Jejunal Crypt Research Articles

Effects of yeast hydrolysate supplementation on intestinal morphology, barrier, and anti-inflammatory functions of broilers.

ObjectiveThis study was conducted to evaluate the effects of dietary yeast hydrolysate (YH) supplementation on intestinal morphology, barrier, and anti-inflammatory functions of broilers.MethodsA total of 320 one day old male broilers were randomly allocated into four groups with eight replicates of ten broilers each. The broilers were supplemented with a basal diet (the control group) or basal diets adding 50, 100, 150 mg/kg YH, respectively. This trial lasted for 42 days. The orthogonal polynomial contrasts were used to determine the linear and quadratic effects of increasing levels of YH.ResultsIn our previous research, supplementing YH improved growth performance by enhancing body weight gain but decreased feed-to-gain ratio. In this study, compared with the control group, dietary YH addition linearly and quadratically decreased serum diamine oxidase activity (p<0.05). Additionally, supplementing YH linearly and/or quadratically decreased jejunal crypt depth (CD), tumor necrosis factor-alpha (TNF-α) concentration as well as mucin 2, interleukin-6 (IL-6), IL-1β, TNF-α, nuclear factor kappa B, and myeloid differentiation factor 88 gene expression levels (p<0.05). Whereas the jejunal villus height (VH), VH/CD, IL-10 concentration as well as zonula occludens-1 and IL-10 gene expression levels were linearly and/or quadratically increased by YH supplementation (p<0.05).ConclusionDietary YH supplementation improved intestinal morphology, barrier and anti-inflammatory functions while decreased intestinal permeability of broilers, which might be related with altering pertinent genes expression. This study provides evidence of YH as a promising feed additive for broilers.

Advantages of sodium butyrate in weaned piglet diet

The aim was to examine production performance, carcass characteristics, and intestinal histomorphology of weaned piglets (28-to 54-day-old) fed diet with 3 or 5 g of sodium butyrate per kg of diet (group II and III). Groups II and III had higher final live weight and total weight gain. The feed to gain ratio was the best in group I. The highest carcass yield was of group III. Significant differences (P<0.05) were observed in Escherischia coli counts in small intestine between control (I) and experimental groups (II and III), and in cecum between control (I) and experimental group II. There were significant differences (P<0.05) between all groups for the intestine length, intestine weight, and both height and width of the ileal villus. The highest villus height/crypt depth ratio of jejunum occurred in group II piglets, while the highest villus height/crypt depth ratio of ileum was in piglets from group III. The significant correlations were determined between amount of sodium butyrate and final live weight, intestinal length, intestinal weight, jejunal and ileal crypt depth, and ileal villus widths.

Dietary palygorskite-based antibacterial agent supplementation as an alternative to antibiotic improves growth performance, intestinal mucosal barrier function, and immunity in broiler chickens

The aim of this study was to evaluate effects of palygorskite-based antibacterial agent (PAA) as an alternative to antibiotic on growth performance, intestinal barrier function, and immunity in broilers. Three hundred and eighty-four mixed-sex 1-day-old Ross 308 broiler chicks were allocated into 6 groups of 8 replicates with 8 birds each. Birds were given a basal diet, an antibiotic diet (50 mg/kg chlortetracycline), and the basal diet supplemented with 250, 500, 1,000, and 2,000 mg/kg PAA for 42 d, respectively. Compared with control group, supplementing 1,000 mg/kg PAA reduced overall feed conversion ratio (P < 0.05), with its value being similar to that of antibiotic group (P > 0.05). However, a higher level of PAA (2,000 mg/kg) increased feed conversion ratio during the late period (P < 0.05). The 1,000 and 2,000 mg/kg PAA decreased plasma endotoxin and D-lactate levels at 42 d (P < 0.05) to comparable values (P > 0.05). The 1,000 mg/kg PAA decreased jejunal crypt depth, while 500 and 1,000 mg/kg PAA increased the ratio between jejunal villus height and crypt depth at 42 d (P < 0.05), with their values being similar to antibiotic group (P > 0.05). The highest level of PAA increased 42-d jejunal mucosal secretory immunoglobulin A and immunoglobulin M concentrations (P < 0.05). The 1,000 and 2,000 mg/kg PAA reduced 21-d interleukin-1β and tumor necrosis factor-α (TNF-α) levels in serum and ileal mucosa and 42-d interferon-γ level in serum and jejunal mucosa (P < 0.05), which did not differ from antibiotic group (P > 0.05). Moreover, PAA administration, regardless of its dosage, reduced 42-d serum TNF-α concentration, and 500 to 2,000 mg/kg PAA decreased 21-d and 42-d jejunal and 42-d ileal mucosal TNF-α levels (P < 0.05), with their values being comparable with antibiotic group (P > 0.05). The results suggested that PAA as an alternative to antibiotic could improve growth performance, intestinal barrier function, and immunity of broilers, and its optimal dosage was 1,000 mg/kg.

