Jejunal Tissue Research Articles

Sequencing approaches to identify distal jejunum microbial community composition and function in broiler chickens fed anti-interleukin-10 during coccidiosis and necrotic enteritis challenge

Strategies to counteract interleukin (IL)-10-mediated immune evasion by Eimeria spp. during coccidiosis- like anti-IL-10 antibodies- may protect broiler chicken health and reduce incidence of secondary necrotic enteritis (Clostridium perfringens) via undetermined mechanisms. Objectives were to use sequencing techniques to evaluate jejunal microbial community composition and function in anti-IL-10-fed broilers during coccidiosis and necrotic enteritis. On d0, Ross 308 chicks were placed in 32 cages (15 chicks/ cage) for a 25-d study and randomly assigned to diets ± 0.03% anti-IL-10. Six chicks/ diet were euthanized for distal jejunum content and tissue collection on d 14 (baseline) before inoculating the remainder with saline or 15,000 E. maxima oocysts (M6 strain). Half the chicks challenged with E. maxima were challenged with C. perfringens (1×108 colony forming units) on d 18 and 19. Follow-up samples (6 chicks/treatment) were collected at 7 and 11 d postinoculation (pi) for the E. maxima-only group, or 3 and 7 dpi for the E. maxima + C. perfringens group with 3/7 dpi being designated as peak and 7/11dpi as postpeak challenge. DNA was extracted from digesta for microbiota composition analysis (16S rRNA gene sequencing) while RNA was extracted from tissue to evaluate the metatranscriptome (RNA sequencing). Alpha diversity and genus relative abundances were analyzed using the diet or challenge main effects with associated interactions (SAS 9.4; P ≤ 0.05). No baseline microbial changes were associated with dietary anti-IL-10. At peak challenge, a diet main effect reduced observed species 36.7% in chicks fed anti-IL-10 vs. control; however, the challenge effect reduced observed species and Shannon diversity 51.2-58.3% and 33.0 to 35.5%, respectively, in chicks challenged with E. maxima ± C. perfringens compared to their unchallenged counterparts (P ≤ 0.05). Low sequencing depth limited metatranscriptomic analysis of jejunal microbial function via RNA sequencing. This study demonstrates that challenge impacted the broiler distal jejunum microbiota more than anti-IL-10 while future research to characterize the microbial metatranscriptome may benefit from investigating other intestinal compartments.

Improvement of in vivo iron bioavailability using mung bean peptide-ferrous chelate

There is an increasing amount of research into the development of a third generation of iron supplementation using peptide-iron chelates. Peptides isolated from mung bean were chelated with ferrous iron (MBP-Fe) and tested as a supplement in mice suffering from iron-deficiency anemia (IDA). Mice were randomly divided into seven groups: a group fed the normal diet, the IDA model group, and IDA groups treated with inorganic iron (FeSO4), organic iron (ferrous bisglycinate, Gly-Fe), low-dose MBP-Fe(L-MBP-Fe), high-dose MBP-Fe(H-MBP-Fe), and MBP mixed with FeSO4 (MBP/Fe). The different iron supplements were fed for 28 days via intragastric administration. The results showed that MBP-Fe and MBP/Fe had ameliorative effects, restoring hemoglobin (HGB), red blood cell (RBC), hematocrit (HCT), and serum iron (SI) levels as well as total iron binding capacity (TIBC) and body weight gain of the IDA mice to normal levels. Compared to the inorganic (FeSO4) and organic (Gly-Fe) iron treatments, the spleen coefficient and damage to liver and spleen tissues were significantly lower in the H-MBP-Fe and MBP/Fe mixture groups, with reparative effects on jejunal tissue. Gene expression analysis of the iron transporters Dmt 1 (Divalent metal transporter 1), Fpn 1 (Ferroportin 1), and Dcytb (Duodenal cytochrome b) indicated that MBP promoted iron uptake. These findings suggest that mung bean peptide-ferrous chelate has potential as a peptide-based dietary supplement for treating iron deficiency.

Relationship between jejunum ATPase activity and antioxidant function on the growth performance, feed conversion efficiency, and jejunum microbiota in Hu sheep (Ovis aries)

BackgroundATPase activity and the antioxidant function of intestinal tissue can reflect intestinal cell metabolic activity and oxidative damage, which might be related to intestinal function. However, the specific influence of intestinal ATPase activity and antioxidant function on growth performance, feed conversion efficiency, and the intestinal microbiota in sheep remains unclear.ResultsThis study analyzed the correlation between ATPase activity and antioxidant function in the jejunum of 92 Hu sheep and their growth performance and feed conversion efficiency. Additionally, individuals with the highest (H group) and lowest (L group) jejunum MDA content and Na+ K+-ATPase activity were further screened, and the effects of jejunum ATPase activity and MDA content on the morphology and microbial community of sheep intestines were analyzed. There was a significant correlation between jejunum ATPase and SOD activity and the initial weight of Hu sheep (P < 0.01). The H-MDA group exhibited significantly higher average daily gain (ADG) from 0 to 80 days old and higher body weight (BW) after 80 days. ATPase and SOD activities, and MDA levels correlated significantly and positively with heart weight. The jejunum crypt depth and circular muscle thickness in the H-ATP group were significantly higher than in the L-ATP group, and the villus length, crypt depth, and longitudinal muscle thickness in the H-MDA group were significantly higher than in the L-MDA group (P < 0.01). High ATPase activity and MDA content significantly reduced the jejunum microbial diversity, as indicated by the Chao1 index and observed species, and affected the relative abundance of specific taxa. Among species, the relative abundance of Olsenella umbonata was significantly higher in the H-MDA group than in the L-MDA group (P < 0.05), while Methanobrevibacter ruminantium abundance was significantly lower than in the L-MDA group (P < 0.05). In vitro culture experiments confirmed that MDA promoted the proliferation of Olsenella umbonata. Thus, ATPase and SOD activities in the jejunum tissues of Hu sheep are predominantly influenced by congenital factors, and lambs with higher birth weights exhibit lower Na+ K+-ATPase, Ca2+ Mg2+-ATPase, and SOD activities.ConclusionsThe ATPase activity and antioxidant performance of intestinal tissue are closely related to growth performance, heart development, and intestinal tissue morphology. High ATPase activity and MDA content reduced the microbial diversity of intestinal tissue and affect the relative abundance of specific taxa, representing a potential interaction between the host and its intestinal microbiota.

