Enterococcus faecium PNC01 isolated from the intestinal mucosa of chicken as an alternative for antibiotics to reduce feed conversion rate in broiler chickens.

BACKGROUND: The development and utilization of probiotics had many environmental benefits for replacing antibiotics in animal production. Bacteria in the intestinal mucosa have better adhesion to the host intestinal epithelial cells compared to bacteria in the intestinal contents. In this study, lactic acid bacteria were isolated from the intestinal mucosa of broiler chickens and investigated as the substitution to antibiotic in broiler production. RESULTS: In addition to acid resistance, high temperature resistance, antimicrobial sensitivity tests, and intestinal epithelial cell adhesion, Enterococcus faecium PNC01 (E. faecium PNC01) was showed to be non-cytotoxic to epithelial cells. Draft genome sequence of E. faecium PNC01 predicted that it synthesized bacteriocin to perform probiotic functions and bacteriocin activity assay showed it inhibited Salmonella typhimurium from invading intestinal epithelial cells. Diet supplemented with E. faecium PNC01 increased the ileal villus height and crypt depth in broiler chickens, reduced the relative length of the cecum at day 21, and reduced the relative length of jejunum and ileum at day 42. Diet supplemented with E. faecium PNC01 increased the relative abundance of Firmicutes and Lactobacillus, decreased the relative abundance of Bacteroides in the cecal microbiota. CONCLUSION: E. faecium PNC01 replaced antibiotics to reduce the feed conversion rate. Furthermore, E. faecium PNC01 improved intestinal morphology and altered the composition of microbiota in the cecum to reduce feed conversion rate. Thus, it can be used as an alternative for antibiotics in broiler production to avoid the adverse impact of antibiotics by altering the gut microbiota.

Comparison of intestinal permeability, morphology, and ileal microbial communities of commercial hens housed in conventional cages and cage-free housing systems.

The gastrointestinal health of poultry can be impacted by a variety of factors including their environment. As egg production moves from conventional cage housing (CC) toward cage-free housing (CF), it is important to understand this impact on intestinal health. This study was conducted to determine if housing type impacted intestinal permeability, morphology, and microbial communities in commercial hens across housing systems. Hens were randomly selected from 2 rooms of CC (n = 25) and CF (n = 25) at a commercial facility. Birds were given fluorescein isothiocyanate dextran (FITC-D) by oral gavage to measure intestinal permeability. Jejunal and ileal samples were collected to evaluate villus height, crypt depth, and their ratio. Ileal contents were collected for bacterial DNA isolation and 16S rRNA gene sequencing. Serum FITC-D was similar between housing type (P = 0.709). Hens housed in the CF had increased jejunal villus height and crypt depth compared with hens from the CC (P < 0.002). Hens from the CC tended to have a greater villus height to crypt depth ratio in both the jejunum and ileum compared with the CF (P = 0.064; P = 0.091, respectively). Microbial community diversity measurements favored hens housed in the CC as ileal contents tended to have increased species richness (P = 0.059), had greater alpha diversity (P = 0.044), and had an increased number of over represented operational taxonomic units (46/64), including Romboutsia sp. (30.80%), Lactobacillus kitasatonis (17.16%), and Lactobacillus aviarius (11.15%). Correlations between microbial communities with intestinal traits identified significant association with the greatest number of correlations with FITC-D and ileal morphology. Many of these correlations identified microbial communities associated with expected traits; thus, providing limited functional data to microbial communities with limited information. The greater number of correlations of ileal morphology with ileal microbial communities suggesting local microbial communities contribute to the intestinal environment distant. In this limited study, several parameters favored hens from CC suggesting an advantage of this system for intestinal health. However, the lower intestinal health parameters observed in CF were not at levels to indicate detrimental effects.

