A comparison of parenteral and enteral feeding in neonatal piglets, including an assessment of the utilization of a glutamine-rich, pediatric elemental diet.

BACKGROUND: The amino acid requirement profile for infants is different than that for adults and needs to be established; this profile also is different for infants receiving total parenteral nutrition. We used the neonatal piglet as a model for the infant to address (1) the metabolic and physiologic changes due to route of feeding and (2) the adequacy of the amino acid pattern in a pediatric elemental diet. METHODS: Diets differed only in their amino acid pattern (modified human milk [MHM] and a commercial pediatric elemental diet [PED]) and were fed continuously for 8 days. Control piglets were fed MHM diet via gastric catheters (IG) and were compared with pigs fed MHM diet via venous catheters (IV) or to pigs IG-fed PED. RESULTS: MHM-IV piglets experienced enlarged livers and gut atrophy, and lower nitrogen retention and body protein content. Higher glutamine (and lower glutamate) in PED-IG, compared with MHM-IG, produced no apparent advantage with respect to gut growth or histology. Proline, histidine, and lysine may have been limiting, and isoleucine and valine excessive, in the PED-IG diet as indicated by plasma concentrations, compared with sow-fed piglets; however, imbalances in the amino acid profile were not excessive because nitrogen retention was not different between MHM-IG or PED-IG pigs. CONCLUSIONS: Therefore, the amino acid profile of MHM needs to be modified to improve nitrogen retention during parenteral feeding and the profile of oral PED could be improved to normalize plasma amino acid concentrations.

Development of the indicator amino acid oxidation technique in chickens: calibration of oxidation system and determination of bicarbonate retention factor.

The indicator amino acid oxidation technique has been widely used as a rapid and sensitive method to determine amino acid requirement. In this technique, the indicator 14C-amino acid is continuously infused and breath 14CO2 is collected to determine oxidation rate, which is used to predict requirement. However, a portion of the 14CO2 produced from amino acid oxidation is retained in the body and must be determined to correct oxidation rates based on breath 14CO2 collection. To develop the indicator amino acid oxidation technique for use in poultry, it is important to accurately measure expired and retained CO2; these measurements were accomplished using intravenous infusion of NaH14CO3. Oxidation chambers were constructed and calibrated to ensure complete collection of breath 14CO2. Hydrochloric acid was infused over 1 h into a beaker containing NaH14CO3 in four closed oxidation chambers, and the 14CO2 released was collected over 2 h. Of the initial radioactivity dose, 99.3 +/- 1.9% was recovered in collected 14CO2. We concluded that the oxidation chamber apparatus is adequate for complete recovery of expired 14CO2. In Experiment 2, we determined bicarbonate retention in five broiler breeder roosters from a primed, constant infusion of NaH14CO3 into the jugular vein for 4 h. At steady state, 86.0 +/- 1.4% of NaH14CO3 infusion was recovered in breath. Because 14% of infused 14C is retained within the body, the present results demonstrate the need to determine bicarbonate retention in chickens during oxidation studies and use it as a correction factor in subsequent experiments.

Development of the indicator amino acid oxidation technique in chickens: L-[1-(14)C]phenylalanine infusion dose and phenylalanine oxidation.

Amino acid requirements of broiler breeder chickens are not well known. The indicator amino acid oxidation (IAAO) technique was adapted for use in broiler breeders as a rapid and sensitive method to determine amino acid requirements. During IAAO, phenylalanine oxidation decreases, inversely to the changes in protein synthesis, as the intake of the limiting test amino acid increases from deficient to adequate. Above the adequate level, phenylalanine oxidation remains constant. Before IAAO can be employed, the optimum priming and constant infusion doses of phenylalanine must be determined. Prelaying catheterized birds aged 20 to 24 wk were placed in closed oxidation chambers attached to a breath collection apparatus. A constant L-[1-(14)C]phenylalanine dose of 3.5 microCi/kg BW/h and priming doses of 4.5, 5.5, and 7.0 microCi/kg BW were used to determine optimal prime:constant dose ratios, minimum time taken for breath 14CO2 excretion to become constant (plateau), and adequate percentage of phenylalanine oxidized. At this constant infusion rate, the optimal priming dose of L-[1-(14)C]phenylalanine was 5.5 microCi/kg BW, resulting in a prime:constant dose ratio of 1.6:1. By using this ratio, the average time taken for breath 14CO2 to reach plateau was 60 min. Average phenylalanine oxidation at plateau, corrected for bicarbonate retention, was 5.5 +/- 1.4% (mean +/- SD), which is adequate for IAAO studies using deficient-to-excess levels of test amino acids. To the authors' knowledge, this study is the first in chickens to establish a primed, constant infusion technique using L-[1-(14)C]phenylalanine. The IAAO technique will be used in future studies to determine amino acid requirements in chickens.

