Jejunal Expression Research Articles

The impact of deoxynivalenol contaminated diet on performance, immune response, intestine morphology and jejunal gene expression in broiler chicken

The aim of this study was to determine the effects of deoxynivalenol (DON) contaminated diet on performance, immune system, gut morphology and jejunal gene expression in broiler chickens. Eighty-one-day old chicks were randomly allotted into two treatments with 4 replicates (10 birds in each replication). Experimental diets were the control diet (maize-soybean meal) and an experimentally contaminated diet with 10 mg/kg DON. The results indicated that DON-challenged birds had decreased (P < 0.05) average feed intake (AFI) during starter period as compared to control group. Also, average daily gain (ADG), AFI and feed conversion ratio (FCR) were not affected (P > 0.05) by inclusion of DON contaminated diet during the whole experimental period. Dietary addition of DON to the basal diet caused Fabricius bursa relative weight reduction, while increased the abdominal fat and serum triglyceride (TG) concentration (P < 0.05). Dietary DON feeding caused an enhancement (P < 0.05) in the blood aspartate aminotransferase (AST) and gamma glutamytransferase (GGT) contents. Moreover, DON decreased the serum total protein (TP) and albumin (ALB) concentrations. Inclusion of DON in diet reduced (P < 0.05) the white blood cell (WBC) count, lymphocyte number and antibody titer against Newcastle disease virus, but increased (P < 0.05) the blood heterophil count. The DON consumption also diminished (P < 0.05) the villus height, villus to crypt ratio, mucosa thickness and villus surface area in the duodenum. Mucin-2 expression was decreased (P < 0.05) by DON consumption, but toll-like receptor-4 (TLR-4) and claudin-5 (CLDN-5) expressions were not affected (P > 0.05) by dietary treatments. In conclusion, although DON could not influence the performance attributes in broiler chickens, it adversely affected the immune response, muc-2 gene expressions in the jejunum and gut morphology, enhanced the liver enzyme indices and lessened the blood protein contents.

Effects of encapsulated cinnamaldehyde on growth performance, intestinal digestive and absorptive functions, meat quality and gut microbiota in broiler chickens.

Essential oils are potential antimicrobial alternatives and their applications in animal feeds are limited due to their fast absorption in the upper gastrointestinal tract. This study investigated the effects of encapsulated cinnamaldehyde (CIN) at 50 mg/kg or 100 mg/kg on the growth performance, organ weights, meat quality, intestinal morphology, jejunal gene expression, nutrient digestibility, and ileal and cecal microbiota. A total of 320 male day-old broiler Cobb-500 chicks were randomly allocated to four treatments with eight pens per treatment (10 birds per pen): 1) basal diet (negative control, NC); 2) basal diet supplemented with 30 mg/kg avilamycin premix (positive control, PC); 3) basal diet with 50 mg/kg encapsulated CIN (EOL); 4) basal diet with 100 mg/kg encapsulated CIN (EOH). Despite birds fed EOH tended to increase (P = 0.05) meat pH at 24 h, all pH values were normal. Similar to PC group, meats from birds fed EOL and EOH showed a reduced (P < 0.05) Warner–Bratzler force shear (WBFS) compared to the NC group. The highest villus to crypt ratios (VH/CD; P < 0.05) were observed in broilers fed either EOL or EOH, with an average of 14.67% and 15.13% in the duodenum and 15.13% and 13.58% in the jejunum, respectively. For jejunal gene expressions, only six out of the 11 studied genes showed statistically significant differences among the dietary treatments. Gene expressions of cationic amino acid transporter 1 (CAT-1) and neutral amino acid transporter 1 (B0AT-1) were upregulated in EOH-fed birds compared to PC and NC-fed birds (P < 0.05), respectively; while the expression of proliferating cell nuclear antigen (PCNA) was downregulated in EOL-fed birds when compared to NC birds (P < 0.05). Nonetheless, the expressions of cadherin 1 (CDH-1), zonula occludens 1 (ZO-1), and maltase-glucoamylase (MG) were all upregulated (P < 0.05) in EOH-fed birds compared to PC-fed birds. The apparent ileal digestibility (AID) of dry matter, crude protein, crude fat and of all 18 tested amino acids increased in EOL-fed birds (P < 0.01). Additionally, relative abundances (%) of ileal Proteobacteria decreased, while ileal and cecal Lactobacillus increased in EOH-fed birds (P < 0.05). In conclusion, dietary encapsulated CIN improved meat quality and gut health by reducing meat WBFS, increasing VH/CD in intestines, jejunal gene expressions, AID of nutrients and beneficial ileal and cecal microbiota composition.

Dietary Lactobacillus fermentum and Bacillus coagulans Supplementation Modulates Intestinal Immunity and Microbiota of Broiler Chickens Challenged by Clostridium perfringens.