The size of intestines in Vietnamese adults

The length of the small intestine of the Westerner is about 5-9 m long, whereas data on the intestinal length of Vietnamese patients is lacking in the liter-ature. This study aims to determine the size of in-testines in Vietnamese and the difference between fixed cadavers, autopsies and in operative patients. There were 130 subjects examined in this study: intestine from 40 formalin fixed cadavers, 30 autopsies and 60 living patients. The cohort includ-ed 91 males and 39 females, with ages ranging from 18 to 75 years-old and origin from various social levels. Subjects were excluded from this study if there was current or prior GI disease, GI surgery, or any other abdominal surgery. The length of the duodenum was 24.3 ± 1.2 cm in for-malin fixed cadavers and 25.60 ± 1.4 cm in autop-sies. The length of the small intestine was 382.5 ± 45.5 cm in preserved cadavers, 442.3 ± 62.5 cm in autopsies and 556.2 ± 74.4 cm in operative pa-tients. The length of the large intestine was meas-ured to be 132.5 ± 17.6 cm in preserved subjects, 149.3 ± 12.1 cm in autopsies and 156.3 ± 14.5 cm in operative patients. The greatest diameter was the jejunum in autopsies, or 4.1 ± 0.37 cm, and the smallest diameter was the ileum in autopsies, or 2.5 ± 0.30 cm. In Vietnamese, the length of the intestine in surgical patients was the longest; in theformalin-preserved group was the shortest, and in autopsies group was in the average range. The length of the Vietnamese small intestine was short-er than that of the European and American sub-jects. Surgeons need to be aware of variations in intestine length so that resection resulting in small bowel syndrome can be anticipated or avoided

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Spatial relationship between telocytes, interstitial cells of Cajal and the enteric nervous system in the human ileum and colon.

Telocytes (TCs) are recently described interstitial cells, present in almost all human organs. Among many other functions, TCs regulate gastrointestinal motility together with the interstitial cells of Cajal (ICCs). TCs and ICCs have close localization in the human myenteric plexus; however, the exact spatial relationship cannot be clearly examined by previously applied double immunofluorescence/confocal microscopy. Data on TCs and submucosal ganglia and their relationship to intestinal nerves are scarce. The aim of the study was to analyse the spatial relationship among these components in the normal human ileum and colon with double CD34/CD117 and CD34/S100 immunohistochemistry and high-resolution light microscopy. TCs were found to almost completely encompass both myenteric and submucosal ganglia in ileum and colon. An incomplete monolayer of ICCs was localized between the TCs and the longitudinal muscle cells in ileum, whereas only scattered ICCs were present on both surfaces of the colonic myenteric ganglia. TC-telopodes were observed within colonic myenteric ganglia. TCs, but no ICCs, were present within and around the interganglionic nerve fascicles, submucosal nerves and mesenterial nerves, but were only observed along small nerves intramuscularly. These anatomic differences probably reflect the various roles of TCs and ICCs in the bowel function.

Impact of dietary chitosan oligosaccharide and its effects on coccidia challenge in broiler chickens.

1. Two experiments were conducted, the first to determine the optimum inclusion of chitosan oligosaccharide (COS) in broiler diets to support growth performance, digestive functions, intestinal morphology, and immune organs. The second experiment evaluated the immune-protective properties of COS on broiler chickens during coccidia challenge (CC).2. Experiment 1 investigated the effect of graded dietary concentration of COS in the diets of broiler chickens using eight cage replicates for each of the six diets. A corn-soybean meal-based diet was used as the basal diet and supplemented with 0.0, 0.5, 1.0, 1.5, 2.0, or 2.5 g of COS/kg feed to form the six treatments.3. The diet supplemented with 1.0 g COS/kg of feed provided the optimal inclusion level for broiler chickens regarding body weight (BW) gain, jejunal villus height, villus height to crypt depth ratio, and ileal energy digestibility at d 22 of age.4. Experiment 2 investigated the immune-protective properties of COS in broiler chickens during CC. A total of 224 male broiler chicks were randomly assigned to eight replicate cages in a 2 × 2 factorial arrangement of treatments with two COS concentrations (0 or 1 g of COS/kg of diet), with or without CC.5. On d 18 of age, birds in the CC group received twice the recommended coccidia vaccine dose of 30 doses/kg BW.6. Coccidia challenge reduced (P < 0.05) and dietary COS increased (P < 0.05) BW gain, and feed intake. Dietary COS mitigated (P < 0.05) the CC-induced effects on gain:feed. Dietary COS supplementation attenuated the CC-induced effects (P < 0.05) on the expression of occludin genes.7. In conclusion, dietary COS improved performance, and the immune-related beneficial impact of COS supplementation was associated with reduced expression of pro-inflammatory cytokine genes.

