Lactobacillus johnsonii 3-1 and Lactobacillus crispatus 7-4 promote the growth performance and ileum development and participate in lipid metabolism of broilers.

Long-term use of antibiotic growth promoter (AGP) in animal production is the main cause of antimicrobial resistance of pathogenic bacteria. Therefore, seeking alternatives to AGP is crucial for animal husbandry. Among all AGP alternatives, probiotics are promising candidates. In this study, two strains of lactic acid bacteria, L. johnsonii 3-1 and L. crispatus 7-4, were isolated from the feces of wild Gallus gallus, which exhibited obvious anti-pathogenic activity and improved the growth performance of broilers. Furthermore, we found that these two strains participated in the lipid metabolism of broilers by reducing the content of TC and TG in ileal epithelial cells and up-regulating the liver AMPKα/PPARα/CPT-1 pathway, which affects abdominal fat deposition. In summary, L. johnsonii 3-1 and L. crispatus 7-4 have the potential to be used as AGP substitutes and participate in the lipid metabolism of broilers to reduce abdominal fat deposition. Importantly, our study reveals for the first time that L. crispatus participates in liver lipid metabolism to reduce abdominal fat deposition in broilers.

Epithelial Regeneration Ability of Crohn's Disease Assessed Using Patient-Derived Intestinal Organoids.

Little is known about the ability for epithelial regeneration and wound healing in patients with inflammatory bowel diseases. We evaluated the epithelial proliferation and wound healing ability of patients with Crohn's disease (CD) using patient-derived intestinal organoids. Human intestinal organoids were constructed in a three-dimensional intestinal crypt culture of enteroscopic biopsy samples from controls and CD patients. The organoid-forming efficiency of ileal crypts derived from CD patients was reduced compared with those from control subjects (p < 0.001). Long-term cultured organoids (≥6 passages) derived from controls and CD patients showed an indistinguishable microscopic appearance and culturing behavior. Under TNFα-enriched conditions (30 ng/mL), the organoid reconstitution rate and cell viability of CD patient-derived organoids were significantly lower than those of the control organoids (p < 0.05 for each). The number of EdU+ cells was significantly lower in TNFα-treated organoids derived from CD patients than in TNFα-treated control organoids (p < 0.05). In a wound healing assay, the unhealed area in TNFα-treated CD patient-derived organoids was significantly larger than that of TNFα-treated control organoids (p < 0.001). The wound healing ability of CD patient-derived organoids is reduced in TNFα-enriched conditions, due to reduced cell proliferation. Epithelial regeneration ability may be impaired in patients with CD.

Feeding colostrum or a 1:1 colostrum:milk mixture for 3 days postnatal increases small intestinal development and minimally influences plasma glucagon-like peptide-2 and serum insulin-like growth factor-1 concentrations in Holstein bull calves.

This study evaluated how feeding colostrum- or a colostrum-milk mixture for 3 d postnatal affects plasma glucagon-like peptide-2 (GLP-2), serum insulin-like growth factor-1 (IGF-1), and small intestinal histomorphology in calves. Holstein bulls (n = 24) were fed colostrum at 2 h postnatal and randomly assigned to receive either colostrum (COL), whole milk (WM), or a 1:1 COL:WM mixture (MIX) every 12 h from 12 to 72 h. A jugular venous catheter was placed at 1 h postnatal to sample blood frequently for the duration of the experiment. Samples were collected at 1, 2, 3, 6, 9, 11, and 12 h. Following the 12-h meal, blood was collected at half-hour intervals until 16 h and then at 1-h intervals from 16 to 24 h. A 27-h sample was taken, then blood was sampled every 6 h from 30 to 60 h. Again, blood was taken at half-intervals from 60 to 64 h, then at 65 and 66 h, following which, a 2-h sampling interval was used until 72 h. Plasma GLP-2 (all time points) and serum IGF-1 (at time points: 1, 6, 12, 18, 24, 36, 48, and 72 h) were both analyzed. Duodenal, jejunal, and ileal tissues were collected at 75 h of age to assess histomorphology and cellular proliferation. Feeding COL, rather than WM, increased plasma GLP-2 by 60% for 2 h and tended to increase GLP-2 by 49.4% for 4 h after the 60-h meal. Insulin-like growth factor-1 area under the curve (from 12 to 72 h) tended to be 27% greater for COL than WM calves but was otherwise unaffected by treatment. Ileal crypts tended to proliferate more with MIX than WM, whereas ileal crypt proliferation did not differ for COL compared with MIX or WM and was not different between treatments in the proximal jejunum. Villi height was increased 1.8 and 1.5× (COL and MIX vs. WM) in the proximal and distal jejunum, respectively, whereas MIX duodenal and ileal villi height tended to be 1.5 and 1.4× that of WM. Crypt depth did not differ in any region. Surface area of the gastrointestinal tract was reduced for WM by 60 and 58% (proximal jejunum) and 38 and 52% (ileum) relative to COL and MIX and was 54% less than MIX in the distal jejunum. Overall, extended COL feeding minimally increased plasma GLP-2 and serum IGF-1 compared with WM feeding. As COL and MIX similarly promoted small intestinal maturation, feeding calves transition milk to promote intestinal development could be a strategy for producers.

