Villous Height Research Articles

Effect of Probiotic E. coli Nissle 1917 Supplementation on the Growth Performance, Immune Responses, Intestinal Morphology, and Gut Microbes of Campylobacter jejuni Infected Chickens.

ABSTRACTCampylobacter jejuni is the most common cause of bacterial foodborne gastroenteritis and holds significant public health importance. The continuing increase of antibiotic-resistant Campylobacter necessitates the development of antibiotic-alternative approaches to control infections in poultry and in humans. Here, we assessed the ability of E. coli Nissle 1917 (EcN; free and chitosan-alginate microencapsulated) to reduce C. jejuni colonization in chickens and measured the effect of EcN on the immune responses, intestinal morphology, and gut microbes of chickens. Our results showed that the supplementation of 3-week-old chickens daily with free EcN in drinking water resulted in a 2.0 log reduction of C. jejuni colonization in the cecum, whereas supplementing EcN orally three times a week, either free or microencapsulated, resulted in 2.0 and 2.5 log reductions of C. jejuni colonization, respectively. Gavaged free and microencapsulated EcN did not have an impact on the evenness or the richness of the cecal microbiota, but it did increase the villous height (VH), crypt depth (CD), and VH:CD ratio in the jejunum and ileum of chickens. Further, the supplementation of EcN (all types) increased C. jejuni-specific and total IgA and IgY antibodies in chicken’s serum. Microencapsulated EcN induced the expression of several cytokines and chemokines (1.6 to 4.3-fold), which activate the Th1, Th2, and Th17 pathways. Overall, microencapsulated EcN displayed promising effects as a potential nonantibiotic strategy to control C. jejuni colonization in chickens. Future studies on testing microencapsulated EcN in the feed and water of chickens raised on built-up floor litter would facilitate the development of EcN for industrial applications to control Campylobacter infections in poultry.

Replacing fishmeal protein with blended alternatives alters growth, feed utilization, protein deposition and gut micromorphology of humpback grouper, Cromileptes altivelis

This research aims to investigate the effects of different dietary fishmeal (FM) protein replacements by blended alternative proteins on growth, feed utilization, protein deposition and gut micromorphology of humpback grouper, Cromileptes altivelis. Five isoenergetic (14.23 kJ/g, dry matter), isoproteic (48.91 %, dry matter) and isolipidic (10 %, dry matter) experimental diets were formulated, with 0 % (R0), 21 % (R21), 42 % (R42), 63 % (R63) and 84 % (R84) FM protein being replaced by blended alternative proteins which consisted of soybean protein isolate (SPI), poultry by-product meal (PBM), hemoglobin powder (HP). Triplicate groups of fish (mean initial body weight: 6.50 ± 0.17 g, 12 fish/group) were manually fed each diet to apparent satiety twice daily for 8 weeks. Results showed that different FM protein replacements by the blended proteins significantly influenced percentage weight gain (WG %), daily feed intake (DFI), feed conversion ratio (FCR), protein efficiency ratio (PER) and protein productive value (PPV) of fish. Fish fed R84 had the lowest WG %, PER and PPV among all experimental groups. The quadratic broken-line regression analysis of WG % indicated that the maximum FM protein replacement level by the blended proteins was 67.0 % of dry matter. Fish fed R84 had lower expression of growth hormone (GH), growth hormone receptor (GHR), insulin like growth factor-I (IGF-1), target of rapamycin (TOR) and S6 kinase 1 (S6K1) than fish fed other diets. General control nonderepressible 2 (GCN2) and CCAAT enhancer binding protein β (C/EBPβ) showed higher expression in the R84 group compared to those in other groups. The villous height (hV), villous width (wV) as well as enterocyte height (hE) of gut were also affected by different experimental treatments, with the R84 group having lower values compared to other groups. Generally, 67.0 % of dietary FM protein could be replaced by a blend of SPI, PBM and HP proteins without negatively influencing the WG % of humpback grouper.

Quantitative histology as a diagnostic tool for celiac disease in children and adolescents

ObjectiveTo measure the villous height, the crypt depth, and the number of intraepithelial lymphocytes/100 enterocytes of the small intestinal mucosa of children and adolescents with celiac disease; and to classify these findings according to Q- Marsh and Q-histology scales. MethodsRetrospective study of a database from the Department of Pathology of biopsies from the second portion of the duodenum of pediatric patients. According to the histological report, three groups were established: celiac disease at diagnosis (n = 50), controls (n = 26), giardiasis (n = 10). In each biopsy, software (cellSens and Image J) evaluated 5 villous heights, 5 crypt depth and the number of intraepithelial lymphocytes/100 enterocytes. ResultsThe celiac group had the lowest mean villous height (197.83 μm) of all three groups (control = 477.70 μm; giardiasis = 397.04 μm. The celiac group's villous:crypt ratio (0.78) was significantly lower than the control group (1.89). The number of intraepithelial lymphocytes ≥25 was exclusive to the celiac group, with a sensitivity and specificity of 100 %. Only celiac patients were included in types 2 and 3 of the Q-histology classification. ConclusionCeliac disease patients showed shorter villous height than other groups, and the number of intraepithelial lymphocytes ≥25 was the best parameter to differentiate celiac from controls and giardiasis groups. Intraepithelial lymphocytes ≥25/100 enterocytes associated with any degree of villous atrophy, the classic Marsh 3 type, set the histological parameters of celiac disease. Quantitative histology is a valuable tool for diagnosing celiac disease, enabling histological changes in a short time, and the Q-histology scale appears to be more suitable than the Q-Marsh scale.

