Assessing seromuscular layer and serosa removal on intestinal permeability measurements in weaned piglet everted sac segments.

The integrity of the intestinal barrier is crucial for regulating the passage of pathogens and toxins, while facilitating nutrient absorption. The everted gut sac technique, an ex-vivo technique, can be used to study interventions on barrier function. This cost-effective approach utilizes relatively large gut segments to study specific intestinal regions. Typically, intact (non-stripped) intestinal segments are used, but their use may underestimate permeability due to the medial positioning of blood vessels relative to the seromuscular layer and serosa. However, removing these layers risks physical damage, resulting in an overestimation of intestinal permeability. Therefore, we investigated the impact of stripping jejunal segments on permeability to fluorescein isothiocyanate-dextran (FITC, 4 kDa) and tetramethylrhodamine isothiocyanate-dextran (TRITC, 40 kDa), and on the absorption of glucose, lysine, and methionine in jejunal segments from 80 piglets at 8 d postweaning. Piglets were subjected to either high or low sanitary housing conditions and diets provoking intestinal protein fermentation or not, expected to influence intestinal permeability. Stripping of the seromuscular layer and serosa increased the passage of 4 kDa FITC-dextran (stripped vs. non-stripped; 1.1 vs. 0.9 pmol/cm2/min, P < 0.001), glucose (40.0 vs. 19.1 pmol/cm2/min, P < 0.001), lysine (2.5 vs. 2.0 nmol/cm2/min, P < 0.001), and methionine (4.1 vs. 2.7 pmol/cm2/min, P < 0.001). As permeability increased, the differences in methionine passage between stripped and non-stripped intestinal segments also increased (slope = 1.30, P = 0.009). The coefficients of variation were comparable between stripped and non-stripped intestines (over all treatments, stripped vs. non-stripped 38% vs. 40%). Stripping, by isolating mucosal processes without introducing additional variation, is thus recommended for studies on intestinal permeability or absorption.

The intestinal barrier is vital for nutrient passage, while impeding pathogen and toxin translocation. The everted gut sac technique is used to study intestinal permeability, incubating an isolated, everted, intestinal segment filled with buffer solution in a medium containing the substances of interest. After incubation, the translocation of the substances into the created intestinal sac can be measured. Typically, intact intestinal segments are used, but under physiological conditions, nutrients do not need to pass the seromuscular layer and serosa to enter the blood flow. Therefore, removing these layers may be preferable, but, on the other hand, also risks physical damage. This study compared the use of non-stripped vs. stripped intestinal segments. Permeability to two markers (FITC-dextran, 4kDa and TRITC-dextran, 40 kDa), and absorption of glucose, lysine, and methionine were measured in non-stripped and stripped jejunal segments obtained from 80 piglets at 8 d postweaning. The piglets were housed under different hygiene and dietary conditions, which were anticipated to alter intestinal permeability. Stripping the seromuscular layer and serosa increased the passage of FITC-dextran, glucose, lysine, and methionine, without reducing assay precision due to physical damage. Thus, removal of the seromuscular layer and serosa is preferred for studying intestinal permeability or absorption.

Effect of adding soluble viscous fibers to diets containing coarse and finely ground insoluble fibers on digesta transit behavior and nutrient digestibility in broiler chickens.