Influence of inclusion level of barley in wheat-based diets and supplementation of carbohydrase on growth performance, nutrient utilisation and gut morphometry in broiler starters.

1. The influence of barley inclusion level and supplementation of a multi-component non-starch polysaccharide degrading enzyme on performance and nutrient utilisation in broilers was investigated. Normal-starch hulled barley was evaluated with five levels of inclusion (0, 141, 283, 424 and 565 g/kg) in a wheat-based diet and two levels of enzyme supplementation (0 and 150 g/tonne of feed; a 5 × 2 factorial arrangement of 10 dietary treatments). All diets were equivalent in metabolisable energy and digestible amino acid contents. A total of 400, one-d old male broilers (five cages/treatment; eight birds/cage) were used in the experiment.2. Regardless of enzyme supplementation, weight gain (WG) increased up to 283 g/kg of barley and was reduced afterwards (P < 0.01). Increasing levels of barley resulted in greater (P < 0.001) gain per feed (G/F). Enzyme addition increased WG (P < 0.05) and G/F (P < 0.001) at each barley inclusion level.3. Birds fed diets with 0 and 565 g/kg barley showed the lowest and highest (P < 0.001to 0.05) digestibility for all nutrients measured, respectively. Digestibility of all nutrients was improved by enzyme supplementation at each barley inclusion level (P < 0.05). The nitrogen-corrected apparent metabolisable energy improved with increasing inclusion of barley (P < 0.001) and supplemental enzyme (P < 0.01). Increasing inclusion of barley increased the relative weight of gizzard (P < 0.001) and reduced jejunal digesta viscosity (P < 0.001). Supplemental enzyme (P < 0.001) reduced digesta viscosity.4. The optimum inclusion level of barley, with respect to growth performance, was 283 g/kg of diet. Increasing barley inclusion improved nutrient and energy utilisation, possibly through lowered digesta viscosity and better function of the gizzard. Feed efficiency and nutrient and energy utilisation can benefit from carbohydrase supplementation in barley-based diets, regardless of barley inclusion level.

4,4'-Diaponeurosporene-Producing Bacillus subtilis Promotes the Development of the Mucosal Immune System of the Piglet Gut.

Gut mucosal immune responses are known to act as the first line of defense against invasion of pathogenic microorganisms. Piglets have an incompletely developed gut mucosal immune system, making them sensitive to intestinal infections. Promoting the development of the mucosal immune system will increase the pathogen resistance of piglets. The aim of the present study was to investigate the effect of carotenoid (4,4'-diaponeurosporene)-producing Bacillus subtilis (B.s-Dia) on intestinal mucosal immunity in piglets. We showed that oral administration to piglets of B.s-Dia remarkably improved the development of Peyer's patches (PPs) (P < 0.01), and increased villus height (P < 0.01) and colon crypt depth (P < 0.01). In addition, B.s-Dia also increased the number of intraepithelial lymphocytes (P < 0.01), while Bacillus subtilis (B.s) had no significant influence on it (P > 0.05). Moreover, B.s-Dia also increased the number of SIgA+ cells (P < 0.01). Oral administration of either B.s or B.s-Dia increased the number of CD4+ and CD8+ cells in ileum lamina propria (P < 0.01). These results indicate that B.s-Dia contributes to a higher extent to porcine mucosal immune system development than B.s, and might serve as an immunopotentiator candidate. Anat Rec, 302:1800-1807, 2019. © 2019 American Association for Anatomy.