Effects of embryo injected with ochratoxins A on hatching quality and jejunum antioxidant capacity of ducks at hatching.

Numerous studies have shown that ochratoxins A (OTA) exerts diverse toxicological effects, namely, hepatotoxicity, nephrotoxicity, genotoxicity, enterotoxicity, and immunotoxicity. The main objective of this study was to investigate the influence of embryonic exposure to OTA by different injection times and OTA doses on hatching quality and jejunal antioxidant capacity of ducks at hatching. In total, 480 fertilized eggs were weighed and randomly assigned into a 4 × 4 factorial design including four OTA doses (0, 2, 4, and 8 ng/g egg) on 8, 13, 18, and 23 of embryonic development (E8, E13, E18, and E23). Each treatment included 6 repeats with 5 eggs per repeat. The results showed that the injection time affected the hatching weight (P < 0.0001). The relative length of the jejunum and ileum on E18 and E23 was lower than on E8 and E13 (P < 0.05). Injection time, doses, and their interaction had no effect on jejunum morphology, namely, villous height (Vh), crypt depth (Cd), and villous height/crypt depth ratio Vh/Cd (P > 0.05). The injection time affected the activities of Superoxide dismutase (SOD) (P < 0.0001), total antioxidant capacity (T-AOC) (P < 0.05) and the malondialdehyde (MDA) content (P < 0.0001). The activity of SOD and T-AOC activities in the jejunum of ducklings injected with OTA at the E8 and E13 was lower than that injected at the E18 (P < 0.05). The highest MDA content was observed in ducklings injected with OTA at the E13 (P < 0.05). The injection time (P < 0.0001), OTA doses and their interaction affected the contents of IL-1β (P < 0.05), which significantly increased especially on E13. In conclusion, the embryo injected with ochratoxins A affected the hatching weight, the relative length of jejunum and ileum, decreased the antioxidant capacity and increased the content of proinflammatory cytokine IL-1β of the jejunum.

Effects of alkaline mineral complex water supplementation on growth performance, inflammatory response, and intestinal barrier function in weaned piglets.

The purpose of the present study was to investigate the effects of drinking water alkaline mineral complex (AMC) supplementation on growth performance, intestinal morphology, inflammatory response, immunity, antioxidant defense system, and barrier functions in weaned piglets. In a 15-d trial, 240 weaned piglets (9.35 ± 0.86 kg) at 28 d of age (large white × landrace × Duroc) were randomly divided into two groups: the control (Con) group and the AMC group. Drinking water AMC supplementation improved (P < 0.01) final body weight (BW) and average daily gain (ADG) in weaned piglets compared to the Con group. Importantly, AMC reduced (P < 0.01) the feed-to-gain (F:G) ratio. AMC water improved the physical health conditions of piglets under weaning stress, as reflected by the decreased (P < 0.05) hair score and conjunctival score. Moreover, there was no significant (P > 0.05) difference in relatively small intestinal length, organ (liver, spleen, and kidney) indices, or gastrointestinal pH value in weaned piglets between the two groups. Of note, AMC significantly promoted the microvilli numbers in the small intestine and effectively ameliorated the gut morphology damage induced by weaning stress, as evidenced by the increased (P < 0.05) villous height (VH) and ratio of VH to crypt depth. Additionally, AMC lessened the levels of lipopolysaccharide (LPS, P < 0.01) and the contents of IL1β (P<0.05), and TNF-α (P<0.05) in the weaned piglet small intestine. Conversely, the gut immune barrier marker, secretory immunoglobulin A (sIgA) levels in serum and small intestine mucosa were elevated after AMC water treatment (P < 0.01). Furthermore, AMC elevated the antioxidant mRNA levels of (P < 0.05) SOD 1-2, (P < 0.01) CAT, and (P < 0.01) GPX 1-2 in the small intestine. Likewise, the mRNA levels of the small intestine tight junction factors Occludin (P < 0.01), ZO-1 (P < 0.05), Claudin 2 (P < 0.01), and Claudin 5 (P<0.01) in the AMC treatment group were notably higher than those in the Con group. In conclusion, drinking water AMC supplementation has an accelerative effect on growth performance by elevating gut health by improving intestinal morphology, the inflammatory response, the antioxidant defense system, and barrier function in weaned piglets.

