Functional and molecular profiling of fasted piglets reveals decreased energy metabolic function and cell proliferation in the small intestine.

The small intestine requires energy to exert its important role in nutrient uptake and barrier function. Pigs are an important source of food and a model for humans. Young piglets and infants can suffer from periods of insufficient food intake. Whether this functionally affects the small intestinal epithelial cell (IEC) metabolic capacity and how this may be associated with an increased vulnerability to intestinal disease is unknown. We therefore performed a 48-h fasting intervention in young piglets. After feeding a standard weaning diet for 2 wk, 6-wk-old piglets (n = 16/group) were fasted for 48 h, and midjejunal IECs were collected upon euthanasia. Functional metabolism of isolated IECs was analyzed with the Seahorse XF analyzer and gene expression was assessed using RNA-sequencing. Fasting decreased the mitochondrial and glycolytic function of the IECs by 50% and 45%, respectively (P < 0.0001), signifying that overall metabolic function was decreased. The RNA-sequencing results corroborated our functional metabolic measurements, showing that particularly pathways related to mitochondrial energy production were decreased. Besides oxidative metabolic pathways, decreased cell-cycle progression pathways were most regulated in the fasted piglets, which were confirmed by 43% reduction of Ki67-stained cells (P < 0.05). Finally, the expression of barrier function genes was reduced upon fasting. In conclusion, we found that the decreased IEC energy metabolic function in response to fasting is supported by a decreased gene expression of mitochondrial pathways and is likely linked to the observed decreased intestinal cell proliferation and barrier function, providing insight into the vulnerability of piglets, and infants, to decreased food intake.NEW & NOTEWORTHY Fasting is identified as one of the underlying causes potentiating diarrhea development, both in piglets and humans. With this study, we demonstrate that fasting decreases the metabolism of intestinal epithelial cells, on a functional and transcriptional level. Transcriptional and histological data also show decreased intestinal cell proliferation. As such, fasting-induced intestinal energy shortage could contribute to intestinal dysfunction upon fasting.

Multi-country metabolic signature discovery for chicken health classification.

INTRODUCTION: To decrease antibiotic resistance, their use as growth promoters in the agricultural sector has been largely abandoned. This may lead to decreased health due to infectious disease or microbiome changes leading to gut inflammation. OBJECTIVES: We aimed to generate a m/z signature classifying chicken health in blood, and obtain biological insights from the resulting m/z signature. METHODS: We used direct infusion mass-spectrometry to determine a machine-learned metabolomics signature that classifies chicken health from a blood sample. We then challenged the resulting models by investigating the classification capability of the signature on novel data obtained at poultry houses in previously unseen countries using a Leave-One-Country-Out (LOCO) cross-validation strategy. Additionally, we optimised the number of mass/charge (m/z) values required to maximise the classification capability of Random Forest models, by developing a novel ranking system based on combined univariate t-test and fold-change analyses and building models based on this ranking through forward and reverse feature selection. RESULTS: The multi-country and LOCO models could classify chicken health. Both resulting 25-m/z and 3784-m/z signatures reliably classified chicken health in multiple countries. Through mummichog enrichment analysis on the large m/z signature, we found changes in amino acid metabolism, including branched chain amino acids and polyamines. CONCLUSION: We reliably classified chicken health from blood, independent of genetic-, farm-, feed- and country-specific confounding factors. The 25-m/z signature can be used to aid development of a per-metabolite panel. The extended 3784-m/z version can be used to gain a deeper understanding of the metabolic causes and consequences of low chicken health. Together, they may facilitate future treatment, prevention and intervention.

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.

Effect of animal activity and air temperature on heat production, heart rate, and oxygen pulse in lactating Holstein cows.

