Jejunal Mucosa Research Articles

Available phosphorus levels modulate gene expression related to intestinal calcium and phosphorus absorption and bone parameters differently in gilts and barrows.

Dietary phytase increases bioavailability of phytate-bound phosphorus (P) in pig nutrition affecting dietary calcium (Ca) to P ratio, intestinal uptake, and systemic utilization of both minerals, which may contribute to improper bone mineralization. We used phytase to assess long-term effects of two dietary available P (aP) levels using a one-phase feeding system on gene expression related to Ca and P homeostasis along the intestinal tract and in the kidney, short-chain fatty acids in stomach, cecum, and colon, serum, and bone parameters in growing gilts and barrows. Growing pigs (37.9±6.2 kg) had either free access to a diet without (Con; 75 gilts and 69 barrows) or with phytase (650 phytase units; n = 72/diet) for 56 days. Samples of blood, duodenal, jejunal, ileal, cecal, and colonic mucosa and digesta, kidney, and metacarpal bones were collected from 24 pigs (6 gilts and 6 barrows per diet). Phytase decreased daily feed intake and average daily gain, whereas aP intake increased with phytase versus Con diet (p<0.05). Gilts had higher colonic expression of TRPV5, CDH1, CLDN4, ZO1, and OCLN and renal expression of TRPV5 and SLC34A3 compared to barrows (p<0.05). Phytase increased duodenal expression of TRPV5, TRPV6, CALB1, PMCA1b, CDH1, CLDN4, ZO1, and OCLN compared to Con diet (p<0.05). Furthermore, phytase increased expression of SCL34A2 in cecum and of FGF23 and CLDN4 in colon compared to Con diet (p<0.05). Alongside, phytase decreased gastric propionate, cecal valerate, and colonic caproate versus Con diet (p<0.05). Phytase reduced cortical wall thickness and index of metacarpal bones (p<0.05). Gene expression results suggested an intestinal adaptation to increased dietary aP amount by increasing duodenal trans- and paracellular Ca absorption to balance the systemically available Ca and P levels, whereas no adaption of relevant gene expression in kidney occurred. Greater average daily gain in barrows related to higher feed intake.

Effect of Different Levels of Niacin on Serum Biochemical Parameters, Antioxidant Status, Cytokine Levels, Inflammatory Gene Expression and Colonic Microbial Composition in Weaned Piglets.

Niacin plays an important role in regulating the gut health of weaned piglets. In this study, 48 25-day-old weaned piglets (7.9 ± 0.20 kg) produced by 14 sows (3 to 4 piglets per sow) were randomly divided into 4 groups with 6 replicates in each group and 2 piglets in each replicate. Each group was fed diets supplemented with 22.5 (N1), 30 (N2), 45 (N3), and 75 (N4) mg/kg of niacin, respectively. Samples were taken at 7 and 14 d, respectively. The study shows that changes in niacin levels significantly affected the content of IgG and IgM in the serum (p &lt; 0.05). Niacin had a significant effect on antioxidant parameters such as MDA, T-SOD, and CuZn-SOD in the jejunal mucosa of weaned piglets (p &lt; 0.05). Moreover, significant differences were observed in the expression of cytokines such as TGF-β, TNF-α, and COX2 in the jejunal mucosa (p &lt; 0.05). The 16S rRNA sequencing analysis showed that there were significant differences in the colonic species composition, which were also accompanied by changes in the isovaleric acid content (p &lt; 0.05). In conclusion, an appropriate increase in niacin dose based on NRC (2012) has an important role in improving the antioxidant status of weaned piglets, alleviating intestinal inflammation in piglets, improving immunity, and regulating the structure of the microbiota.

Dietary supplementation of squalene increases the growth performance of early-weaned piglets by improving gut microbiota, intestinal barrier, and blood antioxidant capacity.

This study aimed to investigate the effects of dietary squalene (SQ) supplementation on the growth performance of early-weaned piglets. Twenty early-weaned piglets were randomly divided into two groups, the squalene group (SQ) and the control group (CON). The CON group was fed a basal diet, and the SQ group was fed a basal diet with 250 mg/kg squalene. The feeding period lasted 21 days. The results showed that SQ significantly increased the final body weight (FWB, P < 0.05), average daily gain (ADG, P < 0.05), and average daily feed intake (ADFI, P < 0.05) and significantly decreased the F/G ratio (feed intake/gain, P < 0.05) and diarrhea index (DI, P < 0.05). In terms of blood biochemical indicators, SQ significantly increased anti-inflammatory factors such as transforming growth factor-β (TGF-β, P < 0.001), interleukin-10 (IL-10, P < 0.001), and interferon-γ (IFN-γ, P < 0.01), and decreased pro-inflammatory factors such as tumor necrosis factor-α (TFN-α, P < 0.001) and interleukin-6 (IL-6, P < 0.001). Furthermore, SQ significantly increased blood antioxidant indexes (P < 0.001) such as superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), catalase (CAT), and total antioxidant capacity (T-AOC) and significantly decreased the level of malondialdehyde (MDA) (P < 0.001). The villus height (P < 0.001) and V/C ratio (villus height/crypt depth, P < 0.001) of the jejunum were significantly increased in the SQ group, while the crypt depth (P < 0.01) was decreased compared to the CON group. The intestinal permeability indexes, namely diamine oxidase (DAO), D-lactic acid (D-Lac), regenerative insulin-derived protein 3 (REG-3), and FITC-Dextran 4 (FD4), significantly decreased the concentrations in the treatment group (P < 0.001), and the antioxidant indexes of the jejunum, such as SOD, GSH-Px, CAT, and MDA, were improved by adding SQ. The qPCR results showed that adding SQ could significantly increase the mRNA expression of jejunal tight-junction proteins, such as zonula occludens-1 (ZO-1, P < 0.001), Occludin (P < 0.001), Claudin (P < 0.001), glucagon-like peptide-2 (GLP-2, P < 0.001), and insulin-like growth factor-1 (IGF-1, P < 0.001). Then, we used Western blotting experiments to further confirm the qPCR results. In addition, it was found that adding SQ increased the abundance of beneficial bacteria such as Gemmiger (P < 0.01) and decreased the abundance of harmful bacteria such as Alloprevotella (P < 0.05), Desulfovibrio (P < 0.05), and Barnesiella (P < 0.05). It was interesting that there was a very close correlation among the fecal microbes, growth performance parameters, intestinal barrier, and blood biochemical indicators. In conclusion, the data suggest that SQ supplementation could effectively improve the growth performance of early-weaned piglets by improving the gut microbiota, intestinal barrier, and antioxidant capacity of the blood and jejunal mucosa.