Effect of common antinutritive factors and fibrous feedstuffs in pig diets on amino acid digestibilities with special emphasis on threonine.

Most feedstuffs contain antinutritive factors (ANF) such as insoluble fibers, lignins, tannins, and lectins. Intake of these ANF has the ability to reduce nutrient digestibility and to increase endogenous protein losses, such as through increased intestinal mucus secretion. The objective of this experiment was to determine the apparent ileal digestibilities (AID) of AA of 6 ANF-enriched diets to estimate endogenous protein loss associated with these ingredients in diets for young pigs. Forty-two 10-kg BW pigs fitted with a simple T-cannula at the distal ileum were randomly assigned to 1 of 7 casein-based diets with: no supplement (control), 100 g/kg of canola meal (CM), 100 g/kg of wheat bran (WB), 150 g/kg of barley (BR), 22.5 g/kg of lignin (LG), 15 g/kg of kidney beans [as a lectin (LE) source], and 15 g/kg of tannins (TN). All diets were formulated to be similar in N, indispensable AA, and caloric contents. After a 7-d adaptation to the test diets, N balance was conducted for 5 d, followed by 24 h of collection of digesta for analyses of AA. Pigs fed BR had 17% lower ADG and 15% lower feed conversion ratio (P < 0.05) compared with control and CM pigs. Pigs fed diets containing WB and BR had lower N retention as a percentage of absorbed N compared with all other groups (P = 0.03). The AID for CP was lower in BR, WB, and LE pigs compared with control. Of the AA, AID of Thr was notably lowest in BR, WB, and TN pigs (P < 0.05). The standardized ileal digestibility was lower in WB and BR pigs for most indispensable AA. Altogether, these data suggest that hemicellulose fiber, at concentrations typical in commercial swine diets, reduces AID of AA by increasing endogenous losses. Understanding the differential effects of ANF on endogenous losses of individual dietary AA will improve the accuracy of diet formulation.

Estimate of the variability of the lysine requirement of growing pigs using the indicator amino acid oxidation technique.

Although AA requirements for the mean in a population of growing pigs are well established, there are no direct estimates of their variability within the population. The indicator AA oxidation method allows repeated measurements in a short period of time so that the AA requirement can be determined for individual pigs. The objective was to determine the Lys requirement in individual pigs to derive a first estimate of the population mean requirement and its variability. Nine individually housed barrows (15 to 18 kg) were surgically implanted with venous catheters for isotope infusion. Pigs were offered, in random order, isonitrogenous and isoenergetic diets with one of seven Lys concentrations (4.8 to 15.5 g of Lys/kg diet, as-fed basis). The pigs were fed twice daily, except for study days when they received one-half of the daily allowance in eight equal hourly meals. After a validated minimum adaptation period, indicator (Phe) oxidation was determined for each dietary Lys level during a 4-h primed, constant infusion of L-[1-(14C)]Phe at a rate of 464 kBq/h. The Lys requirement was calculated using a two-phase linear regression crossover analysis within individual pigs. For each pig, Phe oxidation decreased linearly (P < 0.02) as the dietary Lys concentration increased until the requirement was reached; thereafter, Phe oxidation was not different. The true ileal digestible Lys requirement ranged from 7.5 to 10.6 g/kg of diet (as-fed basis) for the nine animals. The mean requirement for all pigs was 9.1 g/d (CV, 11.6%) or 93.9% (CV, 9.8%) of the predicted (NRC, 1998) requirement based on each pig's mean BW and energy intake. The measured and predicted requirements did not differ. The indicator AA oxidation method gave values for Lys requirement similar to conventional methods. The short (< 3 wk) experimental period allows, for the first time, the estimate of population variability, which provides for more accurate calculation of the effect of altering Lys intake on herd performance and production economics. This method is suitable to use with all dietary indispensable AA.

Organ and plasma amino acid concentrations are profoundly different in piglets fed identical diets via gastric, central venous or portal venous routes.