This paper aimed to study the interactive effects of the addition of soluble arabinoxylans (AX) and the particle size (PS) of soybean hulls (SBH) on digesta mean retention time (MRT) and nutrient digestibility in broiler chickens. A total of 288 one-day old Ross 308 female chicks were assigned to 32 pens (9 birds/pen) and fed a commercial starter diet for 10 d. At 10 d of age, pens were assigned to 1 of 4 dietary treatments (8 pens/diet) containing 120 g/Kg coarse or fine SBH, with or without addition of 50 g/Kg of soluble wheat AX, substituting maize starch. Titanium dioxide (4 g/Kg) and cobalt-EDTA (1 g/Kg) were added as inert markers. Excreta were quantitatively collected from d 22 to 25. Gastrointestinal tract and digesta were collected on d 28, 29, or 30. Arabinoxylans reduced the weight of the gizzard relative to body weight (RW) by 0.07% units (P = 0.005), and increased ceca RW (0.28 vs. 0.34%, P < 0.001) and length (10.45 vs. 11.21 cm/Kg BW, P < 0.001). Arabinoxylans increased digesta MRT in the crop (solids/liquids: +12 min, P < 0.05), small intestine (solids/liquids: +17 min, P < 0.01), and hindgut (liquids: +77.5 min, P < 0.05); and reduced apparent ileal digestibility (AID) and apparent total tract retention (ATTR) of DM (-5.4 and -3.9%, P < 0.001, respectively) and starch (-1.35 and -0.7%, P < 0.001, respectively). Particle size of SBH only affected the ATTR of non-starch polysaccharides, presenting higher retention values with fine SBH (-4.3%-units, P = 0.034). The addition of AX reduced AID of N by 4.3%-units, only in presence of fine SBH (interaction, P < 0.05). In conclusion, arabinoxylans greatly influenced digestion in the chicken GIT, while PS of SBH had marginal effects. Arabinoxylans reduced AID of N only with fine SBH, suggesting coarse SBH counteracted AX effects on N digestion, speculatively by modifying digesta viscosity.

Insoluble fibers affect digesta transit behavior in the upper gastrointestinal tract of growing pigs, regardless of particle size.

Physicochemical characteristics of dietary fibers may modulate digesta transit behavior. The present study was conducted to clarify the effect of level and particle size (PS) of insoluble fibers on digesta mean retention time (MRT) in the proximal gastrointestinal tract (mouth-ileocecal junction). Six ileal-cannulated pigs (26.8 ±â€…2.08 kg) were assigned to 3 dietary treatments in a 3 × 3 replicated Latin-square design. Finely ground (1 mm screen) or coarse (intact) oat husks (OH) and soybean hulls (SBH) were added (50:50, w/w) to a maize-whey protein-wheat-based diet at 50 (low) or 250 g/kg (high) inclusion levels to obtain a low-fine fiber (LF), high-fine fiber (HF), and high-coarse fiber (HC) diet. Markers to follow liquids (Co-EDTA), fine solids (Y3O2), or fibrous particles (Yb-mordanted OH and Cr-mordanted SBH) were given as a single pulse dose and marker concentrations were subsequently measured hourly in digesta for 13 h after administration. Mean retention time values were obtained from the concentration of markers in digesta observed over time by fitting a generalized Michaelis-Menten equation and calculating the time of peak. Fiber addition and fiber particle size neither affected the MRT of liquid nor solid digesta phases (P = 0.903). Segregation between solid and liquid digesta phases was observed for all diets (P < 0.0001), although the extent of segregation was greater for LF compared with HF and HC (P = 0.0220). The MRT of SBH particles, but not of OH-particles was longer for coarse vs fine PS (96 min, P < 0.05). In conclusion, digesta MRT was influenced by the dietary concentration but not by PS of insoluble fibers. The addition of insoluble fibers reduces digesta phase segregation from mouth to distal ileum in growing pigs.

The velocity with which feed passes through the gastrointestinal tract affects the processes of digestion and absorption of nutrients. There is evidence that the presence of insoluble fibers, as well as their physical and chemical characteristics, can modify digesta transit. A better understanding of the impact of insoluble fibers on digestive processes may lead to better feeding strategies for pigs. In this study, we investigated the effect of addition of insoluble fibers, and their particle size, on digesta transit behavior from ingestion to the end of the small intestine of growing pigs. The results showed that there is less segregation between solid and liquid digesta phases when adding fibers, regardless their particle size. Changes in digesta transit behavior mean that the kinetics of digestion and absorption of nutrients, and consequently, the metabolic use of the nutrients absorbed may be altered.

Effect of whole foods on the microbial production of tryptophan-derived aryl hydrocarbon receptor agonists in growing pigs.