Does Deoxynivalenol Affect Amoxicillin and Doxycycline Absorption in the Gastrointestinal Tract? Ex Vivo Study on Swine Jejunum Mucosa Explants.

The presence of deoxynivalenol (DON) in feed may increase intestinal barrier permeability. Disturbance of the intestinal barrier integrity may affect the absorption of antibiotics used in animals. Since the bioavailability of orally administered antibiotics significantly affects their efficacy and safety, it was decided to evaluate how DON influences the absorption of the most commonly used antibiotics in pigs, i.e., amoxicillin (AMX) and doxycycline (DOX). The studies were conducted using jejunal explants from adult pigs. Explants were incubated in Ussing chambers, in which a buffer containing DON (30 µg/mL), AMX (50 µg/mL), DOX (30 µg/mL), a combination of AMX + DON, or a combination of DOX + DON was used. Changes in transepithelial electrical resistance (TEER), the flux of transcellular and intracellular transport markers, and the flux of antibiotics across explants were measured. DON increased the permeability of small intestine explants, expressed by a reduction in TEER and an intensification of transcellular marker transport. DON did not affect AMX transport, but it accelerated DOX transport by approximately five times. The results suggest that DON inhibits the efflux transport of DOX to the intestinal lumen, and thus significantly changes its absorption from the gastrointestinal tract.

Bifidobacterium animalis Promotes the Growth of Weaning Piglets by Improving Intestinal Development, Enhancing Antioxidant Capacity, and Modulating Gut Microbiota.

Probiotics are widely used to promote performance and improve gut health in weaning piglets. Therefore, the objective of this study was to investigate the effects of dietary supplementation with Bifidobacterium animalis subsp. lactis (B. animalis) JYBR-190 on the growth performance, intestine health, and gut microbiota of weaning piglets. The results showed that the dietary addition of B. animalis significantly improved growth performance and decreased diarrhea incidence. B. animalis increased villus height in the duodenum and elevated goblet cell numbers and amylase activity in the jejunum. Additionally, B. animalis supplementation markedly increased total antioxidant capacity in jejunal mucosa but declined the malondialdehyde content. B. animalis treatment did not affect the mRNA expressions associated with the intestinal barrier and inflammatory cytokine in various intestinal segments. Microbiota analysis indicated that a diet supplemented with B. animalis significantly increased the relative abundances of health-promoting bacteria in the lumen, such as Streptococcus, Erysipelotrichaceae, Coprococcus, and Oscillibacter. There was a trend for B. animalis fed piglets to have a higher relative abundance of B. animalis in ileal digesta. Moreover, B. animalis-treated pigs decreased the abundance of Helicobacter and Escherichia-Shigella in ileal mucosa-associated microbiota. In summary, this study showed that B. animalis supplementation stimulated growth performance, improved gut development, enriched beneficial bacteria abundances, and declined intestinal pathogens populations, while B. animalis had limited effects on the intestinal barrier and immune function. IMPORTANCE In the modern swine industry, weaning is a critical period in the pig's life cycle. Sudden dietary, social, and environmental changes can easily lead to gut microbiota dysbiosis, diarrhea, and a decrease in growth performance. To stabilize intestinal microbiota and promote animal growth, antibiotics were widely applied in swine diets during the past few decades. However, the side effects of antibiotics posed a great threat to public health and food safety. Therefore, it is urgent to find and develop antibiotic alternatives. The growing evidence suggested that probiotics can be preferable alternatives to antibiotics because they can modulate microbiota composition and resist pathogens colonization. In this study, our results indicated that dietary supplementation with Bifidobacterium animalis promoted growth in weaning piglets by improving gut development, increasing beneficial bacteria abundances, and declining pathogens populations.

The dynamics of molecular, immune and physiological features of the host and the gut microbiome, and their interactions before and after onset of laying in two hen strains

Aggregation of data, including deep sequencing of mRNA and miRNA data in jejunum mucosa, abundance of immune cells, metabolites, or hormones in blood, composition of microbiota in digesta and duodenal mucosa, and production traits collected along the lifespan, provides a comprehensive picture of lifelong adaptation processes. Here, respective data from two laying hen strains (Lohmann Brown-Classic (LB) and Lohmann LSL-Classic (LSL) collected at 10, 16, 24, 30, and 60 wk of age were analyzed. Data integration revealed strain- and stage-specific biosignatures, including elements indicative of molecular pathways discriminating the strains. Although the strains performed the same, they differed in the activity of immunological and metabolic functions and pathways and showed specific gut-microbiota-interactions in different production periods. The study shows that both strains employ different strategies to acquire and maintain their capabilities under high performance conditions, especially during the transition phase. Furthermore, the study demonstrates the capacity of such integrative analyses to elucidate molecular pathways that reflect functional biodiversity. The bioinformatic reduction of the multidimensional data provides good guidance for further manual review of the data.