Seasonal variation of melatonin secretion across various segments of the gastrointestinal tract in rats

ObjectiveTo investigate whether melatonin (MT) secretion in different parts of the gastrointestinal tract (GIT) exhibits seasonal variations and its correlation with immune regulation. MethodsSixty Sprague-Dawley rats were divided into control and model groups, and the pineal gland was removed in the model group. Stomach, jejunum, ileum, and colon tissues were obtained during the spring equinox, summer solstice, beginning of autumn, autumn equinox, and winter solstice. The levels of MT, MT receptors (MR), arylalkylamine N-acetyltransferase (AANAT), hydroxyindole-O-methyltransferase (HIOMT), interleukin-2 (IL-2), and interleukin-10 (IL-10) in the GIT were measured using enzyme-linked immunosorbent assay. ResultsExcept for the stomach, the jejunum, ileum, and the colon showed seasonal tendencies in MT secretion. In the control group, MT secretion in the jejunum and ileum was the highest in the long summer, and colonic MT secretion was the highest in winter. In the model group, MT levels in the colon were highest in the summer. The seasonal rhythms of the MR, AANAT, HIOMT, IL-2, and IL-10 in the colon were roughly similar to those of MT, and changed accordingly after pinealectomy. ConclusionsGastrointestinal MT secretion is related to seasonal changes, and MT secretion in each intestinal segment is influenced by different seasons. The biological effects of MT in the gut are inextricably linked to the mediation of MR, and a hormone-receptor linkage exists between MT and MR. The effect of seasonal changes on the gastrointestinal immune system may be mediated through the regulation of seasonal secretion of MT.

Anemoside B4 attenuates necrotic enteritis of laying hens induced by Clostridium perfringens via inhibiting NF-κB and PI3K/Akt/mTOR signalling pathways

Poultry necrotic enteritis is an important enteric disease which might be controlled by antibiotics. However, with the excessive use of antibiotics, the phenomenon of drug resistance of Clostridium perfringens is becoming increasingly prominent. Anemoside B4 exhibits important anti-inflammatory, antioxidant and immunomodulatory effects. This study was performed to estimate the effect of Anemoside B4 on chicken necrotic enteritis induced by C. perfringens in vivo and in vitro. In the in vivo experiment we investigated the efficacy of Anemoside B4 on the growth curve, biofilm formation, haemolytic activity, virulence-related gene expression and NF-κB and PI3K/AKT/mTOR activation in Caco-2 cells induced by C. perfringens. The results showed that 12.5–50 μg/mL Anemoside B4 had no antibacterial activity but could inhibit biofilm formation, attenuate haemolytic activity and virulence-related gene expression of C. perfringens and weaken NF-κB and PI3K/Akt/mTOR activation triggered by C. perfringens in Caco-2 cells. In the in vivo experiment, 60 17-day-old healthy White Leghorns were randomly divided into six groups. The growing laying hens of the control group were fed a basic diet, and those of the five challenged groups were fed a basic diet (infection group), added 0.43 g/kg Anemoside B4 (0.43 g/kg Ane group), 0.86 g/kg Anemoside B4 (0.86 g/kg Ane group), 1.72 g/kg Anemoside B4 (1.72 g/kg Ane group) and 40 mg/kg lincomycin (lincomycin group), respectively. All challenged laying hens were infected with 1 × 109 CFU C. perfringens from day 17–20. Blood and intestinal samples were obtained, and the data demonstrated that Anemoside B4 improved the blood biochemical parameters, attenuated jejunum tissue injury, increased the spleen, thymus, bursa of fabricius index, and decreased lesion scores of the jejunum and the ileum. In the jejunum, Anemoside B4 and lincomycin downregulated the expression of IL-1β, IL-6, IL-10, TNF-α and IFN-γ at mRNA levels. Moreover, Anemoside B4 significantly enhanced both mRNA and protein levels of tight junctions ZO-1, Claudin-1 and MUC-2 in the jejunum. Anemoside B4 weakened p-P65, p-PI3K, p-Akt and p-mTOR protein expression in the jejunum infected by C. perfringens. Diets supplemented with Anemoside B4 alleviated C. perfringens-induced necrotic enteritis in laying hens by inhibiting NF-κB and PI3K/Akt/mTOR signalling pathways and improving intestinal barrier functions.