A linear relationship between heart rate (HR) and oxygen consumption (VO2) has been reported in homeothermic animals, indicating that is possible to estimate heat production through HR measurements. This relationship may depend on the animal activity and environmental conditions. The main objective of the present study was to evaluate the effect of the air temperature and animal posture and activity on heat production and VO2 in relation to HR. In addition, as a secondary objective, the energy cost of eating and ruminating versus idling and standing versus lying down was determined. Twelve Holstein lactating cows were housed inside climate-controlled respiration chambers for 8 d, where the air temperature was gradually increased from 7 to 21°C during the night and from 16 to 30°C during the day with daily increments of 2°C for both daytime and nighttime. During the 8-d data collection period, HR and gaseous exchange measurements were performed, and animal posture and activity were recorded continuously. The oxygen pulse (O2P), which represents the amount of oxygen that is consumed by the cow per heartbeat, was calculated as the ratio between VO2 and HR. Results showed that heat production and VO2 were linearly and positively associated with HR, but this relationship largely varied between individual cows. Within the range tested, O2P was unaffected by temperature, but we detected a tendency for an interaction of O2P with the temperature range tested during the night versus during the day. This indicates that the effect of air temperature on O2P is nonlinear. Standing and eating slightly increased O2P (1.0 and 2.5%) compared with lying down and idling, respectively, whereas rumination increased O2P by 5.1% compared with idling. It was concluded that the potential bias introduced by these effects on the O2P for the application of the technique is limited. The energy cost of eating and ruminating over idling was 223 ± 11 and 45 ± 6 kJ/kg0.75 per day, respectively, whereas the energy cost of standing over lying down was 53 ± 6 kJ/kg0.75 per day. We concluded that O2P in dairy cows was slightly affected by both animal posture and activity, but remained unaffected by air temperature within 8 to 32°C. Nonlinearity of the relationship between the O2P and air temperature suggests that caution is required extrapolating O2P beyond the temperature range evaluated in our experiment.

Effect of solid feed level and types of roughage on passage kinetics of milk replacer, concentrate, and roughage in veal calves.

This study aimed (1) to provide estimates of total mean retention times of milk replacer (MR), concentrates, and roughage in veal calves fed a mixed diet; (2) to determine the effect of level and type of solid feed (SF) on passage kinetics of MR, concentrates, and roughages in veal calves; and (3) to compare passage kinetics in veal calves using the fecal excretion curves of indigestible markers and a noninvasive 13C tracer breath test approach to determine whether the latter technique can serve as an alternative. At the start of the trial, 48 Holstein-Friesian calves (6 wk of age; 68 ± 7.7 kg of body weight; BW) were assigned to 1 of 4 dietary treatments (for statistical analysis, only 39 calf observations were used). Three treatments contained chopped wheat straw as roughage in the SF mixture in a concentrate:roughage ratio of 90:10 (dry matter basis). The SF level was 20 g/kg of metabolic BW per day (low straw), 30 g/kg of metabolic BW per day (middle straw), or 40 g/kg of metabolic BW per day (high straw). The fourth treatment (high hay) contained long perennial ryegrass hay as roughage in the SF mixture in a concentrate:roughage ratio of 70:30 (dry matter basis, at 40 g/kg of metabolic BW per day). The quantity of MR was fixed for the high straw treatment, whereas the amount of MR for the other treatments during the adaptation period was adjusted based on a pair gain strategy (i.e., exchanging ration components but keeping similar net energy). At the end of the adaptation period, calves ranged from 12 to 15 wk of age with an average BW of 123 ± 8.6 kg. Passage kinetics of concentrates were estimated by measuring 13C enrichment excess of CO2 in breath from a pulsed-dose of [1-13C]octanoate. Passage kinetics of roughage, concentrates, and MR were also estimated using fecal excretion curves obtained after ingestion of chromium-mordanted roughage, Yb2O3, and Co-EDTA, respectively. We conclude that [1-13C]octanoate cannot serve as a measure for oro-duodenal transit of concentrates because of unrealistic estimates. Based on the fecal excretion curves, we concluded that the total mean retention time of MR (i.e., time to peak; the moment that the excretion curve reaches peak concentration) was, on average, 12.4 h, and that the passage kinetics of MR was not affected by the level or type of SF. The mean retention time of concentrates was shorter (21.4 h) than that of both straw (59.1 h) and hay (36.8 h), and was not affected by the level or type of SF. Also, the mean retention time of the slowest compartment (i.e., the rumen) was shorter for concentrates (39.6 h) than that of straw (110.0 h) and hay (59.2 h). Contrary, the passage of roughage was affected by level and type of SF. Long hay increased time to peak by 22.3 h and decreased ruminal mean retention time by 50.8 h relative to chopped straw, indicating that the passage rate of long hay is faster than that of chopped straw. We conclude that the level and type of SF only affects the passage kinetics of roughage and not that of MR and concentrates.