In a previous study in piglets fed identical diets intravenously, intraportally or intragastrically, we determined that small intestinal atrophy affects nitrogen metabolism to a greater extent than liver by-pass. Because whole-body amino acid homeostasis is dependent on interorgan metabolism, we also measured the free amino acid concentrations in liver, small intestinal mucosa and kidney in order to study alterations in amino acid metabolism within these organs. Piglets (n = 15; 2-4 d old) were fed identical diets continuously for 8 d via gastric (IG), portal (IP) or central venous (IV) catheters. Concentrations of all measured amino acids were affected by route of feeding in one or more organs. In particular, urea cycle amino acid concentrations were altered in plasma and mucosa of IV and IP pigs, suggesting that arginine synthesis by an atrophied gut may have been limited. Furthermore, most indispensable amino acid concentrations were lower in IP pigs for all organs vs. IG pigs; however, except for phenylalanine, plasma concentrations of these amino acids were not different, demonstrating the liver's "smoothing" capability. Gut atrophy in both IV and IP pigs resulted in significantly lower concentrations of all indispensable amino acids compared with IG pigs. Alterations of all amino acids in various organs due to route of feeding suggest that more detailed analyses of regulatory mechanisms and amino acid interactions on interorgan amino acid metabolism are necessary for all amino acids.

Alterations in intestinal uptake and compartmentalization of zinc in response to short-term dexamethasone therapy or excess dietary zinc in piglets.

Premature infants receive high dietary zinc and often glucocorticoids as a treatment for chronic lung disease. A piglet model was developed to investigate intestinal zinc transport and distribution of tissue zinc in response to treatment with short-term (5 d) glucocorticoid therapy or a high zinc diet. Piglets (13-15 d old; n = 21) were randomly allocated to: 1) dexamethasone (DEX) therapy (1.5 mg/kg intramuscularly twice a day), 2) high zinc diet (15.3 mmol/kg), or 3) control group (0.3 mmol dietary zinc/kg and saline intramuscularly twice a day). Pig weight, formula intake, urine volume, and blood glucose were monitored. At necropsy, tissue samples were obtained to measure zinc in plasma and zinc and metallothionein in liver and small intestinal mucosa. Velocity of zinc uptake by intestinal brush border membrane vesicles was measured using 65Zn tracer. Maximum uptake rate and Km for zinc uptake by brush border membrane vesicles were significantly greater (p < 0.05) in DEX compared with control and high zinc groups. DEX-treated piglets had a significantly lower (p < 0.05) zinc efflux rate across brush border membrane vesicles compared with that of the control. The high zinc group had a higher liver (p < 0.05) and mucosal (p < 0.05) zinc content and higher liver metallothionein concentration (p < 0.001) compared with the control and DEX groups. Weight gain over 5 d was not different among groups. Daily blood glucose was higher (p < 0.05) in DEX versus control and high zinc groups.(ABSTRACT TRUNCATED AT 250 WORDS)

Proline ameliorates arginine deficiency during enteral but not parenteral feeding in neonatal piglets.

The indispensability of arginine has not been conclusively established in newborns. Because parenteral feeding bypasses the gut (where de novo synthesis of arginine occurs from proline), a dietary supply of arginine that is sufficient to maintain urea cycle function may be of greater importance during intravenous compared with enteral feeding. Two-day-old piglets (n = 12) were fed nutritionally complete diets for 5 days via either a central vein catheter (IV pigs, n = 6) or a gastric catheter (IG pigs, n = 6). Subsequently, each piglet received three incomplete test diets [arginine free (-ARG/+PRO), proline free (-PRO/+ARG), or arginine and proline free (-ARG/-PRO)] in a randomized crossover design. Plasma ammonia was assayed every 30 min for 8 h or until hyperammonemia was observed. Ammonia increased rapidly in IV pigs receiving -ARG/+PRO and -ARG/-PRO (84 +/- 36 and 74 +/- 37 micromol. l(-1). h(-1), respectively), requiring early diet cessation. A rapid increase was also exhibited by IG pigs receiving the -ARG/-PRO, but not the -ARG/+PRO diet (31 +/- 15 vs. 11 +/- 7 micromol. l(-1). h(-1), respectively, P < 0.05). Plasma arginine and proline were indicative of deficiency (IG and IV groups) when deplete diets were infused. Arginine is indispensable in parenteral and enteral nutrition, independent of dietary proline.