Cassava (Manihot esculenta) root chips inclusion in the diets of broiler chickens: effects on growth performance, ileal histomorphology, and cecal volatile fatty acid production.

Exploration and evaluation of alternative feedstuffs in the poultry diets are critical to the industry when the prices of conventional energy sources (i.e., corn and wheat) fluctuate widely and are limited in supply. Cassava root chips (CRC) is rich in energy and fiber, and thus may serve as an alternative energy source in poultry diets, while the fiber in it will undergo microbial fermentation and improve the gut health of broilers. This study evaluated the effects of dietary inclusion of different levels of CRC on the growth performance and gut health parameters of broiler chickens. A total of 180 day-old chicks were allocated randomly to 5 dietary treatments, each with 6 replicate pens. The 5 dietary treatments were the corn-soybean meal-based diets with 0, 12.5, 25, 37.5, and 50% CRC inclusion in the 42-D feeding trial. Ileal histomorphology and cecal volatile fatty acid (VFA) were evaluated as indicators of gut health. Compared to the control group (0% CRC), broiler body weight was not different (P > 0.05) up to 25% CRC inclusion in the starter period (1 to 21 D) and up to 37.5% in the total study period (1 to 42 D). The average daily gain was significantly lower (P < 0.05) at 50% CRC inclusion in the starter period and for the total study period. Feed conversion ratio did not differ (P > 0.05) up to 37.5% CRC inclusion in the starter diet, and up to 50% CRC inclusion in the finisher diet. Villus height, crypt depth, their ratio, and villus surface area were not different among the treatments (P > 0.05). Total VFA and acetate production increased with the increasing dietary inclusion of CRC from 25 to 50%. In conclusion, CRC can be used to replace conventional energy feedstuff (like corn) up to 25 and 37.5% in starter and finisher diets, respectively. Use of CRC in diets may benefit broiler chickens' production by reducing feed costs and contribute to improving gut health.

Response of broiler chickens to xylanase and butyrate supplementation.

A 2 × 2 factorial experiment was used to evaluate the effect of xylanase and sodium butyrate supplementation on performance, intestinal fermentation, histology, and morphometry in broiler chickens. A total of 384 Ross 308 broiler chicks (1-day-old) were allocated to 4 experimental treatments: CTR (control diet), XYL (CTR diet with 16,000 BXU/kg of xylanase), BUT (CTR diet with 1 kg/t sodium butyrate), and XYL+BUT (CTR diet plus xylanase and sodium butyrate). Each treatment had 8 replicates of 12 animals. Starter and grower diets, based on wheat and soybean meal, and water were available ad libitum. Body weight gain and feed intake were measured from 0 to 42 D, and feed conversion ratio corrected for mortality (FCR) was calculated. The profile of short-chain fatty acids in the duodenum, jejunum, ileum, and ceca digesta on days 21 and 42 was analyzed in addition to the relative weights of the different portions of the gastrointestinal tract (GIT). The villus height (VH), crypt depth (CD), and villus to crypt (VH: CD) ratio from the ileal tissue on day 42 were also evaluated. Statistical comparisons were performed using a 2-way ANOVA. Xylanase supplementation improved 42-D FCR by 5 points (P = 0.006), while butyrate did not affect 42-D FCR. On day 21, birds fed butyrate had heavier total GIT (P = 0.024), duodenum (P < 0.001), and jejunum (P = 0.025). Xylanase did not influence the relative weights in any intestinal section except the crop which was smaller in xylanase supplemented birds. At day 42, the VH: CD ratio was increased with sodium butyrate (P = 0.005). Supplementation of broiler diets with xylanase improved performance but had little effect on intestinal measures, whereas sodium butyrate influenced many of the intestinal indices with no consequence on animal performance.

