Determination of ileal endogenous nitrogen losses and true ileal nitrogen digestibility during non-steady-state conditions of the 15N-isotope dilution technique.

The aim was to determine ileal endogenous nitrogen losses (ENL) and true ileal N-digestibility (TD-N) under non-steady-state conditions of the 15N-isotope dilution technique (15N-IDT), using diets generating low and high ENL and compare results to those obtained under steady-state conditions. Twelve growing pigs (mean LW 22.4 kg) fitted with a post-valve T-caecum cannula were fed an enzyme-hydrolysed casein (EHC)-based diet or an EHC diet + 4% quebracho tannins (QT) and were labelled via continuous 15N-leucine i.v. infusion or twice daily oral 15N-leucine administration. Digesta were collected daily over three consecutive hours with blood plasma sampled on the four consecutive days after cessation of 15N-labelling. There was a significant effect of sampling day on the dilution factor. Endogenous N losses were significantly lower for the EHC than the EHC+QT diet (2.41 vs. 8.69 g/kg DMI), while no significant effect of sampling day was observed. The TD-N of the EHC+QT diet did not differ from the TD-N of the EHC diet (95.1 vs. 92.0%). A significant effect of sampling day was observed for TD-N with day 1 and 2, being higher than day 4. Non-steady-state conditions overestimated ENL by 25-28% as compared to 3 h collections in steady-state conditions, but the relative overestimation was similar for the EHC diet as for the EHC+QT diet. TD-N did not differ significantly compared to 12 h steady-state measurements, but comparison to 3 h steady-state measurements showed that non-steady-state conditions overestimated TD-N for the EHC+QT diet by 9%. However, on day 4 this overestimation disappeared. Using the 15N-IDT during non-steady-state conditions can provide valuable additional data on endogenous N losses and TD-N.

The standardized ileal digestible lysine-to-net energy ratio in the diets of sows to optimize milk nitrogen retention is dynamic during lactation.

Fifty-two multiparous sows (average parity 3.1 ±â€…0.9 and initial BW 245.6 ±â€…32.5 kg) were used to evaluate the effects of dietary standardized ileal digestible (SID) Lys-to-net energy (NE) ratios on nitrogen (N) utilization throughout a 24-d lactation period. Sows were randomly assigned to one of five isoenergetic feeding programs that provided equally spaced and increasing SID Lys-to-NE ratios between 2.79 and 5.50 g SID Lys/Mcal NE. The feeding programs were generated by blending the two extreme diets in varying proportions and were provided to sows immediately after farrowing (day 1) and until weaning at day 24 ±â€…1. Nitrogen balances were conducted between days 4 and 7, 12 and 15, and 20 and 23 ±â€…1 of lactation to represent weeks 1, 2, and 3, respectively, using total urine collection and fecal grab sampling. Contrast statements were used to determine the linear and quadratic effects of increasing Lys-to-NE ratios. Linear and quadratic broken-line and polynomial quadratic (QPM) models were used to determine the optimum dietary Lys-to-NE ratios for N retention in milk. The Bayesian information criterion was used to assess the best fit. Feeding program did not influence sow average daily feed intake (5.8 ±â€…0.1 kg), BW change (-8.2 ±â€…3.1 kg), or change in back fat thickness (-2.6 ±â€…0.7 mm) over the 24-d lactation period, but piglet average daily gain increased with dietary SID Lys-to-NE ratio (linear; P < 0.05). Sow N intake increased with increasing dietary Lys-to-NE ratio in weeks 2 and 3 (linear; P < 0.001). Whole-body N retention (N intake - N output in urine and feces) increased with increasing dietary Lys-to-NE ratio in all weeks (linear; P < 0.05). The N retention in milk tended to increase then decrease with increasing dietary Lys-to-NE ratio in weeks 1 and 2 (quadratic; P = 0.051 and P = 0.081) and the QPM showed optimal milk N retention at 4.28, 4.42, and 4.67 g Lys/Mcal NE for weeks 1, 2, and 3, respectively. Maternal N retention (N intake - N output in urine, feces, and milk) decreased and then increased in week 1 (quadratic; P < 0.01) and increased in weeks 2 and 3 (linear; P < 0.01) with increasing dietary Lys-to-NE ratio. Therefore, the SID Lys-to-NE ratio necessary to optimize milk N output is dynamic throughout lactation. A two-diet feeding program could be created to match optimal weekly or daily SID Lys-to-NE ratios, which could lead to improved piglet ADG and body weights at weaning.

Despite significant changes in nutrient and energy requirements as well as voluntary feed intake during lactation, sows are typically fed a single diet with a static nutrient and energy composition throughout the entire lactation period, which may not optimize milk output. Fifty-two sows were used to explore how various ratios of standardized ileal digestible (SID) Lys to net energy (NE) in lactating sow diets affect the growth of piglets and nitrogen utilization during a 24-d lactation period. Sows were randomly assigned to one of five feeding programs that contained equal amounts of energy and provided equally spaced and increasing Lys-to-NE ratios between 2.79 and 5.50 g SID Lys/Mcal NE immediately after farrowing. The dietary Lys-to-NE ratio did not influence sow daily feed intake, body weight change, or change in backfat thickness over the 24-d lactation period; however, piglet growth rate and body weight at weaning increased with increasing Lys-to-NE ratio. The SID Lys-to-NE ratio necessary to optimize milk nitrogen output was 4.28, 4.42, and 4.67 g SID Lys/Mcal NE during weeks one, two, and three of lactation, respectively. Therefore, it is possible to create a two-diet feeding program offering dynamic SID Lys-to-NE ratios as lactation progresses, which could lead to improved piglet average daily gain and body weights at weaning.