Effects of whole foods on the microbial production of tryptophan-derived aryl hydrocarbon receptor (AhR) ligands in the intestine were investigated in a pig model. Ileal digesta and faeces of pigs after feeding of eighteen different foods were analyzed. Indole, indole-3-propionic acid, indole-3-acetic acid, indole-3-lactic acid, kynurenine, tryptamine, and indole-3-aldehyde were identified in ileal digesta, which were also identified in faeces but at higher concentrations except indole-3-lactic acid, together with skatole, oxindole, serotonin, and indoleacrylic acid. The panel of tryptophan catabolites in ileal digesta and faeces varied across different foods. Eggs induced the highest overall concentration of catabolites in ileal digesta dominated by indole. Amaranth induced the highest overall concentration of catabolites in faeces dominated by skatole. Using a reporter cell line, we observed many faecal samples but not ileal samples retained AhR activity. Collectively, these findings contribute to food selection targeting AhR ligands production from dietary tryptophan in the intestine.

Presence of Unabsorbed Free Amino Acids at the End of the Small Intestine Indicates the Potential for an Increase in Amino Acid Uptake in Humans and Pigs.

BACKGROUND: Unabsorbed free amino acids (AAs) at the end of the small intestine result in a potential preventable nutritional loss. OBJECTIVES: This study aimed to quantify free AAs in terminal ileal digesta of both humans and pigs to investigate its relevance for the nutritional value of food proteins. METHODS: Two studies with three diets were performed: a human study-ileal digesta from eight adult ileostomates were collected over 9 h after ingestion of a single meal unsupplemented or supplemented with 30 g zein or whey; pig study-12 cannulated pigs were fed for 7 d with a diet containing whey or zein or no-protein diet, and ileal digesta were collected on the last 2 d. Digesta were analyzed for total and 13 free AAs. True ileal digestibility (TID) of AAs was compared with and without free AAs. RESULTS: All terminal ileal digesta samples contained free AAs. The TID of AAs in whey was 97% ± 2.4% (mean ± SD) in human ileostomates and 97% ± 1.9% in growing pigs. If the analyzed free AAs would have been absorbed, TID of whey would increase by 0.4%-units in humans and 0.1%-units in pigs. The TID of AAs in zein was 70% ± 16.4% in humans and 77% ± 20.6% in pigs and would increase by 2.3%-units and 3.5%-units, respectively, if the analyzed free AAs would have been fully absorbed. The largest difference was observed for threonine from zein: if free threonine was absorbed, the TID would increase by 6.6%-units in both species (P < 0.05). CONCLUSIONS: Free AAs are present at the end of the small intestine and can potentially have a nutritionally relevant effect for poorly digestible protein sources, whereas the effect is negligible for highly digestible protein sources. This result provides insight into the room for improvement of a protein's nutritional value if all free AAs are to be absorbed. J Nutr 2023;xx:xx-xx. This trial was registered at clinicaltrials.gov as NCT04207372.

Optimization of Capture ELISAs for Chicken Cytokines Using Commercially Available Antibodies.

Cytokines like interferon (IFN)-γ, interleukin (IL)-2, IL-6, IL-10, and IL-12p40 are important biomarkers for characterizing the nature and strength of immune responses. It is important to be able to quantify the cytokines at the protein level in biological samples. Quantification of chicken cytokines is generally performed on the level of messenger RNA (mRNA) by quantitative polymerase chain reaction (qPCR) because very few capture ELISAs for the quantification of chicken cytokine proteins are commercially available. Here, we describe the optimization and validation of capture ELISAs for chicken IL-2, IL-6, IL-10, IL-12p40, and IFN-γ using commercially available antibodies and reagents. First, we determined the optimal concentrations of the antibodies. We then verified the ELISAs' performance and established that the lower limit of detection (LLOD) for all cytokines was below 32 pg/mL. The ELISAs show the same binding characteristics for recombinant and native cytokines (parallelism was <15.2% CV). Values for inter-assay variation were consistently low and mostly <20% CV. Overall, the optimized capture ELISAs are sensitive (<32 pg/mL) and reliable tools to quantify chicken cytokines. These ELISAs can easily and inexpensively be utilized in any immunological lab and may therefore have wide applicability in immunological research for poultry.

Comparison of True Ileal Amino Acid Digestibility between Adult Humans and Growing Pigs.