Unraveling the multi-faceted role of Rosmarinus officinalis L. (rosemary) and diosmetin in managing gut motility

Ethnopharmacological relevanceRosmarinus officinalis L. (Rosemary) is a popular herb with reported effectiveness against diarrhea, anxiety and constipation, albeit with limited pharmacological evidence. Aim of the studyThe current study was aimed at evaluating the therapeutic potential, possible pharmacological mechanisms of action and active constituents of hydro-ethanolic extract of rosemary (Rs.Cr), as potential anti-diarrheal, laxative and anxiolytic agent. MethodRs.Cr was analyzed through reverse-phase high pressure liquid chromatography (RP-HPLC). Laxative, antidiarrheal, and anxiolytic activities were assessed using in vivo models. Spasmogenic and spasmolytic mechanisms were studied on isolated guinea pig ileum and rabbit jejunum tissues, respectively. Possible role of diosmetin, one of the active constituents of Rs.Cr was also evaluated. ResultsRP-HPLC analysis revealed presence of diosmetin, rutin and apigenin in Rs.Cr. Laxative effect was seen at low doses, which was partially reversed in atropinized mice. The spasmogenic mechanism was mediated by cholinergic and histaminergic receptors stimulation. At higher doses, antidiarrheal activity was evident, with reduction in gastrointestinal motility and secretions using charcoal meal and enteropooling assays, respectively. Rs.Cr also showed dose-dependent anxiolytic effect. The antispasmodic mechanisms were mediated by anti-muscarinic and K+ channel opening-like effect (predominant KATP-dependent). Diosmetin exhibited antidiarrheal and antispasmodic activities, but spasmogenic effect was not seen. ConclusionRosemary leaves have dual antidiarrheal and laxative effects, and as well as anxiolytic activity. In addition, the possible modulation of muscarinic and histaminergic receptors, and KATP channels show it as potential herb to be explored for irritable bowel syndrome. Diosmetin is possibly one of its constituents that contributes to its antidiarrheal activity.

Therapeutic potential of probiotics to counter Campylobacter infections: An In vitro and In vivo evaluation in mice

Probiotics, including Lactic Acid Bacteria (LAB), have gained considerable attention due to their potential health benefits for humans and animal livestock. This research aimed to isolate probiotic bacteria of human origin and explore their therapeutic potential against Campylobacter strains, which are recognized as the leading bacterial cause of human gastroenteritis worldwide. A collection of 230 LAB isolates from human stools were screened for the ability of cell free supernatants to inhibit the growth of three strains of C. jejuni and a C. coli. Co-culture and agar-well diffusion assays identified seven isolates with inhibitory activity against Campylobacter ssp., which were characterized as probiotic strains by their tolerance to pH 2 and bile salts, and then identified as Enterococcus spp. using 16S rRNA gene sequencing E. faecalis NM231, E. faecium NM234, E. faecium NM233, E. faecium NM113, E. durans NM232, E. faecalis NM235, and E. faecium NM236. In vivo evaluation was undertaken by the oral administration of selected probiotics to groups of mice prior to challenge with C. jejuni. Histopathological examinations showed challenge groups receiving either mixed probiotics or E. faecium NM234 were protected against C. jejuni-induced jejunal, colonic, ovarian and uterine tissue damage. Immunohistochemical detection of the challenged mice revealed the presence of C. jejuni antigen in jejunal, colonic, ovarian and uterine sections but was absent in mice receiving either mixed probiotics or E. faecium NM234. This study introduces E. faecium NM234 as a promising probiotic against Campylobacter.

Investigation of the nutritional and functional roles of a combinational use of xylanase and β-glucanase on intestinal health and growth of nursery pigs

BackgroundXylanase and β-glucanase combination (XG) hydrolyzes soluble non-starch polysaccharides that are anti-nutritional compounds. This study aimed to evaluate the effects of increasing levels of XG on intestinal health and growth performance of nursery pigs.MethodsForty pigs (6.5 ± 0.4 kg) were assigned to 5 dietary treatments and fed for 35 d in 3 phases (11, 9, and 15 d, respectively). Basal diets mainly included corn, soybean meal, and corn distiller’s dried grains with solubles, contained phytase (750 FTU/kg), and were supplemented with 5 levels of XG at (1) 0, (2) 280 TXU/kg xylanase and 125 TGU/kg β-glucanase, (3) 560 and 250, (4) 840 and 375, or (5) 1,120 and 500, respectively. Growth performance was measured. On d 35, all pigs were euthanized and jejunal mucosa, jejunal digesta, jejunal tissues, and ileal digesta were collected to determine the effects of increasing XG levels and XG intake on intestinal health.ResultsIncreasing XG intake tended to quadratically decrease (P = 0.059) viscosity of jejunal digesta (min: 1.74 mPa·s at 751/335 (TXU/TGU)/kg). Increasing levels of XG quadratically decreased (P < 0.05) Prevotellaceae (min: 0.6% at 630/281 (TXU/TGU)/kg) in the jejunal mucosa. Increasing XG intake quadratically increased (P < 0.05) Lactobacillaceae (max: 40.3% at 608/271 (TXU/TGU)/kg) in the jejunal mucosa. Increasing XG intake quadratically decreased (P < 0.05) Helicobacteraceae (min: 1.6% at 560/250 (TXU/TGU)/kg) in the jejunal mucosa. Increasing levels of XG tended to linearly decrease (P = 0.073) jejunal IgG and tended to quadratically increase (P = 0.085) jejunal villus height to crypt depth ratio (max: 2.62 at 560/250 (TXU/TGU)/kg). Increasing XG intake tended to linearly increase the apparent ileal digestibility of dry matter (P = 0.087) and ether extract (P = 0.065). Increasing XG intake linearly increased (P < 0.05) average daily gain.ConclusionsA combinational use of xylanase and β-glucanase would hydrolyze the non-starch polysaccharides fractions, positively modulating the jejunal mucosa-associated microbiota. Increased intake of these enzyme combination possibly reduced digesta viscosity and humoral immune response in the jejunum resulting in improved intestinal structure, and ileal digestibility of nutrients, and finally improving growth of nursery pigs. The beneficial effects were maximized at a combination of 550 to 800 TXU/kg xylanase and 250 to 360 TGU/kg β-glucanase.