Effects of the algae derivatives on performance, intestinal histomorphology, ileal microflora, and egg yolk biochemistry of laying Japanese quail.

We examined the effect of the Persian Gulf algae derivates, phycocyanin (PC) and fucoidan (FUC), on production performance, egg quality, intestinal histomorphology, ileal microflora, and egg yolk biochemistry of laying Japanese quail. A total of 250 six-wk-old Japanese quails with an average body weight of 215 ± 10 g were allocated to 5 treatments, 5 replicates and 10 birds in each replicate in a completely randomized design. The treatment groups received PC (from Spirulina platensis) and FUC (from brown seaweed) in their drinking water while control groups did not. Treatment groups received PC and FUC at 20 or 40 mg/L levels (denoted as PC20, PC40, FUC20, and FUC40, respectively). All birds were fed the same diet. All treatments significantly improved the percentage of hen day egg production (HDEP) (P = 0.002), egg mass (P = 0.002), and feed conversion ratio (FCR) (P = 0.022) but no difference was noted in egg weight (EW) and feed intake (FI). Different levels of PC and FUC significantly increased the thickness of eggshells (P = 0.022); however, the weight of the digestive tract (liver, spleen, proventriculus, gizzard, and pancreas) and oviduct was not affected. Algal derivates improved the villus height (P = 0.007) and crypt depth (P = 0.007) of the duodenum, as well as, the villus height (P = 0.005) and crypt depth (P = 0.026) of the jejunum. Both algal derivates positively affected the intestinal microflora (populations of Lactobacillus (P = 0.017), Coliform (P = 0.005), and Clostridium (P = 0.000)) whereas aerobic bacteria were unaffected. Yolk cholesterol P = 0.012) and yolk malondialdehyde P = 0.050) content were significantly reduced in experimental treatments compared to the control group. In conclusion, our results showed that the treatment of laying Japanese quails with algal derivates positively affects quail performance, intestinal morphology, intestinal microflora, and yolk cholesterol and malondialdehyde. Additional studies exploring optimal dosages and mechanisms of action is warranted to fully understand the scope of the algae derivates in poultry production.

Ileal phosphorus digestibility of soybean meal for broiler chickens remains consistent across institutions in a collaborative study regardless of non-phytate phosphorus concentration in the pre-experimental starter diet.

The same experimental protocol was used in 4 institutions to evaluate the impact of non-phytate phosphorus (nPP) concentration in the starter diet on regression method-derived ileal P digestibility of soybean meal (SBM) during the subsequent grower phase. A total of 1,536 Ross 308 male broiler chickens on d 0 post hatching were allotted to 2 pre-experimental starter diets that contained 3.5 or 4.5 g nPP/kg (96 replicate cages per diet, 8 birds per cage) for 18 d. Subsequently, 576 birds from each starter diet were selected and allocated to 3 experimental semi-purified grower diets containing 400, 510, or 620 g SBM/kg (32 replicate cages per diet, 6 birds per cage) for 3 d until collection of ileal digesta. Statistical analysis was conducted as a randomized complete block design with the starter period as whole plot and the grower period as split-plot. The only significant 2-way interaction was between grower diet and experimental institution (P < 0.05) on BW gain and gain to feed ratio. The main effect of institution and grower diet impacted (P < 0.05) feed intake, the digestibility of DM, P, and calcium, and disappearance of inositol hexakisphosphate (InsP6) in the grower diets. Birds fed the 3.5 g nPP/kg starter diet had lower (P < 0.05) BW gain and feed intake during the grower period, but presented higher (P < 0.05) digestibility of P and disappearance of InsP6 compared with the birds that were fed the 4.5 g nPP/kg starter diet. Regression method-derived ileal P digestibility of SBM was determined to be 46 or 42% for the respective 3.5 or 4.5 g nPP/kg pre-experimental starter diet and was not affected by the nPP concentration or by the institution. In conclusion, the experimental protocol used in the current study resulted in similar estimates across multiple institutions and is thus endorsed for future application in studies that aim to expand the database of digestible P content in plant source feed ingredients.

Unveiling the influence of adaptation time on xylanase and arabinoxylan-oligosaccharide efficacy: a study on nutrient digestibility, viscosity, and scanning electron microscopy in the small and large intestine of growing pigs fed insoluble fiber.

