Ileal Samples Research Articles

Effect of dietary fructooligosaccharide supplementation on internal organs Salmonella colonization, immune response, ileal morphology, and ileal immunohistochemistry in laying hens challenged with Salmonella enteritidis

A study was conducted to evaluate the efficacy of fructooligosaccharides (FOS) in controlling the infection of Salmonella Enteritidis (SE) in White Leghorns. A total of 30 laying hens (white leghorns W-36) were challenged both orally and cloacally with approximately 108 colony-forming units of nalidxic acid resistant SE (SENAR) and divided into 3 treatments: 1) SENAR challenged + 0.0% FOS, 2) SENAR challenged + 0.5% FOS (Nutraflora), and 3) SENAR challenged + 1.0% FOS. SENAR recovery via fecal shedding was measured at 3- and 6-d post-infection (dpi), whereas in the ceca and internal organs, SENAR recovery was measured at 7-d post-infection. In the first experiment, there was a 1.0 log10 and a 1.3 log10 reduction in cecal SENAR by supplementation of FOS at 0.5 and 1.0%, respectively. In the second experiment, there was a 0.6 log10 and a 0.8 log10 reduction in cecal SENAR by supplementation of FOS at 0.5 and 1.0%, respectively. Fecal shedding was significantly lower in 1.0% FOS supplemented groups compared to SENAR challenge 0.0% FOS. There was no significant difference among the 3 treatments on SENAR recovery in liver with gall bladder and ovaries. However, the frequency of positive SENAR in the ovaries (10 to 40%) in SENAR challenge 0.0% FOS was significantly lower than liver with gall bladder (60 to 80%) in both experiments. There was a significant upregulation of toll-like receptor-4 in 1.0% FOS and interferon gamma in both 0.5 and 1.0% FOS. Histologic measurements of ileal villi height and crypt depth were similar across all treatments. Immunohistochemistry analyses of ileal samples showed that immunoglobulin A positive cells increased as FOS concentration increased reaching significance at 1.0% as well as altered cytokine gene expression in the ileum. Further, FOS supplementation also reduced cecal SENAR and feces SENAR levels. Collectively, the results suggest that dietary supplementation with FOS may impair SE pathogenesis while modulating humoral immunity within the gut-associated lymphoid tissue.

Apparent and standardized ileal digestibility of AA and starch in hybrid rye, barley, wheat, and corn fed to growing pigs1

An experiment was conducted to determine the apparent ileal digestibility (AID) of AA and starch and the standardized ileal digestibility (SID) of AA in three varieties of hybrid rye and in one source of barley, wheat, and corn. Seven growing barrows (initial BW = 26.1 ± 2.4 kg) were randomly allotted to a 7 × 7 Latin square design with seven periods and seven experimental diets. Six diets included one of the grains as the sole source of AA, and an N-free diet was used to determine basal endogenous losses of CP and AA. In each period, ileal digesta were collected for 8 h on days 6 and 7 following a 5-d adaptation period. At the conclusion of the experiment, all ingredients, diets, and ileal digesta samples were analyzed for starch, CP, and AA. The AID of starch was greater (P < 0.05) in wheat and corn than in barley or hybrid rye, but all grains had AID values for starch that were above 95%. Wheat and barley contained more CP and indispensable AA than hybrid rye, but hybrid rye contained more indispensable AA compared with corn. The SID of CP and all indispensable AA was greater (P < 0.05) in barley, wheat, and corn than in the three varieties of rye. However, because of the greater concentration of AA in hybrid rye than in corn, the quantities of standardized ileal digestible CP and AA were not different between corn and hybrid rye. In conclusion, hybrid rye has greater concentrations of most AA than corn, but the digestibility of AA in rye is less than in other cereal grains. It is likely that the reason for the reduced SID of AA in rye is that rye contains more fructans and soluble dietary fiber than other cereal grains, which may increase viscosity and reduce the efficiency of endogenous peptidases.