45 Awardee Talk: Investigation of the nutritional and functional roles of a combinational use of xylanase and β-glucanase on intestinal health and growth of nursery pigs

Abstract The objective of this study was to evaluate the intestinal health and growth performance of nursery pigs affected by dietary supplementation of increasing levels of xylanase and b-glucanase combinations. Pigs [n = 40; initial body weight (BW) = 6.5 ± 0.4 kg] were assigned to 5 dietary treatments and fed for 35 d in 3 phases (11, 9, and 15 d, respectively). Basal diets mainly included corn, soybean meal, and distiller’s dried grains with solubles with phytase (750 FTU/kg) supplemented with 5 levels of xylanase and b-glucanase combinations (NSPase) at 1) 0, 2) 280 TXU/kg xylanase and 125 TGU/kg b-glucanase, 3) 560 and 250, 4) 840 and 375, or 5) 1,120 and 500, respectively. Growth performance and fecal score were measured for each phase. On d 35, all pigs were euthanized and collected jejunal mucosa, jejunal digesta, jejunal tissues, and ileal digesta to determine the effects of increasing NSPase levels (%) and NSPase intake on jejunal mucosa-associated microbiota, jejunal digesta viscosity, immune responses, intestinal morphology, feed digestibility, and growth performance. Increasing NSPase level quadratically decreased (P &amp;lt; 0.05) Bacteroidetes (min: 0.7% at 636 TXU/kg and 284 TGU/kg), Prevotellaceae (min: 0.6% at 630 TXU/kg and 281 TGU/kg) in the jejunal mucosa. Increasing NSPase intake quadratically increased (P &amp;lt; 0.05) Firmicutes (max: 72.7% at 604 TXU/kg and 270 TGU/kg), Lactobacillaceae (max: 40.3% at 608 TXU/kg and 271 TGU/kg), and Lactobacillus (max: 40.3% at 608 TXU/kg and 271 TGU/kg) in the jejunal mucosa. Increasing NSPase intake also quadratically decreased (P &amp;lt; 0.05) Proteobacteria (min: 3.8% at 562 TXU/kg and 251 TGU/kg), Helicobacteraceae (min: 1.6% at 560 TXU/kg and 250 TGU/kg), and Helicobacter (min: 1.6% at 560 TXU/kg and 250 TGU/kg) in the jejunal mucosa. Increasing NSPase intake tended to quadratically decrease (P = 0.059) viscosity of jejunal digesta reaching the minimum (1.74 mPa·s) at 751 TXU/kg and 335 TGU/kg. Increasing NSPase level tended to linearly decrease (P = 0.073) jejunal IgG and tended to quadratically increase (P = 0.085) jejunal villus height to crypt depth ratio reaching the maximum (2.62) at 560 TXU/kg and 250 TGU/kg. Increasing NSPase intake tended to linearly increase the apparent ileal digestibility of dry matter (P = 0.087) and ether extract (P = 0.065). Increasing NSPase intake linearly increased (P &amp;lt; 0.05) average daily gain (ADG). In conclusion, increasing the combinational use of xylanase and β-glucanase positively modulated jejunal mucosa-associated microbiota, reduced jejunal digesta viscosity, reduced intestinal humoral immune response, enhanced villus morphology, increased ileal digestibility of nutrients, and increased growth of nursery pigs. The beneficial effects were maximized when xylanase and β-glucanase were supplemented at a range of 550 to 800 TXU/kg and 246 to 357 TGU/kg, respectively.

69 Effect evaluation of two sources of zinc at EU-authorized level on growth performance and gut health of piglets with a low or normal birth weight

Abstract This study aimed to investigate the effect of ZnSO4 or a porous form of ZnO, at EU-authorized level, on growth performance, gut health and colon microbiota in low and normal birth body weight (BBW) piglets. At birth, 64 piglets were chosen by their BBW and divided into low BBW (LBBW &amp;lt; 1 kg; 32 piglets) and normal BBW (NBBW &amp;gt; 1 kg; 32 piglets). At weaning, 25 d old (d 0), piglets were allocated into four groups (8 piglets/group, 2 pigs/replicate) and fed a standard diet plus 120 ppm of Zn from ZnSO4 or from a porous source of Zn (HZ - HiZox, Animine, France). Groups were balanced for litter of origin and category of BBW. Piglets were weighed weekly until d 21 post-weaning. The prevalence of diarrhea was determined by the fecal index (percentage of animals with fecal score &amp;gt; 3), and feed intake per pen was recorded daily. On d 9 and d 21, one piglet/replicate was slaughtered, and jejunum tissue was collected for morphology, colon content for microbiota analyzes, and distal jejunum pH was measured. Performance, pH, and morphometric parameters were analyzed using a linear mixed model, fecal index using a generalized linear mixed model with a Poisson distribution, including treatment, class of BBW and their interaction as fixed factors, and the litter as random factor. The statistical analysis on alpha, beta diversity and taxonomic composition were carried out using phyloseq, vegan and microbiomeMarker packages. Piglets in the NBBW group had greater BW throughout the study (P &amp;lt; 0.01). The treatment HZ tended to increase the average daily gain (ADG) at d 9-14 (P &amp;lt; 0.10), and significantly increased the gain to feed ratio (G:F) ratio (P &amp;lt; 0.05) at d 0-14. The HZ-fed piglets had a greater G:F in the overall period (P = 0.04). Additionally, the jejunum pH of the HZ-fed piglets was reduced (P = 0.02) at d 21. An interaction was observed between treatment and BBW for fecal index during d 0-14 and d 0-21 (P &amp;lt; 0.01), in which HZ decreased the index in LBBW. An interaction was also observed in villus height (P = 0.05) and absorptive mucosal surface (P = 0.02) on d 21, that resulted greater in NBBW piglets fed ZnSO4. For Beta diversity, using the Adonis test, BBW category tended to affect the microbial structure at d 9 (R2 = 0.04; P = 0.07) and significantly affected the bacterial structure (R2 = 0.05, P = 0.03) at d 21. At d 21, the NBBW piglets fed ZnSO4 had a greater abundance of Succinivibrio which is able to degrade complex carbohydrates, while the NBBW piglets fed HZ had a greater abundance of Ruminococcus gauvreauii which is linked to the production of short chain fatty acids (SCFAs). The use of HZ seems can be a part of the strategy to replace the pharmacological level of ZnO.