The experiment objective was to evaluate the impact of xylanase over time on viscosity and digestibility in growing pigs fed corn-based fiber. Twenty gilts with an initial body weight of 30.6 ±â€…0.2 kg (n = 5 per dietary treatment) were fitted with t-cannulae in the medial jejunum and terminal ileum, housed individually, and randomly assigned to one of four dietary treatments: low-fiber control (LF) with 10.4% total dietary fiber (TDF), 30% corn bran high-fiber control (HF; 26.4% TDF), HF + 100 mg xylanase/kg (XY; Econase XT 25P; AB Vista, Marlborough, UK), and HF + 50 mg arabinoxylan-oligosaccharide/kg (AX). Gilts were limit fed for three 17 d periods (P1, P2, P3); each included 5 d adaptation, 2 d fecal collection, 3 d ileal collection, 3 d jejunal collection, and 4 d related rate of passage study. Data were analyzed as repeated measures using a linear mixed model with surgery date as a random effect, and dietary treatment, period, and their interaction as fixed effects. Jejunal and ileal digesta viscosity did not differ among dietary treatments or periods (P > 0.10). There was a dietary treatment × period interaction for the apparent jejunal digestibility (AJD) of dry matter (DM), gross energy (GE), insoluble dietary fiber (IDF), neutral detergent fiber (NDF), total arabinoxylan (T-AX), total non-starch polysaccharide (T-NSP), and TDF (P≤ 0.05). In P1, LF had the greatest AJD of DM (15.5%), and relative to HF and AX, XY decreased it (9.3%, 10.1 %, and 6.3%, respectively). In P2, the AJD of DM in XY was greater than HF (11.7% vs. 9.1%) but did not differ from AX (10.5%). Relative to HF, in P3, XY increased AJD of DM (11.7 vs 15.3%), and AX decreased it (7.2%). For the AJD of NDF, AX performed intermediately in P1; in P2, relative to HF, XY, and AX increased the AJD of NDF (8.4%, 13.1%, and 11.7%, respectively), and in P3, XY, and LF did not differ (13.6 vs. 14.4%). A similar response was observed for the AJD of IDF and TDF, except for XY having the greatest AJD of IDF, T-AX, T-NSP, and TDF in P3 (P < 0.05). Compared to LF, irrespective of period, HF decreased the apparent ileal digestibility (AID) and apparent total tract digestibility (ATTD) of IDF, TDF, and NDF (P < 0.05). Relative to HF, XY partially mitigated this effect, improving the AID and ATTD of TDF, IDF, and NDF (P < 0.05). Increased corn-based fiber decreased nutrient digestibility, but XY partially mitigated that effect in the small intestine through enhanced fiber digestibility when given sufficient adaptation time.

This study investigated the effects of xylanase and arabinoxylan-oligosaccharide supplementation on viscosity, nutrient and energy digestibility in growing pigs fed a high-fiber diet mainly composed of corn over three time periods. Twenty pigs were surgically fitted with cannula in their jejunum and ileum for sample collection. The pigs were randomly assigned to one of four dietary treatment groups: low-fiber control, high-fiber control, xylanase supplementation, and arabinoxylan-oligosaccharide supplementation. The results showed no significant differences in digesta viscosity among dietary treatments. However, there was an interaction between dietary treatment and time for the digestibility of dry matter, gross energy, and dietary fibers. Initially, xylanase did not impact digestibility, but it improved over time. Conversely, arabinoxylan-oligosaccharide initially improved digestibility but declined by the third period. Findings suggest that the efficacy of xylanase in enhancing nutrient and energy digestibility in pigs fed high-fiber diets may depend on the length of the adaptation period. Xylanase supplementation also demonstrated the potential to counteract the negative effects of high-fiber diets. Further research is needed to optimize the application of xylanase in swine production and determine the optimal conditions for its effectiveness.

Microbial experience through housing in a farmyard-type environment alters intestinal barrier properties in mouse colons.

To close the gap between ultra-hygienic research mouse models and the much more environmentally exposed conditions of humans, we have established a system where laboratory mice are raised under a full set of environmental factors present in a naturalistic, farmyard-type habitat-a process we have called feralization. In previous studies we have shown that feralized (Fer) mice were protected against colorectal cancer when compared to conventionally reared laboratory mice (Lab). However, the protective mechanisms remain to be elucidated. Disruption of the protective intestinal barrier is an acknowledged player in colorectal carcinogenesis, and in the current study we assessed colonic mucosal barrier properties in healthy, feralized C57BL/6JRj male mice. While we found no effect of feralization on mucus layer properties, higher expression of genes encoding the mucus components Fcgbp and Clca1 still suggested mucus enforcement due to feralization. Genes encoding other proteins known to be involved in bacterial defense (Itln1, Ang1, Retnlb) and inflammatory mechanisms (Zbp1, Gsdmc2) were also higher expressed in feralized mice, further suggesting that the Fer mice have an altered intestinal mucosal barrier. These findings demonstrate that microbial experience conferred by housing in a farmyard-type environment alters the intestinal barrier properties in mice possibly leading to a more robust protection against disease. Future studies to unravel regulatory roles of feralization on intestinal barrier should aim to conduct proteomic analyses and in vivo performance of the feralized mice intestinal barrier.

Energy and phosphorus utilization of pulses fed to broiler chickens.