Effect of chronic heat stress on gastrointestinal histology and expression of feed intake-regulatory hormones in broiler chickens

Heat stress (HS) dramatically impairs the growth performance of broiler chickens, mainly as a consequence of reduced feed intake due to the loss of appetite. This study was aimed at evaluating the alterations induced by chronic HS conditions on the morphological and morphometric features of the gastrointestinal (GI) tract and on the expression of some enteroendocrine cells (EECs) involved in the regulation of feed intake in chickens. Three hundred male chickens (Ross 308) were divided into two experimental groups and raised either in thermoneutral environment for the whole fattening period (0–41 days) (TNT group) or subjected to chronic HS conditions (30 °C for 24 h/day) from 35 to 41 days (HS group). Samples of proventriculus, duodenum, jejunum and cecum were collected from 24 broilers (12/group). Haematoxylin-eosin was used for the morphometric evaluations, while immunohistochemistry was applied for the evaluation of EECs expressing ghrelin (GHR), cholecystokinin (CCK), neuropeptide Y (NPY), glucagon-like peptide-1 (GLP-1), and serotonin (5-HT). In the proventriculus, HS reduced total wall thickness and mucous layer height (P ≤ 0.01) as well as mean diameter, circumference, and area of the compound tubular glands (P ≤ 0.001) with respect to TNT. The small intestine of HS birds was characterised by decreased villous height and total thickness (duodenum, P ≤ 0.01; jejunum, P ≤ 0.001), whereas crypt depth and width were reduced only in the jejunum (P ≤ 0.01). HS had negligible effects on the morphological aspects of the cecum. In the proventriculus, an increase in GHR and NPY EECs was observed in response to HS (P ≤ 0.001). Similarly, the small intestine villi of the HS group showed greater GLP-1 (P ≤ 0.05), 5-HT (P ≤ 0.001) and CCK (P ≤ 0.01) EECs. Moreover, the expression of 5-HT EECs was higher in the duodenal (P ≤ 0.01) and jejunal (P ≤ 0.01) crypts of HS birds, whereas GLP-1 and CCK EECs increased only in jejunal crypts (P ≤ 0.05). Finally, 5-HT EEC expression was increased in the cecum of HS group (P ≤ 0.01). In conclusion, these outcomes demonstrate that chronic HS induces morphometric alterations not only in the small intestine but also in a key organ such as the proventriculus. Furthermore, HS conditions affect the presence and distribution of EECs, suggesting that some GI peptides and biogenic amine may be implicated in the regulation of appetite and voluntary feed intake in heat-stressed broiler chickens.

Effects of High Ambient Temperature on Small Intestinal Morphology and Colonic Microbiota in Weaned Piglets.

Simple SummaryThermal stress (TS) is a critical challenge in the swine industry, but knowledge about the influence of TS on intestinal development and microbiota in piglets is still scarce. Thus, the present study investigated the effects of high ambient temperature on growth performance, small intestinal morphology, and colonic microbiota in weaned piglets. Our results showed that high ambient temperature had negative effects on growth performance and colonic microbiota of weaned piglets.A total of 16 crossbred (Duroc × Landrace × Yorkshire) barrows, with an average initial body weight of 8.61 ± 0.24 kg (28 days of age), were randomly allotted into the control group (CON group) and high ambient temperature group (HT group) with 8 replicates per group, 1 pig per replicate. The ambient temperature of the CON group was controlled at 26 ± 1 °C, and the HT group was controlled at 35 ± 1 °C. The study lasted for 21 days. Our results showed that high ambient temperature significantly decreased the average daily feed intake (ADFI) and average daily gain (ADG) of piglets (p < 0.05), and the feed-to-gain ratio was significantly increased (p < 0.05). The liver index, spleen index, and thymus index of piglets in the HT group were significantly decreased (p < 0.05). The villous height (VH) of the duodenum, jejunum, and ileum of piglets in the HT group was significantly decreased (p < 0.05), whereas the crypt depth (CD) of the duodenum was significantly increased (p < 0.05), and the VH-to-CD ratio of the duodenum and ileum was significantly decreased (p < 0.05). The piglets in the HT group showed a higher (p < 0.05) observed-species index, PD whole tree index, and Shannon index, indicating that there was a significant difference in species richness and diversity between the two groups. At the genus level, the piglets in the HT group showed a greater (p < 0.05) percent of Desulfovibrio, Occillibater, and Catenisphaera. HT reduced glycan biosynthesis and metabolism, transport and catabolism, lipid metabolism, amino acids metabolism, secondary metabolites biosynthesis, aging, endocrine system, signaling molecules, and interaction of colon microbiota (p < 0.05), and increased signal transduction, cell motility, transcription, and genetic information processing (p < 0.05).

Albendazole ameliorates inflammatory response in a rat model of acute mesenteric ischemia reperfusion injury