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

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

Mucosal homeostasis is altered in the ileum of gnotobiotic mice.

BACKGROUND: The microbiome of the gastrointestinal tract is a vast collection of microorganisms implicated in numerous aspects of normal physiology and disease pathogenesis. The use of gnotobiotic mouse models, with single or specific communities of microbes comprising the microbiome, can enhance our understanding of the microbiome-host relationship. We hypothesized that gnotobiotic mice would exhibit differences in mucosal homeostasis when compared with mice with conventional flora (CF). MATERIALS AND METHODS: Single-organism gnotobiotic mice were generated containing Escherichia coli MG1655, Akkermansia muciniphila, Bacteroides eggerthii, and Clostridium symbiosum, representing four of the major phyla present in the gastrointestinal tract. Distal ileal segments were harvested from adult mice, and histologic sections were H&E stained and used to measure villus height and crypt depth. Immunohistochemistry was performed with Ki67 and TUNEL as markers of proliferation and apoptosis, respectively. RESULTS: When compared to the ileum from CF mice, the ileum from all groups of gnotobiotic mice had significant increases in nearly all measured parameters. In addition, significant differences were seen among certain gnotobiotic groups for villus height, crypt depth, and apoptosis. CONCLUSIONS: Single-organism gnotobiotic mice demonstrate enhanced morphometric parameters compared with mice with CF and show differences in growth patterns among bacterial species. These findings suggest unique interactions between individual bacteria and the host animal which hold potential for future therapeutic strategies aimed at mucosal restoration. The mechanisms involved in this process therefore warrant further study.

Evaluation of an insect meal of the Black Soldier Fly (Hermetia illucens) as soybean substitute: Intestinal morphometry, enzymatic and microbial activity in laying hens.

This research investigated the ileum morphometry and enzymatic activity, the caecal volatile fatty acid production and the apparent nutrient digestibility in laying hens fed a Hermetia illucens larvae meal (HILM) as a complete replacement of diet soybean meal (SBM). The hens fed HILM exhibited a lower live weight (P<0.05) and a higher incidence of the full digestive tract (P<0.05) than the SBM group. In the duodenum, the maltase exhibited a higher (P<0.05) activity in the HILM group while the intestinal alkaline phosphatase (IAP) had a higher (P<0.05) activity in the SBM group. In the ileum, the maltase and saccarase had a higher activity in the HILM hens (P≤0.01) while the IAP and ɤ glutamil transferase had a higher activity in the SBM group (P<0.05 and P<0.01, respectively). The HILM group showed a higher (P<0.05) villi height in the duodenum, while the opposite happened in the jejunum and the ileum. Only in the ileum the crypt depth resulted higher (P<0.05) in the HIML group than in the SBM. The higher production of acetate (P<0.05) and butyrate (P<0.01) affected the total production of volatile fatty acids of the HILM group. The coefficient of apparent digestibility of dry and organic matter as well as of crude protein was higher (P<0.05) in SBM group. The total replacement of SBM with HILM in laying hens diet from 24 to 45weeks of age resulted in a higher caecal production of butyric acid while the enzymatic activities of brush border membrane were partially reduced.

Feeding value of field beans (Vicia faba L. var. minor) with and without enzyme containing tannase, pectinase and xylanase activities for broilers.

