal colon microbiota, reduced the bacterial predicted capacity to synthetize lipopolysaccharide (LPS), and increased the concentration of succinate in the colon. The colistin-induced disruption of the primocolonizing microbiota was associated with altered gene expression in the colon epithelium including a reduction of toll-like receptor 4 (TLR4) and lysozyme (LYZ). Our data obtained in porcine colonic organoid cell monolayers suggested that these effects were not driven by the variation of succinate or LPS levels nor by a direct effect of colistin on epithelial cells. The disruption of the primocolonizing microbiota imprinted colon epithelial stem cells since the expression of TLR4 and LYZ remained lower in organoids derived from colistin-treated piglet colonic crypts after several passages when compared to control piglets. Finally, the stable imprinting of LYZ in colon organoids was independent of the H3K4me3 level in its transcription start site. Altogether, our results show that disruption of the primocolonizing gut microbiota alters epithelial innate immunity in the colon and imprints stem cells, which could have long-term consequences for gut health.

Influence of Escherichia coli infection on intestinal mucosal barrier integrity of germ-free piglets

AimsWe focused on investigating the influence of Escherichia coli (E. coli) on the intestinal barrier. Material and methodsWe studied changes in the distribution and secretory activities of goblet cells and enteroendocrine cells (EECs), as well as changes in the population of mast cells (MCs) in the jejunal and colonic mucosa of germ-free (GF) piglets as a healthy control group and GF piglets whose intestines were colonised with E. coli bacteria on day 5. Key findingsThe results suggest that the colon of GF piglets is more resistant and less prone to coliform bacterial infection compared to the jejunum. This can be confirmed by a lower degree of histopathological injury index as well as an improvement of the morphometric parameters of the colonic mucosa, together with a significantly increased (p < 0.05) expression of MUC1/EMA, and ZO-3. We also observed a significant decrease in the population of activated MCs (p < 0.001) and EECs (p < 0.001). These findings may indicate a rapid response and better preparation of the intestinal barrier for possible pathological attacks and the subsequent development of mucosal lesions during the development and progression of the intestinal diseases. SignificanceTo date, gut-targeted therapeutic approaches that can modulate bacterial translocation and chronic inflammation are still in their infancy but represent one of the most promising areas of research for the development of new effective treatments or clinical strategies in the future. Therefore, a better understanding of these processes can significantly contribute to the development of these targeted strategies for disease prevention and treatment.

Metagenomics-based inference of microbial metabolism towards neuroactive amino acids and the response to antibiotics in piglet colon.

Gut-derived neuroactive metabolites from amino acids perform a broad range of physiological roles in the body. However, the interaction between microbiota and epithelium in the metabolism of amino acids with neuroactive properties remains unclear in the colon of piglets. To investigate the microbial and epithelial metabolism, metagenomics and mucosal metabolomics were performed using colonic samples from 12 ileum-canulated piglets subjected to a 25-day infusion with saline or antibiotics. We categorized 23 metabolites derived from the metabolism of tryptophan, glutamate, and tyrosine, known as precursors of neuroactive metabolites. Microbial enzymes involved in the kynurenine synthesis via arylformamidase, 4-aminobutyric acid (GABA) synthesis via putrescine aminotransferase, and tyramine synthesis via tyrosine decarboxylase were identified in Clostridiales bacterium, uncultured Blautia sp., and Methanobrevibacter wolinii, respectively. Antibiotics significantly affected the microbiota involved in tryptophan-kynurenine and glutamate-GABA metabolism. An increase in the relative abundance of putrescine aminotransferase and Blautia sp. correlated positively with an increase in luminal GABA concentration. Overall, our findings provide new insights into the microbial ability to metabolize key amino acids that are precursors of neuroactive metabolites.