Preventative effects of Lactobacillus fermentum and Bacillus coagulans against Clostridium perfringens infection in broilers have been well-demonstrated. The present study was conducted to investigate the modulation of these two probiotics on intestinal immunity and microbiota of C. perfringens-challenged birds. The 336 one-day-old broilers were assigned to four groups with six replicates in each group. Birds in the control were unchallenged and fed a basal diet, and birds in the three challenged groups were dietary supplemented with nothing (Cp group), 1 × 109 CFU/kg of L. fermentum (Lf_Cp group), or 1 × 1010 CFU/kg of B. coagulans (Bc_Cp group). Challenge was performed from days 14 to 20, and samples were collected on days 21 and 28. Challenge upregulated interleukin (IL)-1β and transforming growth factor (TGF)-β4 mRNA expression in jejunum on day 21, which was downregulated by B. coagulans and L. fermentum, respectively (P < 0.05). Both probiotic groups upregulated jejunal IL-1β, interferon (IFN)-γ, IL-17, and TGF-β4 on day 28 as well as IFN-γ on day 21 (P < 0.05). The Bc_Cp group increased CD3+ T cell counts in the jejunal crypt on day 21 (P < 0.05). Challenge decreased the ileal ACE index on day 21 and cecal microbial richness on day 28, which were increased by probiotic treatments, and ileal bacterial richness decreased in the Bc_Cp group on day 28 (P < 0.05). Only ileal microbiota on day 21 was distinctly affected with an R-value at 0.3116 by ANOSIM analysis (P < 0.05). Compared with the control, ileal Firmicutes increased on day 21, and ileal Bacteroidetes and cecal Proteobacteria decreased on day 28 in challenged groups (P < 0.05). Challenge increased Romboutsia spp. in the ileum as well as unclassified f_Lachnospiraceae and Ruminococcus_torques group in the cecum, and decreased Lactobacillus spp. in the ileum on day 21, which were all conversely modulated by L. fermentum (P < 0.05). Challenge increased amino acid metabolism of ileal microbiota and membrane transport of cecal microbiota, and decreased amino acid metabolism of cecal microbiota on day 21, which were conversely regulated by both probiotics (P < 0.05). In conclusion, L. fermentum and B. coagulans attenuated the intestinal inflammation and microbial dysbiosis soon after C. perfringens challenge.

Effects of Ursolic Acid on Intestinal Health and Gut Bacteria Antibiotic Resistance in Mice.

Ursolic acid (UA), a natural pentacyclic triterpenoid, has been widely reported to exert anti-oxidant and anti-inflammatory properties. However, the effects of UA on the intestinal homeostasis and gut microbiota were rarely explored. The aim of the present study was to investigate the effects of UA on intestinal health and gut microflora antibiotic-resistance in antibiotic-exposed mice. Kunming mice (n = 80) were randomly allocated into three groups and fed with one of the following diets, respectively: Cont group (n = 20), the basal diet; UA group (n = 20), the basal diet supplemented with 150 mg/kg UA; Tet group (n = 40), the basal diet supplemented with 659 mg/kg chlortetracycline. After 14 days, 10 mice in each group were euthanatized and the remaining 30 mice in the Tet group were randomly allocated into three sub-groups (n = 10 per group) as follows: the Tet group which were kept feeding a Tet diet for 14 days; the Natural Restoration (NatR) group which received a basal diet for 14 days; and the UA therapy (UaT) group which fed a basal diet supplemented with 150 mg/kg UA for 14 days. Throughout the experiment, the weight and the food intake of each mouse were recorded once weekly. Serum LPS and diamine oxidase (DAO), jejunal morphology, jejunal tight junction proteins and nutrient transporters, colonic inflammatory cytokines, gut microbiota and its antibiotic resistance gene (ARG) were examined at euthanasia. The results showed that UA treatment significantly increased average daily food intake (ADFI) of mice. Notably, UA increased the jejunal villi height, decreased the jejunal crypt depth and promoted the expression of jejunum nutrient transporters. UaT group had higher villi height, lower crypt depth and higher nutrient transporter mRNA expression in jejunum than NatR group. Besides, UA decreased serum DAO content, upregulated mRNA expression of ZO-1, claudin-1 and occludin and downregulated TNF-α and IL-6. The mRNA abundances of ZO-1, claudin-1 and occludin and TNF-α and IL-6 in UaT group were, respectively upregulated and downregulated than NatR group. Furthermore, an analysis of 16S rDNA sequences demonstrated that UA increased the abundance of beneficial bacteria in the gut. And the results of ARG test showed that UA downregulated the expression of antibiotic-induced resistance genes. The UaT group inhibited the increase of harmful bacteria abundance and suppressed the mRNA abundances of ARG compared to the NatR group. In conclusion, considering the positive effects of UA on the growth performance and intestinal mucosal barrier, we anticipate that these findings could be a stepping stone for developing UA as a novel substitute of antibiotics.

Effects of Dietary Maltol on Innate Immunity, Gut Health, and Growth Performance of Broiler Chickens Challenged With Eimeria maxima.