44 Investigation of the nutritional and functional roles of mannanase on intestinal health and growth of newly weaned pigs fed a low-cost formulated diet

Abstract This study aimed to investigate the nutritional and functional roles of mannanase on the intestinal health and growth of newly weaned pigs fed a typical or low-cost formulated diet reducing the inclusion of highly digestible protein ingredients. Twenty-four newly weaned pigs at 6.2 ± 0.5 kg initial body weight (BW) were allotted to three dietary treatments based on a randomized complete block design with sex and BW as blocks. Pigs were fed for 37 d in three phases (P1, P2, and P3) and weighed at the end of each phase. Low-cost formulation was achieved by reducing the use of animal protein (fish meal, poultry meal, and blood plasma) and processed soybean meal whereas increasing the use of soybean meal from 13% in typical diets to 28% in P1, from 15% to 32% in P2, and 20.5% to 35% in P3. Treatments were 1) control diet (CON, typical nursery feed), 2) low-cost formulated diet (LCD), and 3) LCD + mannanase inclusion (0 vs. 100 g/t of Natupulse, BASF, providing 800 Thermostable Mannanase Unit, TMU, per ton of feed). On d 37 of feeding, all pigs were euthanized and jejunal mucosal tissues were taken to analyze intestinal health parameters, and gene expression of tight junction proteins and mucins. Pigs fed LCD had reduced (P &amp;lt; 0.05) average daily gain (ADG) and average daily feed intake (ADFI) during the overall period and increased (P &amp;lt; 0.05) fecal score during P3. The LCD decreased (P &amp;lt; 0.05) malondialdehyde and the gene expression of Zona occludens-1 (ZO-) in the jejunum and increased (P &amp;lt; 0.05) serum insulin. The inclusion of mannanase in LCD tended to increase (P = 0.059) BW at d 37, increased (P &amp;lt; 0.05) ADFI during P1, increased (P &amp;lt; 0.05) ADG and gain to feed ratio (G:F) during the overall period, reduced (P &amp;lt; 0.05) fecal score during the overall period, and decreased (P &amp;lt; 0.05) the viscosity of jejunal digesta. However, no effects were observed on intestinal immune response at d 37. In conclusion, feeding LCD negatively affected growth performance and intestinal health of nursery pigs. The inclusion of mannanase to the LCD (800 TMU/t feed) mitigated the negative effects of lowering feed quality observed in growth performance by reducing digesta viscosity and fecal score.

PSI-21 Optimizing sample size for accurate measurements and prediction equations for intestinal morphology in broiler chickens

Abstract Histological measurements of intestinal morphology, including villi height (VH) and crypt depth (CD), are routinely used to evaluate intestinal health and the nutrient absorptive capacity in monogastric animals. These metrics suffer from significant natural variation, and there is no established standard for the number of unique measurements required to robustly estimate these variables for a given animal. Furthermore, the potential utility of additional variables, including villus width (VW), perimeter (VP), or villus area (VA), has not been explored. Therefore, the objective of the current experiment was to estimate the minimum number of measurements required to control the intrasample variance in each variable and determine if more complex or time-consuming metrics, such as VP and/or VA, can be accurately estimated from linear measurements. A set of 1283 intestinal morphology data, including VH, CD, VW, VP, and VA, measured from 68 intestinal tissues (46 ileum and 22 jejunum) of 4 experiments were used in the current study (Figure 1). Retrospective calculation of the CV for all combinations of N measurements per sample was used to determine the impact of measurement number on the quality of data. Data for the jejunum and ileum were pooled together, and the NLIN procedure of SAS was used to estimate the optimal measurement for intestinal morphology measurements using a quadratic broken line. The VA was estimated using geometric calculation with the assumption that VA equals 0.67 × base width × VH. A paired t-test was conducted to compare measured and calculated VA. The optimal number of measurements to minimize within-sample CV for VH, CD, VW, VP, and VA ranged from 10.6 to 10.8. The VH was highly correlated (P &amp;lt; 0.05) with VP (r = 0.95 and 0.87) and VA (r = 0.77 and 0.63) in the jejunum and ileum, respectively (Table 1 and 2). Additionally, there was a significant difference (P &amp;lt; 0.05) between measured (0.110 µm2) and calculated VA (0.086 µm2). In conclusion, this study shows that measuring 11 villi per sample is required for minimizing variations within the jejunal or ileal tissue in broiler chickens. Also, estimating VA based on idealized geometry was found to be inaccurate, necessitating either direct measurement or more complex prediction equations.