Dietary supplementation with synbiotics improves growth performance, antioxidant status, immune function, and intestinal barrier function in broilers subjected to cyclic heat stress.

This study investigated the effects of synbiotics supplementation on growth performance, antioxidant status, immune function, and intestinal barrier function in broilers subjected to cyclic heat stress. One hundred and forty-four 22-day-old male broilers were randomly assigned to one of three treatment groups of six replicates each for a 21-day study, with eight birds per replicate. Broilers in the control group were reared at a thermoneutral temperature and received a basal diet. Broilers in the other two heat-stressed groups were fed a basal diet supplemented without (heat-stressed group) and with 1.5g/kg synbiotic (synbiotic group). One and a half gram of the synbiotic consisted with 3 × 109 colony forming units (CFU) Clostridium butyricum, 1.5 × 109CFU Bacillus licheniformis, 4.5 × 1010CFU Bacillus subtilis, 600mg yeast cell wall, and 150mg xylooligosaccharide. Compared with the control group, heat stress increased rectal temperatures at 28, 35, and 42days of age, respectively (P < 0.05). Birds subjected to heat stress had reduced weight gain, feed intake, and feed efficiency during 22 to 42days (P < 0.05). In contrast, supplementation with the synbiotic decreased rectal temperature at 42days of age and elevated weight gain of heat stress-challenged broilers (P < 0.05). Heat-stressed broilers exhibited a lower superoxide dismutase (SOD) activity in jejunal mucosa and a higher malondialdehyde accumulation in serum, liver and jejunal mucosa (P < 0.05), and the regressive SOD activity was normalized to control level when supplementing synbiotic (P < 0.05). Heat stress increased interleukin-1β (IL-1β) and interferon-γ (IFN-γ) levels in serum and IL-1β content in jejunal mucosa of broilers (P < 0.05). Synbiotic reduced IL-1β level in serum of broilers subjected to heat stress (P < 0.05). Compared with the control group, elevated serum diamine oxidase activity and reduced jejunal villus height were observed in broilers of the heat-stressed group (P < 0.05), and the values of these two parameters in the synbiotic group were intermediate (P > 0.05). Heat stress upregulated mRNA abundance of IL-1β and IFN-γ and downregulated gene expression levels of occluding and zonula occluden-1 (ZO-1) in jejunal mucosa of broilers (P < 0.05). The alterations in the mRNA expression levels of jejunal IL-1β and ZO-1 were reversed by the synbiotic (P > 0.05). In conclusion, dietary synbiotics could improve growth performance, antioxidant capacity, immune function, and intestinal barrier function in heat-stressed broilers.

Effect of an Escherichia coli F4/F18 bivalent oral live vaccine on gut health and performance of healthy weaned pigs

Oral live vaccines stimulate host immunity, but they could also affect intestinal mucosa development and gut microbiota of piglets during the postweaning. The aim of this study was to determine the effect of an oral vaccine against Escherichia coli F4 and F18 (Coliprotec F4/F18®), on gut functionality and integrity, growth performance and health status of postweaning piglets. A total of 96 weaned piglets (23.30 ± 1.85 days of age; 7334 ± 1039 g BW) were divided into two groups (16 replicates/group; three piglets/replicate) as follows: (1) Control (CO), fed a standard diet (prestarter up to 14 days, then starter feed); (2) Treated (TRT): as CO but vaccinated with Coliprotec F4/F18® at weaning (day 0). Piglets were weighed at day 0 and weekly until day 35. Individual faecal score was recorded daily. Piglets were sacrificed at days 10 (1/3 of total) and 35 (2/3). Samples of jejunum mucosa and of cecum content were collected for morphometric, immunohistochemistry analysis and for microbiota profile analysis, respectively. Data were fitted using a linear model including treatment, class of starting BW as fixed factors and litter as random factor. From days 0 to 7, piglets from the TRT group tended to have a higher average daily gain (+22.6%, P = 0.08) and average daily feed intake compared to the CO group (+13.2%, P = 0.022). Gain to feed ratio was lower in the TRT group from days 14 to 35 (−6.6%, P = 0.011). From days 7 to 14, the TRT group had a higher diarrhoea index (−199%, P < 0.001). Crypt depth was higher in the CO group (+10.9%, P = 0.04) at day 10, but not at day 35. Jejunal expression of Claudin-4 (probability of having a score = 3) was higher in the TRT group at day 10 (CO = 1.50% vs TRT = 2.69%, P < 0.0001) and day 35 (CO = 1.29% vs TRT = 1.92%, P = 0.012). Oral vaccine affected beta diversity at day 10 (P = 0.040; R2 = 0.05) and increased the abundance of specific taxa and genera in the cecum at day 10, including Prevotella (lg2FC = 23.2, FDR < 0.001). The results showed how an Escherichia coli-based vaccine supplied to weaned pigs can promote gut health by controlling symptoms of the postweaning perturbation in the first 2 weeks postweaning. In addition, the vaccine strains showed a probiotic-like effect by modulating gut microbiota favouring the establishment of beneficial bacteria, and by promoting gut barrier integrity.

Morphological Adaptation in the Jejunal Mucosa after Iso-Caloric High-Fat versus High-Carbohydrate Diets in Healthy Volunteers: Data from a Randomized Crossover Study.