BackgroundAcute mesenteric ischemia is known as a life threatening condition. Re-establishment of blood flow in this condition can lead to mesenteric ischemia reperfusion (MIR) injury which is accompanied by inflammatory response. Still, clear blueprint of inflammatory mechanism underlying MIR injury has not been provided. Interestingly, Albendazole has exhibited notable effects on inflammation and cytokine production. In this study, we aimed to evaluate outcomes of MIR injury following pretreatment with Albendazole with respect to assessment of mesenteric inflammation and ischemia threshold. MethodsMale rats were randomly divided into sham operated, vehicle treated, Albendazole 100 mg/kg and Albendazole 200 mg/kg groups. MIR injury was induced by occlusion of superior mesenteric artery for 30 min followed by 120 min of reperfusion. Samples were utilized for assessment of epithelial survival and villous height. Immunohistochemistry study revealed intestinal expression of TNF-α and HIF-1-α. Gene expression of NF-κB/TLR4/TNF-α/IL-6 was measured using RTPCR. Also protein levels of inflammatory cytokines in serum and intestine were assessed by ELISA method. ResultsHistopathological study demonstrated that pretreatment with Albendazole could ameliorate decline in villous height and epithelial survival following MIR injury. Also, systemic inflammation was suppressed after administration of Albendazole. Analysis of possible participating inflammatory pathway could demonstrate that intestinal expression of NF-κB/TLR4/TNF-α/IL-6 is significantly attenuated in treated groups. Eventually, IHC study illustrated concordant decline in mesenteric expression of HIF-1-α/TNF-α. ConclusionSingle dose pretreatment with Albendazole could ameliorate inflammatory response and enhance ischemia threshold following induction of MIR injury. More studies would clarify existing causality in this phenomenon.

Gut Barrier Disruption Secondary to Radiofrequency-Assisted Liver Parenchyma Resection in a Porcine Model

BackgroundRadiofrequency-assisted liver resection (RF-LR) techniques minimize intraoperative blood loss, while avoiding the Pringle maneuver. Both surgical excision and radiofrequency ablation of liver parenchyma compromise gut barrier function with subsequent bacterial translocation. The present study sought to investigate in a porcine model the impact of two RF-LR techniques on the integrity and inflammatory response of the gut barrier. MethodsTwenty-four pigs were subjected to either (a) partial hepatectomy (PH) employing the “sequential coagulate-cut” technique using a monopolar electrode (SCC group), the one using the bipolar Habib-4X device (group H), or the “crush-clamp” technique (group CC) or (b) sham operation (group Sham). At 48-h post-operation, ileal tissue was excised to be subjected to histopathologic examination, histomorphometric analysis, and immunohistochemical assessment of the mitotic and apoptotic activities and the expression of interleukin-6 (IL-6), tumor necrosis factor-α (TNFα), and nuclear factor-κB (NFκΒ). ResultsHistopathologic score increased in all PH groups, being higher in group SCC, while lower in group H. Villous height decreased in group SCC only. Mitotic index decreased, while apoptotic index increased in all PH groups. An increase in tissue expression score was noted for IL-6 in group CC, for TNFα in all PH groups, being lower in group H compared to group CC, and for NFκB in all PH groups. ConclusionsThe Habib-4X technique for liver resection proved to preserve the integrity of gut barrier, being less injurious in the intestinal mucosa compared to the SCC and CC techniques.

A Multicenter Cross Sectional Study to Evaluate the Role of Duodenal Bulb Biopsy in the Diagnosis of Celiac Disease

Background and Aim: A systemic immune disorder elicited by dietary gluten in people who are genetically predisposed to it is referred to as a celiac disease. D1 had been ruled out as a biopsy site due to concerns about Brunner's glands, peptic duodenitis, gastric metaplasia, and presumed reduced villous height. Due to histological confounding factors, the duodenal bulb (D1) as a biopsy site was traditionally avoided. The present study aimed to assess the duodenal bulb biopsy role in the diagnosis of celiac disease. Methodology: This multicenter cross sectional study was conducted on 108 celiac disease patients in the department of Gastroenterology, Bacha Khan Medical Complex, Swabi, North West General hospital Peshawar and Pir Abdul Qadir Shah Jilani Institute of Medical Science Gambat Khairpur, Sindh from September 2020 to July 2021. All the celiac disease patients were clinically symptomatic and had anti-tissue transglutaminase (anti TTG IgA) positive antibodies. Endoscopically, six mucosal biopsies were taken from each patient, four each from the distal duodenum and two additional biopsy separately labelled was taken from bulb. Modified Marsh grading was used for morphological grading. Results: Of the total 108 Celiac disease patients, 60 (55.6%) were females and 48 (44.4%) were male patients. the overall mean age was 32.62±6.4 years. Prior to diagnosis of Celiac disease, symptoms median duration was 5.4 years with a range of 6 months to 40 years. The most prevalent clinical presentation were iron deficiency anemia 43% (n=46) and chronic diarrhea 32% (n=35) respectively. Other clinical presentation were chronic pain abdomen, infertility, short stature, and cirrhosis. Elevated TTG of 117.1 ± 49.82 U/ml was measured in all the patients with a range of 20 to 250 U/ml. The incidence of similar and different modified Marsh grade was in 66 (61.1%) and 42 (58.9%) respectively at both site. Out of 42 different modified March grade patients, higher mucosal atrophy in bulb and descending duodenum showed higher grade in 16 and 26 patients respectively. Isolated bulb involvement was seen in nine patients. Due to bulb biopsy, Celiac disease diagnosis was correctly done in these patients. Conclusion: The present study supported the villous atrophy and intraepithelial lymphocytosis patchy nature in biopsy specimens of celiac disease. The diagnosis of celiac disease improved with inclusion of duodenal bulb biopsies in our sampling protocol. Celiac disease patchy nature and distribution demonstrated different Marsh grades in different biopsy sites. The celiac disease diagnostic yield is increased by combining the bulb and the descending duodenum biopsy. Keywords: Celiac Disease, Duodenal bulb biopsy, Modified Marsh grade

