Acceleration of Small Intestine Development and Remodeling of the Microbiome Following Hyaluronan 35 kDa Treatment in Neonatal Mice.

The beneficial effects of human milk suppressing the development of intestinal pathologies such as necrotizing enterocolitis in preterm infants are widely known. Human milk (HM) is rich in a multitude of bioactive factors that play major roles in promoting postnatal maturation, differentiation, and the development of the microbiome. Previous studies showed that HM is rich in hyaluronan (HA) especially in colostrum and early milk. This study aims to determine the role of HA 35 KDa, a HM HA mimic, on intestinal proliferation, differentiation, and the development of the intestinal microbiome. We show that oral HA 35 KDa supplementation for 7 days in mouse pups leads to increased villus length and crypt depth, and increased goblet and Paneth cells, compared to controls. We also show that HA 35 KDa leads to an increased predominance of Clostridiales Ruminococcaceae, Lactobacillales Lactobacillaceae, and Clostridiales Lachnospiraceae. In seeking the mechanisms involved in the changes, bulk RNA seq was performed on samples from the terminal ileum and identified upregulation in several genes essential for cellular growth, proliferation, and survival. Taken together, this study shows that HA 35 KDa supplemented to mouse pups promotes intestinal epithelial cell proliferation, as well as the development of Paneth cells and goblet cell subsets. HA 35 KDa also impacted the intestinal microbiota; the implications of these responses need to be determined.

Immunomodulatory responses in plectasin-supplemented broilers under tropical environmental conditions.

The present study was aimed to determine the immunomodulatory effects of dietary supplementation of the antimicrobial peptide (AMP) plectasin on broiler chickens. The experiment involved 300-day-old Ross chicks reared in a conventional housing system and subjected to ambient temperature and relative humidity. The birds were randomly allocated to five treatment groups: the non-supplemented negative control group (T1), enramycin-supplemented group (T2), and groups supplemented with varying doses of plectasin at 150 ppm, 300 ppm, and 450 ppm (T3, T4, and T5, respectively) from day 1 to 35. The results indicated that plectasin supplementation increased jejunal and ileal goblet cell (GC) counts, serum interferon-gamma (IFN-γ) levels at neonatal age, and serum immunoglobulin Y (IgY) titer on days 7, 21, 28, and 35. These findings confirmed that plectasin induces positive immunomodulatory responses by specifically enhancing gut mucosal barriers, early innate immunity, and humoral immune response. Specifically, supplementation at 150 ppm may be considered as the optimal dose for inclusion in broiler chicken feeds.

Anti-stress effects of rosemary (Rosmarinus officinalis L.) leaf extract on intestinal goblet cells and immobility of forced-swimming test in BALB/c mice.

We investigated the anti-stress effect of rosemary (Rosmarinus officinalis L.) leaf extract (RLE) on restraint-stressed mice and found that RLE alleviated decreases in the number of intestinal goblet cells and amount of hepatic triglycerides. It also decreased the immobility time in the forced-swimming test and activation of microglia in the brain, suggesting that RLE has beneficial effects on stress-induced dysfunctions.

Effect of beta- and alpha-glucans on immune modulating factors expression in enterocyte-like Caco-2 and goblet-like LS 174T cells.