Effect of polyunsaturated fatty acids on postnatal ileum development using the fat-1 transgenic mouse model.

BACKGROUND: Long-chain polyunsaturated fatty acids (LCPUFAs) play a critical role in neonatal health. We hypothesized that LCPUFAs play an essential role in priming postnatal gut development. We studied the effect of LCPUFAs on postnatal gut development using fat-1 transgenic mice, which are capable of converting n-6 to n-3 LCPUFAs, and wild-type (WT) C57BL/6 mice. METHODS: Distal ileum sections were collected from fat-1 and WT mice on days 3, 14, and 28. Fatty acid analyses, histology, RT-qPCR and intestinal permeability were performed. RESULTS: Fat-1 mice, relative to WT mice, showed increased n-3 LCPUFAs levels (α-linolenic acid, docosahexaenoic acid, and eicosapentaenoic acid, p < 0.05) and decreased arachidonic acid levels (p < 0.05) in the ileum. Preweaning fat-1 mice, compared to WT, showed >50% reduced muc2, Tff3, TLR9, and Camp expression (p < 0.05), markers of the innate immune response. There was a >two-fold increased expression of Fzd5 and EphB2, markers of cell differentiation (p < 0.05), and Fabp2 and 6, regulators of fatty acid transport and metabolism (p < 0.05). Despite reduced expression of tight junction genes, intestinal permeability in fat-1 was comparable to WT mice. CONCLUSIONS: Our data support the hypothesis that fatty acid profiles early in development modulate intestinal gene expression in formative domains, such as cell differentiation, tight junctions, other innate host defenses, and lipid metabolism.

Modulating Sterol Concentrations in Infant Formula Influences Cholesterol Absorption and Synthesis in the Neonatal Piglet.

Formula-fed infants present higher cholesterol synthesis rates and lower circulating cholesterol during the postnatal feeding period compared to breast-fed infants, though the mechanisms underlying this phenotype are not fully understood. Typical infant formulas contain vegetable-based fats, inherently including phytosterols (PS), which are structurally similar to cholesterol and may interfere with their absorption. A seven-day old piglets model was used to test the inhibitory effects of PS on cholesterol absorption during postnatal feeding. Following feeding for 21 days with milk-based formulas containing PS and cholesterol levels resembling those in formulas or human-milk, apparent cholesterol digestibility was analyzed in ileal digesta, and cholesterol, PS, and cholesterol synthesis markers were analyzed in plasma and liver samples. Ileal cholesterol digestibility content was increased in the piglets fed low PS formulas and the rate of the hepatic cholesterol synthesis, as determined by the lathosterol-to-cholesterol ratios (L:C), was decreased in the piglets fed LP-formulas and corresponded to reduced nuclear expression of SREBP2 relative to those fed HP-formulas. These results are consistent with the hypothesis that PS in formula can inhibit cholesterol absorption and enhance cholesterol synthesis. Whether or not this leads to entrainment of cholesterol synthesis later in life via early programming awaits further research.

Piglet weight gain during the first two weeks of lactation influences the immune system development.