Background and aims: The conditions for jejunal glucose absorption in healthy subjects have not been thoroughly studied. In this study we investigated differences in the jejunal villi enlargement factor, as well as ultrastructural aspects of the surface enterocytes and mitochondria, comparing 2 weeks of high-carbohydrate (HCD) versus high-fat diets (HFD). We also measured the ketogenesis rate-limiting enzyme 3-hydroxy-3-methylglutaryl-CoA synthase (HMGCS2) in relation to jejunal mitochondria. Methods: A single-centre, randomized, unblinded crossover study in 15 healthy volunteers ingesting strictly controlled equicaloric diets (either HCD or HFD), with 60% energy from the respective source. An enteroscopy was carried out after 2 weeks of each diet and jejunal mucosal biopsies were acquired. Conventional histology, immunofluorescent staining, transmission electron microscopy and confocal microscopy were used. Results: The villi did not demonstrate any change in the epithelial enlargement factor. Despite an increased mitosis, there were no changes in apoptotic indices. However, the ultrastructural analysis demonstrated a significant increase in the enlargement factor at the bases of the villi. The mitochondria demonstrated increased amounts of cristae after the HFD. The confocal microscopy revealed increased HMGCS2 per mitochondrial marker at the top of the villi after the HFD compared to the HCD. Conclusion: There is a morphometric adaption in the jejunal mucosa following the 2-week diets, not only on a histological level, but rather on the ultrastructural level. This study supports the notion that mitochondrial HMGCS2 is regulated by the fat content of the diet and is involved in the expression of monosaccharide transporters.

Characterization of the "gut microbiota-immunity axis" and microbial lipid metabolites in atrophic and potential celiac disease.

IntroductionPotential celiac disease (pCD) is characterized by genetic predisposition, positive anti-endomysial and anti-tissue transglutaminase antibodies, but a normal or almost normal jejunal mucosa (e.g., minor histological abnormalities without villous atrophy). To gain further insights into basic mechanisms involved in the development of intestinal villous atrophy, we evaluated and compared the microbial, lipid, and immunological signatures of pCD and atrophic CD (aCD).Materials and methodsThis study included 17 aCD patients, 10 pCD patients, and 12 healthy controls (HC). Serum samples from all participants were collected to analyze free fatty acids (FFAs). Duodenal mucosa samples of aCD and pCD patients were taken to evaluate histology, tissue microbiota composition, and mucosal immune response.ResultsWe found no significant differences in the mucosa-associated microbiota composition of pCD and aCD patients. On the other hand, in pCD patients, the overall abundance of serum FFAs showed relevant and significant differences in comparison with aCD patients and HC. In detail, compared to HC, pCD patients displayed higher levels of propionic, butyric, valeric, 2-ethylhexanoic, tetradecanoic, hexadecanoic, and octadecanoic acids. Instead, aCD patients showed increased levels of propionic, isohexanoic, and 2-ethylhexanoic acids, and a lower abundance of isovaleric and 2-methylbutyricacids when compared to HC. In addition, compared to aCD patients, pCD patients showed a higher abundance of isobutyric and octadecanoic acid. Finally, the immunological analysis of duodenal biopsy revealed a lower percentage of CD4+ T lymphocytes in pCD infiltrate compared to that observed in aCD patients. The functional characterization of T cells documented a pro-inflammatory immune response in both aCD and pCD patients, but the pCD patients showed a higher percentage of Th0/Th17 and a lower percentage of Th1/Th17.ConclusionThe results of the present study show, for the first time, that the duodenal microbiota of patients with pCD does not differ substantially from that of aCD; however, serum FFAs and local T cells displayed a distinctive profile between pCD, aCD, and HC. In conclusion, our result may help to shed new light on the “gut microbiota-immunity axis,” lipid metabolites, and duodenal immune response in overt CD and pCD patients, opening new paradigms in understanding the pathogenesis behind CD progression.

Effects of lutein on jejunal mucosal barrier function and inflammatory responses in lipopolysaccharide-challenged yellow-feather broilers

Broilers are frequently exposed to various immunological stresses, which lead to intestinal damage, weakened immunity, and even growth retardation. Lutein, as a kind of carotenoid, possesses antioxidant and immunomodulatory functions. Therefore, this study was conducted to investigate the effects of lutein on jejunal mucosal barrier function and inflammatory responses of yellow-feather broilers challenged with lipopolysaccharide (LPS). A total of two hundred eight-eight 1-day-old yellow-feather broilers were randomly allocated to 3 groups with 8 replicate cages containing 12 birds each. Birds were fed broken-rice-soybean basal diet containing 0, 20 and 40 mg/kg lutein (CON, LU20 and LU40) for 26 d. On days 21, 23, and 25 of the trial, broilers were intraperitoneally injected with LPS (1 mg/kg body weight). The results showed that, compared with CON group, LU40 supplementations significantly increased the average daily gain (ADG) of broilers at 1 to 21 and 22 to 26 d of age (P < 0.05), significantly decreased the ratio of feed to gain (F/G) of broilers at 22 to 26 d of age (P < 0.05). LU20 and LU40 supplementations increased goblet cell density in jejunum of broilers under LPS challenge, and LU20 supplementation elevated the villus area (P < 0.05). Scanning electron microscopy of jejunal mucosa revealed significant villi damage, while transmission electron microscopy demonstrated severe enterocyte damage and loss of cellular integrity in CON group. In particular, mitochondria were morphologically altered, appearing irregular or swollen. Apical junctional complexes between adjacent enterocytes were obviously shorter and saccular in CON group. LU20 and LU40 supplementations increased the mRNA expressions of Occludin, Claudin-1, and ZO-1 in the jejunal mucosa of broilers under LPS challenge (P < 0.05), restrained TLR4/MyD88/NF-κB pathway activation in the jejunal mucosa, decreased the mRNA expressions of IL-1β and IL-6, and strengthened the mRNA expressions of IL-4 and IL-10 (P < 0.05). Meanwhile, the protein expressions of p38 and JNK in LU40 group were lower than CON group (P < 0.05). It can be concluded that 40 mg/kg lutein supplementation improved LPS-induced jejunal mucosal barrier function and tamed inflammation of yellow-feather broilers.

