Lessons learned regarding symptoms of tryptophan deficiency and excess from animal requirement studies.

Tryptophan is the precursor for several neurotransmitters and metabolic regulators, which, although quantitatively of little importance in determining the dietary requirement, have major importance for interpreting symptoms of dietary tryptophan deficiency and excess. The quantitative dietary tryptophan requirement appears to vary widely across species, so intakes relative to requirements are more appropriate expressions for comparison of adverse effects across species than daily intake or diet concentration. Symptoms of tryptophan deficiency may occur at intakes as little as 25% below the requirement. Symptoms include reduced feed intake and reduced growth rate but also impaired skeletal development and aberrant behavior. Older animals appear less susceptible than younger animals to tryptophan deficiency and females less than males. Symptoms of excess tryptophan intake include reduced food intake and growth rate. In growing animals, it appears that tryptophan intakes of >10 times the requirement are necessary before there are detrimental effects on growth performance. At still greater intakes, fatty liver and fibrotic changes in muscles, lung, and pancreas and the serotonin syndrome may develop. In pigs, tryptophan intake of 60 times the daily requirement did not cause mortality. The maximal tryptophan oxidation rate, measured in vivo using (13)C universally labeled tryptophan, may be a possible marker of the intake above which increasing intake increases the risk of adverse effects. The advantage of the oxidation technique is that it does not necessarily rely on but still allows the identification and measurement of amino acid metabolites and is therefore simpler and more universally applicable.

The metabolic availability of threonine in common feedstuffs fed to adult sows is higher than published ileal digestibility estimates.

Amino acid (AA) requirements for sows during pregnancy are currently under review. However, requirement recommendations must be accompanied by an estimate of the bioavailability of AA from feeds to ensure adequate supply of AA and to minimize excess nitrogen excretion. Current ileal AA digestibility estimates are based on growing pig data; however, availability of AA in adult pigs may be different from that in growing pigs. The metabolic availability (MA) of threonine (Thr) in corn and barley was determined in 6 pregnant sows using the indicator AA oxidation method and L-[1-(13)C]phenylalanine as the tracer AA. Sows were fed reference diets formulated from 30 to 75% of the breakpoint derived in Expt. 1; all other nutrients were set at 120% of requirement. Test ingredients diets were formulated to supply Thr at 75% of the determined requirement. Tracer phenylalanine was given orally in 8 one-half-hourly meals and expired (13)CO(2) was quantified. The determined MA of Thr from corn and barley fed to pregnant sows was 88.0 and 89.3%, respectively. The determined MA was 7 and 9% greater than the published standard ileal digestibility estimates of Thr in corn (82%) and barley (81%), respectively. Mature animals have a greater capacity to digest and absorb nutrients from feed ingredients than previously assumed based on ileal digestibility studies. Sow diets formulated based on published ileal digestibility estimates are overformulated with respect to available protein and AA and thus increase excess nitrogen excretion and potential environmental concerns.

Application of the indicator amino acid oxidation technique for the determination of metabolic availability of sulfur amino acids from casein versus soy protein isolate in adult men.

Our objective was to determine the metabolic availability (MA) of sulfur amino acids in dietary proteins using the indicator amino acid oxidation (IAAO) technique. Five to seven men received graded levels (20, 40, 60, and 70%) of the mean total sulfur amino acid (TSAA) requirement of 13 mg x kg(-1) x d(-1) as a crystalline AA mixture, casein, and soy protein isolate (SPI) (40, 50, 60, and 70%), respectively. Five of these subjects received 40% of TSAA requirement from SPI supplemented with methionine to the level of 40% of requirement. These 5 subjects also repeated the level of 60% TSAA requirements from both casein and SPI to assess repeatability. The mean MA of TSAA from SPI (71.8 +/- 3.6%) was lower than from casein (87.4 +/- 3.8%, P < 0.05). Supplementation of SPI with methionine decreased the IAAO (11.5 +/- 0.3% administered dose) compared with unsupplemented SPI (12.8 +/- 0.5% administered dose, P < 0.05). IAAO was similar for repeated measurements of casein and SPI, respectively, at the 60% TSAA intake level (10.8 +/- 1.0 vs. 10.7 +/- 1.2% for casein; 12.7 +/- 1.3 vs. 12.9 +/- 2.6% for SPI). In conclusion, the IAAO technique can be used to determine the MA of AA for protein synthesis in test proteins for humans.