The aim of this study was to investigate the effect of the piglet growth during the first week of life on ileal expression of genes and on development of the immune system. Eight litters adjusted to 12 piglets were used. Within each litter, the piglet that showed the lowest weight gain (LWG; n = 8) and the one that showed the highest weight gain (HWG; n = 8) in their first week of life were enrolled. Peripheral blood mononuclear cells (PBMC) were isolated on days 8 and 16 to characterize cellular population profiles and to assess ex-vivo secretion of interleukin-10 (IL-10), IL-6 and tumor necrosis factor-α (TNF-α). On day 16, piglets were euthanized and ileum samples were collected to extract RNA for microarray analysis and gene expression by qPCR. As expected, growth performance of LWG piglet was impaired compared to HWG piglets (P < 0.05). From day 8 to 16, the percentage of CD21+ B cells significantly increased in blood of heavier HWG piglets while the percentage remained constant in smaller LWG piglets (P weight x day = 0.01). For the CD4+CD8α- Th cells, a marked increase was observed in LWG piglets from 8 to 16 days of age (P = 0.002) whereas no significant change occurred in HWG piglets. Percentages of CD14+ monocytes and other MHC-II+ cells were respectively higher and lower on day 8 compared to day 16 for both groups of piglets (P < 0.01). On day 8, LPS-activated PBMC from LWG piglets produced less IL-6 compared to HWG piglets (P < 0.05). Microarray analysis of gene expression in piglets' ileum tissue indicated that several genes involed in defense response and response to oxidative stress were modulated differently in LWG compared to HWG. Gene analysis by Q-PCR confirmed microarray results and revealed that IL-10, SOD1, NOS2, NOD2, TLR4, TLR9, CD40 and CD74 expressions were significantly decreased (P < 0.05) in LWG in comparison to HWG piglets, while MYD88 and NFkBiA showed a tendency to decrease (0.05 ≤ P < 0.07). These results suggest that birth weight and milk intake affect the growth performances and the development of immunity by modulating the expression of genes associated with immunity and oxidative stress in piglets' intestinal tissue, and by affecting the leukocyte populations involved in innate and cell-mediated immunity in nursing piglets. Therefore, impaired development of immune system in LWG piglets might have an impact on their resistance to infections later in life.

Dietary chlorogenic acid supplementation affects gut morphology, antioxidant capacity and intestinal selected bacterial populations in weaned piglets.

Chlorogenic acid (CGA), an ester formed between caffeic acid and quinic acid, is one of the most abundant phenolic acids and is widespread in fruits, vegetables, cereals and tuber crops. Therefore, the present study was conducted to test the hypothesis that dietary supplementation with CGA could improve intestinal health and regulate intestinal selected microbiota in weaned piglets. A total of twenty-four piglets (21 d of age) were randomly assigned to one of four groups according to their initial BW and sex and fed a basal diet (control group) or a basal diet containing 250, 500 and 1000 mg kg-1 CGA, respectively. The whole trial lasted for 28 d. Dietary CGA supplementation increased (P < 0.05) the duodenal villous height and villous height : crypt depth ratio, but decreased (P < 0.05) the F/G ratio and duodenal crypt depth when compared with the control group. Meanwhile, an increase (P < 0.05) in the jejunal villous height and in the ileal villous height : crypt depth ratio were also observed in CGA-fed piglets. Supplementation with CGA significantly increased (P < 0.05) the activity of serum GSH-Px and the activities of duodenal GSH-Px and CAT, upregulated (P < 0.05) the expression of OCLN in the duodenum and jejunum, and decreased (P < 0.05) the ileal MDA content when compared to the control group. In addition, an increase (P < 0.05) in the population of Lactobacillus and a decrease (P < 0.05) in the population of Escherichia coli were observed in the colon of pigs fed CGA diets. Furthermore, pigs fed CGA diets had higher (P < 0.05) propionic and butyric acid concentrations in the colon. Altogether, our results provide evidence that dietary CGA is beneficial for preserving intestinal morphological integrity and selectively regulating intestinal microbiota, which can provide a means to improve gut health and growth performance post-weaning.

Expression profile of some neuronal and glial cell markers in the ovine ileal enteric nervous system during prenatal development.