Two studies were conducted to evaluate the effects of maltol as a postbiotic on innate immunity, gut health, and enteric infection. In the first study, an in vitro culture system was used to evaluate the effects of maltol on the innate immune response of chicken macrophage cells (CMC), gut integrity of chicken intestinal epithelial cells (IEC), anti-parasitic activity against Eimeria maxima, and differentiation of quail muscle cells (QMC) and primary chicken embryonic muscle cells (PMC). All cells seeded in the 24-well plates were treated with maltol at concentrations of 0.1, 1.0, and 10.0 μg. CMC and IEC were stimulated by lipopolysaccharide to induce an innate immune response, and QMC and PMC were treated with 0.5 and 2% fetal bovine serum, respectively. After 18 h of incubation, pro-inflammatory cytokines, tight junction proteins (TJPs), and muscle cell growth markers were measured. In the second study, the dietary effect of maltol was evaluated on disease parameters in broiler chickens infected with E. maxima. Eighty male 1-day-old broiler chickens were allocated into the following four treatment groups: (1) Control group without infection, (2) Basal diet with E. maxima, (3) High maltol (HI; 10.0 mg /kg feed) with E. maxima, and (4) Low maltol (LO; 1.0 mg/kg feed) with E. maxima. Body weights (BW) were measured on days 0, 7, 14, 20, and 22. All chickens except the CON group were orally infected with 104 E. maxima per chicken on day 14. Jejunum samples were collected for gut lesion scoring, and the gene expression of cytokines and TJPs. Data was analyzed using PROC MIXED in SAS. In vitro, maltol not only increased TJPs in IEC and cytokines in the LPS-stimulated CMC but also showed direct cytotoxicity against sporozoites of E. maxima. In vivo, the HI group improved the BW, reduced the gut lesion scores and fecal oocyst shedding, and decreased jejunal TNFSF15 and IL-1β expression in E. maxima-infected chickens. In conclusion, these results demonstrate the beneficial effects of dietary maltol in the enhancement of growth performance, gut health, and coccidiosis resistance and the applicability of maltol as a postbiotic for the replacement of antibiotic growth promoters in commercial poultry production.

The Potential Protective Role of RUNX1 in Nonalcoholic Fatty Liver Disease.

The pathogenic mechanisms underlying nonalcoholic fatty liver disease (NAFLD) are beginning to be understood. RUNX1 is involved in angiogenesis, which is crucial in inflammation, but its role in nonalcoholic steatohepatitis (NASH) remains unclear. The aim of this study was to analyze RUNX1 mRNA hepatic and jejunal abundance in women with morbid obesity (MO) and NAFLD. RUNX1, lipid metabolism-related genes, and TLRs in women with MO and normal liver (NL, n = 28), NAFLD (n = 41) (simple steatosis (SS, n = 24), or NASH (n = 17)) were analyzed by RT-qPCR. The RUNX1 hepatic expression was higher in SS than in NL or NASH, as likewise confirmed by immunohistochemistry. An increased expression of hepatic FAS was found in NAFLD. Hepatic RUNX1 correlated positively with FAS. There were no significant differences in the jejunum RUNX1 expressions in the different groups. Jejunal FXR expression was lower in NASH than in NL, while the TLR9 expression increased as NAFLD progressed. Jejunal RUNX1 correlated positively with jejunal PPARγ, TLR4, and TLR5. In summary, the hepatic expression of RUNX1 seems to be involved in the first steps of the NAFLD process; however, in NASH, it seems to be downregulated. Our findings provide important insights into the role of RUNX1 in the context of NAFLD/NASH, suggesting a protective role.

Early socialization and environmental enrichment of lactating piglets affects the caecal microbiota and metabolomic response after weaning

The aim of this study was to determine the possible impact of early socialization and an enriched neonatal environment to improve adaptation of piglets to weaning. We hypothesized that changes in the microbiota colonization process and in their metabolic response and intestinal functionality could help the animals face weaning stress. A total of 48 sows and their litters were allotted into a control (CTR) or an enriched treatment (ENR), in which piglets from two adjacent pens were combined and enriched with toys. The pattern of caecal microbial colonization, the jejunal gene expression, the serum metabolome and the intestinal physiology of the piglets were assessed before (-2 d) and after weaning (+ 3d). A differential ordination of caecal microbiota was observed after weaning. Serum metabolome suggested a reduced energetic metabolism in ENR animals, as evidenced by shifts in triglycerides and fatty acids, VLDL/LDL and creatine regions. The TLR2 gene showed to be downregulated in the jejunum of ENR pigs after weaning. The integration of gene expression, metabolome and microbiota datasets confirmed that differences between barren and enriched neonatal environments were evident only after weaning. Our results suggest that improvements in adaptation to weaning could be mediated by a better response to the post-weaning stress.

Comparative efficacy of commercially available deoxynivalenol detoxifying feed additives on growth performance, total tract digestibility of components, and physiological responses in nursery pigs fed diets formulated with naturally contaminated corn.