Butyrate Alters Pyruvate Flux and Induces Lipid Accumulation in Cultured Colonocytes.

Butyrate is considered the primary energy source of colonocytes and has received wide attention due to its unique health benefits. Insight into the mechanistic effects of butyrate on cellular and metabolic function relies mainly on research in in-vitro-cultured cells. However, cells in culture differ from those in vivo in terms of metabolic phenotype and nutrient availability. For translation, it is therefore important to understand the impact of different nutrients on the effects of butyrate. We investigated the metabolic consequences of butyrate exposure under various culturing conditions, with a focus on the interaction between butyrate and glucose. To investigate whether the effects of butyrate were different between cells with high and low mitochondrial capacity, we cultured HT29 cells under either low- (0.5 mM) or high- (25 mM) glucose conditions. Low-glucose culturing increased the mitochondrial capacity of HT29 cells compared to high-glucose (25 mM) cultured HT29 cells. Long-term exposure to butyrate did not alter mitochondrial bioenergetics, but it decreased glycolytic function, regardless of glucose availability. In addition, both high- and low-glucose-grown HT29 cells showed increased lipid droplet accumulation following long-term butyrate exposure. Acute exposure of cultured cells (HT29 and Caco-2) to butyrate increased their oxygen consumption rate (OCR). A simultaneous decrease in extracellular acidification rate (ECAR) was observed. Furthermore, in the absence of glucose, OCR did not increase in response to butyrate. These results lead us to believe that butyrate itself was not responsible for the observed increase in OCR, but, instead, butyrate stimulated pyruvate flux into mitochondria. Indeed, blocking of the mitochondrial pyruvate carrier prevented a butyrate-induced increase in oxygen consumption. Taken together, our results indicate that butyrate itself is not oxidized in cultured cells but instead alters pyruvate flux and induces lipid accumulation.

Lactose and Digestible Maltodextrin in Milk Replacers Differently Affect Energy Metabolism and Substrate Oxidation: A Calorimetric Study in Piglets.

BACKGROUND: In recent years, lactose-free infant formulas have been increasingly used. Digestible maltodextrins are commonly used as a substitute for lactose in these formulas, but the effects on energy metabolism are unknown. OBJECTIVE: We aimed to evaluate the differences in energy metabolism and substrate oxidation in piglets fed milk replacers containing lactose compared with maltodextrin as the only source of carbohydrates. METHODS: Piglets (Tempo × Topigs 20) from 8 litters were fed milk replacers containing lactose or maltodextrin (28% w/w, milk powder basis) from 1 to 9 wk of age (n =  4 litters/milk replacer). At 5 wk of age, 4 females and 4 entire males (mean ± SEM bodyweight, 10 ± 0.3 kg) were selected per litter, and housed in 16 groups of 4 littermates, with 2 females and 2 males per pen (n = 8 groups/milk replacer). Between 7 and 9 wk of age, groups were housed for 72 h in climate respiration chambers, and fed their experimental milk replacer in 2 meals per day, at 08:30 and 16:30. Heat production data were calculated from the continuous measurement of gaseous exchanges and analyzed using general linear models in SAS. RESULTS: Resting metabolic rate was 6% less in maltodextrin- than in lactose-fed piglets, notably before the morning meal. The postprandial respiratory quotient was 13% greater in maltodextrin- than in lactose-fed piglets after both meals. Net rates of carbohydrate oxidation were on average 5% greater in maltodextrin- than in lactose-fed piglets, particularly after the afternoon meal, whereas net rates of fat oxidation were 9% less in maltodextrin- than in lactose-fed piglets, particularly after the morning meal. CONCLUSIONS: Compared with lactose, maltodextrin in milk replacers reduced resting metabolic rate in the fasting state, and induced a shift in postprandial substrate oxidation profiles in pigs. Further research is warranted to evaluate the consequences of these metabolic changes for body composition.