The enteric nervous system (ENS) is a network of neurons and glia found in the gut wall and governs this gastrointestinal function independently from the central nervous system (CNS). ENS comprises the myenteric plexus (MP) and the submucous plexus (SP). In this study, we examined the expression profile of neurofilament heavy chain (NF-H), neuron-specific enolase (NSE), calcyclin (S100A6), vimentin and glial fibril acidic protein (GFAP) in ovine ileal enteric neurons and enteric glia cells (EGCs) during prenatal development using an immunohistochemical method. The material of the study consisted of 15 different fetal ileum tissues obtained between days 60 and 150 of pregnancy. NF-H was observed in the majority of ganglion cells in SP and MP throughout the fetal period. It was determined that there was no NF-H reaction in some ganglion cells in Peyer's patches of internal submucosal plexus (ISPF). In the early stage of pregnancy (60-90 days), there was no expression of NSE and S1006 in ileum. After this period, NSE and S1006 were expressed in the ganglion cells of the plexus, indicating an increase in the amount of expression towards the end of pregnancy. In the early period, vimentin expression was only detected in intramuscular interstitial cells (ICs) (60-90 days), but later (90-150 days) it was also seen in the cells around the ganglion cells in the plexus. On days 60-90 of gestation, GFAP expression only occurred in MP, but in later stages, staining was also detected in SP. In the plexus, an immunoreactivity was present in EGCs forming a network around the ganglion cell. During the last period of gestation (120-150 days), the number of GFAP-positive plexus increased, with the majority of these stained cells being observed in MP. Interestingly, weak staining or reaction did not occur in ISPF, unlike other plexuses. In conclusion, this is the first study that demonstrated the expression of NF-H, vimentin, S100A6, NSE and glial fibril acidic protein (GFAP) in ovine ileal ENS in the prenatal period. In the last period of gestation (120-150 days), the expression profile of ENS was similar to that of adult animals. The expression of the used markers increased toward the end of pregnancy. Our results suggest that neurons and EGCs show heterogeneity, and GFAP and NF-H cannot be used as panenteric glial or panneuronal markers, respectively. We also demonstrated, for the first time, the prenatal expression of S100A6 in enteric neurons and the possibility of using this protein for the identification of enteric neurons.

Neuropeptides and lymphocyte populations in the porcine ileum and ileocecal lymph nodes during postnatal life.

The maturation-related changes in the concentrations of galanin (Gal), vasoactive intestinal polypeptide (VIP), substance P (SP) and somatostatin (Som), as well as in subpopulations of lymphocytes expressing antigens CD2 (lymphocytes T), CD4 (T helper), CD8 (T cytotoxic), CD21 (B lymphocytes), CD5-/CD8+ (NK cells) and TCRgamma/delta (gut mucosal/intraepitelial cells) were studied in the ileal Peyer's patches and ileo-cecal lymph nodes in female pigs aged 3 days, 2 weeks, 4 weeks and 4 months. As regards neuropeptide concentrations statistically significant changes in the ileum and lymph nodes were found only in case of Gal and VIP. The concentrations of neuropeptides were significantly higher only in new-born animals. As regards the changes in subpopulations of lymphocytes, statistically significant changes were noticed only in 4-months old animals and were dealing only with CD2+ and TCRgamma/delta cells in the ileum as well as CD4+, CD8+, CD21+ and TCRgamma/delta in lymph nodes. The highest number of CD8+, CD21+ and TCRgamma/delta lymphocytes occurred in 4-months old animals.

Genomic dissection of conserved transcriptional regulation in intestinal epithelial cells.

The intestinal epithelium serves critical physiologic functions that are shared among all vertebrates. However, it is unknown how the transcriptional regulatory mechanisms underlying these functions have changed over the course of vertebrate evolution. We generated genome-wide mRNA and accessible chromatin data from adult intestinal epithelial cells (IECs) in zebrafish, stickleback, mouse, and human species to determine if conserved IEC functions are achieved through common transcriptional regulation. We found evidence for substantial common regulation and conservation of gene expression regionally along the length of the intestine from fish to mammals and identified a core set of genes comprising a vertebrate IEC signature. We also identified transcriptional start sites and other putative regulatory regions that are differentially accessible in IECs in all 4 species. Although these sites rarely showed sequence conservation from fish to mammals, surprisingly, they drove highly conserved IEC expression in a zebrafish reporter assay. Common putative transcription factor binding sites (TFBS) found at these sites in multiple species indicate that sequence conservation alone is insufficient to identify much of the functionally conserved IEC regulatory information. Among the rare, highly sequence-conserved, IEC-specific regulatory regions, we discovered an ancient enhancer upstream from her6/HES1 that is active in a distinct population of Notch-positive cells in the intestinal epithelium. Together, these results show how combining accessible chromatin and mRNA datasets with TFBS prediction and in vivo reporter assays can reveal tissue-specific regulatory information conserved across 420 million years of vertebrate evolution. We define an IEC transcriptional regulatory network that is shared between fish and mammals and establish an experimental platform for studying how evolutionarily distilled regulatory information commonly controls IEC development and physiology.

An investigation into the effects of maternal supplementation with excess iodine on the mechanisms and impacts of reduced IgG absorption in the lamb postpartum.