Comparative efficacy of deoxynivalenol (DON) detoxifying feed additives (FA) was evaluated in growth performance (exp. 1) and apparent total tract digestibility (ATTD; exp. 2) nursery pig studies. Six corn–soybean meal-based diets were used: 1) positive control (PC, formulated with <1.5 ppm DON corn), negative control (NC, formulated with 5.5 ppm DON corn), NC + FA1 (clay plus yeast cell wall extract), NC + FA2 (aluminosilicate), NC + FA3 (aluminosilicate plus fungal extract), and NC + FA4 (sodium metabisulfite, SMB). In exp. 1, 144 pigs (body weight [BW], 10.2 ± 0.1kg) were housed (4 pigs/pen), allocated to diets (n = 6) based on BW, and fed for 4-wk. The BW and feed intake were monitored weekly. On d 7, one pig/pen was bled for plasma and euthanized for organ weight and tissue samples. Assayed DON concentration in PC, NC, NC + FA4 was 0.29, 2.86, and 1.21 ppm, respectively. In wk-1, the average daily gain (ADG) of pigs fed NC + FA4 was not different (P > 0.05) to that of pigs fed PC diet but greater (P = 0.01) than for pigs fed NC without or with other FA. Pigs fed NC and NC + FA2 had lower (P = 0.026) average daily feed intake (ADFI) than pigs fed PC and NC + FA3. Pigs fed NC + FA4 had greater (P = 0.003) G:F than pigs fed the other diets. Diets had no effect (P > 0.05) on ADG, ADFI, and G: F after first week, plasma concentration of urea and creatinine or liver and spleen weight. Pigs fed NC diets had greater (P = 0.01) jejunal mRNA expression of superoxide dismutase 1 relative to pigs fed PC or NC plus FA. Jejunal histomorphology and mRNA expression of nutrient transporters, inflammatory cytokines, and tight junction proteins and ceca digesta concentration of short-chain fatty acids were not affected (P > 0.05) by the diet. In exp. 2, 24 barrows (BW 10.2 ± 0.3 kg) were individually placed in metabolism crates and allocated to four diets: PC, NC, NC + FA3, and NC + FA4 (n = 6) containing TiO2 as digestibility marker. Pigs were adjusted to diets for 5 d, followed by a 2-d grab fecal sample collection. Pigs fed PC and NC + FA4 diets had higher ATTD of dry matter, gross energy, and crude protein than NC fed pigs. The FA3 was intermediate in digestibility response. In conclusion, FA containing sequestering component plus fungal extract or SMB in DON-contaminated feed resulted in commensurate nursery pig performance to PC. The tested FA mitigated intestinal oxidative stress through decreased expression of genes for superoxide dismutase.

Interactions between starch source and gelatinisation degree on performance and small intestinal digestion in broiler chickens

ABSTRACT 1. A 2 × 2 factorial arrangement was used to test the hypothesis that, in pelleted diets, legume starch is digested less rapidly and to a lesser extent than cereal starch, and that increased gelatinisation through extrusion would eliminate the differences between the starch sources. In addition, the trial examined whether a lower ratio of starch to nitrogen disappearance rate (SNDR) could improve feed conversion ratio (FCR). 2. At 17 d of age, male broilers were randomly distributed among four dietary treatments, consisting of either wheat or faba bean starch-rich fraction (FBS) as the sole starch source and pelleting or extrusion as processing methods. Each treatment had 10 replicate pens containing five birds each. 3. Extrusion resulted in a more extensive starch gelatinisation compared to pelleting, as expected. 4. No difference in weight gain at 29 d of age was observed between birds fed starch sources. However, birds fed wheat tended (P = 0.080) to have better FCR than those fed FBS, while the effect of processing methods was insignificant. Thus, there was no interaction between starch source and processing method on FCR. 5. In pelleted diets, FBS had lower and slower starch digestibility compared to wheat in all intestinal segments (P < 0.05). The interaction between starch source and processing method in all intestinal segments (P < 0.001) demonstrated that FBS responded more to gelatinisation through extrusion than did wheat. Thus, differences in starch digestibility between the wheat and FBS were eliminated with extrusion. 6. Feeding extruded diets significantly increased the upper jejunal expression of GLUT1, GLUT2 and SGLT1 compared to pelleted diets, which suggested that glucose absorption was less likely to be a limiting factor for starch utilisation. 7. Pelleting resulted in a lower ratio (P < 0.001) of SNDR compared to extrusion (on average 1.4-fold) but did not improve FCR.