Effects of field beans with various tannin content and exogenous enzyme mixture containing tannase, pectinase and xylanase activities on N-corrected dietary apparent metabolisable energy (AMEn), coefficients of dry matter (DMR) and nitrogen retention (NR), fat digestibility, gastrointestinal tract (GIT) development, jejunal villus morphometry, ileal digesta viscosity and sialic acid were examined. Birds' growth performance and energy conversion ratio (ECR) were also measured. Birds were fed one of eight mash diets. The Control diet contained as major ingredients wheat (400 g/kg) and soybean meal (SBM) (127 g/kg and 221 g crude protein/kg and 12.83 MJ AMEn/kg. To reduce nutrient density, the Control diet also contained washed sand at 119 g/kg. Another three diets containing 300 g/kg of each of three experimental field bean cultivar samples in replacement for SBM and sand were also mixed. Each diet was fed to nine pens with two male Ross 308 broilers. Diets high in tannin had low AMEn, ECR, DMR and NR (p < 0.001). Feeding field beans increased (p < 0.001) the weights of the pancreas and the proventriculus and gizzard (PG) of the birds. Supplementing diets with the enzyme mixture improved (p < 0.001) feed conversion efficiency, AMEn and all nutrient utilisation coefficients despite the tannins in diets. The enzyme mixture reduced ileal digesta viscosity (p < 0.001) and the weight of pancreas, total GIT and PG (p < 0.05) of the birds. It can be concluded that the feeding value of field beans with different tannin contents may vary when fed to broilers. The supplementation of the enzyme mixture improved the feeding value of diets for broilers. The beneficial effect of the addition of the enzyme mixture seems to be mediated through reduced ileal digesta viscosity and improved nutrient availability.

Effects of different dietary tryptophan : lysine ratios and sanitary conditions on growth performance, plasma urea nitrogen, serum haptoglobin and ileal histomorphology of weaned pigs.

A total of 180 mixed-sex pigs (Duroc × (Yorkshire × Landrace); average initial body weight of 7.36 ± 0.2 kg) weaned at 21 ± 1 days were fed corn-soybean meal-wheat-based diets to determine the optimal standardized ileal digestible (SID) tryptophan to lysine ratio (Trp : Lys) in a 2 × 5 factorial arrangement (two sanitary conditions: clean (CL) and unclean (UCL), and five dietary treatments (SID Trp : Lys (16, 18, 20, 22 and 24%)). In each sanitary condition, blood was collected on days 0 and 14 to determine plasma urea nitrogen and on day 14, ileal tissue (one pig per pen) was collected for the measurement of gut morphology. Pigs kept under UCL conditions had lower growth rate (P < 0.05) than under CL conditions. Under CL conditions, the estimated optimal SID Trp : Lys for average daily gain (ADG) was 19.7% whereas under UCL conditions these values were 20.5% and 19.0% for ADG and gain-to-feed ratio, respectively. Under CL conditions, increasing SID Trp : Lys reduced (linear, P = 0.05) plasma urea nitrogen concentration but had no effect (P > 0.10) on villous height (VH), crypt depth ( CD) and VH : CD. In conclusion, an SID Trp : Lys to optimize ADG for pigs raised under UCL conditions was higher (4%) than CL conditions.

Patient-Specific Bacteroides Genome Variants in Pouchitis.

A 2-year longitudinal microbiome study of 22 patients who underwent colectomy with an ileal pouch anal anastomosis detected significant increases in distinct populations of Bacteroides during 9 of 11 patient visits that coincided with inflammation (pouchitis). Oligotyping and metagenomic short-read annotation identified Bacteroides populations that occurred in early samples, bloomed during inflammation, and reappeared after antibiotic treatment. Targeted cultivation of Bacteroides isolates from the same individual at multiple time points and from several patients detected subtle genomic changes, including the identification of rapidly evolving genomic elements that differentiate isogenic strains of Bacteroides fragilis from the mucosa versus lumen. Each patient harbored Bacteroides spp. that are closely related to commonly occurring clinical isolates, including Bacteroides ovatus, B. thetaiotaomicron, B. vulgatus, and B. fragilis, which contained unique loci in different patients for synthesis of capsular polysaccharides. The presence of unique Bacteroides capsular polysaccharide loci within different hosts and between the lumen and mucosa may represent adaptations to stimulate, suppress, and evade host-specific immune responses at different microsites of the ileal pouch. IMPORTANCE: This longitudinal study provides an opportunity to describe shifts in the microbiomes of individual patients who suffer from ulcerative colitis (UC) prior to and following inflammation. Pouchitis serves as a model for UC with a predictable incidence of disease onset and enables prospective longitudinal investigations of UC etiology prior to inflammation. Because of insufficient criteria for predicting which patients will develop UC or pouchitis, the interpretation of cross-sectional study designs suffers from lack of information about the microbiome structure and host gene expression patterns that directly correlate with the onset of disease. Our unique longitudinal study design allows each patient to serve as their own control, providing information about the state of the microbiome and host prior to and during the course of disease. Of significance to the broader community, this study identifies microbial strains that may have genetic elements that trigger the onset of disease in susceptible hosts.