Orientating study about the effects of exocrine pancreatic insufficiency (with or without pancreatic enzyme therapy) on microbiota of ileal and fecal samples – using the animal model of the pancreatic duct ligated, ileo-cecal fistulated minipig

Orientating study about the effects of exocrine pancreatic insufficiency (with or without pancreatic enzyme therapy) on microbiota of ileal and fecal samples – using the animal model of the pancreatic duct ligated, ileo-cecal fistulated minipig

Growth performance and expression of genes encoding enzymes involved in methionine and cysteine metabolism in piglets fed increasing sulphur amino acid to lysine ratio during enterotoxigenic Escherichia coli challenge

A study was conducted to examine the effect of standardised ileal digestible (SID) sulphur amino acids/lysine ratio (SAA/Lys) on performance and expression of methionine adenosyltransferase 1 and 2 alpha (MAT1α and MAT2α), and cystathionine gamma-lyase (CTH) in piglets challenged with an enterotoxigenic Escherichia coli (ETEC). Thirty five [Duroc × (Yorkshire × Landrace)] piglets (6.9 ± 0.5 kg) were randomly assigned to five dietary treatments. The diets were antibiotic free with SID SAA/Lys of 48%, 54%, 60%, 66%, and 72%. Pigs were orally challenged with 6 and 15 mL of ciprofloxacin-resistant ETEC K88+ on days 7 and 10. Blood samples were collected before (BC) and 6, 24, and 48 h after challenge (AC). Body weight gain and feed intake were collected on days 0, 6, and 12 to determine average daily gain (ADG). Gain to feed ratio (G/F) was calculated by dividing ADG by average daily feed intake (ADFI). On day 13, all pigs were euthanized to collect liver and ileal samples to analyse gene expression using real-time polymerase chain reaction. Pigs fed the diet containing SAA/Lys of 66% had the highest ADG, ADFI, and G/F BC. However, ADG, ADFI, and G/F were similar across all ratios AC. Serum tumor necrosis factor alpha concentration at 6 h AC was higher (P &lt; 0.05) than BC and was improved with increasing SAA/Lys. Increasing SAA/Lys quadratically increased (P &lt; 0.01) CTH and MAT1α expression. Ileal expression of CTH and MAT2α were quadratically increased (P &lt; 0.05) with increasing SAA/Lys. In conclusion, SAA/Lys of 60% is suggested to be optimum for piglets to tolerate ETEC pathogenic challenge.

Short communication: Effect of abomasal inorganic phosphorus infusion on phosphorus absorption in large intestine, milk production, and phosphorus excretion of dairy cattle

The objective of the study was to evaluate the effect of inorganic phosphorus (Pi) infusion on P absorption in large intestine, milk production, and phosphorus excretion. Four ruminally and ileally cannulated crossbred cows were used in a 4 × 4 Latin square with 21-d periods. Cows were fed a total mixed ration containing 0.21% P, providing 50% of the cows' P requirement. Cobalt-EDTA was used as marker to measure large intestine digesta flow. On d 13 to 21 of each period, each cow was infused daily with 0, 20.1, 40.2, or 60.3 g of Pi into the abomasum and total collection was conducted on d 18 to 21. Ileal samples were collected every 9 h on d 18 to 21. Feed, digesta, and fecal samples were analyzed for total P and Pi using the molybdovanadate yellow method and blue method, respectively. All data were analyzed using PROC GLIMMIX in SAS 9.3 (SAS Institute Inc., Cary, NC) using contrasts to evaluate linear, quadratic, and cubic effects of Pi infusion dose. Dry matter intake, apparent dry matter digestibility, milk yield, and milk total P were unaffected by Pi infusion. Ileal flow and fecal excretion of total P and Pi increased linearly with increasing infused Pi. In the large intestine, net absorption of TP and Pi was increased linearly with increasing infused Pi. The magnitude of absorption from the large intestine was greater than reflected in current models, raising questions that could be evaluated with longer infusion periods or dietary alteration.

Effect of arabinoxylo-oligosaccharides and arabinoxylans on net energy and nutrient utilization in broilers.