BACKGROUND: It is not feasible to determine the true ileal amino acid (AA) digestibility of protein sources in humans on a routine basis, and the growing pig has been recommended as an animal model for this purpose but requires further validation. OBJECTIVES: To determine and compare true ileal AA digestibility between adult human ileostomates and growing cannulated pigs for a range of food proteins. METHODS: Seven protein sources (black beans, bread, collagen, pigeon peas, wheat bran, whey protein isolate, and zein) that spanned the range of digestibilities typically seen in foods were evaluated. Six female growing pigs received each of the protein sources, as well as a protein-free diet, and digesta were collected via ileal T-cannula. Adult human ileostomates consumed the same protein sources (5-8 ileostomates, depending on the protein source), as well as a protein-free diet, and digesta were collected. Titanium dioxide and celite were included in the diets as indigestible markers. True ileal AA digestibility coefficients were determined. RESULTS: There was a significant effect of protein source (P ≤ 0.001) for all AAs. The effect of species was not significant (P > 0.05) except for total lysine (but not for available lysine). When analyzed within diets, the statistically significant species effect for true lysine digestibility was found for black beans only. Pig and human digestibility values were generally highly and significantly (P ≤ 0.05) correlated. A linear regression equation derived for true ileal AA digestibility (given as coefficients) determined in the human and pig for the overall mean of all AAs was (y = human, x = pig) y = 1.00x - 0.010, with the slope not statistically significant (P > 0.05) from unity and the intercept not different (P > 0.05) from zero. CONCLUSIONS: True ileal AA digestibility values determined in the growing pig can be directly used for predicting digestibility in adult humans.

Reflux of 15N-labeled uric acid after intracloacal infusion in broiler chickens fed low- or high-protein diets.

Reflux of urine from the cloaca into the ceca provides chickens with a mechanism for recycling of urinary-Nitrogen (N) in a way analogous to urea recycling in mammals. However, it is unknown if reflux has substantial relevance in current poultry husbandry, where birds are fed ad libitum and have high protein intake. To evaluate the fate of urinary-N in ad libitum-fed broiler chickens, 15-day-old broilers were assigned to a high (21.9% CP, n = 22) or low (10.2 % CP, n = 22) protein diet. At 25 d of age, 20 broilers per dietary treatment were infused into the cloaca with a pulse dose of 107 mg [1,3-15N]-uric acid. N-contents and 15N-enrichment in digesta, blood plasma, and body tissues were measured at 5, 30, 60, 90, 150, 300, 450, 600, 1,200, or 1,800 min after administration (n = 2 /time-point /diet). Two broilers per dietary treatment were infused with saline and served as control to analyze background 15N-enrichment. The average total recovery (% of infused (w/w)) of 15N from infused uric acid in all body tissues was low (2.9 ± 0.62 %), of which the largest proportion was found in carcass tissue (2.5 ± 0.60%). 15N-enrichment was greatest in intestinal tissues. Even at 1,200 min, 15N-enrichment of ceca (0.46 ± 0.169 APE) and colon (0.13 ± 0.159 APE) digesta was considerably exceeding background enrichment. 15N-enrichment in excess of background enrichment in cecum and colon digesta (10-fold, P < 0.05), and 15N recovery in intestinal tissues (4-fold, P < 0.01) were greater in birds fed the low protein diet compared with the high protein diet, speculatively pointing out differences in the occurrence of reflux, incorporation of uric acid-N derivatives in intestinal tissues by first-pass metabolism, and a prolonged digesta retention time in protein deficient birds. In conclusion, these data confirm that uric acid-N infused in the cloaca can be refluxed and used for body N-deposition, but its contribution to whole body protein metabolism in broilers is probably limited.

Xylan alleviates dietary fiber deprivation-induced dysbiosis by selectively promoting Bifidobacterium pseudocatenulatum in pigs.