An experiment was conducted to determine: (1) the effect of excess maternal I supplementation on the thyroid hormone status of the ewe and her progeny; (2) potential mechanisms underpinning the failure of passive transfer associated with excess I and (3) the growing lambs' response to natural gastrointestinal infection. Twin-bearing ewes received one of two treatments (n 32/treatment group): basal diet (C) or C plus 26·6 mg of iodine/ewe per d (I), supplied as calcium iodate. Ewes were individually fed from day 119 of gestation to parturition. Progeny of I ewes had lower (P<0·01) serum IgG concentrations from 24 h to 28 d postpartum but higher serum IgG concentrations at day 70 postpartum (P<0·05). I supplementation increased the relative expression of Fc receptor, IgA, IgM high affinity and polymeric Ig receptor in the ileum of the lamb at 24 h postpartum; however, thyroid hormone receptor-ß (THRB) and ß-2-microglobulin (B2M) expression declined (P<0·05). Progeny of I ewes had higher growth rates to weaning (P<0·05) and lower faecal egg count (FEC) for Nematodirus battus (P<0·05) between weeks 6 and 10 postpartum. In conclusion, excess maternal I supplementation negatively affected the thyroid hormone status, serum IgG concentration, ileal morphology and the gene expression of THRB and B2M in the ileum and ras-related protein (RAB) RAB25 and the mucin gene (MUC) MUC1 in the duodenum of the lamb postpartum. These effects were followed by an enhancement of average daily gain and lower N. battus FEC in the pre-weaning period of I-supplemented lambs.

[Effect of deoxycholic acid intervention on growth of ileum organoids derived from C57BL/6 mice].

OBJECTIVE: To establish a culture system for mouse intestinal organoids and investigate the effect of deoxycholic acid (DCA) on organoids growth. METHODS: The terminal ileum was collected from 8-month-old C57BL<6 mice. The tissue blocks were treated with EDTA and the crypts were collected and embedded in Matrigel Matrix. Orgnoids growth and buddings were observed in the control group, anhydrous alcohol group, short-term (2 days) 100 µmol

Effects of Curcuma longa rhizome powder on egg quality, performance and some physiological indices of laying hens fed different levels of metabolizable energy.

BACKGROUND: High-energy diets of laying hens may improve roductive performance, although some negative effects may also appear with respect to egg quality and physiological parameters. Curcuma longa rhizome powder (CRP) has beneficial effects on health indices of the birds through antioxidant and antimicrobial properties, especially when the birds experience nutritional stress. RESULTS: Increasing dietary CRP enhanced egg quality by improving eggshell thickness and hardness but decreasing yolk cholesterol content (P < 0.05). The best feed conversion ratio was obtained in birds fed high-apparent metabolizable energy (AME) diets supplemented with 2.0 g kg-1 CRP (P < 0.05). Although increasing dietary AME elevated the serum concentration of triglycerides (P < 0.05) and enzymatic activities of alanine aminotransferase and aspartate aminotransferase (P < 0.05), dietary inclusion of CRP alleviates the blood levels of these enzymes (P < 0.01). Low level of dietary CRP boosted the immune responses to Newcastle virus (P < 0.01) and sheep red blood cells (P < 0.05) antigens but decreased the heterophil to lymphocyte ratio (P < 0.05). Inclusion of at least 2.0 g kg-1 CRP in the diet of laying decreased Escherichia coli enumerations in the ileal content (P < 0.01) and improved villus height, crypt depth and goblet cell numbers (P < 0.05). CONCLUSION: An improvement in the productive performance of laying hens fed high-energy diets might be associated with decreasing health indices and product quality, which could potentially be amended by nutritional modifications such as incorporating medicinal herbs in the feed. © 2016 Society of Chemical Industry.

Wheat bran components modulate intestinal bacteria and gene expression of barrier function relevant proteins in a piglet model.