Arabinoxylo-oligosaccharides (AXOS) are hydrolytic degradation products of arabinoxylans (AX) that can be fermented by the gut microbiota, thus potentially displaying prebiotic properties. This study examined the effects of AX and AXOS on net energy (NE) and nutrient utilization in broilers. Ross 308 broilers (n = 90, 30 birds per treatment) were fed wheat-soybean diets supplemented with pure AX, AXOS produced by exposing the AX to xylanase in vitro (AXOS), or AX with xylanase (AX + E) from d 10 to 21. Performance parameters were measured from d 10 to 21. On d 15, 10 birds per treatment were allocated to closed-circuit net energy chambers to assess the impact of AX and AXOS on dietary energy utilization, through assessment of both metabolisable energy (ME) and NE. Ileal and caecal digesta samples were collected on d 21 to determine the effect of AX and AXOS on ileal and total tract dry matter digestibility, ileal digestible energy, digesta pH, short chain fatty acids (SCFA) and microbiota concentration. Feed conversion ratio was numerically the lowest in birds fed the diet supplemented with AXOS, which is 1.26 compared to 1.37 and 1.30 for AX and AX + E, respectively. Ileal dry matter digestibility was higher in birds fed AXOS than those fed AX (P = 0.047). Ileal digestible energy and total tract dry matter digestibility were higher in birds fed AXOS than those fed AX or AX + E (P = 0.004 and P = 0.001, respectively). Birds fed AXOS had higher ME intake (P = 0.049) and nitrogen retention (P = 0.001) and a strong trend of higher NE (P = 0.056), NE intake (P = 0.057) and retained energy (P = 0.054) compared to those fed AX. Ileal total SCFA, lactic and formic acid concentrations were higher in birds fed AXOS than those fed AX (P = 0.011, P = 0.012 and P = 0.023, respectively). Birds fed AXOS or AX + E had higher caecal total SCFA, acetic, butyric and isovaleric acid concentrations compared to those fed AX (P = 0.001, P = 0.004, P = 0.016 and P = 0.008, respectively), and caecal propionic acid concentration was higher in birds fed AX + E than those fed AX (P = 0.050). Ileal and caecal microbiota concentrations were numerically higher and pH was lower in birds fed AXOS and AX + E than those fed AX. Results from this study indicate that feeding AXOS directly is more efficient than AXOS generation in the gastrointestinal tract, and suggest that AXOS has a potential to be an efficacious prebiotic in broiler diets.

Digestibility index marker type, but not inclusion level affects apparent digestibility of energy and nitrogen and marker recovery in growing pigs regardless of added oat bran.

This study was conducted to investigate whether the apparent ileal digestibility (AID) of GE or nitrogen (N) was influenced by inclusion level and type of digestibility index marker (DIM) and inclusion level of oat bran (OB) and whether the apparent total tract digestibility (ATTD) of GE or DIM recovery was influenced by the 3 aforementioned factors and duration of feces collection. Six diets were formulated as a 2 × 3 factorial arrangement with 2 levels of OB (0 or 100 g/kg) and 3 levels of DIM (2.5, 5.0, or 7.5 g/kg). Chromic oxide and titanium dioxide were used as DIM. Both chromic oxide and titanium dioxide were added to the same diet, and their inclusion levels were consistent in each experimental diet. In Exp. 1, 18 barrows fitted with T-cannulas at the distal ileum were used in a triplicate 6 × 2 incomplete Latin Square design with 6 dietary treatments and 2 periods. The ileal digesta were collected for 3 d after 5-d adaptation, and the AID of GE and N were determined by measuring Cr or Ti in diets and ileal digesta samples. In Exp. 2, a total of 72 barrows were used in a randomized complete block design, and the feces were collected for either 3 or 5 d after a 7-d adaptation according to the assignment. Experimental diets were same as Exp. 1. The ATTD of GE was determined by both total collection and index methods, and DIM recovery was determined by measuring Cr or Ti. In Exp. 1, there was no interaction among OB level, DIM level, and DIM type for AID of GE and N. The AID of GE and N determined by Ti were greater (P < 0.05) than Cr regardless of the OB level or DIM level. Neither the OB level nor the DIM level affected the AID of GE or N. In Exp. 2, no interaction among OB level, DIM level, duration of feces collection, and DIM type for ATTD of GE and DIM recovery was observed. The DIM level and duration of feces collection had no effect on ATTD of GE and DIM recovery. The total collection method resulted in greater (P < 0.05) ATTD of GE than Ti, which was greater (P < 0.05) than that determined by Cr. Similarly, the recovery of Ti was greater (P = 0.007) than Cr. Inclusion of 100 g/kg OB decreased (P = 0.003) ATTD of GE but did not affect the recovery of DIM. In conclusion, the AID of GE and N, the ATTD of GE, and the recovery of Cr or Ti were affected by DIM type, but not DIM level; the inclusion of OB had no effect on AID of GE and N, and DIM recovery; and the duration of feces collection had no effect on ATTD of GE and DIM recovery.

Probiotics Strains Modulate Gut Microbiota and Lipid Metabolism in Mule Ducks.