Intestinal atrophy has a greater impact on nitrogen metabolism than liver by-pass in piglets fed identical diets via gastric, central venous or portal venous routes.

Whole-body nitrogen metabolism is altered during parenteral feeding as a result of gut atrophy and/or lack of splanchnic first-pass metabolism. We developed in vivo models to describe the metabolic and physiologic effects of first-pass metabolism by the small intestine/liver, liver or non-splanchnic tissues. Fifteen 2- to 4-d-old piglets were fed identical diets continuously for 8 d via gastric (IG), portal (IP) or central venous (IV) catheters. Despite similar weight gain, IV and IP pigs had higher nitrogen output and hence lower nitrogen retention (80%) compared with IG pigs (87%) (P = 0.002). Body protein content was also higher in IG pigs (583 mg/g dry matter) compared with IV (550) and IP pigs (534) (P = 0.003). Despite similar intestinal lengths, total small intestinal and mucosal weights were approximately 40% lower in IV and IP pigs than in IG pigs. Free urea cycle amino acids were altered in plasma and mucosa, suggesting that limited arginine synthesis by an atrophied gut may have limited protein deposition. Although villous atrophy was observed in the duodena and jejuna of IV and IP pigs, reduced crypt depth was observed only in IV pigs. Crypt depth was similar in all four gut sections from IG and IP pigs, suggesting that nutrient flux through the liver affects gut growth. Overall, metabolic responses to IV (non-splanchnic) and IP (liver) feeding were similar as a result of gut atrophy, whereas responses to IG (small intestine + liver) and IP (liver) feeding were different, suggesting that small intestinal atrophy affects nitrogen metabolism to a greater extent than liver by-pass.

Calcium competes with zinc for a channel mechanism on the brush border membrane of piglet intestine.

Interactions between Ca(+2) and Zn(+2) at the intestinal brush border membrane occur via unclear mechanisms. We hypothesized that Zn(+2) and Ca(+2) are transported across the brush border membrane via a multidivalent metal channel. Using brush border membrane vesicles (BBMV) prepared from intestines of 8 sow-fed piglets, we sought to determine whether Ca(+2) competes with Zn(+2) for uptake. Extravesicular Zn(+2) was removed with ethylenediamine-tetraacetic acid. Time curves of Zn(+2) and Ca(+2) uptake by BBMV were conducted with increasing concentrations of Ca(+2) and Zn(+2), respectively. Saturation curves compared kinetic parameters of Zn(+2) uptake with and without Ca(+2). In addition, Zn(+2) uptake was measured in the presence of various classical Ca(+2) channel modulators. Over 20 min, a 0.4x concentration of Zn(+2) lowered Ca(+2) uptake by vesicles, whereas a 30x concentration of Ca(+2) was necessary to lower Zn(+2) uptake. These data suggest that Ca(+2) has lower affinity than Zn(+2) for a brush border membrane transport protein. Kinetic parameters showed higher K(m) values with 4 or 15 mM Ca(+2) but unchanged J(max), suggesting competitive inhibition. The Ca(+2) channel blocking agents, La(+3), Ba(+2), verapamil, and diltiazem, inhibited Zn(+2) uptake, whereas calcitriol, trans 1,2 cyclohexanediol, cis/trans 1,3 cyclohexanediol, and the L-type Ca(+2) channel agonist, Bay K8644, induced Zn(+2) uptake. These data were consistent with competition for a common transport mechanism on the brush border membrane, possibly a novel multimetal channel. Copyright 2001 Elsevier Science Inc.

Divalent metals inhibit and lactose stimulates zinc transport across brush border membrane vesicles from piglets.