Blood 15N:13C Enrichment Ratios Are Proportional to the Ingested Quantity of Protein with the Dual-Tracer Approach for Determining Amino Acid Bioavailability in Humans.

BACKGROUND: Assessment of amino acid bioavailability is of key importance for the evaluation of protein quality; however, measuring ileal digestibility of dietary proteins in humans is challenging. Therefore, a less-invasive dual stable isotope tracer approach was developed. OBJECTIVE: We aimed to test the assumption that the 15N:13C enrichment ratio in the blood increases proportionally to the quantity ingested by applying different quantities of 15N test protein. METHODS: In a crossover design, 10 healthy adults were given a semi-liquid mixed meal containing 25 g (low protein) or 50 g (high protein) of 15N-labeled milk protein concentrate simultaneous with 0.4 g of highly 13C-enriched spirulina. The meal was distributed over multiple small portions, frequently provided every 20 min during a period of 160 min. For several amino acids, the blood 15N- related to 13C-isotopic enrichment ratio was determined at t = 0, 30, 60, 90, 120, 180, 240, 300, and 360 min and differences between the 2 meals were compared using paired analyses. RESULTS: No differences in 13C AUC for each of the measured amino acids in serum was observed when ingesting a low- or high-protein meal, whereas 15N AUC of amino acids was ∼2 times larger on the high-protein meal (P < 0.001). Doubling the intake of 15N-labeled amino acids increased the 15N:13C ratio by a factor of 2.04 ± 0.445 for lysine and a factor between 1.8 and 2.2 for other analyzed amino acids, with only phenylalanine (2.26), methionine (2.48), and tryptophan (3.02) outside this range. CONCLUSIONS: The amino acid 15N:13C enrichment ratio in the peripheral circulation increased proportionally to the quantity of 15N-labeled milk protein ingested, especially for lysine, in healthy adults. However, when using 15N-labeled protein, correction for, e.g., α-carbon 15N atom transamination is advised for determination of bioavailability of individual amino acids. This trial was registered at www.clinicaltrials.gov as NCT02966704.

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.

Effect of dietary fiber fermentation on short-chain fatty acid production and microbial composition in vitro.

BACKGROUND: The efficient utilization of fiber-rich co-products is important for optimizing feed resource utilization and animal health. This study was conducted to evaluate the fermentation characteristics of fiber-rich co-products, which had equal quantities of total dietary fiber (TDF), at different time points using batch in vitro methods. It considered their gas production, short-chain fatty acid (SCFA) production, and microbial composition. RESULTS: The fermentation of wheat bran (WB) and oat bran (OB) showed higher and faster (P < 0.05) gas and SCFA production than corn bran (CB), sugar beet pulp (SBP), and soybean hulls (SH). The α-diversity was higher in the CB, SBP, and SH groups than in the WB and OB groups (P < 0.05). At the phylum level, OB and WB fermentation showed lower (P < 0.05) relative abundance of Actinobacteria than the CB, SBP, and SH groups. At the genus level, OB and WB fermentation increased the Enterococcus population in comparison with the CB, SBP, and SH groups, whereas CB and SBP fermentation improved the relative abundance of the Christensenellaceae R-7 group more than the WB, OB, and SH groups (P < 0.05). CONCLUSION: Overall, WB and OB were rapidly fermented by fecal microbiota, in contrast with SBP, SH, and CB. Fermentation of different fiber-rich co-products with an equal TDF content gives different responses in terms of microbial composition and SCFA production due to variations in their physicochemical properties and molecular structure. © 2020 Society of Chemical Industry.

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.

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.

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.