BACKGROUND: Low dietary fiber intake has been shown to disturb the gut microbiome community, damage the mucus barrier, and promote pathogen susceptibility. However, little is known about the temporal response of the gut microbiome to dietary fiber deprivation and the recovery induced by dietary fiber inclusion in pigs. OBJECTIVE: In the present study, temporal responses of ileal and fecal microbiota to dietary fiber deprivation were profiled using an ileum cannulated growing pig model. In addition, the potential of dietary-resistant starch, ß-glucan, and xylan to alleviate gut dysbiosis throughout the gastrointestinal tract, as well as its possible mechanisms were investigated. METHODS: Six cannulated growing pigs were fed a fiber deprivation diet for 35 days. Ileal digesta and feces were collected at days 0, 7, 21, and 35 for 16S rRNA sequencing and short-chain fatty acid (SCFA) determination. Another twenty-four healthy growing pigs were assigned to one of four dietary treatments including (1) fiber-free diet, (2) resistant starch diet, (3) ß-glucan diet, and (4) xylan diet. These twenty-four pigs were fed a corresponding diet for 35 days and slaughtered. Gut microbiome and SCFA concentration were profiled along the gastrointestinal tract. RESULTS: Dietary fiber deprivation-induced consistent microbiota extinction, mainly Bifidobacterium and Lactobacillus, and decreased SCFA concentrations in both ileum and feces. The community structure partially recovered at day 35 compared with baseline while SCFA concentrations remained low. Xylan supplementation alleviated gut dysbiosis by selectively promoting Bifidobacterium pseudocatenulatum within the large intestine. SCFA concentration increased significantly after xylan supplementation and exhibited a positive association with B. pseudocatenulatum abundance. An elevated abundance of xylan degradation-related enzyme genes was also observed in the gut microbiome after xylan supplementation. In vitro growth assay further verified the xylan utilization capacity of B. pseudocatenulatum. CONCLUSIONS: Dietary fiber deprivation could induce probiotic extinction and loss of the SCFA production while potential pathogen was promoted. Xylan intervention could partially restore dietary fiber deprivation-induced gut dysbiosis through selectively promoting B. pseudocatenulatum and therefore normalizing the gut environment. These findings collectively provide evidence that dietary fiber-driven microbiota metabolism bridges the interplay between microbiome and gut health. Video abstract.

ASAS-NANP symposium: digestion kinetics in pigs: the next step in feed evaluation and a ready-to-use modeling exercise.

Growing importance of upcycling agricultural by-products, food waste, and food processing by-products through livestock production strongly increased the variation in the nutritional quality of feed ingredients. Traditionally, feed ingredients are evaluated based on their measured extent of digestion. Awareness increases that in addition to the extent, the kinetics of digestion affects the metabolic fate of nutrients after absorption. Together with a growing body of evidence of complex interactions occurring within the lumen of the digestive tract, this urges the need of developing new approaches for feed evaluation. In a recently developed approach, we propose combining in vitro and in silico methods for feed ingredient evaluation. First steps in the development of such a systems were made by (1) evaluating in vitro the digestion potential of feed ingredients, regarding this as true ingredient properties and (2) predicting in silico the digestive processes like digesta transit, nutrient hydrolysis and absorption using dynamic, mechanistic modeling. This approach allows to evaluate to what extent the digestion potential of each ingredient is exploited in the digestive tract. Future efforts should focus on modeling digesta physicochemical properties and transit, applying in vitro digestion kinetic data of feed ingredients in mechanistic models, and generating reliable in vivo data on nutrient absorption kinetics across feed ingredients. The dynamic modeling approach is illustrated by a description of a modeling exercise that can be used for teaching purposes in digestive physiology or animal nutrition courses. A complete set of equations is provided as an on-line supplement, and can be built in modeling software that is freely available. Alternatively, the model can be constructed using any modeling software that enables the use of numerical integration methods.

Cellulase and Alkaline Treatment Improve Intestinal Microbial Degradation of Recalcitrant Fibers of Rapeseed Meal in Pigs.

The aim of the current study was to investigate whether degradation of rapeseed meal (RSM) by a swine gut microbiota consortium was improved by modifying RSM by treatment with cellulase (CELL), two pectinases (PECT), or alkaline (ALK) compared to untreated RSM and to assess whether microbiota composition and activity changed. The predicted relative abundances of carbohydrate digestion and absorption, glycolysis, pentose phosphate pathway, and pyruvate metabolism were significantly increased upon CELL and ALK feeding, and CELL and ALK also exhibited increased total short-chain fatty acid (SCFA) production compared to CON. Megasphaera, Prevotella, and Desulfovibrio were significantly positively correlated with SCFA production. Findings were validated in ileal cannulated pigs, which showed that CELL and ALK increased fiber degradation of RSM. In conclusion, CELL and ALK rather than PECT1 or PECT2 increased fiber degradation in RSM, and this information could guide feed additive strategies to improve efficiency and productivity in the swine industry.