63 Impacts of dietary myristic acids on mucosa-associated microbiota in relation to intestinal health and growth parameters of nursery pigs

Abstract This study investigated the evaluate the role of dietary myristic acid on mucosa-associated microbiota for their relevance to intestinal health and growth performance of nursery pigs. A total of 36 newly weaned pigs [6.6 ± 0.4 kg body weight (BW)] were assigned to 3 treatments (n = 12/treatment) using a randomized complete block design with initial BW and sex as blocks. Pigs were fed for 35 d in 3 phases (10, 10, and 15 d, respectively). Treatments were 1) NC: basal diet; 2) PC: NC + 0.03% bacitracin in all phases; and 3) MA: NC + myristic acid complex at 0.20% in phases 1 and 2 and 0.12% in phase 3. Pigs were euthanized to collect jejunal mucosa and jejunal tissues for microbiome sequencing and measuring intestinal health parameters. Data were analyzed using preplanned contrasts (NC vs. PC and NC vs. MA) in the Proc MIXED procedure and correlation coefficients between microbiota and the intestinal health parameters were determined by the CORR procedure of SAS. The PC increased (P &amp;lt; 0.05) Lactobacillaceae (22.2 to 55.1%) compared with NC. The MA increased (P &amp;lt; 0.05) Veillonellaceae (1.7 to 5.2%), Lachnospiraceae (0.3 to 3.6%), Coriobacteriaceae (0.6 to 1.5%), and Ruminococcaceae (0.5 to 2.1%) and tended to increase (P = 0.072) Bifidobacteriaceae (6.9 to 18.2%) compared with NC. The PC increased (P &amp;lt; 0.05) Simpson (0.72 to 0.89) index and the MA increased (P &amp;lt; 0.05) Chao1 (124.3 to 221.0), Shannon (3.4 to 5.0), and Simpson (0.7 to 0.9) indexes compared with NC. The relative abundance (RA) of Helicobacteraceae was positively correlated with IL-8 (r = 0.63; P &amp;lt; 0.05) and protein carbonyl (r = 0.65; P &amp;lt; 0.05). The RA of Bifidobacteriaceae was negatively correlated with IgG (r = -0.51; P &amp;lt; 0.05), IL-8 (r = -0.58; P &amp;lt; 0.05), TNF-α (r = -0.46; P &amp;lt; 0.05), and protein carbonyl (r = -0.64; P &amp;lt; 0.05) whereas it was positively correlated with average daily gain (ADG; r = 0.51; P &amp;lt; 0.05) and average daily feed intake (ADFI; r = 0.49; P &amp;lt; 0.05). The RA of Lactobacillaceae negatively correlated with IL-8 (r = -0.59; P &amp;lt; 0.05) and protein carbonyl (r = -0.54; P &amp;lt; 0.05) whereas it was positively correlated with ADFI (r = 0.43; P &amp;lt; 0.05) and MDA (r = 0.44; P &amp;lt; 0.05). The RA of Veilonellaceae and Megasphaera sp were negatively correlated (r = -0.49; r = -0.61; P &amp;lt; 0.05) with IgG, respectively. In conclusion, MA mainly increased RA of Bifidobacterium and Megasphaera, whereas PC increased RA of Lactobacillus and Mitsuokella in the jejunal mucosa. The positive modulations of RA of mucosa-associated microbiota by PC and MA were strongly correlated with improved intestinal health and growth of nursery pigs.

181 Effects of Soy protein concentrate replacing conventional soybean meal on intestinal health and growth of newly weaned pigs challenged with F18+ Escherichia coli.

Abstract This study aims to investigate the impact of various levels of soy protein concentrate (SPC) replacing conventional soybean meal (SBM) on various parameters related to intestinal health and growth performance of nursery pigs under F18+ Escherichia coli challenge. Newly weaned pigs [n = 72; initial body weight (BW) = 6.6 ± 0.3 kg] were randomly allotted to 6 treatment groups based on randomized complete block design with initial BW and sex as blocks and fed for 25 d (P1/2 for 11/14 d, respectively). The treatment groups were 1) NC: a diet with conventional SBM, 2) PC: NC with F18+ E. coli challenge at d 7 (2.1 × 1010 CFU); 3) T1: PC with 5% SPC replacing conventional SBM; 4) T2: PC with 10% SPC replacing conventional SBM; 55) T3: NC with 10% SPC replacing conventional SBM, and 6) AGP: PC with 0.25% bacitracin methylene disalicylate. At the end of study, all pigs were euthanized to collect jejunal tissues to measure parameters related to intestinal health. Data were analyzed using MIXED procedure in SAS. The PC decreased (P &amp;lt; 0.05) average daily gain (ADG) and average daily feed intake (ADFI) on d 7 to d 25 (post-challenge) compared with NC. The T2 decreased (P &amp;lt; 0.05) ADG and ADFI on post-challenge period compared with T3. The ADG tended to linearly decrease (P = 0.067) and ADFI linearly decrease (P &amp;lt; 0.05) on post-challenge period with increasing SPC supplementation replacing SBM in the diets. The E. coli challenge increased (P &amp;lt; 0.05) fecal score in PC, and T2 treatments compared with NC, and T3, respectively. The tumor necrosis factor-alpha (TNF-α) in jejunal mucosa linearly increased (P &amp;lt; 0.05) with increasing SPC supplementation replacing SBM in nursery diets under E. coli challenge. The PC decreased (P &amp;lt; 0.05) interleukin-8 (IL-8) compared with NC. The PC (P = 0.078) and T3 (P = 0.099) tended to decrease immunoglobulin A (IgA) compared with NC. The AGP increased (P &amp;lt; 0.05) IgA compared with PC. The AGP increased (P &amp;lt; 0.05) the expression of occludin (OC) compared with PC. The expression of OC tended to linearly decrease (P = 0.099) with increasing SPC supplementation replacing SBM in nursery diets under E. coli challenge. The T2 increased (P &amp;lt; 0.05) villus height to crypt depth ratio (VH:CD) compared with T3. In conclusion, supplementation of SPC replacing SBM in the diet did not alleviate the deleterious effect of E. coli challenge on growth performance nursery pigs, which might be contributed by reduced feed intake. Additionally, supplementation of SPC seems to exacerbate the negative effect of E. coli challenge with increased intestinal inflammation and intestinal disintegration.