Glucans are complex polysaccharides consisting of repeated units of d-glucose linked by glycosidic bonds. The nutritional contribution in α-glucans is mainly given by starch and glycogen while in ß-glucans by mushrooms, yeasts and whole grains, such as barley and spelt well represented in the Mediterranean Diet. Numerous and extensive studies performed on glucans highlighted their marked anti-tumor, antioxidant and immunomodulatory activity. It has recently been shown that rather than merely being a passive barrier, the intestinal epithelium is an essential modulator of immunity. Indeed, epithelial absorptive enterocytes and mucin secreting goblet cells can produce specific immune modulating factors, driving innate immunity to pathogens as well as preventing autoimmunity. Despite the clear evidence of the effects of glucans on immune system cells, there are only limited data about their effects on immune activity of mucosal intestinal cells strictly related to intestinal barrier integrity. The aim of the study was to evaluate the effects of α and ß glucans, alone or in combination with other substances with antioxidant properties, on reactive oxygen species (ROS) levels, on the expression of ROS-generating enzyme DUOX-2 and of the immune modulating factors Tumor Necrosis Factor (TNF-α), Interleukin 1 ß (IL-1ß) and cyclooxygenase-2 (COX-2) in two intestinal epithelial cells, the enterocyte-like Caco-2 cells and goblet cell-like LS174T. In our research, the experiments were carried out incubating the cells with glucans for 18 h in culture medium containing 0.2% FBS and measuring ROS levels fluorimetrically as dihydrodichlorofluoresce diacetate (DCF-DA) fluorescence, protein levels of DUOX-2 by Western blotting and mRNA levels of, TNF-α, IL-1ß and COX-2 by qRT-PCR. α and ß glucans decreased ROS levels in Caco-2 and LS 174T cells. The expression levels of COX-2, TNF-α, and IL-1ß were also reduced by α- and ß-glucans. Additive effects on the expression of these immune modulating factors were exerted by vitamin C. In Caco-2 cells, the dual oxidase DUOX-2 expression is positively modulated by ROS. Accordingly, in Caco-2 or LS174T cells treated with α and ß-glucans alone or in combination with Vitamin C, the decrease of ROS levels was associated with a reduced expression of DUOX-2. The treatment of cells with the NADPH oxidase (NOX) inhibitor apocynin decrease ROS, DUOX-2, COX-2, TNF-α and IL-1ß levels indicating that NOX dependent ROS regulate the expression of immune modulating factors of intestinal cells. However, the combination of vitamin C, α and ß-glucans with apocynin did not exert an additive effect on COX-2, TNF-α and IL-1ß levels when compared with α-, ß-glucans and Vitamin C alone. The present study showing a modulatory effect of α and ß-glucans on ROS and on the expression of immune modulating factors in intestinal epithelial cells suggests that the assumption of food containing high levels of these substances or dietary supplementation can contribute to normal immunomodulatory function of intestinal barrier.

Consumption of an Oil Palm Fruit Extract Promotes Large Bowel Health in Rats.

Oil palm fruit is widely used for edible oils, but the health benefits of other components are relatively unknown. We examined if consuming a polyphenol-rich extract of the fruit, from a vegetation by-product of oil processing, which also contains fibre, has gastro-intestinal benefits in rats on a Western-type diet (WD). The oil palm preparation (OPP) was added to food (OPP-F) or drinking water (OPP-D) to provide 50 mg of gallic acid equivalents (GAE)/d and compared to effects of high amylose maize starch (HAMS; 30%) in the diet or green tea extract (GT; 50 mg GAE/d) in drinking water over 4 wk. OPP treatments induced some significant effects (P < 0.05) compared to WD. OPP-D increased caecal digesta mass, caecal digesta concentrations of total SCFA, acetate and propionate (OPP-F increased caecal butyrate concentration), the numbers of mucus-producing goblet cells per colonic crypt, and caecal digesta abundance of some bacteria which may provide benefit to the host (Faecalibacterium prausnitzii, Akkermansia muciniphila and Ruminococcus gnavus). HAMS induced similar effects but with greater potency and had a broader impact on microbe populations, whereas GT had minimal impacts. These results suggest dietary OPP may benefit the large bowel.

Dietary fiber sources and non-starch polysaccharide-degrading enzymes modify mucin expression and the immune profile of the swine ileum.