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.

Reduced Feed Intake, Rather than Increased Energy Losses, Explains Variation in Growth Rates of Normal-Birth-Weight Piglets.

Background: Substantial variation in growth rates exists in normal-birth-weight piglets, possibly due to differences in energy efficiency. Within this population, slow growth rates are associated with reduced insulin sensitivity. Slowly digestible starch (SDS) may improve growth efficiency in slowly growing pigs, because it reduces postprandial blood glucose. Objective: The aim of this study was to investigate maintenance energy requirements and efficiency of energy used for growth (incremental energy efficiency) of slow-growing or fast-growing piglets (SG-pigs and FG-pigs, respectively) with equal birth weight that were fed either an SDS or a rapidly digestible-starch (RDS) diet. Methods: Sixteen groups of either five 10-wk-old SG-pigs (mean ± SD: 11.3 ± 1.4 kg) or FG-pigs (15.1 ± 1.7 kg) were housed in climate respiration chambers and fed diets containing 40% RDS or SDS for 2 wk. In week 1, feed was available ad libitum. In week 2, feed supply was restricted to 65% of the observed weekly averaged feed intake [kJ · kg body weight (BW)-0.6 · d-1] in week 1. After week 2, pigs were feed deprived for 24 h, after which heat production was determined. Energy balances, apparent total tract digestibility (ATTD), and incremental energy efficiencies were calculated and analyzed using a general linear model. Results: Gross energy intake (kJ · kg BW-0.6 · d-1) was 4% greater (P = 0.047) for FG-pigs than for SG-pigs. ATTD of fat was 6%-units greater (P = 0.003) for RDS-fed than for SDS-fed pigs. Fasting heat production and incremental energy efficiencies did not differ between pig types or diets. Incremental use of metabolizable energy for fat retention was 2% units (P = 0.054) greater for RDS-fed than SDS-fed pigs. Conclusions: A lower energy intake rather than greater maintenance requirements or lower energy efficiency explains the slow growth of SG-pigs. Incremental RDS intake increased fat deposition more than SDS, whereas energy efficiency was not affected. Thus, feeding SDS instead of RDS does not improve growth efficiency but may result in slightly leaner pigs.

Prenatal, but not early postnatal, exposure to a Western diet improves spatial memory of pigs later in life and is paired with changes in maternal prepartum blood lipid levels.

Maternal obesity and perinatal high-fat diets are known to affect cognitive development. We examined the effects of late prenatal and/or early postnatal exposure to a Western-type diet, high in both fat and refined sugar, on the cognition of pigs (Sus scrofa) in the absence of obesity. Thirty-six sows and their offspring were assigned to 1 of 4 treatments in a 2 × 2 factorial arrangement, with 8 wk prenatal and 8 wk postnatal exposure to a Western diet (enriched in fat, sucrose, and cholesterol) or control diets as factors. Compared to controls, piglets exposed to the prenatal Western diet showed enhanced working and reference memory during the acquisition and reversal phases of a spatial hole-board task. Mothers fed the prenatal Western diet had higher prepartum blood cholesterol and free fatty acid levels. Postnatal exposure to the Western diet did not affect piglet cognitive performance, but it did increase postpartum maternal and postweaning piglet cholesterol levels. The Western diet had no effect on maternal or offspring insulin sensitivity or leptin levels. In conclusion, a prenatal Western diet improved memory function in pigs, which was paired with changes in prepartum maternal blood cholesterol levels. These findings highlight the key role of late fetal nutrition for long-term programming of cognition.-Clouard, C., Kemp, B., Val-Laillet, D., Gerrits, W. J. J., Bartels, A. C., Bolhuis, J. E. Prenatal, but not early postnatal, exposure to a Western diet improves spatial memory of pigs later in life and is paired with changes in maternal prepartum blood lipid levels.

Dietary Amino Acid Deficiency Reduces the Utilization of Amino Acids for Growth in Growing Pigs after a Period of Poor Health.