Phase-specific outcmes of arginine or branched-chain amino acids supplementation in low crude protein diets on performance, nutrient digestibility, and expression of tissue protein synthesis and degradation in broiler chickens infected with mixed Eimeria spp.

A 35-d study investigated the impact of dietary supplementation with Arginine (Arg) or branched-chain amino acids (BCAA) of broilers receiving low-protein diets whilst infected with mixed Eimeria species. All birds were given the same starter (d0-10) and finisher (d28-35) diets. The 4 grower diets used were a positive control (PC) with adequate protein (18.5%), a low protein diet (NC;16.5% CP), or the NC supplemented with Arg or BCAA. Supplemental AA was added at 50% above the recommended levels. The treatments were in a 4 × 2 factorial arrangement, with 4 diets, with or without Eimeria inoculation on d14. Birds and feed were weighed after inoculation in phases: prepatent (d14-17), acute (d18-21), recovery (d22-28), and compensatory (d29-35). Ileal digesta, jejunum, and breast tissue were collected on d21, 28, and 35. There was no diet × Eimeria inoculation on growth performance at any phase. Infected birds weighed less and consumed less feed (P < 0.05) in all phases. In the prepatent and acute phases, birds on the Arg diets had higher weight gain (P < 0.05) and lower FCR, similar to PC, when compared to NC and BCAA-fed ones. Infection reduced AA digestibility on d21 and 28 (Met and Cys). However, birds that received supplemental AA had higher digestibility (P < 0.05) of their respective supplemented AA on d 21 only. Infected birds had lower (P < 0.05) BO + AT and higher PEPT1 expression on d21. There was a diet × Eimeria interaction (P = 0.004) on gene expression at d28; 4EBP1 genes were significantly downwardly expressed (P < 0.05) in birds fed Arg diet, irrespective of infection. Infected birds exhibited an upward expression (P < 0.05) of Eef2 on d21 and d28 but experienced a downward expression on d35. Supplemental Arg and BCAA had variable effects on growth performance, apparent ileal AA digestibility, and genes of protein synthesis and degradation, but the effect of Arg on promoting weight gain, irrespective of the Eimeria challenge, was more consistent.

Deoxynivalenol Induces Intestinal Epithelial Barrier Damage through RhoA/ROCK Pathway-Mediated Apoptosis and F-Actin-Associated Tight Junction Disruption.

Deoxynivalenol (DON) poses a serious global food safety risk due to its high toxicity and contamination rate. It disrupts the intestinal epithelial barrier, allowing exogenous toxins to enter the circulation and resulting in sepsis and systemic toxicity. In this research, 32 male Kunming mice and Porcine Small Intestinal Epithelial (IPEC-J2) cells were treated with DON at 0-4.8 mg/kg (7 d) and 0-12 μM (24 h), respectively. Histopathological results revealed that DON disrupted the intestinal epithelial barrier, causing apoptosis and tight junction (TJ) injury. Immunofluorescence and protein expression results showed that DON-induced p53-dependent mitochondrial pathway apoptosis and fibrillar actin (F-actin)-associated TJ injury and that the RhoA/ROCK pathway were activated in mice jejunal tissue and IPEC-J2 cells. Pretreatment with RhoA or ROCK inhibitors (Rosin or Y-27632) maintained DON-induced apoptosis and F-actin-associated TJ injury in IPEC-J2 cells. Thus, DON induces damage to the intestinal epithelial barrier through the RhoA/ROCK pathway-mediated apoptosis and F-actin-associated TJ disruption.

Comprehensive analysis of differentially expressed mRNA profiles in chicken jejunum and cecum following Eimeria maxima infection

Coccidiosis, a protozoan disease that substantially impacts poultry production, is characterized by an intracellular parasite. The study utilized 48 one-day-old Horro chickens, randomly divided into the infected (I) and control (C) groups. The challenge group of chickens were administered Eimeria maxima oocysts via oral gavage at 21 days old, and each chicken received 2ml containing 7×104 sporulated oocysts. The total RNAs of chicken jejunum and cecum tissues were isolated from three samples, each from I and C groups. Our study aimed to understand the host immune-parasite interactions and compare immune response mRNA profiles in chicken jejunum and cecum tissues at 4 and 7 days post-infection with Eimeria maxima. The results showed that 823 up- and 737 down-regulated differentially expressed mRNA (DEmRNA) in jejunum at 4 days infection and control (J4I verses J4C), and 710 up- and 368 down-regulated DEGs in jejunum at 7 days infection and control (J7I verses J7C) were identified. In addition, DEmRNA in cecum tissue, 1424 up- and 1930 down-regulated genes in cecum at 4 days infection and control (C4I verses C4C), and 77 up- and 191 down-regulated genes in cecum at 7 days infection and control (C7I verses C7C) were detected. The crucial DEmRNAs, including SLC7A5, IL1R2, GLDC, ITGB6, ADAMTS4, IL1RAP, TNFRSF11B, IMPG2, WNT9A, and FOXF1, played pivotal roles in the immune response during Eimeria maxima infection of chicken jejunum. In addition, the potential detection of FSTL3, RBP7, CCL20, DPP4, PRKG2, TFPI2, and CDKN1A in the cecum during the host immune response against Eimeria maxima infection is particularly noteworthy. Furthermore, our functional enrichment analysis revealed the primary involvement of DEmRNAs in small molecule metabolic process, immune response function, inflammatory response, and toll-like receptor 10 signaling pathway in the jejunum at 4 and 7 days post-infection. Similarly, in the cecum, DEmRNAs at 4 and 7 days post-infection were enriched in processes related to oxidative stress response and immune responses. Our findings provide new insights and contribute significantly to the field of poultry production and parasitology.