275 Effects of Reduced Metabolizable Energy by Reducing fat Supplementation on Growth Performance and Intestinal Health of Nursery Pigs

Abstract This study aimed to investigate the effects of reducing metabolizable energy (ME) by reducing fat supplementation on growth performance and intestinal health of nursery pigs. Nursery pigs (n = 120; 7.3 ± 0.6 kg BW) were allotted in a randomized complete block design with sex and initial BW as blocks. Pigs were assigned to 5 dietary treatments with decreasing ME levels (3,400, 3,375, 3,350, 3,325, and 3,300 kcal/kg ME). ME content was gradually reduced by reducing fat supplementation (2%). Pigs were fed for 35 d in 2 phases (P1: 14 d and P2: 21 d). Titanium dioxide was added (0.4%) to P2 diets as an indigestible external marker to measure apparent ileal digestibility (AID) of nutrients. At d 35, a pig with median BW of each pen in 2 treatments (3,400 and 3,300 kcal/kg) was euthanized for sampling jejunal mucosa to analyze oxidative stress and immune status. Jejunal digesta was collected to measure viscosity. Ileal digesta was collected to analyze AID of nutrients. Data were analyzed using SAS. Reducing ME level by 3% (100 kcal/kg) had no effect on growth performance. The 3,300 kcal/kg diet reduced (P &amp;lt; 0.05) AID of DM (59.5 to 47.3%), GE (59.1 to 43.0%), CP (63.4 to 44.6%), and EE (67.7 to 55.2%), increased (P &amp;lt; 0.05) digesta viscosity (1.7 to 2.2 cP) and VH:CD (1.64 to 1.80), and decreased (P &amp;lt; 0.05) VH (467 to 433 µm) when compared with 3,400 kcal/kg diet. Additionally, the 3,300 kcal/kg diet increased (P &amp;lt; 0.05) the concentration of protein carbonyl (0.18 to 0.40 nmol/mg) in the jejunal mucosa. In conclusion, reducing ME by reducing fat supplementation by 2% increased the viscosity of jejunal digesta and protein carbonyl in the jejunal mucosa, resulting in decreased villus height and nutrient digestibility, without affecting growth performance.

391 Young Scholar Award Talk: Supplementation of Oligosaccharide-Based Polymer Enhanced Growth and Disease Resistance of Weaned Pigs by Modulating Intestinal Integrity and Systemic Immunity

Abstract Oligosaccharides have been reported to possess ETEC receptor activity for bacterial adhesions; thus, additional supplementation may efficiently prevent enterotoxin-induced secretory diarrhea. Moreover, grafted polymers that combine multiple substances have been proposed for their potential synergistic effects on preventing human and animal diseases. Therefore, the objective of this experiment was to investigate dietary supplementation of oligosaccharide-based polymer on growth performance, diarrhea severity, intestinal health, and systemic immunity of weaned pigs experimentally infected with an enterotoxigenic Escherichia coli (ETEC) when compared with in-feed antibiotics. Forty-eight pigs (7.23 ± 1.11 kg BW) were individually housed in disease containment rooms and randomly allotted to one of four treatments with 12 replicate pigs per treatment. The four dietary treatments were a nursery basal diet (CON), and 3 additional diets supplemented with 50 mg/kg antibiotics (Carbadox; CAR), 10 or 20 mg/kg of oligosaccharide-based polymer (OBP). The experiment lasted 18 d [7 d before and 11 d after the first inoculation (d 0)]. All pigs were orally inoculated with 3mL of F18 ETEC for 3 consecutive days from d 0 post-inoculation (PI). Growth performance and diarrhea severity were measured throughout the experiment. Blood, lymph nodes, spleen, and intestinal mucosa samples were collected during the peak infection period (d 2 and d 5 PI) and recovery period (d 11 PI). Pigs in CAR or OBP treatment groups had greater (P&amp;lt;0.05) body weight on d 5 or d 11 PI than pigs in the CON group, respectively. Supplementation of CAR or OBP enhanced (P&amp;lt;0.05) feed efficiency from d 0 to 5 PI and reduced (P&amp;lt;0.05) frequency of diarrhea throughout the experiment, compared with pigs in the CON group. Pigs in CAR or OBP groups had reduced (P&amp;lt;0.05) neutrophil counts and serum haptoglobin concentration compared to pigs in the CON group on d 2 and 5 PI. Pigs in OBP reduced (P&amp;lt;0.05) total coliforms in mesenteric lymph nodes on d 5 and 11 PI, whereas pigs in CAR or OBP groups had reduced (P&amp;lt;0.05) total coliforms in the spleen on d 11 PI compared with pigs in the CON group. On d 5 PI, pigs in the OBP group had greater (P&amp;lt;0.05) gene expression of ZO1 in jejunal mucosa, but less (P&amp;lt;0.05) mRNA expression of IL1B, IL6, and TNF in ileal mucosa, in comparison with pigs in the CON group. On d 11 PI, supplementation of CAR or OBP up-regulated (P&amp;lt;0.05) gene expression of CLDN1 in jejunal mucosa, but OBP reduced (P&amp;lt;0.05) IL6 gene expression in ileal mucosa compared to pigs in the CON group. In conclusion, supplementation of oligosaccharide-based polymer improved growth performance, alleviated diarrhea severity, and enhanced gut health in weaned pigs infected with ETEC F18 in a manner similar to in-feed antibiotics.