BACKGROUND: During immune system activation, partitioning of amino acids (AAs) changes between protein gain and use by the immune system. OBJECTIVE: We determined the effects of health status and dietary AA deficiency on nitrogen retention and AA utilization in pigs. METHODS: Barrows (55 d of age) were obtained from good health (GH, n = 14) or poor health (PH, n = 14) status farms and allocated to a diet either adequate in essential amino acids (Adq) or 25% deficient in Met + Cys, Thr, and Trp (Def). Nitrogen balance was measured and AA irreversible loss rates (ILRs) were measured after an intravenous bolus of U-(13)C-labeled L-AAs. RESULTS: On arrival at the experimental facilities, the PH pigs had 14% lower serum albumin and 50% greater serum haptoglobin and blood leukocyte counts than the GH pigs (P < 0.01), but the PH pigs showed signs of recovery during the trial. Total tract nitrogen digestibility was 3 percentage points lower in the PH pigs (P < 0.01). The PH-Adq pigs had compensatory body weight gain after arrival, coinciding with 7% greater nitrogen retention (P < 0.01) in the PH pigs than in the GH pigs. The PH pigs had a 24% greater ILR for Lys. Health status × diet interactions for Lys (P = 0.07), Val (P = 0.03), and Leu (P = 0.10) pool sizes and a greater urea pool size in the PH pigs (P = 0.01) support the observation that the increase in the ILR of Lys in the PH pigs was related to oxidation when feeding the Def diet, but to synthesis when feeding the Adq diet. Feeding Def diets increased monocyte counts by 30% (P < 0.01). CONCLUSIONS: This study illustrates how the competition for AAs between protein synthesis associated with immune system activation and body protein deposition is greater when the dietary supply of Met + Cys, Thr, and Trp is limited in pigs during and after a period of poor health.

Maternal Fish Oil Supplementation Affects the Social Behavior, Brain Fatty Acid Profile, and Sickness Response of Piglets.

BACKGROUND: Maternal dietary docosahexaenoic acid (DHA) intake is thought to affect development in the offspring. OBJECTIVE: We assessed the impact of maternal dietary DHA on behavior, brain fatty acid (FA) profile, and sickness response of offspring in pigs, a pertinent model for human nutrition. METHODS: Sows (n = 24) were fed a diet with DHA-rich fish oil (FO) (20 g/kg) or high-oleic acid sunflower oil (HOSF) (20 g/kg) from day 61 of gestation through lactation. At 4 wk of age, 4 piglets per litter were weaned and mixed with piglets from other litters. Behavior was observed in 4- to 8-wk-old piglets, and brain FA composition was analyzed at 4 (n = 15) and 14 (n = 12) wk. Thirteen-week-old piglets (n = 48) were subjected to a lipopolysaccharide (LPS) challenge. Body temperature, plasma cytokines, and motivation to approach a familiar human, indicative of a sickness response, were measured. RESULTS: FO-fed pigs displayed more social activities (+262%, P = 0.02), played more (+61%, P = 0.03), and tended to show fewer oral manipulative behaviors directed at pen mates (-25%, P = 0.06) than did HOSF-fed pigs up to 4 wk after weaning. Brain DHA concentrations were higher in FO- than in HOSF-fed pigs up to 10 wk after supplementation (+10-50%, P < 0.001), although differences declined with age. Body temperature (P < 0.001) and tumor necrosis factor α and interferon Î³ concentrations (P < 0.05) increased after LPS injection, but no diet effect was found (P > 0.10). LPS-treated pigs were less likely to approach the human than saline-treated pigs in the HOSF-fed (-29%, P = 0.0003), but not in the FO-fed group (-13%, P = 0.11). CONCLUSIONS: Maternal DHA beneficially affected offspring social behavior after weaning and mildly attenuated sickness behavior after an inflammatory challenge in pigs. These behavioral changes may be mediated by increased brain DHA proportions.

Dietary linoleic and α-linolenic acids affect anxiety-related responses and exploratory activity in growing pigs.

