Cornelis F. M. de Lange

Dietary and Endogenous Amino Acids Are the Main Contributors to Microbial Protein in the Upper Gut of Normally Nourished Pigs

Although amino acids (AA) synthesized by enteric microbiota in the upper gut of nonruminants can be absorbed, they do not necessarily make a net contribution to the hosts AA supply. That depends on whether protein or nonprotein nitrogen sources are used for microbial protein production. We determined the contributions of urea, endogenous protein (EP), and dietary protein (DP) to microbial valine (M.VAL) at the distal ileum of growing pigs, based on isotope dilutions after a 4-d continuous infusion of l-[1-(13)C]valine to label EP and of [(15)N(15)N]urea. Eight barrows were assigned to either a cornstarch and soybean meal-based diet with or without 12% added fermentable fiber from pectin. Dietary pectin did not affect (P > 0.10) the contributions of the endogenous and DP to M.VAL. More than 92% of valine in microbial protein in the upper gut was derived from preformed AA from endogenous and DP, suggesting that de novo synthesis makes only a small contribution to microbial AA.

Maintenance Energy Requirements of Growing Pigs and Calves Are Influenced by Feeding Level

The conventional regression method for partitioning heat production (HP) in growing animals between HP associated with either maintenance or growth assumes maintenance HP to be independent of feeding level (FL). However, there are indications that this assumption is not correct and an alternative method is proposed in this study from a reanalysis of 3 trials. In trial 1, 73-, 152-, and 237-kg calves received one milk replacer at 77, 84, 92, and 100% of their ad libitum metabolizable energy (ME) intake. In trial 2, 70-kg barrows received one diet at 60, 80, and 100% of their ad libitum ME intake {2600 kJ ME/[kg body weight (BW)(0.60) · d]}. In trial 3, 60-kg barrows received a basal diet [1700 kJ ME/(kg BW(0.60) · d)] or 4 diets consisting of the basal diet plus 850 kJ ME/(kg BW(0.60)·d) of starch alone or starch with corn gluten, casein, or vegetable oil. In the 3 trials (n = 48, 18, and 28, respectively), HP and activity-related HP were measured on individuals pigs and calves in respiration chambers for 6 d (fed state) and fasting HP (FHP; at zero activity) was calculated as the asymptotic value of HP kinetics on d 7 (feed-deprived state). The FHP changed by 0.22 kJ in calves and 0.14 kJ in pigs/kJ ME intake change during the previous days. The efficiency of using ME for maintenance and growth [k(mg); 1- (HP - FHP)/ME] was not affected by FL (calves: 84%, pigs in trial 2: 74%). In trial 3, k(mg) varied between diets in connection with variations in efficiencies between nutrients (from 55% for corn gluten to 85% for lipid). This new method of representing partitioning of ME intake considers FHP as variable with FL, does not require estimates of maintenance ME requirements, includes efficiencies that depend on diet characteristics, and is not biased by metabolic adaptations of the animal to FL.

Epidermal growth factor containing culture supernatant enhances intestine development of early-weaned pigs in vivo: Potential mechanisms involved

We have previously generated epidermal factor expressing Lactococcus lactis (EGF-LL) using a bioengineering approach, and shown that EGF-LL fermentation supernatant enhanced newly weaned pigs growth. The objective of the current study was to further understand the mechanisms behind this improved performance. Sixty-four piglets were weaned at 3 weeks of age and then fed ad libitum according to a 2-phase feeding program. Four pens with 8 pigs per pen were assigned to each of two treatments for 3 weeks: (1) EGF containing supernatant from EGF-LL culture (SuperEGF) or (2) blank M17GE media (Control). Consistent with previous findings, SuperEGF pigs had an increased average daily gain during week 3 post-weaning (433.4 ± 10.86 vs 388.7 ± 7.76 g; P<0.05) and overall gain:feed ratio (0.757 ± 0.03 vs 0.677 ± 0.01 kg/kg, P < 0.05). Moreover, jejunal structure development was enhanced, and inflammation index was minimized in SuperEGF pigs as indicated by increased villi height (P<0.05), decreased lamina propria width (P<0.05), and higher expression of anti-inflammatory cytokine, IL-13 (P<0.05). Further, goblet cell numbers and Muc2 levels were increased in SuperEGF pigs. Interestingly, the weaning-induced decrease of glucose cotransporter sodium-glucose linked transporter 1 (SGLT1) and glucagon-like peptide-2 (GLP2) levels was reversed by SuperEGF supplementation. Our findings add to our understanding of the mechanisms behind enhancing piglet performance by EGF containing fermentation product.