266 Effects of Lactobacillus Fermentate on Modulation of Mucosa-Associated Microbiota in Relation to Intestinal Health of Nursery Pigs Challenged with F18+ Escherichia Coli

Abstract This study investigated the effects of a Lactobacillus fermentate (LBF) on mucosa-associated microbiota and its correlation with intestinal health of pigs challenged with F18+ Escherichia coli. Newly-weaned pigs (n = 64; 6.6 ± 0.7 kg BW) were allotted in a randomized complete b;ock design (RCBD) to 4 treatments: NC: no-challenge/no-supplement; PC: E. coli/no-supplement; AGP: E. coli/bacitracin (30 g/t feed); and PBT: E. coli/LBF (2 kg/t feed). At d 7, challenge groups were orally inoculated (2.4 x 1010 CFU) with F18+ E. coli whereas NC received sterile solution. Pigs were fed for 28 d until euthanasia to collect jejunal mucosa to evaluate intestinal health and microbiota. PC reduced (P &amp;lt; 0.05) Selenomonas (1.39 to 0.32%). AGP increased (P &amp;lt; 0.05) Bifidobacteriaceae (0.19 to 2.13%), Burkholderiaceae (0.18 to 1.54%), Comamonadaceae (1.13 to 6.97%), Enterobacteriaceae (0.81 to 2.29%), Microbacteriaceae (0.27 to 1.96%), Moraxellaceae (0.28 to 5.57%), and Pseudomonadaceae (0.36 to 2.59%). PBT increased (P &amp;lt; 0.05) Propionibacteriaceae (0.16 to 3.55%). AGP and PBT increased (P &amp;lt; 0.05) the microbiota diversity (Chao1: 48.8 to 76.4 and 71.6, respectively). Burkholderiaceae and Comamonadaceae were negatively correlated with TNF-α (r = -0.31 and -0.30, respectively; P &amp;lt; 0.05), whereas Burkholderiaceae, Comamonadaceae and Microbacteriaceae were positively correlated with protein carbonyl (r = 0.39, 0.33, and 0.37, respectively; P &amp;lt; 0.05). Bifidobacteriaceae and Pseudomonadaceae were positively correlated with VH:CD (r = 0.41 and 0.40, respectively; P &amp;lt; 0.05), whereas Pseudomonadaceae was positively correlated with villus height (r = 0.34; P &amp;lt; 0.05). Enterobacteriaceae was negatively correlated with G:F (r = -0.41; P &amp;lt; 0.05), whereas positively correlated with IL-8 and MDA (r = 0.39 and 0.32; P &amp;lt; 0.05). Moraxellaceae was negatively correlated with IL-6 (r = -0.29, P &amp;lt; 0.05). Collectively, AGP and LBF increased diversity of beneficial microbiota which was related to improved intestinal health in pigs after E. coli F18+ challenge.

261 Effects of soy Protein Concentrate Replacing Animal Protein Supplements on Modulation of Mucosa-Associated Microbiota in Relation to Intestinal Health of Nursery Pigs

Abstract This study aimed to investigate the effects of soy protein concentrate (SPC) replacing animal protein supplements on modulation of mucosa-associated microbiota in relation to intestinal health of nursery pigs. Weaned pigs (n = 56; 6.4 ± 0.6 kg BW) were allotted to 5 treatments in a randomized complete block design (RCBD) with initial BW and sex as blocks and fed for 35 d in 3 phases (P1/2/3 for 10/12/13 d, respectively). Dietary treatments were NC (n = 12): a diet with fish meal 4/2/1%, poultry meal 10/8/4%, and blood plasma 4/2/1% for P1/2/3, respectively; RFM (n = 12): NC with SPC replacing fish meal; RPM (n = 12): NC with SPC replacing poultry meal; RBP (n = 12): NC with SPC replacing blood plasma, and PC (n = 8): NC with SPC replacing all animal protein supplements. All diets were formulated to meet the nutrient requirements by NRC (2012). On d 35, pigs were euthanized to collect jejunal mucosa to evaluate intestinal health and microbiota. PC increased (P &amp;lt; 0.05) Lactobacillaceae (1.04 to 8.15%), and tended to increase (P = 0.073) Prevotellaceae (7.48 to 18.94%), whereas it decreased (P &amp;lt; 0.05) Chao1 index (81.3 to 15.8), Helicobacteraceae (27.95 to 3.02), and tended to reduce (P = 0.099) Campylobacteraceae (2.95 to 0.13). RPM tended to increase (P = 0.090) Prevotellaceae (7.48 to 17.16%) and reduced (P &amp;lt; 0.05) Helicobacteraceae (27.95 to 11.02%) without affecting Chao 1. Decreased Chao 1 was correlated to reduced feed intake and therefore, weight gain of pigs. In conclusion, the use of SPC replacing all animal proteins in nursery diets benefited the composition of the jejunal mucosa-associated microbiota, whereas feed intake and weight gain were reduced. The use of SPC replacing only poultry meal, however, benefitted the composition of the jejunal mucosa-associated microbiota without affecting feed intake and weight gain of nursery pigs.