Due to their complex chemical and physical properties, the effects and mechanisms of action of natural sources of dietary fiber on the intestine are unclear. Pigs are commonly fed high-fiber diets to reduce production costs and non-starch polysaccharide (NSP)-degrading enzymes have been used to increase fiber digestibility. We evaluated the expression of mucin 2 (MUC2), presence of goblet cells, and ileal immune profile of pigs housed individually for 28 days and fed either a low fiber diet based on corn-soybean meal (CSB, n = 9), or two high fiber diets formulated adding 40% corn distillers' dried grains with solubles (DDGS, n = 9) or 30% wheat middlings (WM, n = 9) to CSB-based diet. Pigs were also fed those diets supplemented with a NSP enzymes mix (E) of xylanase, ß-glucanase, mannanase, and galactosidase (n = 8, 10, and 9 for CSB+E, DDGS+E and WM+E, respectively). Feeding DDGS and WM diets increased ileal MUC2 expression compared with CSB diet, and this effect was reversed by the addition of enzymes. There were no differences in abundance of goblet cells among treatments. In general, enzyme supplementation increased gene expression and concentrations of IL-1ß, and reduced the concentrations of IL-4, IL-17A and IL-11. The effects of diet-induced cytokines on modulating intestinal MUC2 were assessed in vitro by treating mouse and swine enteroids with 1 ng/ml of IL-4 and IL-1ß. In accordance with previous studies, treatment with Il-4 induced Muc2 and expansion of goblet cells in mouse enteroids. However, swine enteroids did not change MUC2 expression or number of goblet cells when treated with IL-4 or IL-1ß. Our results suggest that mucin and immune profile are regulated by diet in the swine intestine, but by mechanisms different to mouse, emphasizing the need for using appropriate models to study responses to dietary fiber in swine.

An oil containing EPA and DHA from transgenic Camelina sativa to replace marine fish oil in feeds for Atlantic salmon (Salmo salar L.): Effects on intestinal transcriptome, histology, tissue fatty acid profiles and plasma biochemistry.

New de novo sources of omega 3 (n-3) long chain polyunsaturated fatty acids (LC-PUFA) are required as alternatives to fish oil in aquafeeds in order to maintain adequate levels of the beneficial fatty acids, eicosapentaenoic and docosahexaenoic (EPA and DHA, respectively). The present study investigated the use of an EPA+DHA oil derived from transgenic Camelina sativa in Atlantic salmon (Salmo salar) feeds containing low levels of fishmeal (35%) and fish oil (10%), reflecting current commercial formulations, to determine the impacts on tissue fatty acid profile, intestinal transcriptome, and health of farmed salmon. Post-smolt Atlantic salmon were fed for 12-weeks with one of three experimental diets containing either a blend of fish oil/rapeseed oil (FO), wild-type camelina oil (WCO) or transgenic camelina oil (DCO) as added lipid source. The DCO diet did not affect any of the fish performance or health parameters studied. Analyses of the mid and hindgut transcriptomes showed only mild effects on metabolism. Flesh of fish fed the DCO diet accumulated almost double the amount of n-3 LC-PUFA than fish fed the FO or WCO diets, indicating that these oils from transgenic oilseeds offer the opportunity to increase the n-3 LC-PUFA in farmed fish to levels comparable to those found a decade ago.

Pycnogenol Ameliorates Asthmatic Airway Inflammation and Inhibits the Function of Goblet Cells.