Energy and P utilization in faba beans and peas were evaluated in 3 broiler chicken experiments. In Exp. 1, 240 birds were allotted to 5 diets in a randomized complete block design with BW as a blocking factor on d 18 post hatching to determine the regression-derived energy utilization of faba beans (FB) and field peas (FP). In each of the respective Exp. 2 and 3, regression-derived P utilization in FB and FP were determined with 162 birds assigned to each of 3 diets in a randomized complete block design with BW as a blocking factor on d 19 post hatching. There were 8 replicate cages with 6 birds per cage in Exp. 1, and 6 replicate cages with 9 birds per cage in Exp. 2 and 3. The test ingredients were added to a corn-soybean meal-based diet at 15% or 30% in Exp. 1, whereas FB was included at 21%, 42%, or 63% and FP at 16%, 32%, or 48% in Exp. 2 and 3, respectively. In Exp. 1, the apparent ileal digestibility (AID) of gross energy (GE) and the ileal digestible energy (IDE) in the diets decreased linearly (P < 0.01). There was a quadratic response or a linear decrease (P < 0.05) with increasing concentrations of FB or FP, respectively, on the apparent total tract utilization (ATTU) of GE, metabolizable energy (ME), and nitrogen-corrected ME (MEn). The respective IDE, ME, and MEn determined were 2,541, 2,628, and 2,394 kcal/kg DM in FB and 2,254, 2,540, and 2,331 kcal/kg DM in FP. In Exp. 2 and 3, the ileal digestible and retainable P intake were linearly increased (P < 0.01). The estimated true ileal digestibility and true total tract utilization of P in FB were 66.5% and 66.7%, respectively. The respective corresponding values for FP were 73.4% and 73.8%. In conclusion, the information on utilization of energy and P in FB and FP provided could enhance proper diet formulation when using these ingredients.

The bioelectrical conduction system around the ileocecal junction defined through in vivo high-resolution mapping in rabbits.

Coordinated contractions across the small and large intestines via the ileocecal junction (ICJ) are critical to healthy gastrointestinal function and are in part governed by myoelectrical activity. In this study, the spatiotemporal characteristics of the bioelectrical conduction across the ICJ and its adjacent regions were quantified in anesthetized rabbits. High-resolution mapping was applied from the terminal ileum (TI) to the sacculus rotundus (SR), across the ICJ and into the beginning of the large intestine at the cecum ampulla coli (AC). Orally propagating slow wave patterns in the SR did not entrain the TI. However, aborally propagating patterns from the TI were able to entrain the SR. Bioelectrical activity was recorded within the ICJ and AC, revealing complex interactions of slow waves, spike bursts, and bioelectrical quiescence. This suggests the involvement of myogenic coordination when regulating motility between the small and large intestines. Mean slow wave frequency between regions did not vary significantly (13.74-17.16 cycles/min). Slow waves in the SR propagated with significantly faster speeds (18.51 ± 1.57 mm/s) compared with the TI (14.05 ± 2.53 mm/s, P = 0.0113) and AC (9.56 ± 1.56 mm/s, P = 0.0001). Significantly higher amplitudes were observed in both the TI (0.28 ± 0.13 mV, P = 0.0167) and SR (0.24 ± 0.08 mV, P = 0.0159) within the small intestine compared with the large intestine AC (0.03 ± 0.01 mV). We hypothesize that orally propagating slow waves facilitate a motor-brake pattern in the SR to limit outflow into the ICJ, similar to those previously observed in other gastrointestinal regions.NEW & NOTEWORTHY Competing slow wave pacemakers were observed in the terminal ileum and sacculus rotundus. Prevalent oral propagation in the sacculus rotundus toward the terminal ileum potentially acts as a brake mechanism limiting outflow. Slow waves and periods of quiescence at the ileocecal junction suggest that activation may depend on the coregulatory flow and distention pathways. Slow waves and spike bursts in the cecum impart a role in the coordination of motility.

TLR2 and TLR4 Modulate Mouse Ileal Motility by the Interaction with Muscarinic and Nicotinic Receptors.

Irritable bowel syndrome (IBS) is a chronic functional bowel disorder characterized by intestinal dysmotility. Changes in intestinal microbiota (dysbiosis) can lead to alterations in neuro-muscular functions in the gut. Toll-like receptors (TLRs) 2 and 4 recognize intestinal bacteria and are involved in the motor response induced by gastrointestinal (GI) neurotransmitters. Acetylcholine (ACh) is a well-known neurotransmitter involved in the regulation of GI motility. This study aimed to evaluate the role of TLR2 and TLR4 in the intestinal motor-response induced by ACh in the mouse ileum, as well as the expression and function of the muscarinic and nicotinic ACh receptors. Muscle contractility studies showed that the contractions induced by ACh were significantly lower in TLR2-/- and TLR4-/- with respect to WT mice. In WT mice, the contractions induced by ACh were reduced in the presence of AF-DX AF-DX 116 (a muscarinic ACh receptor (mAChR) M2 antagonist), 4-DAMP (a mAChR M3 antagonist), mecamylamine (a nicotinic AChR receptor (nAChR) α3ß4 antagonist) and α-bungarotoxin (a nAChR α7 antagonist). In TLR2-/- mice, the contractions induced by ACh were increased by AF-DX 116 and mecamylamine. In TLR4-/- mice, the contractions induced by ACh were reduced by α-bungarotoxin and 4-DAMP. The mRNA and protein expressions of M3 and α3 receptors were diminished in the ileum from TLR2-/- and TLR4-/- with respect to WT mice. However, the levels of mRNA and protein of ß4 were diminished only in TLR4-/- but not in TLR2-/- mice. In conclusion, our results show that TLR2 and TLR4 modulates the motor responses to ACh in the mouse ileum. TLR2 acts on muscarinic M2 and M3 and nicotinic α3ß4 ACh receptors, while TLR4 acts on muscarinic M3 and nicotinic α3ß4 and α7 ACh receptors.

Impact of increasing standardized ileal digestible valine: lysine in diets containing 30% dried distiller grains with solubles on growing pig performance.