Growth performance of early-weaned pigs is enhanced by feeding epidermal growth factor-expressing Lactococcus lactis fermentation product.

We have previously generated epidermal growth factor expressing Lactococcus lactis (EGF-LL) using bioengineering approach, and shown that feeding newly-weaned piglets EGF-LL improves digestive function. To address concerns over the use of genetically modified organisms (GMO), the objective of the current study was to investigate the effect of feeding the EGF-LL fermentation product, after removal of the genetically modified EGF-LL, on growth performance and intestine development of newly-weaned piglets. One hundred and twenty newly-weaned piglets were fed ad libitum according to a 2-phase feeding program. Four pens were assigned to each of three treatments: (1) complete EGF-LL fermentation product (Ferm), (2) supernatant of EGF-LL fermentation product, after removal of EGF-LL (Supern), or (3) blank M17GE media (Control). EGF-LL or its fermented supernatant was administrated to piglets in the first 3 weeks post-weaning; their growth performance was monitored throughout treatment, and for the following week. Daily body weight gain (254.8g vs. 200.5g) and Gain:Feed (0.541kg/kg vs. 0.454kg/kg) of pigs on the Supern group were significantly improved compared to that of Control, although no difference was observed between the Ferm and Control pigs. Intestinal sucrase activity was increased in Supern- compared to Control group (166.3±62.1 vs. 81.4±56.5nmol glucose released/mg protein; P<0.05). The lack of growth response with Ferm pigs may be attributed to an overload of bacteria (daily dose included 4.56×10(10)CFU/kg BW/day EGF-LL). These results suggest that GMO-free EGF-LL fermentation product is effective in increasing growth performance of early-weaned piglets.

Nitrogen Absorbed from the Large Intestine Increases Whole-Body Nitrogen Retention in Pigs Fed a Diet Deficient in Dispensable Amino Acid Nitrogen

BACKGROUND Nitrogen absorption from the large intestine is considered of limited value for supporting body protein synthesis in animals and humans, but it may be of benefit when the dietary supply of nitrogen for synthesis of dispensable amino acids (DAAs) is deficient. OBJECTIVE A whole-body nitrogen balance study was conducted to evaluate the impact of nitrogen absorption from the large intestine of pigs fed a diet deficient in DAA nitrogen. METHODS Nine cecally cannulated barrows were fed a cornstarch and casein-based diet with a high indispensable amino acid (IAA) nitrogen to total nitrogen ratio (IAA:TN; 0.75). Pigs were randomly assigned to saline or 1 of 2 urea nitrogen infusion rates into the cecum (low and high, 1.5 and 3.0 g/d, respectively) following a 3 × 3 Latin square design. At the high urea nitrogen infusion rate, IAA:TN was 0.55. At slaughter, liver samples were taken to measure activity of carbamoyl phosphate synthetase I (CPS-I), glutamate dehydrogenase (GDH), and Gln synthetase (Gln-S). RESULTS Whole-body nitrogen retention improved with urea infusion (4.86 ± 0.20 g/d, 6.40 ± 0.21 g/d, and 7.75 ± 0.19 g/d for saline and low and high infusion rates, respectively; P < 0.05), as well as body weight gain. The marginal efficiency of using nitrogen absorbed from the large intestine for improving nitrogen retention was not affected by urea nitrogen infusion rate (P > 0.10). Enzyme activity of CPS-I or Gln-S was not different between treatments (P > 0.10), but GDH showed a trend for a positive linear response with increasing urea nitrogen infusion rate (P = 0.06). CONCLUSION These results indicate that urea nitrogen absorbed from the large intestine is efficiently used for increasing body protein deposition when feeding pigs a diet deficient in DAA nitrogen.

Nonprotein Nitrogen Is Absorbed from the Large Intestine and Increases Nitrogen Balance in Growing Pigs Fed a Valine-Limiting Diet