Impact of acute undernutrition on growth, ileal morphology and nutrient transport in a murine model.

Undernutrition represents a major public health challenge for middle- and low-income countries. This study aimed to evaluate whether a multideficient Northeast Brazil regional basic diet (RBD) induces acute morphological and functional changes in the ileum of mice. Swiss mice (∼25 g) were allocated into two groups: i) control mice were fed a standard diet and II) undernourished mice were fed the RBD. After 7 days, mice were killed and the ileum collected for evaluation of electrophysiological parameters (Ussing chambers), transcription (RT-qPCR) and protein expression (western blotting) of intestinal transporters and tight junctions. Body weight gain was significantly decreased in the undernourished group, which also showed decreased crypt depth but no alterations in villus height. Electrophysiology measurements showed a reduced basal short circuit current (Isc) in the undernourished group, with no differences in transepithelial resistance. Specific substrate-evoked Isc related to affinity and efficacy (glutamine and alanyl-glutamine) were not different between groups, except for the maximum Isc (efficacy) induced by glucose. Transcription of Sglt1 and Pept1 was significantly higher in the undernourished group, while SN-2 transcription was decreased. No changes were found in transcription of CAT-1 and CFTR, while claudin-2 and occludin transcriptions were significantly increased in the undernourished group. Despite mRNA changes, SGLT-1, PEPT-1, claudin-2 and occludin protein expression showed no difference between groups. These results demonstrate early effects of the RBD on mice, which include reduced body weight and crypt depth in the absence of significant alterations to villus morphology, intestinal transporters and tight junction expression.

Quantitative morphometric analysis of the myenteric nervous plexus ganglion structures along the human digestive tract.

BACKGROUND/AIM: All the functions of the digestive system are controlled, guided and initiated by the autonomic nervous system. A special part of this system placed in the wall of the gastrointestinal tract is known as the enteric or metasympathetic nervous system. The aim of this study was to analyse myenteric nervous plexus in different parts of the digestive tract. METHODS: We examined the myenteric nervous plexus of the esophagus, stomach, duodenum, jejunum, ileum, transverse colon and rectum in tissue samples taken from 30 cadavers of persons aged 20-84 years. After standard histological processing sections were stained with hematoxylin-eosin, cresyl violet (CV) and AgNO3 method. Multipurpose test system M42 was used in morphometric analysis. The results were analyzed by t-test and analysis of variance. RESULTS: The number of neurons per cm² surface was the lowest in the esophagus (2.045 ± 310.30) and the largest in the duodenum (65,511 ± 5,639). The statistical processing showed significant differences (P < 0.001) in the number of neurons between the esophagus and all other parts of the digestive tract. The maximal value of the average surface of the myenteric nervous plexus neurons was observed in the esophagus (588.93 ± 30.45 µm²) and the lowest in the stomach (296.46 ± 22.53 µm²). CONCLUSION: There are differences in the number of ganglion cells among different parts of the human digestive tract. The differences range from a few to several tens of thousands of neuron/cm2. The myenteric nervous plexus of the esophagus was characterized by a significantly smaller number of neurons but their bodies and nuclei are significantly larger compared to other parts of the digestive tract.

6ß-N-Heterocyclic Substituted Naltrexamine Derivative BNAP: A Peripherally Selective Mixed MOR/KOR Ligand.