Microencapsulated probiotic Lactiplantibacillus plantarum and/or Pediococcus acidilactici strains ameliorate diarrhoea in piglets challenged with enterotoxigenic Escherichia coli

Lactiplantibacillus plantarum (strains 22F and 25F) and Pediococcus acidilactici (strain 72N) have displayed antibacterial activity in vitro, suggesting that they could be used to support intestinal health in pigs. The aim of this study was to determine if microencapsulated probiotics could reduce the severity of infection with enterotoxigenic Escherichia coli (ETEC) in weaned pigs. Sixty healthy neonatal piglets were cross-fostered and separated into five groups. Piglets to be given the microencapsulated probiotics received these orally on days 0, 3, 6, 9, and 12. Only piglets in groups 1 and 5 did not receive probiotics: those in groups 2 and 4 received the three microencapsulated probiotic strains (multi-strain probiotic), and piglets in group 3 received microencapsulated P. acidilactici strain 72N. After weaning, the pigs in groups 3-5 were challenged with 5 mL (at 109 CFU/mL) of pathogenic ETEC strain L3.2 carrying the k88, staP, and stb virulence genes. The multi-strain probiotic enhanced the average daily gain (ADG) and feed conversion ratio (FCR) of weaned piglets after the ETEC challenge (group 4), whilst supplementing with the single-strain probiotic increased FCR (group 3). Piglets in groups 3 and 4 developed mild to moderate diarrhoea and fever. In the probiotic-fed piglets there was an increase in lactic acid bacteria count and a decrease in E. coli count in the faeces. By using real-time PCR, virulence genes were detected at lower levels in the faeces of pigs that had received the probiotic strains. Using the MILLIPLEX MAP assay, probiotic supplementation was shown to reduce pro-inflammatory cytokines (IL-1α, IL-6, IL-8, and TNFα), while group 4 had high levels of anti-inflammatory cytokine (IL-10). Challenged piglets receiving probiotics had milder intestinal lesions with better morphology, including greater villous heights and villous height per crypt depth ratios, than pigs just receiving ETEC. In conclusion, prophylactic administration of microencapsulated probiotic strains may improve outcomes in weaned pigs with colibacillosis.

134 Water- and Feed-Based Arginine Impacts on Gut Integrity in Weanling Pigs

Abstract Two hundred and forty newly weaned pigs (PIC, Hendersonville, TN) were used to determine if supplementing with additional arginine either in the water or in the feed, and the combinations thereof, improved intestinal integrity and growth performance in the nursery. Three pigs (21 ± 2 d) per pen for a total of 80 pens, within 4 water lines, were randomly assigned to 1 of 3 arginine-dietary treatments across the 20 pens for a 4×3 factorial. With 4 water treatments: 0, 4, 8 and 12% Arg and 3 dietary treatments: basal level, 1.55 and 1.75% SID Arg. Pigs and feeders were weighed at the d0, d7 (water and diet change), d21 (diet change), and d41. Eighty pigs were euthanized at d7, for ileum evaluation of villus height and crypt depth. Remaining pigs were then taken off the arginine-water treatment portion of the experiment, and fed the diets formulated to contain 3 different levels of dietary arginine. Data were analyzed by pen as repeated measures (SAS 9.4). Feeding additional arginine resulted in an improvement (basal: 20.35, 1.55: 22.22, 1.75: 20.93 kg; P = 0.04) in pig body weight compared with basal. No interaction was detected when using arginine in the water and feed simultaneously, on piglet performance. The use of 8% arginine in the water resulted in a reduction in crypt depth (0:132.54, 4:140.70, 8:117.28, 12:131.99; P &amp;lt; 0.01) and 4% Arg reduced total villous height:crypt depth ratio (0:2.50, 4:2.09, 8:2.56, 12:2.43; P &amp;lt; 0.02). The feeding of arginine resulted in an improvement in final body weight of the pigs; however, there were no differences in intestinal villi when Arg is added to the feed. However, the use of Arg in the water resulted in an improvement in intestinal villi, but no phenotypical change in piglet growth in the nursery.

Potential Effects of Anthocyanin-Rich Roselle (Hibiscus sabdariffa L.) Extract on the Growth, Intestinal Histomorphology, Blood Biochemical Parameters, and the Immune Status of Broiler Chickens.