Standardized ileal digestible tryptophan to lysine ratios affect performance and regulate intestinal mRNA expression of amino acid transporters in weaning pigs fed a low crude protein diet

The objective of this study was to estimate the effect of standardized ileal digestible (SID) tryptophan (Trp) to lysine (Lys) ratios on growth performance and jejunal expression of amino acid transporters in weaning pigs. A total of 240 hybrid [(Duroc, male) x (Large White x Landrace, female)] piglets (6.52 ± 0.42 kg) were used in a 28-day growth trial and randomly allotted to one of four diets providing SID Trp to Lys ratios of 0.15, 0.18, 0.21, or 0.24 and were allocated in 6 pens per treatment with 10 pigs per pen. The average daily gain (ADG), average daily feed intake (ADFI), and serum levels of Trp, serine, isoleucine and leucine in pigs significantly increased when the SID Trp to Lys ratios increased from 0.15 to 0.18, 0.21, or 0.24 (P < 0.05). The pigs fed the diet with the 0.24 ratio had a reduced serum urea nitrogen (SUN) concentration compared with the pigs fed the 0.15 ratio diet (P < 0.05). The villus heights of the duodenum and jejunum in pigs fed the diet with a 0.21 ratio were greater than those of pigs fed the 0.15 ratio diet (P < 0.05). The increase in the SID Trp to Lys ratio from 0.15 to 0.18 decreased the intestinal ASCT2 mRNA level in weaning pigs (P < 0.05). The increase in the SID Trp to Lys ratio from 0.15 to 0.21 or 0.24 increased the mRNA abundance of intestinal B°AT1 (P < 0.05). The increase in the SID Trp to Lys ratio from 0.15 to 0.18 increased the mRNA abundances of intestinal CAT-1 and rBAT (P < 0.05). The intestinal b0,+AT mRNA level in the pigs fed the 0.21 ratio diet was higher than that in pigs in other three treatments (P < 0.05). The mRNA abundances of y+LAT1 and 4F2hc in the pigs fed the 0.15 ratio diet were lower than those in pigs in other three treatments (P < 0.05). The intestinal PepT-1 mRNA level in the pigs fed the 0.15 or 0.24 ratio diet was lower than that of the pigs fed the 0.21 ratio diet (P < 0.05). In conclusion, the optimal SID Trp to Lys ratio was determined to be 0.18 for weaning pigs that were fed a low crude protein diet using ADG and jejunal amino acid transporter mRNA expression as the response criteria.

The effects of multispecies probiotic formulations on growth performance, hepatic metabolism, intestinal integrity and fecal microbiota in growing-finishing pigs

Probiotics have received attention as alternatives to antibiotics due to their relative biosafety. Despite the health-promoting effects of multispecies probiotic formulation (MPF) in weaning pigs were well-documented, the health-promoting potential of MPF in growing-finishing pigs remain limited. Therefore, we conducted a six-week experiment to investigate the beneficial effects of MPF supplementation (the formulations containing Lactobacillus plantarum CJLP243, L. fermentum LF21, L. salivarius E4101, Leuconostoc paramesenteroides KJP421, Bacillus subtilis CJMPB957 and B. licheniformis CJMPB283) on intestinal environments and hepatic lipid metabolism in growing-finishing pigs. The results showed that the growth performance of MPF group was improved compared to control group during the experimental period without affecting feed intake. The dietary MPF supplementation also significantly increased the jejunal mRNA expression levels of zonula occludens-1 (ZO-1) and decreased that of interleukin-12 (IL-12). Dietary MPF supplementation was found to not only reduce the hepatic triglyceride concentration but down-regulate hepatic mRNA expression levels of lipid metabolism-related markers, such as as sterol regulatory element-binding transcription factor 1c (SREBP-1c), CCAAT/enhancer binding protein α (CEBPα), acetyl coA carboxylase (ACC) and carnitine palmitoyl transferase 1β (CPT1β). Nonetheless, MPF supplementation also had the capability to modulate gut microbiota populations, including Clostridiaceae, Lachnospiraceae, and Turicibacter. Collectively, our results suggest that dietary MPF inclusion could be a promising approach to promote the growth performance and the overall health of growing-finishing pigs via modulation of gut microbiota.

Modulation of Intestinal Phosphate Transport in Young Goats Fed a Low Phosphorus Diet.

The intestinal absorption of phosphate (Pi) takes place transcellularly through the active NaPi-cotransporters type IIb (NaPiIIb) and III (PiT1 and PiT2) and paracellularly by diffusion through tight junction (TJ) proteins. The localisation along the intestines and the regulation of Pi absorption differ between species and are not fully understood. It is known that 1,25-dihydroxy-vitamin D3 (1,25-(OH)2D3) and phosphorus (P) depletion modulate intestinal Pi absorption in vertebrates in different ways. In addition to the apical uptake into the enterocytes, there are uncertainties regarding the basolateral excretion of Pi. Functional ex vivo experiments in Ussing chambers and molecular studies of small intestinal epithelia were carried out on P-deficient goats in order to elucidate the transepithelial Pi route in the intestine as well as the underlying mechanisms of its regulation and the proteins, which may be involved. The dietary P reduction had no effect on the duodenal and ileal Pi transport rate in growing goats. The ileal PiT1 and PiT2 mRNA expressions increased significantly, while the ileal PiT1 protein expression, the mid jejunal claudin-2 mRNA expression and the serum 1,25-(OH)2D3 levels were significantly reduced. These results advance the state of knowledge concerning the complex mechanisms of the Pi homeostasis in vertebrates.