Pycnogenol® (PYC) is utilized in the treatment of various diseases ranging from chronic inflammation to circulatory diseases, but its efficacy and functional mechanism in pediatric asthma continue to remain obscure. Therefore, the purpose of this study was to investigate the effectiveness and molecular mechanism of PYC on regulation of asthmatic airway inflammation. We found that PYC with tail intravenous injection of 50 mg/kg or intragastric administration of 100 mg/kg all reduced ovalbumin (OVA)-induced airway injury. Pharmacokinetics of PYC was evaluated by high-performance liquid chromatography assay, indicating that PYC was quickly absorbed into the blood after intragastric administration, and PYC metabolism was later improved gradually with increase of time after PYC administration. PYC has a higher bioavailability of 71.96%, and it was more easily absorbed by the body. PYC inhibited the number of total inflammatory cells and levels of interleukin (IL)-4, IL-5, IL-9, and IL-13 in bronchoalveolar lavage fluid of OVA-induced mice. PYC inhibited IL-13 secretion from the Th2 cells, thereby causing a reduction in expression of the signaling molecules in JAK/STAT6 pathway in airway epithelial cells. STAT6 silence suppressed IL-13-increased acetylcholine level. STAT6 overexpression promoted expression of goblet cell metaplasia-associated molecules (FOXA3, SPDEF, and Muc5ac). PYC suppressed OVA-induced expression of FOXA3, SPDEF, and Muc5ac in lung. Our findings indicate that PYC has a higher bioavailability and it prevents emergence of OVA-induced airway injury and airway inflammation in mice by inhibiting IL-13/JAK/STAT6 pathway and blocking release of acetylcholine to reduce goblet cell metaplasia.

Short-Term Omega 3 Fatty Acids Treatment for Dry Eye in Young and Middle-Aged Visual Display Terminal Users.

OBJECTIVE: To evaluate the effect of an omega 3 fatty acid (O3FA) oral supplement (2,400 mg/day) for 45 days on dry eye symptoms, tear production, stability, and conjunctival cytology in young and middle-aged visual display terminal (VDT) users. METHODS: Institutional review board approval was obtained, and a randomized, double-blind, interventional study was done; eyes of 256 VDT users were randomized to receive 4 capsules twice daily for 45 days (O3FA group), each containing 180 mg of eicosapentaenoic acid and 120 mg docosahexaenoic acid. The O3FA group was compared with another group (n=266) who received 8 capsules of a placebo (olive oil). Patients were evaluated at baseline, 30 days, and 45 days. The primary outcome measure was an improvement in dry eye symptoms. Secondary outcome measures were improvement in the Nelson grade on conjunctival impression cytology, Schirmer test values, and tear film breakup time (TBUT). Means of groups (pretreatment, day 30, and day 45) were compared with repeated-measure analysis of variance. The relation between the outcome variables and VDT time was evaluated using linear regression. RESULTS: In the O3FA group, the mean symptom score differed significantly (P<0.005) (pretreatment, 30 days, and 45 days); the TBUT and Nelson grade also improved significantly but only after 45 days of intervention. Schirmer test values did not differ significantly after adjustment for multiple comparisons (P=0.010). The change was not significant in the placebo group. CONCLUSION: Consumption of 2,400 mg/day of O3FA supplement improves symptoms, tear stability, and conjunctival cytology but not tear production in symptomatic VDT users.

Favourable effects of grape seed extract on intestinal epithelial differentiation and barrier function in IL10-deficient mice.

The impairment in the rate of cell proliferation and differentiation leads to a negative consequence on the renewal of the intestinal epithelium, which is the aetiological factor of a number of digestive diseases. Grape seed extract (GSE), a rich source of proanthocyanidins, is known for its beneficial health effects. The present study evaluated the beneficial effects of GSE on colonic cell differentiation and barrier function in IL10-deficient mice. Female mice aged 6 weeks were randomised into two groups and given drinking-water containing 0 or 0.1 % GSE (w/v) for 12 weeks. GSE supplementation decreased serum TNF-α level and intestinal permeability, and increased the colonic goblet cell density that was associated with increased mRNA expression of mucin (Muc)-2. Immunohistochemical analyses showed lower accumulation of ß-catenin in the crypts of colon tissues of the GSE-supplemented mice, which was associated with a decreased mRNA expression of two downstream effectors of Wingless and Int (Wnt)/catenin signalling, myelocytomatosis oncogene protein (Myc) and cyclin D1 (Ccnd1). Consistently, GSE supplementation decreased the number of colonic proliferating cell nuclear antigen-positive cells, a well-known cell proliferation marker, and a weakened extracellular signal-regulated kinases 1 and 2 (ERK1/2) signalling. In summary, these data indicate that supplementation of 0.1 % GSE for 12 weeks improved gut barrier function and colonic cell differentiation in the IL10-deficient mice probably via inhibiting Wnt/ß-catenin pathway.