Differently Pre-treated Rapeseed Meals Affect in vitro Swine Gut Microbiota Composition.

The aim of the study was to investigate the effect of untreated and processed rapeseed meal (RSM) on fiber degradability by pig gut microbiota and the adaptation of the microbiota to the substrate, by using the Swine Large Intestine in vitro Model (SLIM). A standardized swine gut microbiota was fed for 48 h with pre-digested RSM which was processed enzymatically by a cellulase (CELL), two pectinases (PECT), or chemically by an alkaline (ALK) treatment. Amplicons of the V3-V4 region of the 16S rRNA gene were sequenced to evaluate the gut microbiota composition, whereas short chain fatty acids (SCFA) were measured to assess fiber degradation. Adaptive gPCA showed that CELL and ALK had larger effects on the microbiota composition than PECT1 and PECT2, and all substrates had larger effects than CON. The relative abundance of family Prevotellaceae was significantly higher in CELL treatment compared to other treatments. Regardless of the treatments (including CON), the relative abundance of Dorea, Allisonella, and FamilyXIIIUCG_001 (in the order of Clostridiales) were significantly increased after 24 h, and Parabacteroides, Mogibacterium, Intestinimonas, Oscillibacter, RuminococcaceaeUCG_009, Acidaminococcus, Sutterella, and Citrobacter were significantly higher in abundance at time point 48 compared to the earlier time points. Prevotella 9 had significant positive correlations with propionic and valeric acid, and Mogibacterium positively correlated with acetic and caproic acid. There was no significant difference in SCFA production between untreated and processed RSM. Overall, degradability in the processed RSM was not improved compared to CON. However, the significantly different microbes detected among treatments, and the bacteria considerably correlating with SCFA production might be important findings to determine strategies to shorten the fiber adaptation period of the microbiota, in order to increase feed efficiency in the animal, and particularly in pig production.

Determination of True Ileal Amino Acid Digestibility in the Growing Pig for Calculation of Digestible Indispensable Amino Acid Score (DIAAS).

Digestible indispensable amino acid score (DIAAS) has been recommended by the FAO for the evaluation of protein quality in human foods, but the application of DIAAS is currently limited because of a lack of published data on the true ileal amino acid (AA) digestibility of AAs in foods. The importance of DIAAS is highlighted. To calculate DIAAS, it is necessary to determine the true ileal AA digestibility of human foods using the growing pig as an animal model for the human based on previous FAO recommendations. A method is described in detail in Supplemental Methods to determine the true ileal AA digestibility of foods for humans using the pig as a model for the adult human. Adoption of the method will enable consistency in the development of databases on predicted true ileal AA digestibility in human foods for the calculation of DIAAS.

Circadian misalignment imposed by nocturnal feeding tends to increase fat deposition in pigs.

Misalignment of day/night and feeding rhythms has been shown to increase fat deposition and the risk for metabolic disorders in humans and rodents. In most studies, however, food intake and intake patterns are not controlled. We studied the effects of circadian misalignment on energy expenditure in pigs while controlling for food intake as well as intake patterns. Twelve groups of five male pigs were housed in respiration chambers and fed either during the day (10.00-18.00 hours; DF) or night (22.00-06.00 hours; NF), bihourly the same sequential meals, representing 15, 10, 25, 30 and 20 % of the daily allowance. Paired feeding was applied to ensure equal gross energy intake between treatments. Apparent total tract digestibility, energy balances and heat partitioning were measured and analysed using a mixed linear model. Apparent total tract energy and DM digestibility tended to be lower for NF-pigs than DF-pigs (P < 0·10). Heat production was 3 % lower for NF-pigs than DF-pigs (P < 0·026), increasing fat retention by 7 % in NF-pigs (P = 0·050). NF-pigs were less active than DF-pigs during the feeding period, but more active during the fasting period. RMR was greater for DF-pigs than NF-pigs during the fasting period. Methane production was 30 % greater in NF-pigs than DF-pigs (P < 0·001). In conclusion, circadian misalignment has little effect on nutrient digestion, but alters nutrient partitioning, ultimately increasing fat deposition. The causality of the association between circadian misalignment and methane production rates remains to be investigated.