Clostridium butyricum improves the intestinal health of goats by regulating the intestinal microbial community.

Clostridium butyricum, as a probiotic with a variety of active products, has been widely used to improve the intestinal health of humans and animals. Previous studies had demonstrated that Clostridium butyricum exhibited potential protective and positive effects in human disease research and animal production by producing a variety of beneficial substances, such as intestinal inflammation, the intestinal epithelial barrier, metabolic diseases, and regulation of the gut microbiota. Therefore, we hypothesized that dietary Clostridium butyricum supplementation could improve gut health in fattening goats by modulating gut microbiota. However, it is unclear whether Clostridium butyricum can reach the intestine through the rumen, so 15 healthy Albas goats were selected and randomly divided into 3 treatments with 5 replicates in each group. The groups were divided as follows: control group (CON: basal diet), rumen-protected Clostridium butyricum group (RPCB: basal diet plus 1.0 × 109 CFU/kg Clostridium butyricum coated with hydrogenated fat), and Clostridium butyricum group (CB: basal diet plus 1.0 × 109 CFU/kg Clostridium butyricum). The experiment was slaughtered after a 70-day growth test, and the jejunal mucosa and intestinal contents of the goats were collected to determine tight junction proteins related genes expression and 16S rDNA microbial sequencing analysis to evaluate the intestine health. The results showed that dietary supplementation with Clostridium butyricum significantly increased the expression of the Claudin-4 gene of the jejunal mucosa (P < 0.05) and had a trend toward a significant increase in the Occludin gene (0.05 < P < 0.10). However, Clostridium butyricum had no significant effect on the expression of intestinal inflammatory factors (P > 0.10). In addition, the relative fractionation of Clostridium and Clostridiaceae_unclassified in the gut microbiota at the genus level decreased significantly compared with controls (P < 0.05). The results of the analysis of the level of Clostridium species showed that Clostridium butyricum only existed in the treatment group. And the correlation results showed that Occludin and Claudin-4 genes were positively correlated with Sharppea and Clostridium butyricum, and negatively correlated with Clostridium (P < 0.05). Supplementing Clostridium butyricum in the diet did not significantly affect the intestinal immune function of goats, while regulation of the intestinal microbiota was associated with improving the intestinal epithelial barrier.

Grape seed procyanidins improve intestinal health by modulating gut microbiota and enhancing intestinal antioxidant capacity in weaned piglets

Grape seed procyanidins (GSPs) are polyphenolic compounds extracted from grape seeds and are known for their antioxidant, anti-inflammatory, antimicrobial, and immune modulating properties. As weaning stress in piglets can cause dysbacteriosis and oxidative damage to the intestine, resulting in diarrhea, the present study was conducted to determine whether administering GSPs can enhance intestinal antioxidant capacity and modulate gut microbiota to improve the overall gut health in weaned piglets. A total of 96 crossbred piglets were allocated randomly to four treatment groups; each treatment group comprised six replicate pens with four piglets per pen. Each group was fed one of the four dietary treatments for 28 days: the basal diet (control group) or the basal diet supplemented with 50, 100, or 150 mg/kg of GSPs. One piglet from each pen was euthanized to collect the jejunal mucosae for evaluating the intestinal antioxidant capacity and to collect the digesta of the cecum, colon, and rectum for assessing the succession of the gut microflora and determining the variations in the production of short chain fatty acid (SCFA). Dietary supplementation with GSPs enhanced the jejunal antioxidant capacity in weaned piglets, as evidenced by enhanced superoxide dismutase (SOD) activity (P < 0.001), increased mRNA abundance of superoxide dismutase (Sod1) and catalase (Cat) (P = 0.001 and P = 0.014, respectively), and decreased malondialdehyde (MDA) concentration (P < 0.001). The 100 mg/kg GSPs supplemented group showed enhanced Ace and Chao1 indices of the cecal, colonic, and rectal microflora when compared with those of the control group (P < 0.05), reflecting a rich bacterial community. Supplementing piglet diets with different concentrations of GSPs reduced the relative abundances of Proteobacteria and Anaerovibrio in various intestinal segments. A 100 mg/kg GSPs supplementation raised the abundances of probiotic Prevotellaceae_NK3B31_group and Prevotella_1 in colon and rectum. Dietary GSPs supplementation increased the concentrations of the following SCFAs: acetic acid, propionic acid, and butyric acid in the gut of weaned piglets. In particular, piglets supplemented with 100 mg/kg GSPs presented remarkable increases in the concentrations of acetic acid and butyric acid in the cecum and colon when compared to the control group (P < 0.05). In summary, GSPs might exert salutary effects on the intestine of weaned piglets by enhancing the intestinal antioxidant capacity, modulating gut microbiota composition, and increasing the concentration of microbial metabolites (SCFAs) in the gut.

Gastrointestinal digestibility insights of different levels of coated complex trace minerals supplementation on growth performance of yellow-feathered broilers.