A total of 2,430 pigs (DNA 600 × Topigs Norsvin 70, initially 39.4 kg) were used in a 28-d trial to determine the standardized ileal digestible (SID) Val:Lys requirement for pigs fed with diets containing 30% DDGS. Treatments included five diets containing 30% DDGS with SID Val:Lys ratios of 60%, 65%, 70%, 75%, and 80%, plus a corn soybean meal (SBM) diet, for a total of six dietary treatments. Diets were formulated to be isocaloric through the inclusion of fat and to contain equal amounts of SID Lys within phase. Pens were assigned to dietary treatment in a randomized complete block design with initial body weight (BW) as the random blocking factor. Each dietary treatment was replicated 15 times and pens contained 27 pigs, balanced for sex. Increasing the SID Val:Lys ratio in diets containing 30% DDGS increased (Quadratic; P ≤ 0.007) 14-d BW, final BW, average daily gain (ADG), average daily feed intake (ADFI), and gain:feed (G:F) for all periods. Providing an SID Val:Lys ratio of 75% resulted in the heaviest 14-d BW and final BW as well as greater ADG and ADFI of pigs fed with 30% DDGS for all periods. The G:F response was maximized when an SID Val:Lys ratio of 70% was provided from day 0 to 14 while an SID Val:Lys ratio of 75% maximized G:F from day 14 to 28 and for the cumulative period. The SID Val:Lys requirement was estimated at 66.6% (95% CI [65.9, 67.4]), 65.7 (95% CI: [64.8, 66.5]), and 68.4% (95% CI [66.0, 70.8]) for ADG, ADFI, and G:F, respectively, using the straight broken line (SBL) method and 69.9% (95% CI [68.2, 71.5]), 67.6 (95% CI [65.4, 69.8]), and 72.8% (95% CI [69.8, 75.8]) for the quadratic broken line (QBL) method. Pigs fed the corn-SBM diet had heavier 14-d BW, final BW, and greater ADG, ADFI, G:F (P ≤ 0.032) compared to pigs fed diets containing 30% DDGS, except for cumulative ADFI compared to pigs receiving 75% SID Val:Lys (P = 0.167). In conclusion, these results suggest that when feeding 30% DDGS during the growing period, an SID Val:Lys ratio of 68% would yield more than 99% and 97% of the maximum ADG and G:F response for the 39 to 68 kg pigs. However, growth performance of pigs fed diets containing 30% DDGS did not equate to pigs consuming the corn-SBM diet regardless of the SID Val:Lys ratio.

Understanding the nutrient requirements of pigs across various conditions is important to determine the most economical method of feeding pigs. When feeding pigs corn byproducts from the ethanol industry, such as dried distiller grains with solubles (DDGS), the growth performance of pigs decreases. This is thought to be due to the composition of the dietary protein, more specifically the amino acid concentration of the diet. In the present study, one specific amino acid, valine, was incrementally increased in diets containing 30% DDGS. The growth performance of pigs increased when digestible valine increase until a ratio of 68% was reached between digestible valine and the amino acid lysine. These results indicate that the digestible valine requirement for pigs between 39 and 68 kg of body weight is 68% of the lysine requirement. However, when growth performance of pigs fed with 30% DDGS was compared to a standard corn soybean meal diet, pigs fed with the standard corn soybean meal diet had greater performance, regardless of digestible valine level. More research is required to understand why the addition of 30% DDGS in pig diets negatively impacts the growth performance relative to a standard corn soybean meal diet.

Protective Effects of Selenium Nanoparticle-Enriched Lactococcus lactis NZ9000 against Enterotoxigenic Escherichia coli K88-Induced Intestinal Barrier Damage in Mice.

Composite microecological agents have received widespread attention due to their advantageous properties, including safety, multiple effects, and low cost. This study was conducted to evaluate the protective effects of selenium (Se) nanoparticle (SeNP)-enriched Lactococcus lactis NZ9000 (L. lactis NZ9000-SeNPs) against enterotoxigenic Escherichia coli (ETEC) K88-induced intestinal barrier damage in C57BL/6 mice. The oral administration of L. lactis NZ9000-SeNPs significantly increased the villus height and the number of goblet cells in the ileum; reduced the levels of serum and ileal interleukin-1ß (IL-1ß), tumor necrosis factor alpha (TNF-α), and interferon gamma (IFN-γ); and increased the activities of thioredoxin reductase (TrxR) and glutathione peroxidase (GSH-Px) compared with the ETEC K88-infected group not treated with L. lactis NZ9000-SeNPs. In addition, L. lactis NZ9000-SeNPs significantly attenuated the reduction of the expression levels of occludin and claudin-1, dysbiosis of the gut microbiome, and activation of the Toll-like receptor (TLR)/nuclear factor kappa B (NF-κB)-mediated signaling pathway induced by ETEC K88. These findings suggested that L. lactis NZ9000-SeNPs may be a promising and safe Se supplement for food or feed additives. IMPORTANCE The beneficial effects of microecological agents have been widely proven. Se, which is a nutritionally essential trace element for humans and animals, is incorporated into selenoproteins that have a wide range of pleiotropic effects, ranging from antioxidant to anti-inflammatory effects. However, sodium selenite, a common addition form of Se in feed and food, has disadvantages such as strong toxicity and low bioavailability. We investigated the protective effects of L. lactis NZ9000-SeNPs against ETEC K88-induced intestinal barrier injury in C57BL/6 mice. Our results show that L. lactis NZ9000-SeNPs effectively alleviate ETEC K88-induced intestinal barrier dysfunction. This study highlights the importance of developing a promising and safe Se supplement for the substitution of sodium selenite applied in food, feed, and biomedicine.