Changes in progenitors and differentiated epithelial cells of neonatal piglets.

This study aimed to assess the changes of small intestinal morphology, progenitors, differentiated epithelial cells, and potential mechanisms in neonatal piglets. Hematoxylin and eosin staining of samples from 36 piglets suggested that dramatic changes were observed in the jejunum crypts depth and crypt fission index of neonatal piglets (P < 0.001). The number of intestinal stem cells (ISC) tended to increase (P < 0.10), and a decreased number of enteroendocrine cells appeared in the jejunal crypt on d 7 (P < 0.05). Furthermore, the mRNA expression of jejunal chromogranin A (ChgA) was down-regulated in d 7 piglets (P < 0.05). There was an up-regulation of the adult ISC marker gene of SPARC related modular calcium binding 2 (Smoc2), and Wnt/β-catenin target genes on d 7 (P < 0.05). These results were further verified in vitro enteroid culture experiments. A mass of hollow spheroids was cultured from the fetal intestine of 0-d-old piglets (P < 0.001), whereas substantial organoids with budding and branching structures were cultured from the intestine of 7-d-old piglets (P < 0.001). The difference was reflected by the organoid budding efficiency, crypt domains per organoid, and the surface area of the organoid. Furthermore, spheroids on d 0 had more Ki67-positive cells and enteroendocrine cells (P < 0.05) and showed a decreasing trend in the ISC and goblet cells (P < 0.10). Moreover, the mRNA expression of spheroids differed markedly from that of organoids, with low expression of intestinal differentiation gene (Lysozyme; P < 0.05), epithelial-specific markers (Villin, E-cadherin; P < 0.05), and adult ISC markers (leucine-rich repeat-containing G protein-coupled receptor 5 [Lgr5], Smoc2; P < 0.001), and up-regulation of fetal marker (connexin 43 [Cnx43]; P < 0.05). The mRNA expression of relevant genes was up-regulated, and involved in Wnt/β-catenin, epidermal growth factor (EGF), Notch, and bone morphogenetic protein (BMP) signaling on d 7 organoids (P < 0.05). Spheroids displayed low differentiated phenotype and high proliferation, while organoids exhibited strong differentiation potential. These results indicated that the conversion from the fetal progenitors (spheroids) to adult ISC (normal organoids) might largely be responsible for the fast development of intestinal epithelial cells in neonatal piglets.

Effect of dietary folic acid and energy density on immune response, gut morphology, and oxidative status in blood and breast muscle of broiler chickens

Folic acid (FA) plays essential roles in many metabolic functions and has been reported to have antioxidant effects. Therefore, dietary supplementation with high levels of FA may improve gut health and prevent potential oxidative stress caused by feeding a high energy density diet to broiler chickens. Broiler chickens were assigned into eight treatments, consisting of either a normal energy (NE) or high energy (HE) density diet, and four FA levels (2.2, 5, 10, and 15 ppm). Data were analyzed by SAS 16 GLM procedure. Birds-fed HE diets had increased (P &lt; 0.05) plasma concentrations of calcium and albumin but reduced (P &lt; 0.005) weights of ceca and bursa compared with those fed NE diets. Dietary supplementation with 10 ppm FA significantly increased (P &lt; 0.05) birds’ heart weight and bile acid concentration. Folic acid and energy density interactions were significant for jejunal villus height (VH; P = 0.0226), villus width (VW; P &lt; 0.0001), and crypt depth (CD; P = 0.0332). Among the NE group, birds fed 5–15 ppm FA had reduced (P &lt; .0001) VW, while in the HE groups, 15 ppm FA supplementation resulted in an increased jejunal VH (P = 0.0317) compared with other treatments. In conclusion, dietary supplementation with increased levels of FA in HE diets could be beneficial for the intestinal health of broiler chickens.

Pharmacologic Ascorbate Enhances the Therapeutic Index of ATM-Inhibitor Based Chemoradiation for Colorectal Cancer