Oral omega-3 fatty acid treatment for dry eye in contact lens wearers.

PURPOSE: The aim of this study was to evaluate the effect of dietary omega-3 fatty acid (O3FA) supplementation on dry eye symptoms, tear film tests, and conjunctival impression cytology in patients with contact lens wear-associated dry eye. METHODS: In this randomized, double-blind, multicentric trial, contact lens wearers (n = 496) were randomized to receive either O3FAs or placebo capsules (corn oil) twice daily for 6 months. Subjects underwent examinations at baseline, 3 months, and 6 months. At each visit, a questionnaire of dry eye symptoms and lens wear comfort was administered. Subjects further underwent measurement of tear film break-up time (TBUT) and a Schirmer test. Conjunctival impression cytology was performed by the transfer method. Improvement in symptoms and lens wear comfort were primary outcome measures. Changes from baseline in TBUT, Schirmer, and Nelson grade at 6 months were secondary outcome measures. RESULTS: The mean improvement in symptom score in the O3FA group was 4.7 ± 2 (2.0) as compared with 0.5 ± 2 (0.9) in the placebo group (P < 0.0001). Lens wear comfort levels improved significantly (P < 0.0001) from baseline. There was a significant increase in TBUT [3.3 ± 2 (1.5)] and Nelson grade [0.7 ± 2 (0.6)] in the O3FA group (P < 0.0001) as compared with 0.3 ± 2 (0.6) and 0.1 ± 2 (0.4) in the placebo group (P = 0.164 and 0.094, respectively). However, the magnitude of increase in Schirmer score [2.0 ± 2 (1.5)] was relatively small (P = 0.08). CONCLUSIONS: The results of this study point toward benefits of orally administered O3FAs in alleviating dry eye symptoms, improving lens wear comfort, and cytological changes in contact lens wearers.

Effect of dietary supplementation of prebiotics and probiotics on intestinal microarchitecture in broilers reared under cyclic heat stress.

This study was designed to evaluate the effect of dietary supplementation of prebiotics, mannanoligosaccharides (MOS) and/or probiotics (LBP) on intraepithelial lymphocytes (IEL) count, goblet cells (GC) count and differentiation and intestinal micro-architecture in broilers reared under cyclic heat stress. Day-old broilers (n = 250) were randomly divided into five groups. Fifty birds were reared within the thermoneutral zone (TNZ). Remaining birds were subjected to cyclic heat stress from day 21 to 42 (35° C, 75% RH, 8 h/d). The birds were fed corn-soy-based basal diet or the same diet supplemented with 0.5% MOS (HS-MOS), or 0.1% LBP (HS-LBP), or their combination (HS-SYN). The birds were slaughtered on day 42. Tissue samples were collected from mid-duodenum, jejunum and ileum, and stained with haematoxylin and eosin or combined Alcian blue and PAS technique. All differences were considered significant at p < 0.05. The IEL count increased in all intestinal segments of the HS group compared with the TNZ group and decreased in all supplemented groups compared with the HS group. Compared with the TNZ, heat stress reduced villus height, crypt depth and surface area in duodenum and ileum, and increased crypt depth in ileum. Villus width decreased in duodenum and jejunum compared with the TNZ group. Supplementation of LBP, MOS and SYN reversed all these changes in duodenum, while only increased villus height and surface area in ileum. In jejunum, the villus height and surface area increased with HS-LBP, and crypt depth increased with HS-MOS. The number of GC containing acid mucins (duodenum and ileum) and mixed mucins (ileum) were increased in the HS compared with the TNZ. Supplementation of MOS, LBP and SYN maintained the enhanced activity of goblet cells. In conclusion, dietary supplementation of MOS and/or LBP may be helpful in alleviating some of the detrimental effects of heat stress on microstructure of the broiler gut.