This study was designed to evaluate the optimum additional level of coated complex trace minerals (TMs) and its impacts on the growth performance of broilers through measurement of digestibility of nutrients and intestinal development. In a 56-day trial, a total of 360 one-day-old male yellow-feathered broilers were randomly divided into six dietary treatment groups. Each treatment contained six replicates, with 10 birds. The control group was supplemented with 1,000 mg/kg of uncoated complex TMs in the basal diet (UCCTM1000). The remaining 5 treatments were degressively supplemented with coated complex TMs from 1,000 to 200 mg/kg in the basal diet, which were considered as (CCTM1000), (CCTM800), (CCTM600), (CCTM400), (CCTM200), respectively. Results: On comparing the UCCTM1000 supplementation, the CCTM1000 supplementation decreased the feed to gain ratio (F/G) (P < 0.05), increased digestibility of crude protein (CP) (P < 0.05), crude fat (CF) (P < 0.05), villus height (VH) of duodenum (P < 0.05), and the mRNA expression level of occludin in jejunal mucosa (P < 0.05). In addition, the F/G was lower in the CCTE600 group than that in the CCTE200 group (P < 0.05). The VH to crypt depth (CD) ratio (V/C) of jejunum and ileum in the CCTM400 and CCTM600 groups was higher (P < 0.05) than that in the CCTM1000 group. The serum endotoxin and D-lactate level and CP digestibility were increased by dietary coated complex TMs addition level. The mRNA expression levels of claudin-1 and ZO-1 in the CCTM600 group were higher (P < 0.05) than that in the CCTM1000 group. In conclusion, adding 600 mg/kg of coated complex TMs showed the minimum F/G and the maximum crude protein digestibility and intestine development of yellow-feathered broilers compared with other treatments. This supplementation level of coated complex TMs could totally replace 1,000 mg/kg of uncoated complex TMs to further decrease the dose of TMs and raise economic benefit.

Effect of Dietary Lactose Supplementation on Growth Performance and Intestinal Epithelium Functions in Weaned Pigs Challenged by Rotavirus.

Simple SummaryLactose is a kind of carbohydrate that exists in mammal milk. It has some physiological functions, such as providing energy, regulating gut microbiota, and affecting immunity. Rotavirus (RV) is the main pathogen that induces severe diarrhea in piglets, which impairs their growth and development. In this study, we investigated whether different levels (4% and 6%) of dietary lactose supplementation alleviates RV-induced diarrhea in weaned piglets. The results showed that lactose administration relieved the negative effect of RV on growth, which was derived from the improvement of nutrient utilization, gut barrier function, and immunity. Moreover, supplementing 6% lactose in the diets had a tendency to alleviate diarrhea in RV-infected piglets. Thus, we suggest that the diet of weaned piglets should be supplemented with more than 4% lactose (especially in the early period of weaning) if the cost of feed can be afforded.The purpose of this study was to investigate whether dietary lactose supplementation relieves rotavirus (RV)-induced diarrhea and gut dysfunction. Thirty-six crossbred weaned piglets were randomly allocated into three groups and fed diets containing 0, 4%, and 6% lactose for 20 days. On Day 15, half of the piglets in each group were orally infused with RV. RV infection impaired growth performance; induced severe diarrhea; decreased serum D-xylose concentration and morphology and sIgA level of jejunal mucosa; downregulated MUC1, MUC2, occludin, Bcl-2, IL-4, pBD3, pBD2, and pBD1 mRNA expression of jejunal mucosa and/or mesenteric lymph nodes; upregulated Bax, caspase-3, IL-2, IFN-γ, and IFN-β mRNA expression of jejunal mucosa and/or mesenteric lymph nodes; and damaged microbiota and metabolites of cecal digesta in weaned piglets (p < 0.05). Dietary lactose supplementation improved nutrient digestibility and growth performance and relieved the negative influence of RV challenge on intestinal barrier function, mRNA expression of cytokines, and host defense peptides of jejunal mucosa and/or mesenteric lymph nodes in weaned piglets (p < 0.05). Dietary administration of 6% lactose tended to relieve diarrhea (p = 0.07). These results suggest that lactose in feed increases growth performance and has a tendency to alleviate RV-induced diarrhea, derived from the improvement of nutrient utilization, gut barrier function, and immunity.

Agrobacterium sp. ZX09 β-Glucan Attenuates Enterotoxigenic Escherichia coli-Induced Disruption of Intestinal Epithelium in Weaned Pigs.

To explore the protective effect of dietary β-glucan (BGL) supplementation on intestinal epithelium exposure to enterotoxigenic Escherichia coli (ETEC), thirty-two weaned pigs were assigned to four groups. Pigs were fed with a basal diet or basal diet containing 500 mg/kg BGL, and were orally infused with ETEC or culture medium. Results showed BGL supplementation had no influence on growth performance in weaned pigs. However, BGL supplementation increased the absorption of D-xylose, and significantly decreased the serum concentrations of D-lactate and diamine oxidase (DAO) in the ETEC-challenged pigs (p < 0.05). Interestingly, BGL significantly increased the abundance of the zonula occludens-1-(ZO-1) in the jejunal epithelium upon ETEC challenge (p < 0.05). BGL supplementation also increased the number of S-phase cells and the number of sIgA-positive cells, but significantly decreased the number of total apoptotic cells in the jejunal epithelium upon ETEC challenge (p < 0.05). Moreover, BGL significantly increased the duodenal catalase (CAT) activity and the ileal total superoxide dismutase (T-SOD) activity in the ETEC-challenged pigs (p < 0.05). Importantly, BGL significantly decreased the expression levels of critical inflammation related proteins such as the tumor necrosis factor-α (TNF-α), interlukin-6 (IL-6), myeloid differentiation factor 88 (MyD88), and nuclear factor-κB (NF-κB) in the jejunal and ileal mucosa upon ETEC challenge (p < 0.05). BGL also elevated the propanoic acid content and the abundance of Lactobacillus and Bacillus in the colon upon ETEC challenge (p < 0.05). These results suggested BGL could alleviate the ETEC-induced intestinal epithelium injury, which may be associated with suppressed inflammation and improved intestinal immunity and antioxidant capacity, as well as the improved intestinal macrobiotic.