Regulation of serotonin production by specific microbes from piglet gut

BackgroundSerotonin is an important signaling molecule that regulates secretory and sensory functions in the gut. Gut microbiota has been demonstrated to affect serotonin synthesis in rodent models. However, how gut microbes regulate intestinal serotonin production in piglets remains vague. To investigate the relationship between microbiota and serotonin specifically in the colon, microbial composition and serotonin concentration were analyzed in ileum-cannulated piglets subjected to antibiotic infusion from the ileum when comparing with saline infusion. Microbes that correlated positively with serotonin production were isolated from piglet colon and were further used to investigate the regulation mechanisms on serotonin production in IPEC-J2 and a putative enterochromaffin cell line RIN-14B cells.ResultsAntibiotic infusion increased quantities of Lactobacillus amylovorus (LA) that positively correlated with increased serotonin concentrations in the colon, while no effects observed for Limosilactobacillus reuteri (LR). To understand how microbes regulate serotonin, representative strains of LA, LR, and Streptococcus alactolyticus (SA, enriched in feces from prior observation) were selected for cell culture studies. Compared to the control group, LA, LR and SA supernatants significantly up-regulated tryptophan hydroxylase 1 (TPH1) expression and promoted serotonin production in IPEC-J2 cells, while in RIN-14B cells only LA exerted similar action. To investigate potential mechanisms mediated by microbe-derived molecules, microbial metabolites including lactate, acetate, glutamine, and γ-aminobutyric acid were selected for cell treatment based on computational and metabolite profiling in bacterial supernatant. Among these metabolites, acetate upregulated the expression of free fatty acid receptor 3 and TPH1 while downregulated indoleamine 2,3-dioxygenase 1. Similar effects were also recapitulated when treating the cells with AR420626, an agonist targeting free fatty acid receptor 3.ConclusionsOverall, these results suggest that Lactobacillus amylovorus showed a positive correlation with serotonin production in the pig gut and exhibited a remarkable ability to regulate serotonin production in cell cultures. These findings provide evidence that microbial metabolites mediate the dialogue between microbes and host, which reveals a potential approach using microbial manipulation to regulate intestinal serotonin biosynthesis.

Dietary bile acid supplementation in weaned piglets with intrauterine growth retardation improves colonic microbiota, metabolic activity, and epithelial function

BackgroundIntrauterine growth retardation (IUGR) is one of the major constraints in animal production. Our previous study showed that piglets with IUGR are associated with abnormal bile acid (BA) metabolism. This study explored whether dietary BA supplementation could improve growth performance and colonic development, function, microbiota, and metabolites in the normal birth weight (NBW) and IUGR piglets. A total of 48 weaned piglets (24 IUGR and 24 NBW) were allocated to four groups (12 piglets per group): (i) NBW group, (ii) NBW + BA group, (iii) IUGR group, and (iv) IUGR + BA group. Samples were collected after 28 days of feeding.ResultsThe results showed that dietary BA supplementation increased the length and weight of the colon and colon weight to body weight ratio, while decreased the plasma diamine oxidase (DAO) concentration in the NBW piglets (P < 0.05). Dietary BA supplementation to IUGR piglets decreased (P < 0.05) the plasma concentrations of D-lactate and endotoxin and colonic DAO and endotoxin, suggesting a beneficial effect on epithelial integrity. Moreover, dietary BA supplementation to NBW and IUGR piglets increased Firmicutes abundance and decreased Bacteroidetes abundance (P < 0.05), whereas Lactobacillus was the dominant genus in the colon. Metabolome analysis revealed 65 and 51 differential metabolites in the colon of piglets fed a diet with/without BA, respectively, which was associated with the colonic function of IUGR piglets. Furthermore, dietary BA supplementation to IUGR piglets upregulated the expressions of CAT, GPX, SOD, Nrf1, IL-2, and IFN-γ in colonic mucosa (P < 0.05).ConclusionsCollectively, dietary BA supplementation could improve the colonic function of IUGR piglets, which was associated with increasing proportions of potentially beneficial bacteria and metabolites. Furthermore, BA shows a promising application prospect in improving the intestinal ecosystem and health of animals.