The 6ß-N-heterocyclic naltrexamine derivative, NAP, has been demonstrated to be a peripherally selective mu opioid receptor modulator. To further improve peripheral selectivity of this highly potent ligand, its pyridal ring was quaterinized with benzyl bromide to produce BNAP. In radioligand binding assay, the Ki of BNAP for MOR was 0.76 ± 0.09 nM and was >900-fold more selective for MOR than DOR. The Ki for KOR was 3.46 ± 0.05 nM. In [(35)S]GTPγS ligand stimulated assay, BNAP showed low agonist efficacy with 14.6% of the maximum response of DAMGO with an EC50 of 4.84 ± 0.6 nM. However, unlike its parent compound NAP, BNAP displayed partial agonist activity at KOR with % maximum response at 45.9 ± 1.7% of U50,488H. BNAP did not reverse morphine-induced antinociception when administered subcutaneously but did antagonize when administered intracerebroventricularly. BNAP antagonized morphine-induced contractions of the circular muscle in mice colon. BNAP inhibition of field-stimulated contractions in longitudinal muscle strips for the guinea-pig ileum were also blocked by nor-BNI, a kappa opioid receptor antagonist. BNAP induced inhibition of acetic acid induced abdominal stretching in chronic morphine treated mice. These findings suggest that BNAP is a dual MOR antagonist/KOR agonist and may have functional use in irritable bowel patients.

Effects of low-protein diets supplemented with indispensable amino acids on growth performance, intestinal morphology and immunological parameters in 13 to 35 kg pigs.

The objective of this study was to determine if a moderate or high reduction of dietary CP, supplemented with indispensable amino acids (IAA), would affect growth, intestinal morphology and immunological parameters of pigs. A total of 40 barrows (initial BW=13.50±0.50 kg, 45±2 day of age) were used in a completely randomized block design, and allocated to four dietary treatments containing CP levels at 20.00%, 17.16%, 15.30% and 13.90%, respectively. Industrial AA were added to meet the IAA requirements of pigs. After 4-week feeding, blood and tissue samples were obtained from pigs. The results showed that reducing dietary CP level decreased average daily gain, plasma urea nitrogen concentration and relative organ weights of liver and pancreas (P<0.01), and increased feed conversion ratio (P<0.01). Pigs fed the 13.90% CP diet had significantly lower growth performance than that of pigs fed higher CP at 20.00%, 17.16% or 15.30%. Moreover, reducing dietary CP level decreased villous height in duodenum (P<0.01) and crypt depth in duodenum, jejunum and ileum (P<0.01). The reduction in the dietary CP level increased plasma concentrations of methionine, alanine (P<0.01) and lysine (P<0.05), and decreased arginine (P<0.05). Intriguingly, reducing dietary CP level from 20.00% to 13.90% resulted in a significant decrease in plasma concentration of IgG (P<0.05), percentage of CD3+T cells of the peripheral blood (P<0.01), also down-regulated the mRNA abundance of innate immunity-related genes on toll-like receptor 4, myeloid differentiation factor 88 (P<0.01) and nuclear factor kappa B (P<0.05) in the ileum. These results indicate that reducing dietary CP level from 20.00% to 15.30%, supplemented with IAA, had no significant effect on growth performance and had a limited effect on immunological parameters. However, a further reduction of dietary CP level up to 13.90% would lead to poor growth performance and organ development, associated with the modifications of intestinal morphology and immune function.

Effect of different doses of coated butyric acid on growth performance and energy utilization in broilers.

We recently applied four dietary treatments in experiments I and II to determine the effect of protected calcium butyrate (BP) on growth performance and nutrient digestibility in broiler chickens. A group of one-day-old male Ross 308 broiler chicks (total 960, 480 per trial) were used in the study. In experiment I, the basal diets were fed with protected BP inclusion (0.2, 0.3, or 0.4 g/kg of finished feed) (BP) or without (C). In experiment II, 4 different diets were tested: 1) basal diet with no supplementation (C), 2) basal diet supplemented with protected BP (0.3 g/kg) (BP), 3) basal diet supplemented with avilamycin (6 mg/kg, active substance) a common antibiotic growth promoter (AGP) (Av), and 4) basal diet supplemented with the combination of both avilaymicin and BP. In experiment I, considering the entire study period, the use of BP improved feed conversion ratio (P<0.05) irrespective of the dose. Apparent total tract crude fat digestibility and apparent metabolizable energy corrected for nitrogen (AMEN) were improved after BP supplementation (P<0.05). In experiment II, A or AB diets improved (P<0.05) body weight gain compared to the control treatment. The diets Av, BP, and AvB improved (P<0.05) feed conversion ratio compared to the control treatment. Birds from the treatment diet were characterized by having the thickest mucosa (P<0.05). On days 14, 35, and 42, the use of AB diets improved AMENcontent compared to the control treatment (P<0.05). The apparent ileal digestibility of amino acid data showed that Av or AvB treated birds were characterized by higher Asp, Glu, Cys, Gly, and Ala ileal digestibility than the control animals (P<0.05). The use of Av, BP, or AvB increased ileal digestibility of Thr, Ser, and Pro (P<0.05). There is an indication that BP, alone or in combination with avilamycin, improve the digestion and absorptive processes and consequently birds performance results.