Low-methoxyl pectin stimulates small intestinal mucin secretion irrespective of goblet cell proliferation and is characterized by jejunum Muc2 upregulation in rats.

Generally, soluble fibers increase small intestinal mucin secretion by increasing the number of goblet cells in a viscosity-dependent manner. The present study aimed to examine the mechanism by which low-methoxyl pectin (LPC) affects mucin secretion in the small intestine. First, diets containing 50 g/kg of low-viscosity fiber (LPC, gum arabic, guar gum, low-molecular konjac mannan, arabinogalactan, sodium alginate) or high-molecular konjac mannan (KMH) were fed to Wistar rats for 10 d. Luminal mucin was greater in the LPC and KMH groups than in the fiber-free control group, but only the KMH group had more goblet cells in the ileum compared with the other groups. Next, Sprague-Dawley rats were fed LPC, KMH, or high-methoxyl pectin (HPC) diets (50 g/kg) for 10 d. The KMH and LPC groups, but not the HPC group, had greater luminal mucin than the control group, whereas jejunum Muc2 expression was higher only in the LPC group. Sprague-Dawley rats fed the LPC diet for 1 or 3 d had greater luminal mucin and jejunum Muc2 expression than those fed the control diet. In vitro studies using HT-29MTX cells showed that, of the various fibers studied, only LPC and HPC affected mucin secretion. Finally, Wistar rats were fed the LPC diet with or without neomycin in drinking water for 10 d; neomycin treatment did not compromise the effect of LPC on mucin secretion. We conclude that LPC does not affect the number of goblet cells but can interact directly with the epithelium and stimulate small intestinal mucin secretion.

Influence of L-threonine supplementation on goblet cell numbers, histological structure and antioxidant enzyme activities of laying hens reared in a hot and humid climate.

1. A previous experiment in our laboratory found that L-threonine supplementation at 0.2 and 0.3% increased egg production and the concentration of serum IgG, respectively. The objective of this current trial was to determine if both supplementation levels can positively influence histological structure, goblet cell numbers, or antioxidant enzyme activities. 2. Babcock Brown layers (n=576), 40 weeks of age, were allocated to three treatment groups, each of which included 6 replicates of 32 hens. Each group received the same basal diet formulated with maize, peanut meal and crystalline amino acids. L-threonine was added to the basal diet at 0 (control), 0.2, and 0.3%, respectively for 8 weeks. Chemical analysis of the diets for threonine values were 0.47, 0.66 and 0.74 %, respectively. 3. The numbers of goblet cells did not change due to L-threonine supplementation. Also, L-threonine had no affect on the villus height and mucosal thickness. No differences were found due to treatments among groups in the activity of alkaline phosphatase (ALP) in jejunum or ileum. L-threonine supplementation at 0.2% maximised the concentration of superoxide dismutases (SOD) in both serum and liver. 4. In conclusion, L-threonine supplementation had no affect on gut morphology but may have an antioxidant function at 0.2%.

Theaflavins extracted from black tea inhibit airway mucous hypersecretion induced by cigarette smoke in rats.