Effects and Underlying Mechanisms of Zearalenone Mycotoxin at Concentrations Close to the EC Recommendation on the Colon of Piglets after Weaning

Zearalenone (ZEN) is a mycotoxin produced by Fusarium fungi that contaminates food and feed, affecting both human and animal health. Among farm animals, the pig is a great consumer of grains and has a native sensitivity to mycotoxins. As the main route of contamination is oral, the intestine is the first defense barrier that plays an important role in the immune response being able to secrete effector molecules (cytokines). At the European level, there are no regulations regarding the amount of ZEN that can be present in the feed of piglets, only recommendations for piglets 0.100 mg ZEN/kg feed (100 ppb). In this study, the effects of ZEN in concentrations below (75 ppb) and above (290 ppb) EU recommendation on the level of some key markers involved in the oxidative and inflammatory response, as well as the mechanisms and signaling pathways through which ZEN could produce its toxicity, were monitored in the colon of weaned piglets. The exposure of the piglets to the lower concentration of ZEN (75 ppb) did not lead to changes in stress and inflammation markers or in the signaling pathways associated with these processes.

Maternal Magnolol Supplementation during Pregnancy and Lactation Promotes Antioxidant Capacity, Improves Gut Health, and Alters Gut Microbiota and Metabolites of Weanling Piglets.

Maternal nutrition exerts a profound effect on the postnatal performance of offspring, especially during the weaning period. The multifunctional bioactive component magnolol (MAG) has shown promise as a dietary supplement. This study aimed to explore the effects of maternal MAG supplementation on the antioxidant capacity, gut health, gut microbiome, and metabolome composition of weanling piglets. Fifty pregnant sows were randomly divided into two equally sized groups, the control group and the group supplemented with 100 g/t MAG during the gestation and lactation periods, and 7 days postweaning, the pups were euthanized. The microbiome and metabolome features of weanling piglet colons were compared. Our results revealed that maternal MAG supplementation modified the serum redox status of weanling piglets by decreasing malondialdehyde concentration and increasing superoxide dismutase activity and total antioxidant capacity. Moreover, the decreased indicators of diarrhea were accompanied by improved gut barrier function, in which serum diamine oxidase concentration was decreased, and expressions of zona occludens-1, claudin-1, and intestinal alkaline phosphatase were increased in the colon of weanling piglets from sows supplemented with MAG. Further analysis of the gut microbiota indicated that maternal MAG supplementation significantly increased the relative abundance of beneficial bacteria in the colon of weanling piglets, including Faecalibacterium prausnitzii and Oscillospira. Metabolome analysis identified 540 differential metabolites in the colon of piglets from MAG-fed dams, of which glycerophospholipid classes were highly correlated with progeny gut health and key beneficial bacteria. Our findings indicated that maternal MAG supplementation can improve the oxidative status and gut health of weanling piglets, possibly due to alterations in the gut microbiota and metabolites.

The Early Life Microbiota Is Not a Major Factor Underlying the Susceptibility to Postweaning Diarrhea in Piglets.

Postweaning diarrhea (PWD) in piglets impair welfare, induce economic losses and lead to overuse of antibiotics. The early life gut microbiota was proposed to contribute to the susceptibility to PWD. The objective of our study was to evaluate in a large cohort of 116 piglets raised in 2 separate farms whether the gut microbiota composition and functions during the suckling period were associated with the later development of PWD. The fecal microbiota and metabolome were analyzed by 16S rRNA gene amplicon sequencing and nuclear magnetic based resonance at postnatal day 13 in male and female piglets. The later development of PWD was recorded for the same animals from weaning (day 21) to day 54. The gut microbiota structure and α-diversity during the suckling period were not associated with the later development of PWD. There was no significant difference in the relative abundances of bacterial taxa in suckling piglets that later developed PWD. The predicted functionality of the gut microbiota and the fecal metabolome signature during the suckling period were not linked to the later development of PWD. Trimethylamine was the bacterial metabolite which fecal concentration during the suckling period was the most strongly associated with the later development of PWD. However, experiments in piglet colon organoids showed that trimethylamine did not disrupt epithelial homeostasis and is thus not likely to predispose to PWD through this mechanism. In conclusion, our data suggest that the early life microbiota is not a major factor underlying the susceptibility to PWD in piglets. IMPORTANCE This study shows that the fecal microbiota composition and metabolic activity are similar in suckling piglets (13 days after birth) that either later develop post-weaning diarrhea (PWD) or not, which is a major threat for animal welfare that also causes important economic losses and antibiotic treatments in pig production. The aim of this work was to study a large cohort of piglets raised in separates environments, which is a major factor influencing the early life microbiota. One of the main findings is that, although the fecal concentration of trimethylamine in suckling piglets was associated with the later development of PWD, this gut microbiota-derived metabolite did not disrupt the epithelial homeostasis in organoids derived from the pig colon. Overall, this study suggests that the gut microbiota during the suckling period is not a major factor underlying the susceptibility of piglets to PWD.