Background:Livestock production should respond to societal, environmental and economic changes. Since 2006 and the ban on antibiotics as growth factors in European Union, the use of probiotics has become widespread and has demonstrated the effect of intestinal microbiota on the performance of farm animals.Objective:The aim of this study was to investigate the effect of supplementation with Lactobacillus salivarius (as a probiotics strain or combined with other strains) on zootechnical performance, metabolic and immune gene expression and intestinal microbiota diversity in mule ducks using high-throughput sequencing and real-time PCR.Method:The mule ducks were reared for 79 days and overfed for 12 days with or without probiotics. Samples were collected at 14 (starting period) and 91 days (end of overfeeding period), 3 hours post feeding.Results:Irrespective of digestive content, age, level of feed intake or supplementation with probiotics, Firmicutes, Proteobacteria and Bacteroidetes were the dominant phyla in the bacterial community in mule ducks. At 14 days, both the ileal and cecal samples were dominated by Firmicutes (in particular the Clostridiales order). Overfeeding induced a shift between Clostridiales and Lactobacillales in the ileal samples whereas in the cecal samples, the relative abundance of Firmicutes decreased. Overfeeding also induced hepatic over-expression of Fatty Acid Synthase (FAS) and of the lipid transporter gene Fatty Acid Binding Protein 4 (FABP4). This increase in lipid metabolism genes is associated with a decrease in inflammatory response.Conclusion:Finally, probiotic supplementation had only a slight impact on gene expression and microbiota diversity, both at 14 days and after overfeeding.

Disruption of Epithelial HDAC3 in Intestine Prevents Diet-Induced Obesity in Mice

Intestinal microbiota modulate metabolism and associate closely with epithelial cells in the intestine. In intestinal epithelial cells (IECs), histone deacetylase 3 (HDAC3) integrates microbiota-derived signals to control intestinal homeostasis. We investigated whether HDAC3 in IECs regulates metabolism and the development of obesity in mice. Adult C57BL/6 (control) mice and mice with constitutive or inducible IEC-specific disruption of Hdac3 (HDAC3ΔIEC mice) were placed on a standard chow or high-fat diet (HFD, 60% kcal from fat). We measured body composition, weight, glucose tolerance, and energy expenditure. IECs were isolated from small intestine and gene expression, and lipid levels were analyzed. HDAC3 levels were determined in 43 pediatric patient ileal biopsy samples and compared with body weight. Control mice fed an HFD gained weight, became obese, and had reduced glucose tolerance with increased serum insulin, whereas HFD-fed HDAC3ΔIEC mice did not develop obesity. Serum levels of triglycerides were reduced in HDAC3ΔIEC mice, and these mice had less liver fat and smaller adipocytes, compared with HFD-fed control mice. HDAC3ΔIEC mice had similar food intake and activity as control mice, but higher energy expenditure because of increased catabolism. IECs from HDAC3ΔIEC mice had altered expression levels of genes that regulate metabolism in response to the microbiota (such as Chka, Mttp, Apoa1, and Pck1) and accumulated triglycerides compared with IECs from control mice. The microbiota-derived short-chain fatty acid butyrate was decreased in obese mice. Butyrate significantly reduced the activity of HDAC3 and increased Pck1 expression in only control IECs. Administration of butyrate to control mice with diet-induced obesity, but not HDAC3ΔIEC mice, led to significant weight loss. Disruption of HDAC3 in IECs of mice after they became obese led to weight loss and improved metabolic profile. Levels of HDAC3 in intestinal biopsy samples correlated with patient weight. We found that epithelial HDAC3 promotes development of diet-induced obesity in studies of mice and that butyrate reduces activity of HDAC3 in IECs to prevent diet-induced obesity. This pathway might be manipulated to prevent or reduce obesity-associated disease.

The Effect of Hydrated Sodium Calcium Aluminosilicate on Fatty Liver and the Composition of the Intestinal Microbiota in Overfed Landes Geese