The objective of this study was to determine the impact of wheat bran and its main polysaccharides on intestinal bacteria and gene expression of intestinal barrier function relevant proteins. Thirty freshly weaned male piglets were assigned randomly to five dietary treatment groups with six piglets per group. Accordingly, five synthetic diets including a basal control diet without fiber components (CON), wheat bran diet (10% wheat bran, WB), arabinoxylan diet (AX), cellulose diet (CEL) and combined diet of arabinoxylan and cellulose (CB) were studied. The piglets were fed ad libitum for 30 d. Lower Escherichia coli (E. coli) populations in WB group and higher probiotic (Lactobacillus and Bifidobacterium) populations in groups fed diets containing arabinoxylan (WB, AX and CB) were observed and compared with CON group. Compared with CON group, the gene expressions of cystic fibrosis transmembrane conductance regulator (CFTR), calcium-activated chloride channel regulator 1 (CLCA1) and voltage-gated chloride channel 2 (CIC2) were suppressed in the WB group. And wheat bran down-regulated gene expression of pro-inflammation (TNF-α, IL-1ß, IL-6) and TLRs/MyD88/NF-κB pathway compared with CON group. In conclusion, wheat bran and its main polysaccharides could change intestinal microflora and down-regulate the gene expression of intestinal barrier function relevant proteins in the distal small intestinal mucosa.

Intestinal barrier analysis by assessment of mucins, tight junctions, and α-defensins in healthy C57BL/6J and BALB/cJ mice.

The intestinal barrier is gaining increasing attention because it is related to intestinal homeostasis and disease. Different parameters have been used in the past to assess intestinal barrier functions in experimental studies; however most of them are poorly defined in healthy mice. Here, we compared a number of barrier markers in healthy mice, established normal values and correlations. In 48 mice (24 C57BL/6J, 24 BALB/cJ background), we measured mucus thickness, and expression of mucin-2, α-defensin-1 and -4, zonula occludens-1, occludin, junctional adhesion molecule-A, claudin-1, 2 and -5. We also analyzed claudin-3 and fatty acid binding protein-2 in urine and plasma, respectively. A higher expression of mucin-2 protein was found in the colon compared to the ileum. In contrast, the α-defensins-1 and -4 were expressed almost exclusively in the ileum. The protein expression of the tight junction molecules claudin-1, occludin and zonula occludens-1 did not differ between colon and ileum, although some differences occurred at the mRNA level. No age- or gender-related differences were found. Differences between C57BL/6J and BALB/cJ mice were found for α-defensin-1 and -4 mRNA expression, and for urine and plasma marker concentrations. The α-defensin-1 mRNA correlated with claudin-5 mRNA, whereas α-defensin-4 mRNA correlated with claudin-3 concentrations in urine. In conclusion, we identified a number of murine intestinal barrier markers requiring tissue analyses or measurable in urine or plasma. We provide normal values for these markers in mice of different genetic background. Such data might be helpful for future animal studies in which the intestinal barrier is of interest.

Transcriptome analysis reveals regional and temporal differences in mucosal immune system development in the small intestine of neonatal calves.

BACKGROUND: Postnatal development of the mammalian mucosal immune system is crucial for responding to the rapid colonization by commensal bacteria and possible exposure to pathogens. This study analyzed expression patterns for mRNAs and their relationship with microRNAs (miRNAs) in the bovine small intestine during the critical neonatal period (0 to 42 days). This analysis revealed molecular mechanisms regulating the postnatal development of the intestinal mucosal immune system. RESULTS: Small intestine samples (jejunum and ileum) were collected from newborn male, Holstein calves immediately post-partum (n = 3) and at 7 (n = 5), 21 (n = 5), and 42 (n = 5) days of age and the transcriptomes were profiled using RNA-Seq. When analyzing all time points collectively, greater expression of genes encoding the complement functional pathway, as well as lower expression of genes encoding Toll-like receptors and NOD-like receptors were observed in the jejunum when compared to the ileum. In addition, significant changes in the expression of immune-related genes were detected within the first week post-partum in both jejunum and ileum. For example, increased expression of genes encoding tight junction proteins (claudin 1, claudin 4 and occludin), an antimicrobial peptide (Regenerating Islet-Derived 3-γ), NOD-like receptors (NACHT, LRR and PYD domain-containing protein 3), regulatory T cell marker (forkhead box P3), and both anti-inflammatory (interleukin 10) and pro-inflammatory (interleukin 8) cytokines was observed throughout the small intestine of 7-day-old calves when compared to newborn calves. Moreover, the expression of mucosal immune-related genes were either positively or negatively correlated with total bacterial population depending on both intestinal region and age. The integrated analysis of miRNAs and mRNAs supported the conclusion that miRNAs may regulate temporal changes in the expression of genes encoding tight junction proteins (miR-335), cytokines (miR-335) and bacterial recognition (miR-100) during the first week of small intestine development. CONCLUSION: The rapid development of transcriptional differences between jejunum and ileum reveal that these two intestinal regions make distinct contributions to the intestinal mucosal immune system during the early neonatal period. In addition, transcriptome analysis indicates that the first week after birth is a very dynamic developmental period for the intestinal mucosal immune system and these changes may be regulated by both miRNAs and microbial colonization. Findings from this study indicate that a detailed analysis of both the abundance and diversity of the colonizing microbiome may be necessary to understand factors regulating the rapid development of the mucosal immune system during the first week of life.