Whole digesta properties as influenced by feed processing explain variation in gastrointestinal transit times in pigs.

Physicochemical properties of diets are believed to play a major role in the regulation of digesta transit in the gastrointestinal tract. Starch, being the dominant nutrient in pig diets, strongly influences these properties. We studied transport of digesta solids and liquids through the upper gastrointestinal tract of ninety pigs in a 3 × 3 factorial arrangement. Dietary treatments varied in starch source (barley, maize and high-amylose maize) and form (isolated starch, ground cereal and extruded cereal). Mean retention times (MRT) of digesta solids ranged 129-225 min for the stomach and 86-124 min for the small intestine (SI). The MRT of solids consistently exceeded that of liquids in the stomach, but not in the SI. Solid digesta of pigs fed extruded cereals remained 29-75 min shorter in the stomach compared with pigs fed ground cereals (P < 0·001). Shear stress of whole digesta positively correlated with solid digesta MRT in the stomach (r 0·33, P < 0·001), but not in the SI. The saturation ratio (SR), the actual amount of water in stomach digesta as a fraction of the theoretical maximum held by the digesta matrix, explained more variation in digesta MRT than shear stress. The predictability of SR was hampered by the accumulation of large particles in the stomach. In addition, the water-holding capacity of gelatinised starch leads to a decreased SR of diets, but not of stomach digesta, which was caused by gastric hydrolysis of starch. Both of these phenomena hinder the predictability of gastric retention times based on feed properties.

Pigs Ferment Enzymatically Digestible Starch when it Is Substituted for Resistant Starch.

BACKGROUND: Feeding behavior is controlled by satiety mechanisms, which are affected by the extent of starch digestion, and thus resistant starch (RS) intake. Alterations in feeding behavior to changes in RS intake may depend on the adaptation of processes involved when shifting from starch digestion to fermentation or vice versa. OBJECTIVES: The aim of this study was to investigate how growing pigs adapt their feeding behavior in response to increasing and decreasing dietary RS concentrations. METHODS: Thirty-six groups of 6 pigs (25.4 ± 2.8 kg; Hypor Libra × Hypor Maxter; male:female, 1:1) were fed diets containing 50% high-amylose maize starch (high RS; HRS) or waxy maize starch (low RS; LRS). Over 28 d, diets were exchanged following a 5-step titration (25% per step) that was executed in the upward (LH) or downward direction (HL). Twelve groups received a control diet to correct for changes over time. Individual feeding behavior and total tract starch digestion and fermentation were evaluated. The response in each parameter to increasing dietary HRS inclusion was estimated through the use of linear regression procedures, and tested for titration direction and sex effects. RESULTS: Complete substitution of LRS with HRS increased the proportion of starch fermented, which was greater in LH pigs than in HL pigs (17.6% compared with 8.18%; P < 0.001), and decreased the feed intake (106 g/d; P = 0.021) and meal size (12.6 g; P < 0.001) of LH pigs, but not of HL pigs. In LH pigs, the size of the starch fermentation response positively correlated with the size of the feed intake response (r = 0.90, P < 0.001). CONCLUSIONS: The attenuated response in starch fermentation in HL pigs indicates that pigs adapt more slowly to dietary supply of digestible starch than to RS, consequently resulting in fermentation of enzymatically digestible starch. Feed intake and feeding behavior only changed in pigs poorly adapting to RS, indicating that adequacy of adaptation, rather than RS itself, drives feed intake. These findings stress the importance of diet history for nutrient digestion and feeding behavior.

Starch digestion kinetics and mechanisms of hydrolysing enzymes in growing pigs fed processed and native cereal-based diets.