Transcriptome Analysis Reveals the Genes Involved in Growth and Metabolism in Muscovy Ducks.

Muscovy ducks are among the best meat ducks in the world. The objective of this study was to identify genes related to growth metabolism through transcriptome analysis of the ileal tissue of Muscovy ducks. Duck ileum samples with the highest (H group, n = 5) and lowest (L group, n = 5) body weight were selected from two hundred 70-day-old Muscovy ducks for transcriptome analysis by RNA sequencing. In the screening of differentially expressed genes (DEGs) between the H and L groups, a total of 602 DEGs with a fold change no less than 2 were identified, among which 285 were upregulated and 317 were downregulated. Gene Ontology (GO) enrichment analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis revealed that glutathione metabolism, pyrimidine metabolism, and protein digestion and absorption processes played a vital role in regulating growth and metabolism. The results showed that 7 genes related to growth and metabolism, namely, ANPEP, ENPEP, UPP1, SLC2A2, SLC6A19, NME4, and LOC106034733, were significantly expressed in group H, which was consistent with the phenotype results. The validation of these 7 genes using real-time quantitative PCR results indicated that the expression level of ENPEP was significantly different between the H and L groups (P < 0.05). This study provides a theoretical basis for exploring the influence of the ileum on growth and metabolism in ducks.

Detection of acute stress by smooth muscle electromyography: A translational study on rat and human.

AIMS: Non-invasive and simultaneous recording of gastrointestinal (GI) activity during stress induction is still an unexplored field. In our previous investigation, the stress-induced alteration of the gastrointestinal tract was explored in rats. Our aims were to expand our previous rat experiment and to induce stress response in rats (Study 1) and humans (Study 2) to detect the GI tract activity, heart rate and body temperature. MATERIALS AND METHODS: In the preclinical sample, acute stress was induced by immobilization in Sprague-Dawley rats (N = 10). Acute stress response was generated by the Trier Social Stress Test among healthy volunteers (N = 16). Detection of acute stress was measured by using smooth muscle electromyography, which recorded the myoelectric waves of the gastrointestinal tract (stomach, ileum and colon) simultaneously with heart rate and body temperature in rats and humans. KEY FINDINGS: The myoelectric waves of the stomach, the cecum and the ileum increased during immobilization in rats, rising in parallel with heart rate and the dermal temperature of the abdominal surface. The same alterations were found during the stress period among humans, except in the case of the colon, where no change was detected. SIGNIFICANCE: The crucial role of the GI tract in stress response was revealed by translating the outcome of basic research into human results. The similar GI alterations during stress in rats and humans underpin the robustness of our findings. In summary, our preliminary translational-based study can serve as an appropriate basis for further human studies.

Effects of spray dried yeast (Saccharomyces cerevisiae) on growth performance and carcass characteristics, gut health, cecal microbiota profile and apparent ileal digestibility of protein, amino acids and energy in broilers.

The aim of present study was to determine the effects of supplementation of either synbiotic or probiotic on growth performance and carcass characteristics, gut health, cecal microbiota prolife and apparent ileal digestibility of protein, amino acids, and energy in broilers. Two hundred and forty-day-old straight-run broilers (Ross 308) were allotted randomly to 1 of 5 dietary treatments including basal diet (control), supplemented with either synbiotic (Nutromax P) or probiotic (Actera), each at 0.5 and 1 g/kg of the diet for 5 weeks. The overall findings of the study indicated better (p < 0.05) growth performance of broilers by synbiotic supplementation (1 g/kg) compared with those fed probiotic (1 g/kg) supplemented and control diets. The broilers consuming diet supplemented with 1 g/kg synbiotic has an increased carcass yield in comparison with those fed control diet. The findings of gut health indicated significantly increased villus height and goblet cells, by synbiotic supplementation (1 g/kg), compared with control diet in broilers. The broilers fed 1 g/kg synbiotic supplemented diets had 18% increased protein, 9 to 31% higher amino acid, and 34% better energy digestibility, whereas 8.4% decreased protein digestibility in broilers fed probiotic (1 g/kg) supplemented compared with control diet in broilers. The broilers fed synbiotic (1 g/kg) supplemented diets had increased cecal Lactobacillus and decreased Salmonella, E. coli, and Clostridium count compared with those fed control diet. In conclusion, synbiotic supplementation (1 g/kg) resulted in improved production performance, balanced cecal microbial composition, and better digestibility of nutrients in broilers compared with those fed control and diets supplemented with probiotics.

Porcine colonoids and enteroids keep the memory of their origin during regeneration.