Use of larvae meal as protein source in broiler diet: Effect on growth performance, nutrient digestibility, and carcass and meat traits.

The aim of this research was to study the effect of insect meal from larvae ( larvae meal [TML]) as complete replacement of soybean meal (SBM) on growth performance, nutrient digestibility, and carcass and meat traits of broilers. A total of eighty 30-d-old male Shaver brown broilers were homogenously divided into 2 groups (each consisting of 8 replicates of 5 birds). Up to 62 d of age, the groups were fed 2 isoproteic and isoenergetic diets differing for the ingredient used as the main protein source: the control group was fed a corn-SBM-based diet, whereas in the TML group, the SBM was completely replaced by TML. Broiler growth performance was measured during the trial. At 62 d of age, 2 broilers per replicate (16 per group) were slaughtered and apparent ileal digestibility coefficients and carcass and meat traits were determined. The use of TML as the main protein source in the broiler diet had no significant effect on most growth performance and carcass traits and chemical and physical properties of meat, the latter being important for marketing purposes. The feed conversion ratio in the entire experimental period (from 30 to 62 d) was improved in the TML group compared with the SBM group ( < 0.05). The apparent ileal digestibility coefficients of DM, OM, and CP in broilers fed the SBM diet were greater ( < 0.01) than the other group. The full digestive system in broilers fed SBM had a lower ( < 0.05) absolute and relative weight than that of broilers fed TML. Also, the weight and the percentage of the spleen in the SBM group were lower ( < 0.05) than those in the TML group. The length of the entire intestine in the group fed TML was greater ( < 0.05) than the other group and the same happened when intestinal length was expressed as percentage of broiler BW ( < 0.05). Among the different intestinal tracts, the ileum and ceca of broilers fed TML had a greater ( < 0.05) length than that of broilers fed SBM. Also, ceca weight (as an absolute value or percentage on live weight) in broilers fed TML was greater ( < 0.05) than that of broilers fed SBM. Breast pH and cooking losses in broilers fed TML were greater ( < 0.05) than those of broilers fed SBM. Chemical characteristic of meat obtained from right breast of slaughtered broilers were unaffected by dietary treatments. larvae meal can be a suitable alternative protein source for growing broilers and also when used as principal protein contributor to the diet.

The several elements of intestinal innate immune system at the beginning of the life of broiler chicks.

Functional capacity of digestive system and intestinal adaptive immunity are immature at hatching of broiler chicks. Therefore, intestinal innate immunity after hatching is vital to young chicks. The purpose of this study was to investigate expression and tissue distributions of several elements of the innate immune system (i.e., TLR2, TLR4, CD83, and MHC class II expressing cells) in the intestine of one-day-old chicks. For this purpose, ileum and cecum were examined the under different conditions, which included the control and 1, 3, 6, 12, or 24 h after injection of lipopolysaccharide (LPS) and phosphate buffered saline. The findings indicated that regardless of the antigenic stimulation, Toll-like receptor (TLR) 2 and TLR4 expressing cells were present in the intestinal tissues of one-day-old chicks. We noticed that the intestinal segments have different TLR expression levels after LPS stimulation. Dendritic cells were identified, and they left the intestinal tissue after LPS treatment. MHC class II molecules were diffusely present in both the ileum and cecum. This study demonstrates that the intestinal tissue of one-day-old chicks has remarkable defensive material, including histological properties and several elements of the innate immune system. Microsc. Res. Tech. 79:604-614, 2016. © 2016 Wiley Periodicals, Inc.