Influence of Maternal Nutrition and One-Carbon Metabolites Supplementation during Early Pregnancy on Bovine Fetal Small Intestine Vascularity and Cell Proliferation.

To investigate the effects of nutrient restriction and one-carbon metabolite (OCM) supplementation (folate, vitamin B12, methionine, and choline) on fetal small intestine weight, vascularity, and cell proliferation, 29 (n = 7 ± 1 per treatment) crossbred Angus beef heifers (436 ± 42 kg) were estrous synchronized and conceived by artificial insemination with female sexed semen from a single sire. Then, they were allotted randomly to one of four treatments in a 2 × 2 factorial arrangement with the main factors of nutritional plane [control (CON) vs. restricted feed intake (RES)] and OCM supplementation [without OCM (-OCM) or with OCM (+OCM)]. Heifers receiving the CON level of intake were fed to target an average daily gain of 0.45 kg/day, which would allow them to reach 80% of mature BW by calving. Heifers receiving the RES level of intake were fed to lose 0.23 kg/heifer daily, which mimics observed production responses in heifers that experience a diet and environment change during early gestation. Targeted heifer gain and OCM treatments were administered from d 0 to 63 of gestation, and then all heifers were fed a common diet targeting 0.45 kg/d gain until d 161 of gestation, when heifers were slaughtered, and fetal jejunum was collected. Gain had no effect (p = 0.17) on the fetal small intestinal weight. However, OCM treatments (p = 0.02) displayed less weight compared to the -OCM groups. Capillary area density was increased in fetal jejunal villi of RES - OCM (p = 0.02). Vascular endothelial growth factor receptor 2 (VEGFR2) positivity ratio tended to be greater (p = 0.08) in villi and was less in the crypts (p = 0.02) of the RES + OCM group. Cell proliferation decreased (p = 0.02) in villi and crypts of fetal jejunal tissue from heifers fed the RES + OCM treatment compared with all groups and CON - OCM, respectively. Spatial cell density increased in RES - OCM compared with CON + OCM (p = 0.05). Combined, these data show OCM supplementation can increase expression of VEGFR2 in jejunal villi, which will promote maintenance of the microvascular beds, while at the same time decreasing small intestine weight and crypt cell proliferation.

Lotus Leaf Extract Alleviates Lipopolysaccharide-Induced Intestinal Injury in Mice by Regulating Oxidative Stress and Inflammation.

Inflammatory bowel disease, a disease featured by intestinal epithelial barrier destruction and dysfunction, has been a constant threat to animal health. The primary objective of this research was to assess the impact of the extract derived from lotus leaves (LLE) on lipopolysaccharide (LPS) induced damage to the intestines in mice, as well as to investigate the fundamental mechanism involved. The LLE was prepared using ultrasonic extraction in this experiment, and the LLE total flavonoid content was 117.02 ± 10.73 mg/g. The LLE had strong antioxidant activity in vitro, as assessed by 2, 2-diphenyl-1-picrylhydrazyl, ferric reducing antioxidant power, and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) methods. In the vivo experiment, different doses of LLE (50, 100, and 200 mg/kg) were administered for 2 weeks before LPS treatment in mice. The results revealed that LLE alleviates intestinal tissue damage in LPS-induced mice. In the jejunum tissue, LLE significantly upregulated mRNA and protein expression levels of tight junction proteins, such as ZO-1, occludin, and claudin-1, and decreased the contents of the inflammatory cytokines, interleukin (IL)-1β, IL-6, and tumor necrosis factor-α. Furthermore, the malondialdehyde and lactate dehydrogenase contents increased by LPS in the liver were significantly reduced after administration of LLE, and the total antioxidant capacity, superoxide dismutase, and reduced glutathione decreased by LPS were remarkably increased by LLE. It was found that LLE could relieve LPS-induced oxidative stress by upregulating mRNA and protein expression of Nrf2 and HO-1 in jejunum tissue. In conclusion, LLE alleviates LPS-induced intestinal damage through regulation of the Nrf2/HO-1 signal pathway to alleviate oxidative stress, reducing inflammatory factors and increasing the expression of tight junction proteins in mice.

Integrated Transcriptome and Proteome Analyses of β-Conglycinin-Induced Intestinal Damage in Piglets.

β-conglycinin (β-CG) induces intestinal damage in piglets; however, its regulatory mechanisms are not fully understood. This study aimed to investigate the molecular mechanisms by which β-CG regulates intestinal injury in piglets through downstream genes and proteins. Our findings revealed that β-CG significantly reduced villus height while increasing the crypt depth. In addition, we analyzed the transcriptome and proteome of jejunum tissues after the β-CG treatment. In total, 382 differentially expressed genes (DEGs) and 292 differentially expressed proteins (DEPs) were identified between the treatment and the control groups. The expression levels of DEGs and DEPs were validated by using quantitative reverse transcription polymerase chain reaction (qRT-PCR) and Western blotting, respectively. The findings revealed a consistent correlation between their expression levels and transcriptomic and proteomic data. In addition, Gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses of DEGs and DEPs revealed their enrichment in oxidation-related GOs, as well as in lysosome-related pathways. A protein-protein interaction (PPI) regulatory network was constructed based on the DEPs. The integration of transcriptomic and proteomic analyses identified six genes that were significantly different at both the transcript and the protein levels. This study provides valuable insights into the molecular mechanisms underlying β-CG-induced intestinal injury in piglets.