Nitrogen absorption from the large intestine, largely as ammonia and possibly as amino acids (AAs), is generally thought to be of little nutritional value to nonruminant animals and humans. Ammonia-nitrogen absorbed from the large intestine, however, may be recycled into the small intestine as urea and incorporated into microbial AAs, which may then be used by the host. A cecal infusion study was performed to determine the form in which nitrogen is absorbed from the large intestine and the impact of large intestine nitrogen supply on nitrogen balance in growing pigs. Eighteen cecally cannulated barrows (initial body weight: 22.4 ± 1.2 kg) were used to determine the effect of supplying nitrogen into the large intestine from either casein or urea on whole-body nitrogen retention and urea kinetics. Treatments were cecal infusions of saline (control), casein, or urea with nitrogen infused at a rate of 40% of nitrogen intake. In a subsample of 9 pigs, (15)N(15)N-urea was infused via i.v. during the nitrogen-balance period to determine urea kinetics. All pigs were fed a valine-limiting cornstarch-soybean meal-based diet. More than 80% of infused nitrogen was apparently absorbed. Urea flux and urinary nitrogen excretion increased (P ≤ 0.05) by the same amount for both nitrogen sources, but this increase did not fully account for the increase in nitrogen absorption from the large intestine. Whole-body nitrogen retention improved with nitrogen infusions (129 vs. 114 g/d; P < 0.01) and did not differ (P > 0.05) between nitrogen sources. Absorption of nitrogen from the large intestine appears to be in the form of nonprotein nitrogen, which appears to be returned to the small intestine via urea and used there for microbial AA production and should therefore be considered when determining nitrogen and AA supply and requirements.

Evidence for validity of ileal digestibility coefficients in monogastrics.

Measures of amino acid (AA) digestibility are used widely to estimate bioavailability of AA in feed and food ingredients for monogastric animals. In principle, the digestibility assay is simpler than in vivo assessments of AA bioavailability and allows for simultaneous estimation of the bioavailability of all AA in an experimental diet. It is generally assumed that absorption of intact AA in the hindgut of monogastrics is minimal, even though colonocytes do contain AA transporters and have been shown to absorb AA. This assumption is supported by the observation that infusion of AA into the hindgut does not improve nitrogen balance in monogastrics. In addition, growth performance of monogastrics is more highly correlated with ileal than faecal AA digestibility. Therefore, ileal digestibility coefficients provide better estimates of AA bioavailability than faecal digestibility coefficients. Measures of apparent ileal digestibility (AID) of AA are confounded with endogenous gut AA losses (EAAL). The curvilinear increase in AID of AA with increasing dietary AA level has been attributed to the relatively large contribution of EAAL to total ileal AA flows at low dietary AA levels. Subtracting basal EAAL from total ileal AA flows yields standardized ileal digestibility (SID) coefficients that appear to be more additive than AID coefficients in mixtures of feed ingredients. An implicit assumption when using SID AA coefficients in diet formulation is that the post-absorptive utilization of AA is not influenced by the dietary protein source. This assumption appears inappropriate when using feed or food ingredients that have been over-heated, induce large amounts of EAAL, or contain substantial amounts of fermentable fibre. Improved understanding of processes that contribute to the discrepancy between bioavailability and ileal digestibility will allow a more effective use of AA digestibility coefficients in diet formulation.

Nutrient composition and seasonal availability of local feedstuffs for pigs in western Kenya

Carter, N. A., Dewey, C. E., Lukuyu, B., Grace, D. and de Lange, C. F. M. 2015. Nutrient composition and seasonal availability of local feedstuffs for pigs in western Kenya. Can. J. Anim. Sci. 95: 397-406. In this study, nutritional values and seasonal availability were estimated for 25 local feedstuffs for pigs in western Kenya, based on analyzed nutrient contents and a review of the literature. Characteristics considered included: crude protein (CP), neutral detergent fibre (NDF), acid detergent fibre (ADF), ether extract (EE), Ca, P, standardized total tract digestible (STTD) P, total Lys, standardized ileal digestible (SID) Lys (all as g kg-1 of DM), and digestible energy (DE) (kcal kg-1 DM). Contents of total Lys, SID lysine, STTD P, and DE were estimated. Overripe improved-variety avocado (Persea americana) had the greatest estimated DE (5280 kcal kg-1 DM) and EE content (521 g kg-1 of DM). Cattle rumen content had the greatest NDF content (664 g kg-1 of DM). Sun-dried fish (Rastrineobola argentea) had the greatest estimated STTD P content (13.5 g kg-1 of DM). Amaranthus spinosus L. had the greatest Ca content (26 g kg-1 of DM). Fresh cattle blood had the greatest CP and estimated SID Lys content (944 and 83.0 g kg-1 of DM, respectively). Feedstuffs availability periods are: March through May plus September and October; June through August; and November through February. Local feedstuffs of sufficient nutritional value for pigs are seasonally available. Estimated nutritional values may assist in diet formulation.