Theaflavins isolated from black tea have been used in studies on the prevention of tumor growth. The aim of this study was to investigate whether treatment with theaflavins influences the mucus hypersecretion induced by cigarette smoke in the lungs of experimental rats. Firstly, cigarette smoke was aerosolized using a machine designed for inhalation by rats. The rats were divided into the negative control group, the cigarette smoke inhalation group, the theaflavins (TFs) treatment group, and the TFs +cigarette smoke inhalation group. The animals were sacrificed on day 60 of the experiment. Secondly, the rats were treated with theaflavins at different doses via a gastric tube and sacrificed on day 30. The changes in the levels of mucin 5AC (MUC5AC) and epidermal growth factor receptor (EGFR) in the airway were evaluated. Cigarette smoke induced a significant increase in the levels of MUC5AC and EGFR in all groups. These increases could be reversed by intragastric administration of theaflavins. The effect was more pronounced with the duration of treatment and coincided with a decrease in the expression of both targets. The rats showed various degrees of reduction in the expression of these parameters, which correlated with the theaflavin dose. TFs could inhibit the activation of EGFR, decrease the level of MUC5AC, and relieve airway mucous hypersecretion via the EGFR signaling pathway. These effects correlated directly with the duration of action and the dosage. In the future, oral theaflavins might be valuable in the treatment of chronic airway inflammation.

Treatment of severe keratoconjunctivitis sicca by parotid duct transposition after tympanic neurectomy in rabbits.

PURPOSE: To investigate the feasibility of parotid duct transposition after tympanic neurectomy to treat severe keratoconjunctivitis sicca (KCS) in rabbits. METHODS: Thirty rabbits were divided into three groups in experiment 1. One eye was operated on, and the contralateral eye served as the control. In the KCS group, the lacrimal gland, harderian gland, and nictitating membrane were removed. In the group with parotid duct transposition (DT), the parotid duct was transposed into the lower conjunctival fornix. In the group with parotid duct transposition after tympanic neurectomy (DTTN), the tympanic nerve was resected in addition to parotid duct transposition. Schirmer test was performed and density of corneal staining was determined monthly after surgery, and goblet cell density was measured at postoperative month 3. In experiment 2, the tympanic nerve was resected on one side in 12 rabbits. Both sides of the parotid gland were resected for histopathology at intervals of 2 months to 1 year after surgery. RESULTS: Tear secretion from operated eyes at rest increased significantly after surgery in the treatment groups compared with the KCS group. Tear secretion from operated eyes after chewing was significantly lower in the DTTN than in the DT group. The corneal staining scores were higher in the operated than in the control eyes of the three groups, without significant difference among the operated eyes. Parotid gland atrophy on the operated side occurred at postoperative month 4 and recovered to normal 1 year after surgery. CONCLUSIONS: Parotid duct transposition after tympanic neurectomy could effectively reduce gustatory epiphora but may be insufficient to promote ocular surface health.

Effects of dietary enzymes on performance and intestinal goblet cell number of broilers exposed to a live coccidia oocyst vaccine.

An experiment was conducted to evaluate the effects of dietary enzymes on performance, tibia ash, and intestinal goblet cells of broilers administered a live coccidia oocyst vaccine (Coccivac B, Schering Plough, Kenilworth, NJ). Cobb 500 straight-run broilers were obtained and one-half of the chicks were sprayed with the live coccidia oocyst vaccine. Chicks were weighed and placed in battery brooders with respect to nonvaccinated or vaccinated group according to dietary treatment. The 8 dietary treatments were a positive control (0.90% Ca and 0.45% available P), a negative control (NC; 0.80% Ca and 0.35% available P), NC + phytase (PHY), NC + protease (PRO), NC + xylanase (XYL), NC + PHY+ PRO, NC + PHY + XYL, and NC + PHY + PRO + XYL. A diet × vaccination interaction (P > 0.05) was not observed for feed intake or BW gain. Feed conversion ratio was improved (P ≤ 0.05) in birds fed NC + PHY + XYL compared with NC. Vaccination reduced (P ≤ 0.05) feed intake and BW gain from d 0 to 18. Tibia ash was reduced (P ≤ 0.05) in the NC and PRO or XYL diets. Vaccination increased goblet cell numbers in the duodenum of birds fed XYL, whereas no differences were found in goblet cell numbers between nonvaccinated and vaccinated birds in other dietary treatments, which resulted in a diet × vaccination interaction (P ≤ 0.05). Protease decreased and NC + PHY+ PRO increased goblet cells in the jejunum at d 7, which resulted in a diet × vaccination interaction (P ≤ 0.05). At d 18, NC + PHY + XYL was the only diet in which vaccination decreased goblet cells in the jejunum, resulting in a diet × vaccination interaction (P ≤ 0.05). The data indicate that NC + PHY + XYL improved the feed conversion ratio in broilers fed corn-soybean meal diets. The vaccination × dietary enzyme interaction altered the number of goblet cells in the small intestine. Dietary enzyme supplementation did not alleviate reductions in growth performance associated with the use of a live coccidia oocyst vaccine.