Probiotic Bacillus subtilis 29,784 improved weight gain and enhanced gut health status of broilers under necrotic enteritis condition

The study investigated the benefit of a Bacillus subtilis probiotic (Bs 29,784) in necrotic enteritis (NE)–challenged broilers. Four treatments were performed with 312 male day-old Ross 308 reared in floor pens from day 0 to day 35: 2 groups fed control diet without or with NE challenge (CtrlNC and CtrlNE); 2 groups fed probiotic and antibiotic supplements in the control diet with NE challenge (ProNE and AntNE). Necrotic enteritis challenge procedures commenced with inoculation of Eimeria spp 1 mL/bird per os at day 9 and Clostridium perfringens EHE-NE18 (approximately 108 cfu/mL) 1 mL/bird per os at day 14 and day 15. Performance parameters were measured on day 16 and day 35. Lesion, cecal microbiota, and jejunal gene expression were analyzed on day 16. Necrotic enteritis challenge significantly suppressed the performance parameters compared with CtrlNC: 27% weight gain reduction, 11 points feed conversion ratio (FCR) increase at day 16, and 12% weight gain reduction, 5-point FCR increase at day 35. By day 35, ProNE and AntNE treatments enabled significantly higher weight gain (4 and 9%, respectively) than CtrlNE. Compared with CtlrNE and contrary to AntNE, ProNE treatment exhibited upregulation of genes coding for tight junctions proteins (CLDN1, JAM2, TJP1), cytokines (IL12, interferon gamma, TGFβ), and Toll-like receptors (TLR5, TLR21) suggesting enhanced immunity and intestinal integrity. 16S NGS analysis of cecal microbiota at day 16 showed a decreased alpha diversity in challenged groups. Principal component analysis of operational taxonomic unit (OTU) abundance revealed that ProNE and AntNE grouped closely while both distantly from CtrlNC and CtrlNE, which were separately grouped, indicating the similar effects of ProNE and AntNE on the OTU diversity that were however different from both CtrlNC and CtrlNE. Microbiota analysis revealed an increase of genera Faecalibacterium, Oscillospira, and Butyricicoccus; and a decrease of genera Ruminococcus, Lactobacillus, and Bacteroides; and an increase of the Firmicutes-to-Bacteroidetes ratio in ProNE and AntNE groups compared with the CtlrNE group. It is concluded that Bs 29,784 may enable improved health of broiler chickens under NE conditions thus performance implications.

Yeast-based nucleotide supplementation in mother sows modifies the intestinal barrier function and immune response of neonatal pigs.

In the present study, we aimed to evaluate the effects of maternal yeast-based nucleotide (YN) supplementation on the intestinal immune response and barrier function in neonatal pigs, as well as the diarrhoea rate and growth performance in suckling piglets. Sixty-four late-gestation sows were assigned to the following groups: the CON (fed a basal diet) and YN groups (fed a basal diet with 4 g YN/kg diet). The experiment started on d 85 of gestation and ended on d 20 of lactation. Diarrhoea rate and average daily gain of the piglets were recorded, and samples of blood and intestines from neonatal piglets were collected before they consumed colostrum during farrowing. Compared with the CON group, maternal YN supplementation increased the weaning weight of litter and decreased the diarrhoea rate (P < 0.01). In addition, maternal YN supplementation promoted the ileal villus development in the neonates compared with that in the CON group (P < 0.01). Maternal YN supplementation also increased the ileal secretory immunoglobulin A (sIgA) level compared with that in the CON group (P < 0.05). The real-time PCR results showed that maternal dietary YN supplementation increased the jejunal and ileal expression of interleukin (IL)-17, IL-8, IL-1β, IL-10 and tumor necrosis factor (TNF)- α in the neonates compared with that in the CON group (P < 0.05). Overall, maternal nucleotide supplementation improved the villus development and innate immunity of neonatal piglets during late pregnancy. This may be associated with the decrease in diarrhoea and the increase in weaning weight of the litter of suckling piglets.

Combination of Lactobacillus species and yeast ameliorates adverse effect of deoxynivalenol contaminated diet on immune system, gut morphology and jejunal gene expression in broiler chickens

The aim of this study was to examine the effect of different levels of Lactobacillus species and yeast (Saccharomyces cerevisiae) as a toxin binder in deoxynivalenol (DON) diet on immune parameters, gut morphology and jejunal gene expression in broiler chickens. Three hundred sixty-one-day-old female broilers were assigned into nine treatments with four replicates each in a completely randomised design (3 × 3 factorial arrangement) including three levels of Lactobacillus (0, 0.20 and 0.40 g/kg) and yeast (0, 0.75 and 1.50 g/kg) in DON contaminated diets. The results showed that DON challenged birds showed suppressed spleen relative weight and reduced white blood cell and lymphocyte percentage, while inclusion the highest level of Lactobacillus sp. and yeast to DON diet improved these parameters (p < .05). Newcastle antibody titre was increased (p < .05) by increasing the level of Lactobacillus sp. and yeast in DON included diet. Increasing the level of Lactobacillus sp. and yeast in DON diet was linearly enhanced (p < .05) the villus height, muscular layer and absorptive surface area in ileum. Combination of 0.40 g/kg Lactobacillus sp. and 1.50 g/kg yeast caused an enhancement (p < .05) in mucosa and muscular layer thicknesses of jejunum and ileum, respectively. The expressions of toll like receptor-4 and claudin-5 were down-regulated and up-regulated respectively by administration of the highest level of Lactobacillus sp. and yeast. In conclusion, the results indicated that dietary inclusion of the highest level of Lactobacillus sp. and yeast improved the spleen weight, some immunological parameters, villus height, muscular layer thickness and gene expressions in DON- challenged broilers. HIGHLIGHTS Deoxynivalenol (DON) consumption (10 mg/kg) induce intestinal and immunological changes in broilers. Combination of Lactobacillus sp. and yeast reduce DON toxicity. Inclusion 0.4 g/kg lactobacillus and 1.5 g/kg yeast improve villus height and humoral immunity.