Ataxia telangiectasia mutated protein (ATM) is one of the key sensors of DNA damage and specific inhibitors of ATM are potent radiosensitizers. However, their clinical utility with radiation (RT) is limited because they lack tissue specificity and increase normal tissue injury. Pharmacologic (high dose) ascorbate (P-AscH-) selectively increases oxidative stress in tumors while functioning as a donor antioxidant and reducing RT damage in normal tissues. We hypothesized that P-AscH- could enhance the therapeutic index of ATM-inhibitor based chemoradiation (CRT) for colorectal cancer (CRC) by simultaneously enhancing efficacy and reducing RT bowel injury.Human HCT116, SW480, and HT29 and murine CT26 and MC38 CRC models were used. Clonogenic survival was assessed following single-fraction RT (2-8 Gy) +/- P-AscH- (5 pM/cell) +/- veliparib (PARP), VE821 (ATR), or KU60019 (ATM). Catalase expression was induced using HCT116 cells expressing a doxycycline inducible catalase transgene. DNA double strand breaks (DSBs) were quantified using neutral comet assays 0-24 hours post RT. Cell cycle phases were assessed using flow cytometry. ATM and pATM localization were assessed using IF. Jejunal toxicity was assessed using IHC in fixed tissues following single fraction (10 Gy) whole abdominal RT in c57bl/6 mice. Tumor growth delay was assessed following RT (5 Gy x 3) +/- drug treatment in unilateral flank tumors.Veliparib, VE821, and KU60019 were potent radiosensitizers in HCT116, SW480, HT29, MC38, and CT26 CRC tumor models and P-AscH- further reduced clonogenic survival with DRIs in all lines except for HT29. In contrast, P-AscH- enhanced survival of cultured HUVEC and FHs-74 cells exposed to RT. Enhanced cell kill with P-AscH- is H202 mediated as it is completely attenuated by inducible catalase expression. P-AscH- significantly increased the number of DNA DSBs in tumors after RT in vitro. Despite the increase in DNA DSBs, P-AscH-significantly decreased nuclear localization of activated pATM after RT and significantly decreased the fraction of cells in G2/M phases of the cell cycle. In vivo, RT + P-AscH- + KU60019 induced more tumor growth delay/clearance than all other combinations in unilateral MC38 or HCT116 flank tumors. Finally, P-AscH- significantly reduced loss of jejunal crypt cell density, epithelial architecture, and markers of lipid and protein oxidation following whole abdominal RT.P-AscH- selectively enhances the efficacy of ATM-based CRT in CRC tumor models while simultaneously decreasing RT-mediated small bowel toxicity. In tumors, P-AscH- enhances DNA DSBs by stimulating an H202 flux and prevents activation of DNA repair pathways and cell cycle checkpoints by inhibiting RT-induced activation of ATM. Selective radioprotectors like P-AscH- could facilitate the clinical translation ATM inhibitors as radiosensitizers.

Dietary Supplementation With Didancao (Elephantopus scaber L.) Improves Meat Quality and Intestinal Development in Jiaji Ducks.

Didancao (Elephantopus scaber L.) has been used as a traditional herbal medicine and has exhibited a beneficial role in animal health. This study aimed to investigate the effects of dietary supplementation with E. scaber on growth performance, meat quality, intestinal morphology, and microbiota composition in ducks. A total of 480 Jiaji ducks (42 days old, male:female ratio = 1:1) were randomly assigned to one of four treatments. There were six replicates per treatment, with 20 ducks per replicate. The ducks in the control group (Con) were fed a basal diet; the three experimental groups were fed a basal diet supplementation with 30 (T1), 80 (T2), and 130 mg/kg (T3) of E. scaber. After a 48-day period of supplementation, growth performance, meat quality, intestinal morphology, and microbiota composition were evaluated. The results showed that no differences were observed in the final body weight, average daily feed intake, and average daily gain among the four groups. Compared with that in the Con group, the feed conversion in the T1 and T2 groups was increased significantly; the T2 group was shown to decrease the concentration of alanine aminotransferase in serum; the T3 group was lower than the Con group in the concentration of aspartate aminotransferase and was higher than the Con group in the concentration of high-density lipoprotein-cholesterol. The highest concentration of creatinine was observed in the T1 group. The T2 group was higher than the Con group in the contents of Phe, Ala, Gly, Glu, Arg, Lys, Tyr, Leu, Ser, Thr, Asp, and total amino acids in the breast muscle. Moreover, the T2 group was higher than the Con group in the contents of meat C18:2n−6 and polyunsaturated fatty acid. The concentration of inosinic acid in the T1, T2, and T3 groups was significantly higher than that in the Con group. However, the Con group was higher than the T2 or T3 group in the Zn content. The T2 group was lower than the Con group in the jejunal crypt depth. The T3 group was higher than the Con group in the ileal villus height and the ratio of villus height to crypt depth. In addition, the T3 group had a trend to significantly increase the abundance of Fusobacteria. Compared with the Con group, the T1 and T2 groups displayed a higher abundance of Subdoligranulum. Collectively, dietary supplementation with 80 mg/kg of E. scaber improves meat quality and intestinal development in ducks.