The development of alternative in vitro culture methods has increased in the last decade as three-dimensional organoids of various tissues, including those of the small and large intestines. Due to their multicellular composition, organoids offer advantages over traditionally used immortalized or primary cell lines. However, organoids must be accurate models of their tissues of origin. This study compared gene expression profiles with respect to markers of specific cell types (stem cells, enterocytes, goblet, and enteroendocrine cells) and barrier maturation (tight junctions) of colonoid and enteroid cultures with their tissues of origin and colonoids with enteroids. Colonoids derived from three healthy pigs formed multilobed structures with a monolayer of cells similar to the crypt structures in colonic tissue. Colonoid and enteroid gene expression signatures were more similar to those found for the tissues of their origin than to each other. However, relative to their derived tissues, organoids had increased gene expression levels of stem cell markers Sox9 and Lgr5 encoding sex-determining region Y-box 9 and leucine-rich repeat-containing G protein-coupled rector 5, respectively. In contrast, expression levels of Occl and Zo1 encoding occludin and zonula occludens 1, respectively, were decreased. Expression levels of the cell lineage markers Atoh1, Cga, and Muc2 encoding atonal homolog 1, chromogranin A, and mucin 2, respectively, were decreased in colonoids, whereas Sglt1 and Apn encoding sodium-glucose transporter 1 and aminopeptidase A, respectively, were decreased in enteroids. These results indicate colonoid and enteroid cultures were predominantly comprised of undifferentiated cell types with decreased barrier maturation relative to their tissues of origin.

Goat milk protein digestibility in relation to intestinal function.

BACKGROUND: Milk is an important high-quality animal protein source in low- and middle-income countries (LMICs). Although the true ileal digestibility and absorption of milk has been shown to be high in French adults, this may be lower in individuals from LMICs who are at risk of environmental enteropathy. OBJECTIVE: To determine the true ileal indispensable amino acid (IAA) digestibility of intrinsically labeled goat milk protein in South Indian women of reproductive age (WRA), using the dual-isotope tracer technique, and to measure intestinal absorption of amino acid and inert sugar in the same participants using L-allo-isoleucine and a dual-sugar assay. METHODS: Milk with 2H-labeled protein collected from a lactating goat fed intrinsically 2H-labeled fodder (maize and cowpea) was spray dried. Labeled milk protein was administered in a plateau feeding protocol to WRA with normal BMI, in whom urinary lactulose and mannitol recovery and the lactulose/mannitol ratio (LMR) were measured, to determine its true ileal IAA digestibility by the dual-isotope tracer technique with a reference U-13C-amino acid mixture. A phenylalanine absorption index was calculated from the plasma to meal ratio of 13C9 phenylalanine within the digestibility protocol. On a separate day, the allo-isoleucine absorption index was estimated from the ratio of plasma allo-isoleucine enrichments after oral 13C6-15N-L- and intravenous 2H10-L-allo-isoleucine administration. RESULTS: The means ± SDs of true ileal IAA digestibility of goat milk protein, lactulose and mannitol recovery, LMR, allo-isoleucine and phenylalanine absorption index were 94.0 ± 2.9%, 0.09 ± 0.03%, 7.9 ± 2.3%, 0.012 ± 0.004, 88.4 ± 3.8% and 24.5 ± 1.6%, respectively. The LMR correlated with the allo-isoleucine absorption index (rs = -0.93, P = 0.008). CONCLUSION: The true ileal digestibility of goat milk protein in South Indian WRA with normal intestinal absorptive function and integrity was comparable to earlier estimates in healthy French adults.

Effects of limestone particle size, phytate, calcium source, and phytase on standardized ileal calcium and phosphorus digestibility in broilers.

Work was carried out to determine the effects of limestone (LM) geometric mean diameter (GMD), phytate, Ca source, and phytase on standardized ileal digestibility (SID) of Ca and P in broilers. Twelve treatments (TRT) were tested. One basal corn and corn germ-based diet was prepared without adding inorganic Ca or P (TRT1, 0.02% Ca). Limestone from the same source (800 or 151 µm GMD) and bone (151 µm GMD) were added to TRT 2-4, (0.7% Ca). TRT5 was a corn/soybean meal (SBM) diet with 800 µm GMD LM (0.77% Ca). Buttiauxella sp. phytase was added (1,000 U/kg) to TRT 1-5 to prepare TRT 6-10, respectively. In addition, 800 or 150 µm GMD LM and monosodium P were added to a nitrogen-Ca-phytate-free diet (TRT 11 and 12, respectively). Treatments were analyzed as a complete block design using SAS mixed procedures and with factorial analysis on subsets of data: 1) LM GMD (151 vs. 800 µm GMD) with or without phytase: TRT 2, 3, 7, and 8; 2) phytate (corn or corn-free) with the same LM at 2 different sizes: TRT 2, 3, 11, and 12; 3) Ca source (LM vs. bone) with or without phytase: TRT 2, 4, 6, 8, and 9; 4) phytate source (corn vs. corn/SBM) with or without phytase: TRT 2, 5, 7, and 10. Broilers (4/pen) were fed mash diets ad lib for 36 h (20-22 d of age). At the end of the trial, distal ileal digesta were collected. Fine LM had lower SID Ca (38.09%) vs. coarse LM (49.18%), irrespective of phytase (P < 0.05). Standardized ileal digestibility of P was lower when the smaller LM was used vs. coarse LM with either 0 or 1,000 phytase U/kg (P < 0.05). Both SID Ca and P were higher without phytate vs. when phytate from corn was present (P < 0.05). Ca from bone was more digestible and its impact on SID P smaller vs. LM regardless of phytase (P < 0.05). In addition, phytate from SBM was more digestible and responsive to phytase as compared to phytate from corn (P < 0.05). In conclusion, dietary factors affected Ca and P digestibility and their response to phytase inclusion.