Interactions between metals of similar coordination chemistry are of relevance to infant nutrition due to the highly variable metal:metal ratios found in formulas. Using ratios similar to those found in infant formulas, our objectives were to determine the effects of metals and of lactose and other saccharides on Zn(+2) transport across intestinal brush border membranes. Brush border membrane vesicles prepared from intestines of 5 preweaned piglets were used to determine whether Ca(+2), Mg(+2), Fe(+2), Cu(+2), Cd(+2), or Mn(+2) would antagonize Zn(+2) uptake. (65)Zn(+2) uptake by brush border membrane vesicles was measured over 20 min with metal concentrations constant, and at 1 min with increasing metal concentrations. Zn(+2) bound to the external surface of vesicles was removed with ethylenediamine-tetraacetic acid. Lactose induced Zn(+2) uptake to a greater extent than glucose polymer, whereas maltose, galactose, or galactose/glucose had no effect. Over 20 min, a 10:1 concentration of Fe(+2), Cd(+2), Cu(+2), and Mn(+2) lowered Zn(+2) uptake significantly (P < 0.05). Higher concentrations of divalent cation significantly lowered Zn(+2) (0.2 or 0.1 mM) uptake for all metals tested (P < 0.05), except for Mn(+2) (0.1 mM Zn(+2)). Inhibition constant determination quantified relative competitive potential with Mg(+2) < Ca(+2) << Mn(+2) < Fe(+2) < Zn(+2) << Cu(+2). Relative amounts of Ca(+2), Mg(+2), and Fe(+2) similar to those found in infant formulas reduced Zn(+2) uptake by at least 40%. Our data demonstrate that dietary minerals compete during brush border membrane transport, and may help explain antagonistic mineral interactions observed in vivo. Divalent metal concentrations and lactose content of milk affect zinc absorption in neonates and must be carefully considered in formula design.

Threonine requirement of neonatal piglets receiving total parenteral nutrition is considerably lower than that of piglets receiving an identical diet intragastrically.

Evidence is accumulating that the amino acid requirements for neonates receiving total parenteral nutrition (TPN) are significantly different than those for oral feeding and need to be determined. The parenteral threonine requirement was determined in 3-d-old male Yorkshire piglets (n = 25) by examining the effect of varying dietary threonine intakes [0.05-0.6 g/(kg.d)] on phenylalanine oxidation. The diet included adequate energy, total amino acids and phenylalanine, with excess tyrosine. Phenylalanine kinetics were determined from a primed, continuous intravenous infusion of L-[1-14C]phenylalanine. Phenylalanine oxidation, estimated from the rate of 14CO2 released in expired air during isotope infusion, decreased (P < 0.05) as threonine intake increased from 0.05 to 0.15 g/(kg.d) and was low and constant for threonine intakes >0.15 g/(kg.d). Using breakpoint analysis with 95% confidence interval (CI), mean requirement and safe level of parenteral threonine intake were estimated to be 0.19 and 0.21 g/(kg. d), respectively (equivalent to 13 and 14 mg/g amino acids, respectively). To compare these data with those of orally fed controls, we then repeated the experiment by infusing identical diets intragastrically to piglets (n = 25); the varying dietary threonine intakes were 0.1-1.2 g/(kg.d). Employing identical kinetics and analyses, the mean requirement and safe level of oral threonine intake were estimated to be 0.42 and 0.51 g/(kg.d), respectively (equivalent to 28 and 34 mg/g amino acids, respectively). These data demonstrate that the threonine requirement of neonates during TPN is approximately 45% of the mean oral requirement.

Steady state is not achieved for most plasma amino acids during 12 hours of fasting in the neonatal piglet.

Kinetics studies in neonates are important to establish the requirement for amino acids and to understand the mechanisms of normal and altered metabolism. During kinetics experiments, plasma amino acid concentrations should be in steady state. Our objective was to determine whether 12 h of fasting, after parenteral or enteral feeding, resulted in a steady state in concentrations of amino acids. Two-day-old piglets were implanted with catheters (d 0), and randomly assigned to either intragastric (i.g., n = 6) or i.v. (n = 6) feeding. On d 5, piglets were fasted for 12 h. During the first 2 h, plasma concentrations of almost all amino acids declined except asparagine (i.g. and i.v.), tyrosine (i.v.), and glycine (i.v.), which increased. Only i.g. glycine did not change. Between 2 and 12 h, the only indispensable amino acids that did not change were phenylalanine (i.v.) and histidine (i.g. and i.v.). The branched-chain amino acids increased during this period (i.v. and i.g.). The greatest change was tyrosine, increasing 13% (i.v.) and 32% (i.g.) per hour. After 12 h of refeeding, glycine, serine, threonine, and asparagine concentrations were lower than baseline (p<0.05) in the i.v. group. In i.g. fed piglets, only threonine remained below baseline (p<0.05), and arginine was greater than baseline (p<0.05). Differences between i.v. and i.g. may be the result of impaired small intestinal metabolism secondary to parenteral feeding. In neonatal pigs, most plasma amino acids were unstable during 12 h of fasting. Thus, kinetics studies that require a steady state must be conducted in the fed state.