The potential effects of anthocyanin-rich roselle, Hibiscus sabdariffa L. extract (ARRE) on the growth, carcass traits, intestinal histomorphology, breast muscle composition, blood biochemical parameters, antioxidant activity, and immune status of broiler chickens were evaluated. In the present study, Hibiscus acidified ethanolic extract was reported to have a total anthocyanin content of about 359.3 mg cyanidin 3-glucoside/100 g DW, total polyphenol concentration (TPC) of about 598 mg gallic acid equivalent (GAE)/100 g DW, and total flavonoids (TFs) of about 100 mg quercetin equivalent (QE)/100 g DW. Two-hundred-fifty one-day-old chicks (Ross 308 broiler) (87.85 gm ± 0.32) were randomly allotted to five experimental groups and fed on basal diets supplemented with five levels of ARRE: 0, 50, 100, 200, and 400 mg Kg−1 for 35 days. Dietary ARRE addition did not improve the birds’ growth and carcass traits. Supplemental ARRE increased the n-3 polyunsaturated fatty acids (PUFA) (ω-3) percentage in the breast muscle. Dietary ARRE increased the villous height, and the ARRE100 group raised the villus height to crypt depth ratio. Dietary ARRE increased the immunoexpression of immunoglobulin G (IgG) in the spleen. The serum thyroxine hormone (T4) level was higher in the ARRE200 group. The serum growth hormone level was increased by ARRE addition in a level-dependent manner. According to the broken-line regression analysis, the optimum inclusion level of ARRE was 280 mg Kg−1. All levels of supplemental ARRE decreased the serum triglyceride level. The serum total antioxidant capacity (TAC) was increased in the ARRE100-ARRE400 groups, the serum superoxide dismutase (SOD) level was increased in the ARRE200 group, and the serum malondialdehyde (MDA) level was decreased by increasing the ARRE level. Supplemental ARRE significantly increased the serum levels of lysozymes and IL10. The serum complement 3 (C3) level was increased in ARRE200 and ARRE400 groups. It can be concluded that dietary ARRE addition had many beneficial effects represented by the improvements in the bird’s metabolic functions, blood biochemistry, intestinal morphology, antioxidant activity, immune status, and higher ω-3 content in the breast muscles. However, it had no improving effect on the birds’ growth.

Effects of Exopolysaccharides from Lactiplantibacillus plantarum JLAU103 on Intestinal Immune Response, Oxidative Stress, and Microbial Communities in Cyclophosphamide-Induced Immunosuppressed Mice.

This study investigated the effects of the exopolysaccharide from Lactiplantibacillus plantarum JLAU103 (EPS103) on the intestinal immune response, oxidative stress, intestinal mucosal barrier, and microbial community in cyclophosphamide-induced immune-suppressed mice. The results showed that EPS103 promoted the secretion of cytokines and the generation of secretory immunoglobulin A and mucin-2 in the small intestine of mice, which might be related to the activation of the MAPK pathway. Additionally, EPS103 protected against oxidative stress by activating antioxidation enzymes and Nrf2/Keap1 pathways. It also improved the intestinal physical barrier functions via regulating the ratio of villous height to crypt depth and upregulating the expression of tight-junction proteins. Meanwhile, EPS103 promoted the generation of short-chain fatty acids (SCFAs) and modulated the constituents of gut microbiota. These results suggested that EPS103 may modulate the intestinal immunoresponse relying on the regulation of SCFA production and gut microbiota in immunosuppressed mice, resulting in the activation of systemic immunity.

Dietary supplementation of free or two fat-coated sodium butyrate with varying release times on gastrointestinal development and tight junctions in preweaning Holstein calves

The aim of this study was to determine the effects of free and two fat-coated sodium butyrate (SB) on performance, diarrhea incidence, gastrointestinal tract (GIT) development, tight junctions (TJ), and adherens junctions in preweaning calves. Forty male calves were divided into four treatment groups: control starter (CON), control starter supplemented with free SB (USB), rapid-release SB (RSB), and slow-release SB (SSB) at a SB addition level of 3.0 g/kg (as-fed basis) in a high starch starter (380 g/kg). The calves were fed calf starter for 9 weeks and then the GIT of four calves from each group was harvested. There was no difference in the growth performance and feed efficiency under SB treatments (P > 0.05), although RSB decreased starter intake for the entire period as compared to the CON and USB (P < 0.05). Diarrhea rates and frequencies were lower with the USB and RSB treatments as compared with CON (P < 0.05). The weight and papillae surface area of the rumen in USB were higher than the CON, RSB, and SSB (P < 0.05), although RSB increased the mucosa thickness of the abomasum as compared to the other three groups (P < 0.05). The mucosa thickness of the duodenum in SSB was thicker than USB and RSB (P < 0.05). The villous height and villus height:crypt depth ratio of the ileum was higher in USB than the CON, RSB, and SSB (P < 0.05). Muscularis thickness of the cecum was higher in USB than the RSB and SSB (P < 0.05), and the parameter of the colon was higher in RSB than SSB (P < 0.05). Claudin-1 was increased in the rumen by USB (P < 0.05) and claudin-1 was increased by RSB, and occludin and zonula occludens protein-1 were increased by USB and SSB in the jejunum (P < 0.05). However, occludin was decreased by SSB, and zonula occludens protein-1 was impaired by both RSB and SSB in the colon (P < 0.05). SB treatments had no effect (P > 0.05) on the expression of e-cadherin and β-catenin in the GIT. The incidence of diarrhea was negatively correlated with the protein expression of claudin-1 (P < 0.05) within the jejunum. In conclusion, USB showed more benefits than the RSB and SSB in high starch starter on the diarrhea incidence, GIT development, and TJ in calves, and diarrhea incidence was associated with claudin-1 expression in the jejunum.