Research Note: The effect of sequential displacement of dietary dextrose with myo-inositol on broiler chicken growth performance, bone characteristics, ileal nutrient digestibility, and total tract nutrient retention.

A total of 480 male Cobb 500 broiler chicks were assigned to one of 6 dietary treatments to explore the energy equivalence of myo-inositol compared with dextrose. The 6 dietary treatments included a corn and soy-based control ration formulated with 5% anhydrous dextrose and 5 further diets that were generated by the sequential displacement of increments of 1% dextrose with myo-inositol. Each diet was fed to 8 replicate cages of 10 chicks per cage from day 8 to day 18 after hatch. The BW gain, feed intake, and feed conversion ratio (FCR) were measured, and on day 15 to day 17, excreta were collected to estimate the total tract nutrient retention. Ileal digestibility of nutrients and tibia mineral content was assessed on day 18. The displacement of dextrose with myo-inositol generated a significant linear reduction in the FCR that did not reach a plateau at 5% dietary inclusion of myo-inositol. There was no effect of the displacement of dextrose with myo-inositol on bone mineral concentration. However, supplemental myo-inositol linearly reduced ileal digestibility of DM, calcium, and ileal digestible energy. Myo-inositol addition resulted in a significant linear increase in the total tract retention of CP. It can be concluded that myo-inositol has an energy equivalence equal to approximately 78% of that of dextrose for young broiler chicks but exerts a range of extra caloric effects that improve feed efficiency and may influence nitrogen (N) retention and the uric acid cycle. Future work should focus on the role of phytase and myo-inositol on uric acid, creatine kinase, and other metabolites involved in renal function and biochemical flows of N in urine and feces in nonruminants.

Electroacupuncture accelerates the delayed intestinal transit in POI by suppressing M1 like muscularis macrophages and IL6 secretion.

BACKGROUND: Electroacupuncture (EA) at ST-36 could accelerate the delayed gastrointestinal (GI) motility in many GI motility dysfunction models, but the definite effect and mechanisms are unclear. In this study, we intended to investigate the effects of EA on intestinal manipulation (IM) mice model and involved mechanisms. METHODS: Male C57BL/6 mice were randomized into five groups: normal control, intestinal manipulation (IM), IM with sham EA (SEA), IM with high-frequency EA (HEA), and IM with low-frequency EA (LEA). The GI transit was evaluated. The infiltration of muscularis macrophages (MMφ) and its phenotype were analyzed with flow cytometry. Magnetic-activated cell sorting was applied to isolate MMφ, and the relationship between the MMφ and interstitial cells of Cajal (ICCs) was further investigated. RESULTS: (1) Compared with the IM group, HEA and LEA attenuated the delayed intestinal transit. (2) Both the HEA and LEA obviously reduced the MMφ and suppressed the M1 activation of the MMφ in the ileum. (3) EA restored the disrupted ICC networks through inhibiting the release of IL6 by the MMφ. CONCLUSION: (1) Electroacupuncture at acupoint ST-36 could accelerate the delayed intestinal transit in the IM murine model by restoring the ICC networks. (2) EA protected the ICCs through reducing the MMφ, inhibiting its M1 polarization and its IL6 secretion.

Pharmacological screening of the phenolic compound caffeic acid using rat aorta, uterus and ileum smooth muscle.

Caffeic acid is a phenolic compound widely found in commonly consumed foods such as pears, apples and coffee, and is pharmacologically known for its antioxidant, anti-inflammatory and anti-asthmatic properties. However, its relaxant activity in the aorta, uterus and ileum smooth muscle has not been investigated. This study aimed to comparatively evaluate the effect of caffeic acid on smooth muscle from different organs (aorta, uterus and ileum), and the contractions of this different organ were induced by different agonists. The organ bath technique was used, where the organs were placed in different cuvettes with 10 mL of Tyrode solution for 1 h to stabilize, then, myometrial, intestinal strip and aortic ring contractions were evoked using different contractile agonists (KCl 60 mM, PHE 0.1 µM, OT 10-2 IU/mL, CCh 10-6 M and BaCl2 0.1-30 mM); increasing concentrations of caffeic acid (0.03-7 mM) were administered in the experimental preparations. In the presence of KCl (60 mM), caffeic acid caused relaxations with the following EC50 values: 2.7 ± 0.26 mM/mL (aorta), 5.7 ± 0.71 mM/mL (uterus) and 2.1 ± 0.39 mM/mL (ileum). When in the presence of different agonists, PHE (0.1 µM) for the aorta, OT (10-2 IU/mL) for the uterus and CCh (10-6 M) for the ileum, caffeic acid caused relaxations with EC50 values of: 2.7 ± 0.31 mM/mL; 2.2 ± 0.34 mM/mL and 2.0 ± 0.28 mM/mL, respectively. The inhibitory effect of caffeic acid on serotonergic (aorta and uterus) and muscarinic receptors (uterus and ileum), as well as its possible involvement with L-type Ca2+ channels, was also observed. This study reports the pharmacological characterization of caffeic acid on smooth muscle from different organs, for which caffeic acid was more potent in the ileum. A diverse understanding of its performance as a possible therapeutic product is attributed to its relaxant effect.