516 Late-Breaking: Iturin a Protects the Intestinal Mucosal Integrity Against E. coli Containing the Gene Coding for Heat-stable Enterotoxin B

Abstract Enterotoxigenic E. coli causes severe infectious diarrhea with high morbidity and mortality in weanling pigs mainly through the production of heat-stable enterotoxins. Iturin A, originally isolated from the insect Hyalophora cecropia, exerts strong antibiotic activity against Gram-positive and Gram-negative bacteria. Thus, our study investigated whether iturin A could protect the intestinal epithelium from the insults of E. coli Rosetta containing the gene coding for heat-stable enterotoxin b (STb-Rosetta). Twenty-eight piglets were divided into four groups and were orally administered PBS (control), 0.16 g/kg BW iturin A, 2 × 109 CFU STb-Rosetta, and 0.16 g/kg BW iturin A + 2 × 109 CFU STb-Rosetta respectively for 8 days. The intestinal epithelial morphology and barrier function were assessed by H&amp;E staining and Ussing chamber, and the growth advantage in enteroids derived from jejunal crypts of piglets in each group was used to evaluate the intestinal stem cell (ISC) activity. The results showed that iturin A increased the average daily gain (ADG) and average daily feed intake (ADFI) of piglets exposed to STb-Rosetta (P &amp;lt; 0.05). Meanwhile, STb-Rosetta-induced decrease in jejunum weight and damage to jejunal morphology and barrier function were significantly improved after iturin A supplementation (P &amp;lt; 0.05). Furthermore, the jejunal crypt cells from piglets in the iturin A + STb-Rosetta group had greater growth advantages of ISCs compared with the STb-Rosetta group, including enteroid forming efficiency, surface area, budding efficiency, and branching coefficient (P &amp;lt; 0.05). Our results indicate that iturin A protects against STb-Rosetta-induced intestinal mucosal injury, which provides an intervention strategy that regulates the function of ISCs in the high turnover-rate crypt-villous axis under ETEC infection. This study was supported by the National Natural Science Foundation of China (31872389, 32072777) and Basic and Applied Basic Research Foundation of Guangdong Province (2019B1515210021).

Lauric acid alleviates deoxynivalenol-induced intestinal stem cell damage by potentiating the Akt/mTORC1/S6K1 signaling axis

Intestinal stem cell (ISC)-driven intestinal homeostasis is subjected to dual regulation by dietary nutrients and toxins. Our study investigated the use of lauric acid (LA) to alleviate deoxynivalenol (DON)-induced intestinal epithelial damage. C57BL/6 mice in the control, LA, DON, and LA + DON groups were orally administered PBS, 10 mg/kg BW LA, 2 mg/kg BW DON, and 10 mg/kg BW LA + 2 mg/kg BW DON for 10 days. The results showed that LA increased the average daily gain and average daily feed intake of the mice exposed to DON. Moreover, the DON-triggered impairment of jejunal morphology and barrier function was significantly improved after LA supplementation. Moreover, LA rescued ISC proliferation, inhibited intestinal cell apoptosis, and promoted ISC differentiation into absorptive cells, goblet cells, and Paneth cells. The jejunum crypt cells from the mice in the LA group expanded into enteroids, resulting in a significantly greater enteroid area than that in the DON group. Furthermore, LA reversed the DON-mediated inhibition of the Akt/mTORC1/S6K1 signaling axis in the jejunum. Our results indicated that LA accelerates ISC regeneration to repair intestinal epithelial damage after DON insult by reactivating the Akt/mTORC1/S6K1 signaling pathway, which provides new implications for the function of LA in ISCs.

The way insoluble fiber incorporated in the diet changes its physiological response

The physiological effect of insoluble fibers may change with diet composition. The present study aimed to determine the difference in the physiological responses of broiler chickens fed diet incorporated with rice hulls in different ways. Two hundred and forty broiler chicks at two-days old were randomly placed in 30 cages and fed: control diet (based on corn-soybean) (CO), rice hull supplementation diet (CO + 4% of rice hulls) (RS), and rice hull inclusion diet (inclusion of 4% rice hulls in diet) (RI). The significant effects were mostly found in the starter phase. The RI increased ADG (P&lt;0.05) but reduced empty ceca weight (P&lt;0.05) in comparison to CO and RS. The RS increased the jejunal crypt depth (P&lt;0.05) and reduced the small intestinal content (P&lt;0.05). Birds fed RS and RI had higher villus height (P&lt;0.05) and thicker jejunal mucosa (P&lt;0.05) than those fed the CO. In the finisher phase, birds fed the RS had the narrower width of the upper and lower jejunal villi than the others. Supplementation or inclusion of insoluble fiber in the diet will lead to different physiological responses due to changes in diet composition.