ABSTRACT Goose fatty liver is a delicious food product and the overfeeding will cause the abnormal physiology of the geese. The objective of this study was to investigate the effect of supplementation with hydrated sodium calcium aluminosilicate (HSCAS) on the fatty liver, ileal and cecal microbiota of Landes geese during overfeeding. Sixty 70-day-old Landes geese (body weight= 3.0 ± 0.05 kg) were randomly divided into three groups, two of which were overfed with whole corn supplemented with or without HSCAS for 20 days when the fatty liver reaches to the maximum size and the negative control group was ad libitum access to the corn basal diet. The intestinal contents of the ileum and cecum from three geese per group were used for high-throughput sequencing. As a result of this study, the HSCAS-treatment led to an increase in relative liver weight (p<0.05) of geese compared with the overfeeding control group. The richness and diversity of the bacterial communities decreased in the ileum and ceca after overfeeding. Overfeeding increased the relative abundance of Firmicutes, especially Lactobacillus, in ileal samples. HSCAS supplementation increased the relative abundance of Lactobacillus, and decreased the relative abundance of Actinobacillus in the ileum and the relative abundance of Erysipelotrichi, Bacteroides and Escherichia in the ceca. Bacterial richness indicators were also increased in samples from ileum and ceca after HSCAS supplementation. In conclusion, dietary HSCAS supplementation promoted liver performance in overfed Landes geese. HSCAS treatment had a beneficial effect on the intestinal microbiota composition in geese during the overfeeding.

Short-chain arabinoxylans prepared from enzymatically treated wheat grain exert prebiotic effects during the broiler starter period

Carbohydrate-degrading multi-enzyme preparations (MEP) are used to improve broiler performances. Their mode of action is complex and not fully understood. In this study, we compared the effect of water-soluble fractions isolated at the pilot scale from wheat grain incubated with (WE) and without (WC) MEP. The fractions were incorporated in a wheat-based diet (0.1% w/w) to feed Ross PM3 broilers and compared with a non-supplemented control group (NC). The body weight gain (BWG), feed intake (FI), and feed conversion ratio (FCR) until d 14 were determined. At d 14, ileal and cecal contents and tissue samples were collected from euthanized animals. The intestinal contents were used to measure the short-chain fatty acids (SCFA) concentration using gas chromatography and to determine the abundance and composition of microbiota using 16S sequencing. Villi length of ileal samples was measured, while L-cell and T-cell densities were determined using immuno-histochemistry. The MEP treatment increased the amount of water-soluble arabinoxylans (AX) and reduced their molecular weight while retaining their polymer behavior. The WE fraction significantly (P < 0.05) increased FI by 13.8% and BWG by 14.7% during the first wk post hatch when compared to NC. No significant effect on FCR was recorded during the trial. The WE increased the abundance of Enterococcus durans and Candidatus arthromitus in the ileum and of bacteria within the Lachnospiraceae and Ruminococcaceae families, containing abundant butyrate-producing bacteria, in the ceca. It also increased the concentration of SCFA in the ceca, decreased the T-lymphocyte infiltration in the intestinal mucosa, and increased the glucagon-like-peptide-2 (GLP-2)-producing L-cell density in the ileal epithelium compared with WC and NC. No significant effects were observed on villi length. These results showed that AX present in the WE fraction altered the microbiota composition towards butyrate producers in the ceca. Butyrate may be responsible for the reduction of inflammation, as suggested by the decrease in T-lymphocyte infiltration, which may explain the higher feed intake leading to improved animal growth.

Lipopolysaccharide-induced maternal inflammation induces direct placental injury without alteration in placental blood flow and induces a secondary fetal intestinal injury that persists into adulthood.

Premature birth complicates 10%-12% of deliveries. Infection and inflammation are the most common etiologies and are associated with increased offspring morbidity and mortality. We hypothesize that lipopolysaccharide (LPS)-induced maternal inflammation causes direct placenta injury and subsequent injury to the fetal intestine. Pregnant C57Bl6 mice were injected intraperitoneally on day 15.5 with 100μg/kg LPS or saline. Maternal serum, amniotic fluid, placental samples, and ileal samples of offspring were obtained assessed for inflammation and/or injury. Maternal placental ultrasounds were performed. Placental DNA was isolated for microbiome analysis. Maternal injection with LPS caused elevated IL-1β, IL-10, IL-6, KC-GRO, and TNF. Placental tissue showed increased IL-1β, IL-6, and KC-GRO and decreased IL-10, but no changes were observed in amniotic fluid. Placental histology demonstrated LPS-induced increases in mineralization and necrosis, but no difference in placental blood flow. Most placentas had no detectable microbiome. Exposure to maternal LPS induced significant injury to the ilea of the offspring. Lipopolysaccharide causes a maternal inflammatory response that is mirrored in the placenta. Placental histology demonstrates structural changes; however, placental blood flow is preserved. LPS also induces an indirect intestinal injury in the offspring that lasts beyond the neonatal period.