This study aimed to examine in vivo starch digestion kinetics and to unravel the mechanisms of starch hydrolysing enzymes. Ninety pigs (23 (sd 2·1) kg body weight) were assigned to one of nine treatments in a 3×3 factorial arrangement, with starch source (barley, maize, high-amylose (HA) maize) and form (isolated, within cereal matrix, extruded) as factors. We determined starch digestion coefficients (DC), starch breakdown products and digesta retention times in four small-intestinal segments (SI1-4). Starch digestion in SI2 of pigs fed barley and maize, exceeded starch digestion of pigs fed HA maize by 0·20-0·33 DC units (P<0·01). In SI3-4, barley starch were completely digested, whereas the cereal matrix of maize hampered digestion and generated 16 % resistant starch in the small intestine (P<0·001). Extrusion increased the DC of maize and HA maize starch throughout the small intestine but not that of barley (P<0·05). Up to 25 % of starch residuals in the proximal small intestine of pigs was present as glucose and soluble α(1-4) maltodextrins. The high abundance of glucose, maltose and maltotriose in the proximal small intestine indicates activity of brush-border enzymes in the intestinal lumen, which is exceeded by α-amylase activity. Furthermore, we found that in vivo starch digestion exceeded our in vitro predictions for rapidly digested starch, which indicates that the role of the stomach on starch digestion is currently underestimated. Consequently, in vivo glucose release of slowly digestible starch is less gradual than expected, which challenges the prediction quality of the in vitro assay.

Increasing intake of dietary soluble nutrients affects digesta passage rate in the stomach of growing pigs.

The passage rate of solids and liquids through the gastrointestinal tract differs. Increased dietary nutrient solubility causes nutrients to shift from the solid to the liquid digesta fraction and potentially affect digesta passage kinetics. We quantified: (1) the effect of three levels of dietary nutrient solubility (8, 19 and 31 % of soluble protein and sucrose in the diet) at high feed intake level (S) and (2) the effect of low v. high feed intake level (F), on digesta passage kinetics in forty male growing pigs. The mean retention time (MRT) of solids and liquids in the stomach and small intestine was assessed using TiO2 and Cr-EDTA, respectively. In addition, physicochemical properties of digesta were evaluated. Overall, solids were retained longer than liquids in the stomach (2·0 h, P<0·0001) and stomach+small intestine (1·6 h, P<0·001). When S increased, MRT in stomach decreased by 1·3 h for solids (P=0·01) and 0·7 h for liquids (P=0·002) but only at the highest level of S. When F increased using low-soluble nutrients, MRT in stomach increased by 0·8 h for solids (P=0·041) and 0·7 h for liquids (P=0·0001). Dietary treatments did not affect water-binding capacity and viscosity of digesta. In the stomach of growing pigs, dietary nutrient solubility affects digesta MRT in a non-linear manner, while feed intake level increases digesta MRT depending on dietary nutrient solubility. Results can be used to improve predictions on the kinetics of nutrient passage and thereby of nutrient digestion and absorption in the gastrointestinal tract.

ß-Glucans and Resistant Starch Alter the Fermentation of Recalcitrant Fibers in Growing Pigs.

Interactions among dietary ingredients are often assumed non-existent when evaluating the nutritive value and health effects of dietary fiber. Specific fibers can distinctly affect digestive processes; therefore, digestibility and fermentability of the complete diet may depend on fiber types present. This study aimed to evaluate the effects of readily fermentable fibers (ß-glucans and resistant starch) on the degradation of feed ingredients containing more persistent, recalcitrant, fibers. Six semi-synthetic diets with recalcitrant fibers from rapeseed meal (pectic polysaccharides, xyloglucans, and cellulose) or corn distillers dried grain with solubles (DDGS; (glucurono)arabinoxylans and cellulose) with or without inclusion of ß-glucans (6%) or retrograded tapioca (40%) substituted for corn starch were formulated. Six ileal-cannulated pigs (BW 28±1.4 kg) were assigned to the diets according to a 6×6 Latin square. ß-glucan-extract increased apparent total tract digestibility (ATTD) of non-glucosyl polysaccharides (accounting for ~40% of the fiber-fraction) from rapeseed meal (6%-units, P<0.001), but did not affect non-glucosyl polysaccharides from DDGS. Retrograded tapioca reduced ATTD of non-glucosyl polysaccharides from rapeseed meal and DDGS (>10%-units, P<0.001), indicating that the large amount of resistant starch entering the hindgut was preferentially degraded over recalcitrant fibers from rapeseed meal and DDGS, possibly related to reduced hindgut-retention time following the increased intestinal bulk. Fermentation of fiber sources was not only dependent on fiber characteristics, but also on the presence of other fibers in the diet. Hence, interactions in the gastrointestinal tract among fibrous feed ingredients should be considered when evaluating their nutritive value.