Association of sodium butyrate and phytase on the performance, bone quality and intestinal development in broilers

This study assessed the effect of the combination of microencapsulated sodium butyrate (SB) and phytase (PH) on the performance, intestinal integrity, and bone development of broilers. The experiment comprised 736 chicks distributed in a completely randomised design in a 2 × 2 factorial scheme (with and without the inclusion of SB and with the inclusion of PH at the recommended dose and superdosing) totalling four treatments and eight repetitions of 23 birds each. SB was added at 0.750 kg/tonne and PH was included at 750 phytase units (FTU)/kg and 1 500 FTU/kg of diet. The live weight (LW), feed intake (FI) and feed conversion (FC) were evaluated weekly. Bone measurements of the tibia and femur, densitometry, length, the Seedor index, and diameter were performed at seven and 28 days. In the same ages, fragments of the jejunum and ileum segments were collected and subjected to a morphometry analysis. At seven days of age, the birds supplemented with the phytase superdosing showed a better FI and LW, and the microencapsulated SB in the diet showed a better LW and FC. In the period from one to 28 days, the treatment supplemented with SB provided a higher FI and LW. The SB supplementation resulted in greater bone measurements at seven and 28 days. The use of the superdosing phytase increased the villus length and width of the jejunum at seven days and at 28 days. There was an increase in the villus length, villus-to-crypt ratio, absorption area, and muscular layer of the jejunum and a decrease in the crypt width of the jejunum. The SB supplementation increased the ileum crypt width at seven days and there was no effect on any intestinal segment at 28 days. The supplementation of SB or PH 1 500 FTU/kg in the broiler diets’ growth promoter or antibiotic-free resulted in a better performance and bone measurements, while the association of the additional PH 1 500 FTU/kg and SB supplementation resulted in the greater depth and width of the jejunum crypt and villus : ileum crypt at seven days and greater villus length and ileum absorption area at 28 days. The association of SB and PH may be a strategy to improve the performance and bone quality and intestinal integrity in broilers.

Effect of stocking density on growth performance, feather quality, serum hormone, and intestinal development of geese from 1 to 14 days of age

This study was conducted to investigate the effect of stocking density on growth performance, feather quality, serum hormone, and intestinal development of geese from 1 to 14 d of age. A total of 450 one-day-old geese were randomly allotted to 45 battery cage (0.65 m × 0.62 m) pens according to 5 stocking densities (15, 20, 25, 30, and 35 birds/m2). The results showed that ADG and ADFI were reduced (P < 0.05) as stocking density increased from 15 to 35 birds/m2, but increasing stocking density did not influence (P > 0.05) feed conversion ratio (FCR) and body measurement traits. High stocking density significantly decreased (P < 0.05) the feather quality of back, thoracoabdominal, wing, and tail. No significant difference (P > 0.05) was found in serum concentration of adrenocorticotrophic hormone, cortisol, corticosterone, triiodothyronine, and thyroxine. The weight of cecum and intestine decreased (P < 0.05) as the stocking density increased. Increasing stocking density decreased (P < 0.05) jejunal villus height and villus height-to-crypt depth ratio, and increased (P < 0.05) jejunal crypt depth and ileal crypt depth in geese. Consequently, the high stocking density could depress the growth and impaired feather quality and intestinal development of geese. Under our experimental conditions, we recommend that the stocking density of geese from 1 to 14 d of age should not more than 20 birds/m2.

Maternal obesity (MO) programs morphological changes in aged rat offspring small intestine in a sex dependent manner: Effects of maternal resveratrol supplementation

Maternal obesity (MO) leads to offspring metabolic problems. The mechanisms involved are multifactorial. The small intestine plays an important role in the absorption of nutrients and is modified as we age. Few studies have explored MO programming effects on offspring (F1) small intestine morphology. The aim of this study was to investigate MO effects on old adult F1 intestinal morphology, and whether any F1 intestinal changes due to MO were modified by maternal resveratrol supplementation. From weaning throughout pregnancy and lactation, female Wistar rats (F0) ate standard chow (controls, C: 5%-fat) or high-fat diet (MO: 25%-fat). One month before mating at postnatal day (PND) 120 through lactation half of each group received 20 mg/kg/day of resveratrol orally (Cres or MOres). After weaning F1 were fed with chow diet until the end of the study at PND 650. Body weight, percent of fat, glucose, cholesterol and triglyceride serum concentrations were determined. F1 small intestinal samples were collected for histological analysis. Male F1 body weight was higher in MO and MOres compared with C and Cres. Female F1 body weight and percent of fat was higher in MO than C and MOres. Triglyceride concentrations were higher in MO and MOres male F1 compared with C and Cres. There were no differences among groups in female triglyceride concentrations. Male F1 duodenal villus height was smaller in MO compared with MOres. Female F1 duodenal and jejunal crypt depth was smaller in MO compared with C and was greater compared with MOres. Female F1 villus height in jejunum was greater in MO compared with MOres. In conclusion, exposure to the developmental challenge of MO changed the aged F1 intestinal morphological and metabolic profiles. Maternal resveratrol supplementation ameliorated these effects in an F1 sex dependent manner.

L-Carnosine Protects Against Deoxynivalenol-Induced Oxidative Stress in Intestinal Stem Cells by Regulating the Keap1/Nrf2 Signaling Pathway.