Time-course responses of ileal and fecal microbiota and metabolite profiles to antibiotics in cannulated pigs.

We investigated the time-course effects of therapeutic antibiotics on intestinal microbial composition and metabolism in an ileal-cannulated pig model. Sixteen ileal-cannulated piglets (12 ± 0.5kg) were assigned to two groups (n = 8) and fed standard diets with or without antibiotics. At 4days before, and at days 2, 7, and 13 after antibiotic administration, ileal and fecal samples were collected for analysis of microbiota composition via 16S rRNA MiSeq sequencing and metabolites (short-chain fatty acids, biogenic amines, and indole). It was found that Lactobacillus and Bifidobacterium had decreased by an average 2.68-fold and 508-fold in ileum on days 2-13, and by an average 45.08-fold and 71.50-fold in feces on days 7-13 (P < 0.05). Escherichia/Shigella had increased by an average 265-fold in ileum on days 2-13, and by an average 36.70-fold in feces on days 7-13 (P < 0.05). Acetate concentration had decreased in ileum by an average 2.88-fold on days 2-13, and by 1.83-fold in feces on day 7 (P < 0.05). Cadaverine concentration had increased by an average 7.03-fold in ileum on days 2-13, and by an average 9.96-fold in feces on days 7-13 (P < 0.05), and fecal indole concentration had increased by an average 2.51-fold on days 7-13 (P < 0.05). Correlation analysis between significant microbes and metabolites indicated that the antibiotic-induced microbiota shift appeared to result in the changes of intestinal metabolism. In conclusion, antibiotic administration led to dynamic changes in microbial communities and metabolism in ileum and feces, with ileal microbiota being more prone to shift than fecal microbiota.

Tissular growth factors profile after teduglutide administration on an animal model of intestinal anastomosis.

Teduglutide is an enterotrophic analogue of glucagon-like peptide-2, with an indirect and poorly understood mechanism of action, approved for the rehabilitation of short-bowel syndrome. This study aims to analyze the response of tissue growth factors to surgical injury and teduglutide administration on an animal model of intestinal anastomosis. Wistar rats (n = 59) were distributed into four groups: "ileal resection" or "laparotomy", each one subdivided into "postoperative teduglutide administration" or "no treatment"; and sacrificed at the third or the seventh day, with ileal sample harvesting. Gene expression of insulin-like growth factor 1 (Igf1), vascular endothelial growth factor a (Vegfa), transforming growth factor β1 (Tgfβ1), connective tissue growth factor (Ctgf), fibroblast growth factor 2 (Fgf2), fibroblast growth factor 7 (Fgf7), epidermal growth factor (Egf), heparin-binding epidermal-like growth factor (Hbegf), platelet-derived growth factor b (Pdgfb) and glucagon-like peptide 2 receptor (Glp2r)was studied by real-time polymerase chain reaction. Upregulation of Fgf7, Fgf2, Egf, Vegfaand Glp2rat the third day and of Pdgfat the seventh day was verified in the perianastomotic segment. Teduglutide administration was associated with higher fold-change of relative gene expression of Vegfa(3.6 ± 1.3 vs.1.9 ± 2.0, p = 0.0001), Hbegf(2.2 ± 2.3 vs. 1.1 ± 0.9, p = 0.001), Igf1(1.6 ± 7.6 vs. 0.9 ± 0.7, p = 0.002) and Ctgf(1.1 ± 2.1 vs. 0.6 ± 2.0, p = 0.013); and lower fold-change of Tgfβ1, Fgf7and Glp2r. Those results underscore the recognized role of Igf1and Hbegfas molecular mediators of the effects of teduglutide and suggest that other humoral factors, like Vegfand Ctgf, may also be relevant in the perioperative context. Induction of Vegfa, Igf1and Ctgfgene expressions might indicate a favorable influence of teduglutide on the intestinal anastomotic healing.