Stimulation of ileal epithelium growth and regeneration by dietary nucleotide extracts.

The gastrointestinal tract epithelium plays an important role not only in digestion and absorption of nutrients, but also in antigen and pathogen signal translocation toward the gut associated lymphoid tissue. Malnutrition in various degrees is recognized as the most common cause of the immune system dysfunction. Research done in the past several years has revealed that dietary nucleotides (dNT) represent an essential compound of nutrition because of their importance in metabolic pathways, energetic processes and nucleic acid synthesis during tissue renewal. Much evidence accumulated suggests that dNT are essential for the growth and maturation of the gut epithelia. In previous experiments we have documented immunoregulative properties of dNT-containing extracts. In this study Balb/c female mice were fed (1) standard diet, (2) dNT-supplemented diet, and (3) dNT-supplemented water for 4 weeks. The supplement in dose of 100 mg/kg/l comprised original extract (Imuregen, Uniregen Ltd., Náchod, Czech Republic). Samples of terminal ileum in each dietary group were removed for histological examination. The length of villi was evaluated by computer morphometry. The highest growth of intestinal villi was observed in group administered dNT-supplemented water. We have found no pathological changes of intestinal epithelium in any experimental group.

Capsaicin-sensitive nerve fibres induce epithelial cell proliferation, inflammatory cell immigration and transforming growth factor-alpha expression in the rat colonic mucosa in vivo.

BACKGROUND: Capsaicin-sensitive nerve fibres protect gastrointestinal mucosa in animal models of mucosal injury by modulation of mucosal blood flow and mucus secretion. The aim of our study was to evaluate the effects of capsaicin-sensitive nerve fibres in rat colonic mucosa on epithelial cell proliferation and transforming growth factor-alpha (TGFalpha) expression, which is important in mucosal defence, protection and repair. METHODS: Male Wistar rats received either a capsaicin enema with or without giving antagonists to calcitonin-gene-related-peptide (CGRP) or substance P (SP) i.v. immediately prior to the capsaicin enemas; a capsaicin enema after sensory desensitization as described previously; or a vehicle enema. In all experiments, animals received 50 mg/kg BrdU i.v. and were killed at 2, 4, 8, 12, 24 and 48 h after the various treatments. Colonic mucosal specimens were evaluated microscopically for mucosal damage, changes in the numbers of inflammatory cells and BrdU-immunoreactive epithelial cell nuclei. In the same specimens, TGFalpha-mRNA and -protein expression were evaluated by RT-PCR and Western blot analysis using standardized procedures. RESULTS: A significant increase in the number of mucosal inflammatory cells and an increase in BrdU-immunoreactive nuclei were detected following mucosal exposure to capsaicin. A 2-fold increase of TGFalpha mRNA and a 10-fold increase of TGFalpha protein expression were obtained 2-12 h after capsaicin enemas. The effects on the invading number of inflammatory cells and on the increase in BrdU immunoreactive epithelial cell nuclei were significantly reduced by both CGRP and SP antagonists and were abolished in rats previously sensory-desensitized. CONCLUSION: Capsaicin-sensitive nerve fibres modulate epithelial cell proliferation and TGFalpha expression in colonic mucosa as well as a migration of inflammatory cells into the colonic mucosa. These effects are mediated by the neurotransmitters CGRP and SP.