199 Supplemental arginine affects intestinal gene expression and serum concentration of amino acids of heat stress pigs

Abstract The exposure of pigs to heat stress (HS) reduces feed intake and can damage their intestinal epithelia, affecting the amino acid (AA) absorption. Because Arg is involved in the restoration of intestinal epithelia cells and HS reduces Arg intake, the effect of dietary supplementation with Arg above requirement on the expression of specific AA transporters and the serum concentration (SC) of free AA in HS pigs was analyzed. Twenty pigs (25.3 ± 4.4 kg BW) were randomly assigned to one of two dietary treatments: control wheat-soybean meal diet supplemented with L-Lys and L-Thr (CON), and the CON diet added with 0.20% L-Arg (ARG). Pigs had at libitum access to feed and water. At the end of a 21-d trial, 6 pigs per treatment were sacrificed; blood and intestinal mucosa samples from each segment of the small intestine were collected to analyze the SC of AA and the expression of AA transporters, respectively. Ambient temperature (AT) ranged from 28.2 to 38.7 °C. Supplementation of L-Arg increased approx. 5x the abundance of mRNA coding for the synthesis of AA transporter b0,+ (P&amp;lt; 0.05) and tended to increase that of the AA transporter B0 (P&amp;lt; 0.10) in duodenum, but no effect on gene expression in jejunum and ileum was observed. The SC of AA (mg/dL) for CON and ARG pigs were: Arg, 27.40, 38.32; His, 9.64, 12.9; Ile, 14.33, 15.73; Leu, 23.32, 29.97; Lys, 31.67, 39.72; Met, 5.14, 8.22; Phe, 15.18, 18.07; Thr, 20.82, 31.55; Trp, 8.50, 11.32; Val, 22.47, 33.35, respectively. Supplementation of L-Arg increased serum Arg, His, Met, Thr, Trp, and urea (P&amp;lt; 0.05); tended to increase Val (P&amp;lt; 0.10), but did not affect Ile, Leu, Lys, and Phe (P &amp;gt;0.10). These results indicate that supplementation of L-Arg may help to recover the intestinal epithelia, improve the absorptive function of the small intestine, and increase the availability of some indispensable AA in HS pigs.

Effects of cLFchimera peptide on intestinal morphology, integrity, microbiota, and immune cells in broiler chickens challenged with necrotic enteritis

Three hundred and sixty 1-day-old male broiler chicks were randomly allocated to 4 treatments of 6 replicates to evaluate the effects of cLFchimera, a recombinant antimicrobial peptide (AMP), on gut health attributes of broiler chickens under necrotic enteritis (NE) challenge. Treatments were as follows: (T1) unchallenged group fed with corn-soybean meal (CSM) without NE challenge and additives (NC); (T2) group fed with CSM and challenged with NE without any additives (PC); (T3) PC group supplemented with 20 mg cLFchimera/kg diet (AMP); (T4) PC group supplemented with 45 mg antibiotic (bacitracin methylene disalicylate)/kg diet (antibiotic). Birds were sampled for villi morphology, ileal microbiota, and jejunal gene expression of cytokines, tight junctions proteins, and mucin. Results showed that AMP ameliorated NE-related intestinal lesions, reduced mortality, and rehabilitated jejunal villi morphology in NE challenged birds. While the antibiotic non-selectively reduced the count of bacteria, AMP restored microflora balance in the ileum of challenged birds. cLFchimera regulated the expression of cytokines, junctional proteins, and mucin transcripts in the jejunum of NE challenged birds. In conclusion, cLFchimera can be a reliable candidate to substitute growth promoter antibiotics, while more research is required to unveil the exact mode of action of this synthetic peptide.

Deregulated Serotonin Pathway in Women with Morbid Obesity and NAFLD.