Human Endogenous Retroviruses: Residues of Ancient Times Are Differentially Expressed in Crohn’s Disease

Background: Eight percent of the human genome consists of human endogenous retroviruses (HERV). These genetic elements are remnants of ancient retroviral germ-line infections. Altered HERV expression is associated with several chronic inflammatory diseases. A physiological role of the HERV-derived proteins syncytin-1 and -2 has been described for the integrity of the human placental cell layer in terms of maintaining feto-maternal tolerance. The aim of this project was to investigate HERV expression in Crohn’s disease (CD) with a further focus on syncytins in the gut. Material and Methods: Seventy-four ileal and colonic tissue samples of CD patients and healthy controls have been investigated for mRNA expression of major HERV groups by a comprehensive microarray screening. The most prominent differences have been validated by qRT-PCR. Immunohistochemistry (IHC), Western Blot (WB) and qRT-PCR were performed for syncytin-1 and -2. Results: HERV microarray screening revealed a distinct expression profile in ileal and colonic tissue, as well as differential expression in CD compared to healthy controls. qRT-PCR validated differential expression of at least 3 HERV-groups in CD. qRT-PCR, IHC and WB showed a tissue-dependent diminished epithelial expression of syncytins in inflamed CD. Conclusion: For the first time, HERV expression has been comprehensively studied in the gut. Between CD and healthy controls we could show a tissue dependent differential HERV expression profile. Notably, we could show that syncytin-1 and -2 are expressed in the epithelial layer in ileal and colonic tissue samples, whereas their diminished tissue-dependent expression in inflamed CD might modulate inflammatory processes at the gut barrier.

Effect of Emblica officinalis(amla) Fruit Powder on Morphometry of the Intestinal Mucosa in Broiler Chicken

To study the effect of dietary supplementation of amla fruit powder on morphometry of intestinal mucosa of broiler chicken. A total of 300 day old commercial broiler chicks were randomly distributed into six treatments having five replicates consisting of ten birds each. The chicks fed with standard basal diet in two different growth phases i.e. starter (0-28d) and finisher (29-42 d). The first group was kept as control (T1) and given basal diet without antibiotic, while second group (T2) was given basal diet with antibiotic. In third (T3), fourth (T4) , fifth (T5) and sixth (T6) groups, basal diet was supplemented with amla fruit powder @ 0.25%, 0.50%, 0.75% and 1%, respectively. Then one bird from each replicate was slaughtered and ileal sample were collected for morphometric analysis. Mean villus height of the birds under different dietary treatments ranged from 984.56 µm to 1196.72 µm. Dietary supplementation of amla fruit powder increased the villi height and significant increment was observed in T6 followed by T5 and T4. Height: Depth of intestinal crypts of the birds under different dietary treatments ranged from 5.44 to 8.12 and significant effect on the ratio of Height: Depth of intestinal crypts was recorded in the 0.75% amla powder supplemented group (T5). The morphometric analysis results in the present study showed that the supplementation of antioxidants increased villus height but decreased crypt depth in broilers.

Development of a clinical prediction score for detection of suspected cases of equine grass sickness (dysautonomia) in France.

Equine grass sickness (EGS) (equine dysautonomia) is a neurodegenerative condition of grazing equines. Pre-mortem diagnosis of EGS is a challenge for practitioners as definitive diagnosis requires ileal/myenteric lymph node biopsies. This study aimed to develop a clinical score that could be used by practitioners to improve the detection of acute or subacute EGS cases in the field. Suspected EGS cases were declared by veterinary practitioners. A case was classified as confirmed positive if ileal or rectal biopsy samples showed neuronal degeneration typical of EGS. A semi-quantitative scoring system, including epidemiological and clinical data, was created to attempt to classify suspected EGS horses into confirmed positive or negative cases. Each variable was weighted based on a boosted regression trees model, while taking into account its clinical relevance. Twenty-eight EGS cases were confirmed by biopsy during the entire study period. The best cut-off value for the score to have a high sensitivity while maximizing specificity was 8, with a sensitivity of 100% and a specificity of 53%. In our dataset, 77% of animals would be correctly classified with this cut-off value of 8. Highest sensitivity was chosen in order to detect the highest number of potential cases. Our score represents an inexpensive and useful tool to aid in the identification of suspected EGS cases in the field and selection for further diagnostics procedures to confirm or rule out the disease. Application of the score to larger populations of animals would be required to further adapt and refine the score.

Intestinal microbiota profiles associated with low and high residual feed intake in chickens across two geographical locations.