The intestinal epithelium is nourished by various nutrients and subjected to persistent and widespread feed-derived mycotoxin stress. l-Carnosine (LC) possesses robust antioxidant activity; however, its role in protecting intestinal mucosa against deoxynivalenol (DON) is still unclear. In this study, 300mg kg-1 BW LC and 3mg kg-1 BW DON are orally administered to mice either alone or in combination for 10 days to investigate the role of LC in protecting the intestine against DON. This study found that LC alleviates the growth retardation of mice and repairs the damaged jejunal structure and barrier functions under DON exposure. LC rescues the intestinal stem cells (ISCs), increases the growth advantage in enteroids derived from jejunal crypts of mice in each group ex vivo, improves the proliferation and apoptosis of intestinal cells, and promotes ISC differentiation into absorptive cells, goblet cells, and Paneth cells. Furthermore, LC activates Nrf2 signaling by binding to Keap1 to reverse the striking DON-induced increase in ROS levels. The study findings unveil that LC potentiates the antioxidant capacity of ISCs by regulating the Keap1/Nrf2 signaling pathway, which contributes to the intestinal epithelial regeneration response to DON insult.

Dietary Lactobacillus fermentum and Bacillus coagulans Supplementation Modulates Intestinal Immunity and Microbiota of Broiler Chickens Challenged by Clostridium perfringens.

Preventative effects of Lactobacillus fermentum and Bacillus coagulans against Clostridium perfringens infection in broilers have been well-demonstrated. The present study was conducted to investigate the modulation of these two probiotics on intestinal immunity and microbiota of C. perfringens-challenged birds. The 336 one-day-old broilers were assigned to four groups with six replicates in each group. Birds in the control were unchallenged and fed a basal diet, and birds in the three challenged groups were dietary supplemented with nothing (Cp group), 1 × 109 CFU/kg of L. fermentum (Lf_Cp group), or 1 × 1010 CFU/kg of B. coagulans (Bc_Cp group). Challenge was performed from days 14 to 20, and samples were collected on days 21 and 28. Challenge upregulated interleukin (IL)-1β and transforming growth factor (TGF)-β4 mRNA expression in jejunum on day 21, which was downregulated by B. coagulans and L. fermentum, respectively (P < 0.05). Both probiotic groups upregulated jejunal IL-1β, interferon (IFN)-γ, IL-17, and TGF-β4 on day 28 as well as IFN-γ on day 21 (P < 0.05). The Bc_Cp group increased CD3+ T cell counts in the jejunal crypt on day 21 (P < 0.05). Challenge decreased the ileal ACE index on day 21 and cecal microbial richness on day 28, which were increased by probiotic treatments, and ileal bacterial richness decreased in the Bc_Cp group on day 28 (P < 0.05). Only ileal microbiota on day 21 was distinctly affected with an R-value at 0.3116 by ANOSIM analysis (P < 0.05). Compared with the control, ileal Firmicutes increased on day 21, and ileal Bacteroidetes and cecal Proteobacteria decreased on day 28 in challenged groups (P < 0.05). Challenge increased Romboutsia spp. in the ileum as well as unclassified f_Lachnospiraceae and Ruminococcus_torques group in the cecum, and decreased Lactobacillus spp. in the ileum on day 21, which were all conversely modulated by L. fermentum (P < 0.05). Challenge increased amino acid metabolism of ileal microbiota and membrane transport of cecal microbiota, and decreased amino acid metabolism of cecal microbiota on day 21, which were conversely regulated by both probiotics (P < 0.05). In conclusion, L. fermentum and B. coagulans attenuated the intestinal inflammation and microbial dysbiosis soon after C. perfringens challenge.

Effects of Ursolic Acid on Intestinal Health and Gut Bacteria Antibiotic Resistance in Mice.