Impact of acute undernutrition on growth, ileal morphology and nutrient transport in a murine model

Undernutrition represents a major public health challenge for middle- and low-income countries. This study aimed to evaluate whether a multideficient Northeast Brazil regional basic diet (RBD) induces acute morphological and functional changes in the ileum of mice. Swiss mice (∼25 g) were allocated into two groups: i) control mice were fed a standard diet and II) undernourished mice were fed the RBD. After 7 days, mice were killed and the ileum collected for evaluation of electrophysiological parameters (Ussing chambers), transcription (RT-qPCR) and protein expression (western blotting) of intestinal transporters and tight junctions. Body weight gain was significantly decreased in the undernourished group, which also showed decreased crypt depth but no alterations in villus height. Electrophysiology measurements showed a reduced basal short circuit current (Isc) in the undernourished group, with no differences in transepithelial resistance. Specific substrate-evoked Isc related to affinity and efficacy (glutamine and alanyl-glutamine) were not different between groups, except for the maximum Isc (efficacy) induced by glucose. Transcription of Sglt1 and Pept1 was significantly higher in the undernourished group, while SN-2 transcription was decreased. No changes were found in transcription of CAT-1 and CFTR, while claudin-2 and occludin transcriptions were significantly increased in the undernourished group. Despite mRNA changes, SGLT-1, PEPT-1, claudin-2 and occludin protein expression showed no difference between groups. These results demonstrate early effects of the RBD on mice, which include reduced body weight and crypt depth in the absence of significant alterations to villus morphology, intestinal transporters and tight junction expression.

Colostrum quality affects immune system establishment and intestinal development of neonatal calves.

The first meal of a neonatal calf after birth is crucial for survival and health. The present experiment was performed to assess the effects of colostrum quality on IgG passive transfer, immune and antioxidant status, and intestinal morphology and histology in neonatal calves. Twenty-eight Holstein neonatal male calves were used in the current study, 24 of which were assigned to 1 of 3 treatment groups: those that received colostrum (GrC), transitional milk (GrT, which was obtained after the first milking on 2-3 d after calving), and bulk tank milk (GrB) only at birth. The 4 extra neonatal calves who were not fed any milk were assigned to the control group and were killed immediately after birth to be a negative control to small intestinal morphology and histology detection. Calves in GrC gained more body weight than in GrT, whereas GrB calves lost 0.4 kg compared with the birth weight. Serum total protein, IgG, and superoxide dismutase concentrations were highest in GrC, GrT was intermediate, whereas GrB was the lowest on d 2, 3, and 7. Apparent efficiency of absorption at 48 h, serum complement 3 (C3), and complement 4 (C4) on d 2, 3, and 7 in GrB was low compared with GrC and GrT. On the contrary, malondialdehyde on d 7 increased in GrB. Calves in GrC had better villus length and width, crypt depth, villus height/crypt depth (V/C) value, and mucosal thickness in the duodenum, jejunum, and ileum, whereas GrT calves had lower villus length and width, crypt depth, and mucosal thickness than those fed colostrum. Villi of calves in GrB were nonuniform, sparse, severely atrophied, and apically abscised, and Peyer's patches and hydroncus were detected. Overall, colostrum is the best source for calves in IgG absorption, antioxidant activities, and serum growth metabolites, and promoting intestinal development. The higher quality of colostrum calves ingested, the faster immune defense mechanism and the more healthy intestinal circumstances they established.