Intestinal microbe-host interactions can affect the feed efficiency (FE) of chickens. As inconsistent findings for FE-associated bacterial taxa were reported across studies, the present objective was to identify whether bacterial profiles and predicted metabolic functions that were associated with residual feed intake (RFI) and performance traits in female and male chickens were consistent across two different geographical locations. At six weeks of life, the microbiota in ileal, cecal and fecal samples of low (n = 34) and high (n = 35) RFI chickens were investigated by sequencing the V3-5 region of the 16S rRNA gene. Location-associated differences in α-diversity and relative abundances of several phyla and genera were detected. RFI-associated bacterial abundances were found at the phylum and genus level, but differed among the three intestinal sites and between males and females. Correlation analysis confirmed that, of the taxonomically classifiable bacteria, Lactobacillus (5% relative abundance) and two Lactobacillus crispatus-OTUs in feces were indicative for high RFI in females (P < 0.05). In males, Ruminococcus in cecal digesta (3.1% relative abundance) and Dorea in feces (<0.1% relative abundance) were best indicative for low RFI, whereas Acinetobacter in feces (<1.5% relative abundance) related to high RFI (P < 0.05). Predicted metabolic functions in feces of males confirmed compositional relationships as functions related to amino acid, fatty acid and vitamin metabolism correlated with low RFI, whereas an increasing abundance of bacterial signaling and interaction (i.e. cellular antigens) genes correlated with high RFI (P < 0.05). In conclusion, RFI-associated bacterial profiles could be identified across different geographical locations. Results indicated that consortia of low-abundance taxa in the ileum, ceca and feces may play a role for FE in chickens, whereby only bacterial FE-associations found in ileal and cecal digesta may serve as useful targets for dietary strategies.

Microbial diversity and composition in different gut locations of hyperlipidemic mice receiving krill oil.

Low-dose (LD, 100mgkg-1day-1), moderate-dose (MD, 200mgkg-1day-1), and high-dose (HD, 600mgkg-1day-1) krill oil treatments have a stepwise, enhanced effect on alleviating hyperlipidemia, and 16S rRNA sequencing of the fecal samples demonstrates that krill oil treatment alters microbial communities. Feces may not represent all microbial communities in the gastrointestinal (GI) tract. Therefore, in this study, the stored ileal and colon samples collected from LD and HD groups were sequenced, and the location-specific modulations of microbial communities were observed after krill oil treatments. The 16S rRNA sequencing of the ileal samples showed that the LD and HD groups have similar patterns between control and high-fat diet (HFD) treatments, and six most abundant genera and 40 operational taxonomic units that respond to krill oil treatment were identified. However, the 16S rRNA sequencing of the colon samples showed that LD krill oil shifts the structure from the HFD to that of the control, whereas the HD group was distributed between the control and HFD groups. The corresponding most abundant genera and responsive OTUs totaled 4 and 45, respectively. In conclusion, different gastrointestinal tract locations contain different microbial communities. These results will help to provide a comprehensive understanding of the role of dietary krill oil in modulating the gut microbiota and alleviating hyperlipidemia.

Gene expression and morphological changes in the intestinal mucosa associated with increased permeability induced by short-term fasting in chickens.

Short-term fasting for 4.5 and 9hr has been demonstrated to increase intestinal permeability (IP) in chickens. This study aimed to investigate the effects of 0, 4.5, 9 and 19.5hr fasting on intestinal gene expression and villus-crypt architecture of enterocytes in jejunal and ileal samples. On day 38, Ross-308 male birds were fasted according to their group and then euthanised. Two separate intestinal sections (each 2cm long, jejunum and ileum) were collected. One section was utilised for villus height and crypt depth measurements. The second section was snap-frozen in liquid nitrogen for quantitative polymerase chain reaction (qPCR) analysis of tight junction proteins (TJP) including claudin-1, claudin-3, occludin, zonula occludens (ZO-1, ZO-2), junctional adhesion molecules (JAM) and E-cadherin. Additionally genes involved in enterocyte protection including glucagon-like peptide (GLP-2), heat-shock protein (HSP-70), intestinal alkaline phosphatase (IAP), mammalian target of rapamycin (mTOR), toll-like receptors (TLR-4), mucin (MUC-2), cluster differentiation (CD-36) and fatty acid-binding protein (FABP-6) were also analysed. Normally distributed data were analysed using one-way analysis of variance ANOVA. Other data were analysed by non-parametric one-way ANOVA. Villus height and crypt depth were increased (p<.05) only in the ileum after fasting for 4.5 and 9hr compared with non-fasting group. mRNA expression of claudin-3 was significantly reduced in the ileum of birds fasted for 9 and 19.5hr, suggesting a role in IP modulation. However, all other TJP genes examined were not statistically different from control. Nevertheless, ileal FABP-6 of all fasted groups was significantly reduced, which could possibly be due to reduced bile acid production during fasting.