Ursolic acid (UA), a natural pentacyclic triterpenoid, has been widely reported to exert anti-oxidant and anti-inflammatory properties. However, the effects of UA on the intestinal homeostasis and gut microbiota were rarely explored. The aim of the present study was to investigate the effects of UA on intestinal health and gut microflora antibiotic-resistance in antibiotic-exposed mice. Kunming mice (n = 80) were randomly allocated into three groups and fed with one of the following diets, respectively: Cont group (n = 20), the basal diet; UA group (n = 20), the basal diet supplemented with 150 mg/kg UA; Tet group (n = 40), the basal diet supplemented with 659 mg/kg chlortetracycline. After 14 days, 10 mice in each group were euthanatized and the remaining 30 mice in the Tet group were randomly allocated into three sub-groups (n = 10 per group) as follows: the Tet group which were kept feeding a Tet diet for 14 days; the Natural Restoration (NatR) group which received a basal diet for 14 days; and the UA therapy (UaT) group which fed a basal diet supplemented with 150 mg/kg UA for 14 days. Throughout the experiment, the weight and the food intake of each mouse were recorded once weekly. Serum LPS and diamine oxidase (DAO), jejunal morphology, jejunal tight junction proteins and nutrient transporters, colonic inflammatory cytokines, gut microbiota and its antibiotic resistance gene (ARG) were examined at euthanasia. The results showed that UA treatment significantly increased average daily food intake (ADFI) of mice. Notably, UA increased the jejunal villi height, decreased the jejunal crypt depth and promoted the expression of jejunum nutrient transporters. UaT group had higher villi height, lower crypt depth and higher nutrient transporter mRNA expression in jejunum than NatR group. Besides, UA decreased serum DAO content, upregulated mRNA expression of ZO-1, claudin-1 and occludin and downregulated TNF-α and IL-6. The mRNA abundances of ZO-1, claudin-1 and occludin and TNF-α and IL-6 in UaT group were, respectively upregulated and downregulated than NatR group. Furthermore, an analysis of 16S rDNA sequences demonstrated that UA increased the abundance of beneficial bacteria in the gut. And the results of ARG test showed that UA downregulated the expression of antibiotic-induced resistance genes. The UaT group inhibited the increase of harmful bacteria abundance and suppressed the mRNA abundances of ARG compared to the NatR group. In conclusion, considering the positive effects of UA on the growth performance and intestinal mucosal barrier, we anticipate that these findings could be a stepping stone for developing UA as a novel substitute of antibiotics.

Pharmacologic ascorbate enhances the therapeutic index of ATM-inhibitor based chemoradiation for colorectal cancer.

3609 Background: Ataxia telangectasia mutated protein (ATM) is one of the key sensors of DNA damage and specific inhibitors of ATM are potent radiosensitizers. However, their clinical utility with radiation (RT) is limited because they lack tissue specificity and increase normal tissue injury. Pharmacologic (high dose) ascorbate (P-AscH-) selectively increases oxidative stress in tumors while functioning as a donor antioxidant and reducing RT damage in normal tissues. We hypothesized that P-AscH- could enhance the therapeutic index of ATM-inhibitor based chemoradiation (CRT) for colorectal cancer (CRC) by simultaneously enhancing efficacy and reducing RT bowel injury. Methods: Human HCT116, SW480, and HT29 and murine CT26 and MC38 CRC models were used. Clonogenic survival was assessed following single-fraction RT (2-8 Gy) +/- P-AscH- (5 pM/cell) +/- veliparib (PARP), VE821 (ATR), or KU60019 (ATM). Catalase expression was induced using HCT116 cells expressing a doxycycline inducible catalase transgene. DNA double strand breaks (DSBs) were quantified using neutral comet assays 0-24 hours post RT. Cell cycle phases were assessed using flow cytometry. ATM and pATM localization were assessed using IF. Jejunal toxicity was assessed using IHC in fixed tissues following single fraction (10 Gy) whole abdominal RT in c57blj/6 mice. Tumor growth delay was assessed following RT (5 Gy x 3) +/- drug treatment in unilateral flank tumors. Results: Veliparib, VE821, and KU60019 were potent radiosensitizers in HCT116, SW480, HT29, MC38, and CT26 CRC tumor models and P-AscH- further reduced clonogenic survival with DRIs in all lines except for HT29. In contrast, P-AscH- enhanced survival of cultured HUVEC and FHs-74 cells exposed to RT. Enhanced cell kill with P-AscH- is H202 mediated as it is completely attenuated by inducible catalase expression. P-AscH- significantly increased the number of DNA DSBs in tumors after RT in vitro. Despite the increase in DNA DSBs, P-AscH-significantly decreased nuclear localization of activated P-ATM after RT and significantly decreased the fraction of cells in G2/M phases of the cell cycle. In vivo, RT + P-AscH- + KU60019 induced more tumor growth delay/clearance than all other combinations in unilateral MC38 or HCT116 flank tumors. Finally, P-AscH- significantly reduced loss of jejunal crypt cell density, epithelial architecture, and markers of lipid and protein oxidation following whole abdominal RT. Conclusions: P-AscH- selectively enhances the efficacy of ATM-based CRT in CRC tumor models while simultaneously decreasing RT-mediated small bowel toxicity. In tumors, P-AscH- enhances DNA DSBs by stimulating an H202 flux and prevents activation of DNA repair pathways and cell cycle checkpoints by inhibiting RT-induced activation of ATM. Selective radioprotectors like P-AscH- could facilitate the clinical translation ATM inhibitors as radiosensitizers.