P026 Inflammatory bowel disease does not preclude the presence of environmental enteropathy in Zambian adults

Abstract Background Although data is scarce, inflammatory bowel disease (IBD) seems to be increasing in incidence in newly industrialised countries.(1) The environmental factors driving this change are not entirely clear, although it seems likely that changing diet and improving hygiene and sanitation are involved. In China, incidence of IBD is positively associated with gross domestic product.(2) Environmental enteropathy (EE) is a lesion of the small bowel with villous blunting, chronic inflammation and increased gut permeability seen in people living in insanitary conditions in resource-limited settings. We set out to examine whether EE would be present in patients with IBD in Zambia, as these individuals are likely to come from higher socioeconomic strata hence would be expected to live in more sanitised environments. Here we report preliminary data. Methods IBD cases were recruited at time of diagnosis or follow-up. Two groups of controls were recruited: individuals of higher socioeconomic status attending the GI clinic for investigation of symptoms of dyspepsia who had normal upper GI endoscopy were recruited to control group A, and healthy individuals from Misisi compound where studies on EE have been conducted over the past two decades to control group B (low socioeconomic status). All participants gave informed consent and underwent upper GI endoscopy and collection of duodenal biopsies. Haematoxylin- and eosin-stained sections of paraffin-embedded formalin-fixed biopsies were imaged on an Olympus VS120 scanning microscope then assessed for villous height, crypt depth, and epithelial surface area. Demographic information was collected via questionnaire. Results Biopsies were collected from 13 cases, 7 controls in group A and 10 controls in group B. Demographics are shown in Table 1. Cases and controls in group A had a higher number of rooms in the house than controls in group B but there was no significant difference in other environmental factors. Other than crypt depth, which was greater in controls from the lower socioeconomic group, there was no significant difference in morphometric parameters between IBD cases and controls (Table 2). The most severe enteropathy is seen in the biopsy from control group B (Figure 1c) which is made up of participants recruited a socioeconomically deprived district of Lusaka. Figure 1 Conclusion EE can coexist alongside IBD, suggesting that immunological drivers of these conditions are not distinct, and do not support the hygiene hypothesis. There is evidence of some degree of EE in small bowel biopsies from all cases and controls. Normal villous height to crypt depth ratio is likely to be between 3 and 5 (3) whereas medians for villous height:crypt depth in all groups in this study were less than 2.

Age-dependent development in protein digestibility and intestinal morphology in weaned pigs fed different protein sources

Today, weaner diets are optimised using digestibility coefficients obtained from grower-finisher pigs, which may overestimate the digestibility in weaners. The aim of this study was to evaluate the standardised ileal digestibility (SID) of CP and amino acids (AAs), and the intestinal morphology in pigs 0–4 weeks postweaning when fed different protein sources. The experiment included 128 pigs weaned at day 28 and the protein sources were wheat, soybean meal (SBM), enzyme-treated soybean meal (ESBM), hydrothermally treated rapeseed meal (HRSM) and casein. The experiment was conducted as a difference method study including wheat in all diets. Eight pigs were slaughtered on the day of weaning (day 0) and six pigs/treatment were slaughtered at days 7, 14, 21, and 28 postweaning. The SID of CP and AA, as average over the four weeks, was lowest for ESBM and highest for wheat and casein, with SBM and HRSM being intermediate. The SID of CP and AA increased (both linear and quadratic, P < 0.05) over time after weaning. The average SID of CP for all protein sources postweaning was 0.38, 0.59, 0.76, and 0.71 on days 7, 14, 21, and 28, respectively. These differences were significant (P < 0.05) between days 7 and 21, and between days 7 and 28 (P < 0.05), whereas there tended to be a difference between days 7 and 14 (P = 0.06). Protein source did not affect the small intestinal morphology response parameters, whereas time after weaning did. Villous height and villous height to crypt depth ratio differed (P < 0.05) between the days 0 and 7, with shorter villi and a higher ratio at day 7. Crypt depth was not altered between days 0 and 7, or between days 7 and 14. For villi density, crypt density and small intestinal length, a significant increase from days 7 to 14 was observed, but there was no further increase to or difference between days 21 and 28. In conclusion, the low SID of CP in casein on day 7 (0.50) illustrates the challenges related to protein digestion in weanling pigs. The SID of CP and AA is very low during the first two weeks postweaning and time after weaning is more important for protein digestibility, than the source of protein. Fewer mature epithelial cells and less absorptive area in the small intestine in the early postweaning period may partly explain the poor protein digestibility.

Prevalence and pathologic study of porcine epidemic diarrhea in Jeju

From April 2014 to September 2015, 153 piglets from 52 farms in Jeju were diagnosed with porcine epidemic diarrhea (PED). The major PED cases were focused on suckling piglets (144 piglets, 94.1%), particularly in 1-7-day-old piglets. Histopathologically, severe villous atrophy was observed in the small intestine, especially in the jejunum and ileum. The mean villous height to crypt depth ratios of the jejunum and ileum were 1.4:1 and 1.5:1, respectively. The major histopathologic findings of the small intestine were cytoplasmic vacuolation, cuboidalization, squamation, and exfoliation of the mucosal enterocytes in the villi. The cytoplasmic vacuolations in the enterocytes were the most prevalent lesions in the small intestine and were more severe in the ileum than in the jejunum. According to immunohistochemistry methods, the PED virus (PEDV) antigens were presented in the cytoplasms of the enterocytes, and were distributed more prevalently in the ileum than in the jejunum. PEDV antigens were also detected in the colon of 29 piglets (19.5%). Sequence comparison and phylogenetic analysis indicated that 12 PEDV had more than a 98.9% homology with each other. These PEDV strains were highly homologous with the genogroup 2 North American group.