Whole-body retention of α-linolenic acid and its apparent conversion to other n -3 PUFA in growing pigs are reduced with the duration of feeding α-linolenic acid

In the present study, fifteen growing pigs were used to determine the whole-body oxidation, retention efficiency (RE) and apparent conversion (AC) of α-linolenic acid (18 : 3n-3) to n-3 highly unsaturated fatty acids (HUFA), including EPA (20 : 5n-3) and DHA (22 : 6n-3). The pigs were fed a diet containing 10% flaxseed for 30 d. Whole-body fatty acid composition was determined at initial (27.7 (SE 1.9) kg), intermediate (day 15; 39.2 (SE 1.4) kg) and final (45.7 (SE 2.2) kg) body weight. On day 12, four pigs were fed 10 mg/kg of uniformly labelled (13)C-18 : 3n-3 (single-bolus dose) to determine the oxidation of 18 : 3n-3. Expired CO2 samples were collected for 24 h thereafter. The whole-body content of n-3 PUFA increased linearly (P< 0.0001) with time; however, the content of 22 : 6n-3 exhibited a quadratic response (P< 0.01) with a peak occurring at 15 h. As a proportion of intake, the RE of 18 : 3n-3 tended to reduce with time (P = 0.098). The AC of ingested 18 : 3n-3 to the sum of n-3 HUFA was reduced with time (P< 0.05; 12.2 v. 7.53 % for days 0-15 and days 15-30, respectively). The AC of 18 : 3n-3 to 20 : 5n-3 or 22 : 6n-3 was lower than that to 20 : 3n-3, both for days 0-15 (P < 0.05; 1.14 or 1.07 v. 7.06 %) and for days 15-30 (P< 0.05; 1.51 or 0.33 v. 4.29 %). The direct oxidation of 18 : 3n-3 was 7.91 (SE 0.98) % and was similar to the calculated disappearance of 18 : 3n-3 between days 0 and 30 (8.81 (SE 5.24) %). The oxidation of 18 : 3n-3 was much lower than that reported in other species. The AC of 18 : 3n-3 to n-3 HUFA was reduced over time and that to 20 : 3n-3 in the present study was much higher than that reported in other species and should be explored further.

Ammonia Nitrogen Added to Diets Deficient in Dispensable Amino Acid Nitrogen Is Poorly Utilized for Urea Production in Growing Pigs

Background: Including ammonia in low-crude protein (CP) diets deficient in dispensable amino acid (DAAs) increases nitrogen retention in growing pigs.Objective: We investigated the absorption and metabolism of dietary ammonia nitrogen in the portal-drained viscera (PDV) and liver of pigs fed a diet deficient in DAA nitrogen.Methods: Eight pigs with an initial mean ± SD body weight (BW) of 26.5 ± 1.4 kg were surgically fitted with 4 catheters each (portal, hepatic and mesenteric veins, and carotid artery). The pigs were fed (2.8 × 191 kcal/kg BW0.60), for 7 d and every 8 h, a diet deficient in DAA nitrogen supplemented with increasing amounts of ammonia nitrogen (CP: 7.76%, 9.27%, and 10.77%; indispensable amino acid nitrogen:total nitrogen ratio: 0.71, 0.59, and 0.50 for control and low- and high-ammonia diets, respectively). The treatment sequence was based on a Latin square design with 3 consecutive periods. On the last day of each period, blood flows in the portal and hepatic veins were determined with a continuous infusion of ρ-amino hippuric acid into the mesenteric vein. Serial blood samples were taken to determine ammonia and urea nitrogen concentration. Net balances of ammonia and urea nitrogen were calculated for the PDV and liver.Results: Cumulative (8 h) ammonia nitrogen appearance in the portal vein increased (P ≤ 0.05) with ammonia intake (433, 958, and 1629 ± 60 mg ammonia nitrogen/meal for control and low- and high-ammonia diets, respectively). The cumulative hepatic uptake of ammonia nitrogen increased (P ≤ 0.05) with ammonia nitrogen supply. The cumulative urea nitrogen appearance in the hepatic vein tended to increase (P ≤ 0.10) only in high-ammonia treatment (-92.5, -59.4, and 209.7 ± 92 mg urea nitrogen/meal for control and low- and high-ammonia diets, respectively) and, relative to the control diet, represented -6.0% and 11% of ammonia nitrogen intake.Conclusion: Dietary ammonia nitrogen is poorly utilized for urea production across splanchnic organs when pigs are fed diets deficient in DAA nitrogen.