Influence of high- and low-fermentable dietary fibres in sows' diet on the colostrum potential against Clostridioides difficile toxin-induced effects in IPEC-J2 cells.

Sow colostrum has been reported to protect the IPEC-J2 cells and piglet colon tissues from detrimental effect of Clostridioides difficile toxins. Since dietary fibre can influence the colostrum composition in sows, we hypothesised that it can also differentially affect the colostrum potential against C. difficile toxin-induced effects in IPEC-J2. IPEC-J2 were incubated with colostrum from sows fed either high-fermentable sugar beet pulp (SBP) or low-fermentable lignocellulose (LNC) fibres and in combination with the toxins and analysed by trans-epithelial electrical resistance (TEER) and cell viability using propidium iodide in flow cytometry. Toxins drastically decreased the integrity of IPEC-J2. Colostrum from the sows fed either SBP or LNC exerted protective effect against toxins on IPEC-J2 integrity and this effect was numerically superior in the SBP group. Differences in the percentages of TEER between different treatments were noted after 2 h (p = 0.043), 3 h (p = 0.017) and 4 h (p = 0.017) of incubation and a tendency for differences was noted after 5 h of incubation (p = 0.071). Colostrum from either SBP- or LNC-fed sows did not protect the IPEC-J2 from toxin-induced death. Colostrum of the sows fed either high-fermentable or low-fermentable fibres has a potential to protect IPEC-J2 from the loss of integrity, which may be important in protection from C. difficile-infection development in neonatal piglets.

Grape seed meal by-product is able to counteract oxidative stress induced by lipopolysaccharide and dextran sulphate in IPEC cells and piglets after weaning.

Oxidative stress is a pivotal factor in the pathogenesis of intestinal inflammation, leading to cellular damage and tissue injury. Natural antioxidants compounds found in agro-industrial by-products have proven their effectiveness in treatment of intestinal inflammation and oxidative stress, exhibiting many favourable effects. The aim of this study was to evaluate the capacity of a grape seed meal byproduct (GSM) to counteract the effects induced by E. coli lipopolysaccharide (LPS, 5μg/ml) in vitro on IPEC-1 cells and by dextran sulphate sodium (DSS, 1g/b.w./day) in vivo on piglets after weaning. Reactive oxygen species (ROS), pro-oxidant markers (malondialdehyde MDA, thiobarbituric acid reactive substances TBARS, protein carbonyl, DNA oxidative damage) antioxidant enzymes (catalase -CAT, superoxide dismutase -SOD, glutathione peroxidase -GPx, endothelial and inducible nitric oxide synthases -eNOS and iNOS) and several important components of Keap1/Nrf2 signalling pathway were analysed in IPEC-1 cells as well as in piglet's colon and lymph nodes. Our results demonstrated that GSM extract or 8% dietary GSM showed anti-oxidant properties counteracting the pro-oxidant response (ROS, MDA-TBARS, protein carbonyl, DNA/RNA damage) induced by LPS or DSS and restoring the levels of endogenous antioxidant enzymes, including CAT, SOD, GPx, eNOS and iNOS in colon and mesenteric lymph nodes. These beneficial effects were modulated via Nrf2 signalling pathway in both in vitro and in vivo studies.