BACKGROUND: Growing evidence suggests that the dietary ratio of linoleic acid (LA) to α-linolenic acid (ALA), the precursors of arachidonic acid (AA) and docosahexaenoic acid (DHA), respectively, may affect behavior in mammals. OBJECTIVE: This study aimed at evaluating the impact of dietary LA and ALA intake on behaviors of growing pigs, a pertinent model for human nutrition. METHODS: At 7 wk of age, 32 pigs were allocated to 4 dietary treatments varying in daily intake of LA (1.3 and 2.6 g · kg body weight(-0.75) · d(-1) for low- and high-LA groups, respectively) and ALA (0.15 and 1.5 g · kg body weight(-0.75) · d(-1) for low- and high-ALA groups, respectively) for 4 wk. Between days 12 and 18, general behavior in the home pen was observed and pigs were subjected to an open field and novel object test. At 11 wk of age, brain fatty acid composition was analyzed. RESULTS: Compared with high LA intake, low LA intake increased the time spent on exploration, particularly nosing in the home pen (P < 0.05) and the open field (P < 0.05), and tended to reduce the time spent lying with eyes open in the home pen (P = 0.09). Time spent lying with eyes open also tended to be affected by the interaction between LA and ALA (P = 0.08). A high-LA/high-ALA intake (ratio of 2; P < 0.05) and a low-LA/high-ALA intake (ratio of 1; P = 0.06) decreased the latency to approach the novel object compared with a low-LA/low-ALA intake (ratio of 9). DHA in the frontal cortex was positively correlated with exploratory behaviors in the home pen (rs = 0.56, P < 0.01), whereas AA was negatively correlated with time spent lying with eyes closed (rs = -0.48, P < 0.01). CONCLUSIONS: Low LA intake and a low dietary LA:ALA ratio increased exploration and decreased anxiety-related behaviors in pigs. It is suggested that changes in brain DHA and AA induced by dietary LA and ALA intake mediate these behavioral changes.

Fermentation in the small intestine contributes substantially to intestinal starch disappearance in calves.

BACKGROUND: The proportion of starch disappearing from the small intestinal lumen is generally lower in ruminants than in monogastric animals, and there are indications that the starch digestion capacity in ruminants is limited. OBJECTIVES: Milk-fed calves were used to study the rate-limiting enzyme in starch hydrolysis and to quantify starch fermentation in ruminants. METHODS: Forty male Holstein-Friesian calves were fed milk replacer containing either lactose (control) or 1 of 4 corn starch products. The following starch products differed in the enzyme ratios required for their complete hydrolysis to glucose: gelatinized starch [α-amylase and (iso)maltase], maltodextrin [(iso)maltase and α-amylase], maltodextrin with α-1,6-branching (isomaltase, maltase, and α-amylase), and maltose (maltase). In the adaptation period, calves were stepwise exposed to an increasing dose of the starch product for 14 wk to allow maximal adaptation of all enzyme systems involved. In the experimental period, apparent total tract and ileal starch product disappearance, total tract starch product fermentation, and α-amylase, maltase, and isomaltase activities were determined at 18% inclusion of the starch product. RESULTS: Maltase and isomaltase activities in the brush border did not increase for any of the starch product treatments. Luminal α-amylase activity was lower in the proximal (3.9 ± 3.2 and 2.7 ± 1.7 U/mg Co for control and starch product calves, respectively) but greater in the distal small intestine of starch-fed calves than in control calves (0.0 ± 0.0 and 6.4 ± 1.5 U/mg Co for control and starch product calves, respectively; means ± SEs for control and means ± pooled SEMs for starch product treatments). Apparent ileal (61.6% ± 6.3%) and total tract (99.1% ± 0.4%) starch product disappearance did not differ between starch product treatments, suggesting that maltase activity limits starch digestion in ruminants. Total tract starch product fermentation averaged 414 ± 43 g/d, corresponding to 89% of intake, of which half was fermented before the terminal ileum, regardless of starch product treatment. CONCLUSION: Fermentation, rather than enzymatic digestion, is the main reason for small intestinal starch disappearance in milk-fed calves.