Non-alcoholic fatty liver disease (NAFLD) extends from simple steatosis (SS) to non-alcoholic steatohepatitis (NASH). Peripheral serotonin (5-HT) has become as an important regulator of different metabolic pathways. 5-HT has been related to obesity and lipid accumulation in the liver. The objective of this study was to assess the relationship between the 5-HT signaling pathway and the degree of NAFLD, as well as to investigate whether peripheral 5-HT levels are related to the hepatic and jejunal mRNA abundance of serotonin receptors (HTR) in a cohort of women with morbid obesity (MO) and NAFLD. ELISA was used to quantify the serum 5-HT from normal-weight subjects (n = 26) and patients with MO (n = 58). We used RTq-PCR analysis to evaluate the relative expression of HTR in women with MO with normal liver (n = 22), SS (n = 21), and NASH (n = 15). The 5-HT was diminished in women with MO under a hypocaloric diet, regardless of the presence of NAFLD. Additionally, we report a negative correlation of 5-HT levels with metabolic syndrome criteria, suggesting that serotonin may have a protective role in obesity. Additionally, the hepatic expression of HTR2A and HTR2B were decreased in women with MO and NAFLD, but no significant differences in the HTR jejunal expression according to the presence of NAFLD were found.

Reduced phosphorus intake throughout gestation and lactation of sows is mitigated by transcriptional adaptations in kidney and intestine

BackgroundThe environmental impact of pig farming need to be reduced, with phosphorus (P) being of particular interest. Specified dietary regimens and management systems contribute to meet environmental concerns and reduce economic constrains. However, pregnant and lactating sows represent vulnerable individuals, whose reproductive potential and metabolic health status relies on adequate supply of macro- and micronutrients. The aim of this study was to investigate, whether sows fed with a dietary P content that is below or above current recommendations are capable to maintain mineral homeostasis during the reproduction cycle and which endogenous mechanisms are retrieved therefore in kidney and jejunum. Nulliparous gilts were fed iso-energetic diets with recommended (M), reduced (L), or high (H) amounts of mineral P supplements throughout gestation and lactation periods. Blood metabolites and hormones referring to the P homeostasis were retrieved prior to term (110 days of gestation) and at weaning (28 days of lactation). Transcriptional responses in kidney cortex and jejunal mucosa were analyzed using RNA sequencing.ResultsThe variable dietary P content neither led to an aberration on fertility traits such as total weaned piglets nor to an effect on the weight pattern throughout gestation and lactation. Serum parameters revealed a maintained P homeostasis as reflected by unaltered inorganic P and calcium levels in L and H fed groups. The serum calcitriol levels were increased in lactating L sows. The endocrine responses to the dietary challenge were reflected at the transcriptional level. L diets led to an increase in CYP27B1 expression in the kidney compared to the H group and to an altered gene expression associated with lipid metabolism in the kidney and immune response in the jejunum.ConclusionsOur results suggest that current P requirements for gestating and lactating sows are sufficient and over supplementation of mineral P is not required. Shifts in renal and jejunal expression patterns between L and H groups indicate an affected intermediate metabolism, which long-term relevance needs to be further clarified.

Ezetimibe Enhances Macrophage-to-Feces Reverse Cholesterol Transport in Golden Syrian Hamsters Fed a High-Cholesterol Diet

The aim of this work was to evaluate reverse cholesterol transport (RCT) in hamster, animal model expressing CETP under a high cholesterol diet (HF) supplemented with Ezetimibe using primary labelled macrophages. We studied three groups of hamsters (n=8/group) for 4 weeks: 1) chow diet group: Chow, 2) High cholesterol diet group: HF and 3) HF group supplemented with 0.01% of ezetimibe: HF+0.01%Ezet. Following intraperitoneal injection of 3H-cholesterol-labelled hamster primary macrophages, we measured the in vivo macrophage-to-feces RCT. .HF group exhibited an increase of triglycerides (TG), cholesterol, glucose in plasma and higher TG and cholesterol content in liver (p<0.01) compared to Chow group. Ezetimibe induced a significant decrease in plasma cholesterol with a lower LDL and VLDL cholesterol (p<0.001) and in liver cholesterol (p<0.001) and TG (p<0.01) content compared to HF. In vivo RCT essay showed an increase of tracer level in plasma and liver (p<0.05) but not in feces in HF compared to Chow group. The amount of labelled total sterol and cholesterol in liver and feces was significantly reduced (p<0.05) and increased (p=0.05) respectively with Ezetimibe treatment. No significant increase was obtained for labelled feces bile acids in HF+0.01%Ezet compared to HF. Ezetimibe decreased SCD1 gene expression and increased SR-B1 (p<0.05) in liver but did not affect NPC1L1 nor ABCG5 and ABCG8 expression in jejunum. In conclusion, ezetimibe exhibited an atheroprotective effect by enhancing RCT in hamster and decreasing LDL cholesterol. Ours findings showed also a hepatoprotective effect of ezetimibe by decreasing hepatic fat content. Significance Statement This work was assessed to determine the effect of ezetimibe treatment on high cholesterol diet induced disturbances and especially the effect on reverse cholesterol transport in animal model with CETP activity and using labelled primary hamster macrophages. We were able to demonstrate that ezetimibe exhibited an atheroprotective effect by enhancing RCT and by decreasing LDL cholesterol in hamster. We showed also a hepatoprotective effect of ezetimibe by decreasing hepatic fat content.