Fermented rye with Agaricus subrufescens and mannan-rich hydrolysate based feed additive to modulate post-weaning piglet immune response

BackgroundThe process of weaning in piglets is often associated with an increased inflammation response in the intestine and compromised intestinal integrity and morphology, favoring a delay in intestinal maturation and a predisposal to diseases. Research has shown the potential of different nutritional strategies to reduce the production of pro-inflammatory cytokines, with the main goal to manipulate health and performance of pigs. Promising examples of nutritional strategies are fungal fermented products and their derivatives which are described to contain several compounds that may play a role in gastrointestinal health and pathogenic bacteria control. Products from Agaricus subrufescens mushroom are reported to contain prophylactic and therapeutic properties including antimicrobial and immunomodulatory properties.ResultsThis study analysed the post-weaning immune status in intestinal tissue and blood of piglets, with the objective to evaluate the gastrointestinal health and immune modulation response induced by a blend of mannan-rich hydrolyzed copra meal and fermented rye with A. subrufescens. Intestinal histomorphology demonstrated a villus height reduction in jejunum and increase in ileum on day 15, while increased villous height in jejunum and ileum on day 30. The results showed that in post-weaning piglets, the feed additive stimulates an immunomodulation effect most evident at 15 days post-weaning, with significant lower expression of cytokines Interferon (IFN) γ, Interleukin (IL) 1α, IL-1β, IL-6, IL-8, IL-10 and Transforming Growth Factor (TGF) β in jejunum, accompanied with an increase in peripheral blood mononuclear cells (PBMC) cytokine gene expression of IL-1β, IL-6, IL-8, IL-10, IL-12p35 (IL-12α), IL-12p40 (IL-12β), Tumor Necrosis Factor (TNF) α, IFN-α, and TGF-β. In piglets fed the feed additive, the quantity of Immunoglobulin (Ig) A producing cells in jejunum, ileum was reduced on day 15 and 30 post-weaning, and on day 30 and 45 post-weaning in colon tissue. Natural Killer (NK) cells count in blood were increased on day 15 post-weaning in the piglets fed the feed additive.ConclusionThis study implies the potential of the blend including mannan-rich hydrolyzed copra meal and fermented rye with A. subrufescens on immune modulation in the intestine of post-weaning piglets.

The Effect of Exogenous Bile Acids on Antioxidant Status and Gut Microbiota in Heat-Stressed Broiler Chickens.

Bile acids are critical for lipid absorption, however, their new roles in maintaining or regulating systemic metabolism are irreplaceable. The negative impacts of heat stress (HS) on growth performance, lipid metabolism, and antioxidant status have been reported, but it remains unknown whether the bile acids (BA) composition of broiler chickens can be affected by HS. Therefore, this study aimed to investigate the modulating effects of the environment (HS) and whether dietary BA supplementation can benefit heat-stressed broiler chickens. A total of 216 Arbor Acres broilers were selected with a bodyweight approach average and treated with thermal neutral (TN), HS (32°C), or HS-BA (200 mg/kg BA supplementation) from 21 to 42 days. The results showed that an increase in average daily gain (P < 0.05) while GSH-Px activities (P < 0.05) in both serum and liver were restored to the normal range were observed in the HS-BA group. HS caused a drop in the primary BA (P = 0.084, 38.46%) and Tauro-conjugated BA (33.49%) in the ileum, meanwhile, the secondary BA in the liver and cecum were lower by 36.88 and 39.45% respectively. Notably, results were consistent that SBA levels were significantly increased in the serum (3-fold, P = 0.0003) and the ileum (24.89-fold, P < 0.0001). Among them, TUDCA levels (P < 0.01) were included. Besides, BA supplementation indeed increased significantly TUDCA (P = 0.0154) and THDCA (P = 0.0003) levels in the liver, while ileal TDCA (P = 0.0307), TLCA (P = 0.0453), HDCA (P = 0.0018), and THDCA (P = 0.0002) levels were also increased. Intestinal morphology of ileum was observed by hematoxylin and eosin (H&E) staining, birds fed with BA supplementation reduced (P = 0.0431) crypt depth, and the ratio of villous height to crypt depth trended higher (P = 0.0539) under the heat exposure. Quantitative RT-PCR showed that dietary supplementation with BA resulted in upregulation of FXR (P = 0.0369), ASBT (P = 0.0154), and Keap-1 (P = 0.0104) while downregulation of iNOS (P = 0.0399) expression in ileum. Moreover, 16S rRNA gene sequencing analysis and relevance networks revealed that HS-derived changes in gut microbiota and BA metabolites of broilers may affect their resistance to HS. Thus, BA supplementation can benefit broiler chickens during high ambient temperatures, serving as a new nutritional strategy against heat stress.