Effects of red-osier dogwood extracts on growth performance, intestinal digestive and absorptive functions, and meat quality of broiler chickens

A total of 320 one-day-old Cobb 500 chicks with an initial weight of 48.3 ± 3.3 g·pen−1 were assigned to four dietary treatments with eight replicates provided in three phases for 46 d. The treatments were fed as mash diets and included (1) negative control (NC) corn-soybean basal diet, (2) positive control (PC) basal diet with 30 ppm avilamycin, (3) basal diet supplemented with 1000 ppm red-osier dogwood extracts (RDE1), and (4) basal diet with 3000 ppm red-osier dogwood extracts (RDE2). Results showed reduced jejunal crypt depth in RDE1 and increased villus:crypt ratio in groups (either RDE1 or RDE2) (P &lt; 0.05). Cationic amino acid (AA) transporter mRNA abundance was decreased (P &lt; 0.05) in RDE1, RDE2, and PC treatments, but peptide and neutral AA transporter mRNA abundance were higher (P &lt; 0.05) in RDE2 compared with NC. Apparent ileal digestibility of crude fat was increased in RDE2 and PC compared with NC, whereas AA digestibility was greater in RDE1, RDE2, and PC (P &lt; 0.05). In conclusion, red-osier dogwood had no effect on growth performance, improved the intestinal health and function of broiler chickens, and had no detrimental effects on meat quality.

Growth performance, carcass traits and gut health of broiler chickens fed diets incorporated with single cell protein.

ObjectiveThis study was conducted to evaluate single cell protein (SCP), produced from Methylococcus species, as a protein source on the growth performance, carcass traits and gut health of broiler chickens.MethodsTen iso-nitrogenous and iso-caloric diets containing 0 (Control), 2.5%, 5%, 7.5%, and 10% SCP replacing either soybean meal (T1 to T5) or fish meal (T6 to T10) were formulated. Each diet prepared for starter (0 to 14 days), grower (15 to 21 days), and finisher (22 to 42 days) phases was offered to four replicates of 10 chicks each (n = 400). Growth performance at different phases and carcass characteristics and intestinal morphology on 42nd day of trial were measured.ResultsBody weight gain in groups fed 2.5% and 5% SCP diets were comparable to control during different phases and cumulatively, however lower (p<0.01) in 7.5% and 10% SCP diets. Feed conversion ratio was better (p<0.01) in 2.5% and 5% SCP diets. Dressing percentage, abdominal fat percentage and meat:bone ratio were not affected (p>0.05) by SCP inclusion in the diets. However, breast percentage was higher (p<0.01) in 2.5% and 5% SCP groups and thigh percentage higher in 7.5% and 10% SCP groups. Total microbial count in duodenum, jejunum and ileum were not affected (p>0.05) by SCP inclusion up to 10% in diets. Duodenal villi length and crypt depth were highest (p<0.01) in group fed 5% SCP diets and lowest in group fed 10% SCP diets. Jejunal villi length and crypt depth as well as ileal villi length were lowest (p<0.01) in group fed 10% SCP diets. Body weight gain, feed consumption, feed conversion ratio and gut health were better (p<0.01) in broilers fed fish meal based diets compared to soybean meal based diets.ConclusionIt was concluded that inclusion of SCP up to 5% replacing soybean meal in broiler diets is beneficial in improving growth rate, breast yield and gut health status.

Decreased nutrient digestibility due to viscosity is independent of the amount of dietary fibre fed to growing pigs.

Fibre content and its effect on chyme viscosity are associated with changes in the digestive system of humans and pigs. It is unclear if fibre content and viscosity affect digestive function independently or interactively. We evaluated apparent ileal digestibility (AID) of nutrients and intestinal function in thirty-six ileal-cannulated barrows fed for 29 d either maize-soyabean meal (MSBM) or high-fibre MSBM + 30 % distillers dried grains with solubles (MSBM+DDGS) modified to three levels of viscosity by adding 5 % non-viscous cellulose (CEL), 6·5 % medium-viscous carboxymethylcellulose (MCMC) or 6·5 % high-viscous CMC (HCMC). Digesta were collected on days 27 and 28 and intestinal samples on day 29. Feeding CMC, regardless of fibre content, increased viscosity of whole digesta (P = 0·003) and digesta supernatant (P < 0·0001) compared with CEL. Feeding MSBM+DDGS or CMC decreased AID of DM (P = 0·003; P < 0·0001) and crude protein (P = 0·02; P<0·0001) compared with MSBM or CEL. Feeding CMC regardless of fibre content increased jejunal crypt depth (P = 0·02) and ileal goblet cell area (P = 0·004) compared with CEL. Adding DDGS or CMC did not affect villus height and gene expression of jejunal monosaccharide and amino acid transporters. Feeding HCMC, regardless of fibre content, elevated amylase activity by 46 and 50 % in jejunal (P = 0·03) and ileal digesta (P = 0·01) compared with CEL. In summary, diets with increased viscosity decreased nutrient digestibility and induced intestinal changes that were independent of the amount of fibre fed.