Caffeic acid improves intestinal barrier functions by regulating colonic bacteria and tight junction protein expression and alleviating inflammation in weaning piglets

The experiment investigated the effect of caffeic acid on bacteria, short-chain fatty acids (SCFA), and the expression of tight junction protein and inflammation related genes in the colon of weaning piglets. Thirty-six weaning piglets were allocated to three treatment groups, which were fed with a basal diet, a basal diet supplemented with 250 mg/kg or 500 mg/kg caffeic acid for 28 days. The results showed that caffeic acid treatment increased the contents of acetate acid, propionate acid and total SCFA. Moreover, real-time quantitative PCR showed that the number of Bifidobacterium (p < 0.05) and Lactobacillus (p < 0.05) were increased and the number of Escherichia coli (p < 0.05) was decreased by caffeic acid in colonic mucosa. Real-time quantitative PCR also showed that the mRNA levels of zonula occludens-1 (p < 0.01), claudin-1 (p < 0.01), occludin (p < 0.01), mucin 1 (MUC1) (p < 0.01), MUC2 (p < 0.01), interleukin 4 (IL-4) (p < 0.01) and IL-10 (p < 0.05) were increased, while the mRNA expression levels of histone deacetylases (p < 0.01), IL-1 (p < 0.01), IL-6 (p < 0.01) and tumor necrosis factor-α (TNF-α) (p < 0.01) were decreased, by caffeic acid in colonic mucosa. These results suggested that caffeic acid could improve intestinal barrier function in weaned pigs, which might be mediated by regulating colonic bacteria and tight junction protein expression and alleviating inflammation.

Effects of different starch diets on growth performance, intestinal health andfaecal microbiota of growing pigs.

This experiment was conducted to investigate the effects of different starch source diets on growth performance, intestinal health, and, microbiota of growing pigs. Eighteen healthy "Duroc × Landrace × Yorkshire" pigs (50 ± 0.61 kg, Castrated boar) were randomly divided into three groups with six replicates and one pig per replicate. The pigs in the three treatments were fed diets prepared with cassava flour (CF), rice bran (RB)andsorghum flour (SF), respectively, and the nutritional levels of the three treatments were the same. The experiment lasted for 28 days. The results showed that pigs in the RB group had significantly increased average daily gain (ADG, p < 0.05) compared with pigs in CF and SF groups. Compared with pigs in the CF group, the final body weight (FBW) of growing pigs in the RB group was increased and the ratio of feed to gain (F: G) was decreased (p < 0.05). There was no significant difference between FBW and F: G between the SF group and the other two groups. Compared with the CF group, the RB group significantly increased the jejunum amylase activity (p < 0.05), and there was no significant difference between the SF group and the other two groups. Compared with growing pigs in the CF group and SF group, the duodenal villus height and villus height/crypt depth ratio of growing pigs in the RB group were significantly increased (p < 0.05). The concentrations of acetic acid, propionic acid, and total VFA in the colon and caecum of piglets in the SF group were significantly increased (p < 0.05) compared to piglets in CF and RB groups, and there was no significant difference between the CF group and RB group. Compared with the RB group, caecal butyric acid concentration was significantly increased in SF and CF groups (p < 0.05). Seven dominant phyla were identified at the phylum level, among which Firmicutes, Bacteroidotaand Spirochaetota were dominant phyla, accounting for 74.18%, 14.87% and6.56% of the RB grouprespectively. Cassava flour group accounted for 80.22%, 9.64% and3.71%; Accounting for 65.33%, 17.34% and13.07% of the SF group. Through the comparative analysis of microbial differences among the treatment groups, it was found that at the phylum level, compared with the SF group, the abundance of Synergistota in the diet of the CF group and the diet of the RB group was significantly increased (p < 0.05). The abundance decreased significantly (p < 0.05). The quantity of Desulfobacterota in the RB group was significantly higher than that in the CF group (p < 0.05). In conclusion, compared with sorghum starch and cassava starch, RB starch can improve the activity of digestive enzymes and villus height in the small intestine of growing pigs and promote the growth of pigs by protecting the intestinal health of growing pigs.