Development of the indicator amino acid oxidation technique to determine the availability of amino acids from dietary protein in pigs.

Standardized ileal ("true") digestibility is currently the best estimate of amino acid digestibility, but it does not measure bioavailability. Growth assays to determine amino acid bioavailability are expensive and laborious; thus, a rapid method is needed. Applying the principle of slope-ratio assay to the indicator amino acid oxidation (IAAO) method, we hypothesized that the reduction in indicator oxidation per gram of lysine in feedstuffs relative to that per gram of free lysine represented the bioavailability of lysine, here termed "metabolic availability." Indicator oxidation in pigs was linear over increasing lysine intakes (r = 0.90, P = 0.001) when the dietary lysine contents were 2 SD below the mean lysine requirement of the pigs. Peas were treated (raw, heated to reduce lysine availability, or heated with added lysine) to test the responsiveness of the IAAO to differing lysine availability. Free lysine reduced indicator oxidation by 3.16% of dose oxidized per gram added lysine, whereas the addition of protein lysine as raw (-2.81%) and heated peas (-1.73%) reduced oxidation to a lesser degree. Adding free lysine to heated peas decreased indicator oxidation, evidence that heating had worsened the utilization of pea protein for protein synthesis by reducing the bioavailability of lysine alone. Pea diets differed only in the availability of lysine; therefore IAAO detected differences in lysine bioavailability. Because the IAAO technique responds to lysine available at the sites of protein synthesis, the metabolic availability covers all losses during digestion, absorption, and utilization of lysine. This method can determine the metabolic availability of amino acids of a feedstuff within 2 wk.

Manipulation of dietary protein and nonstarch polysaccharide to control swine manure emissions.

Odor and greenhouse gas (GHG) emissions from stored pig (Sus scrofa) manure were monitored for response to changes in the crude protein level (168 or 139 g kg(-1), as-fed basis) and nonstarch polysaccharide (NSP) content [i.e., control, or modified with beet pulp (Beta vulgaris L.), cornstarch, or xylanase] of diets fed to pigs in a production setting. Each diet was fed to one of eight pens of pigs according to a 2 x 4, full-factorial design, replicated over three time blocks with different groups of animals and random assignment of diets. Manure from each treatment was characterized and stored in a separate, ventilated, 200-L vessel. Repeated measurements of odor, carbon dioxide (CO2), methane (CH4), and nitrous oxide (N2O) emissions from the vessels were taken every two weeks for eight weeks. Manure from high-protein diets had higher sulfur concentration and pH (P < or = 0.05). High-NSP (beet pulp) diets resulted in lower manure nitrogen and ammonia concentrations and pH (P < or = 0.05). Odor level and hedonic tone of exhaust air from the storage vessel headspaces were unaffected by the dietary treatments. Mean CO2 and CH4 emissions (1400 and 42 g d(-1) m(-3) manure, respectively) increased with lower dietary protein (P < or = 0.05). The addition of xylanase to high-protein diets caused a decrease in manure CO2 emissions, but an increase when added to low-protein diets (P < or = 0.05). Nitrous oxide emissions were negligible. Contrary to other studies, these results do not support the use of dietary protein reduction to reduce emissions from stored swine manure.

Growth potential, but not body weight or moderate limitation of lysine intake, affects inevitable lysine catabolism in growing pigs.

Inevitable catabolism contributes to the inefficiency of using dietary lysine intake for body protein deposition (PD). This study was conducted to determine the effects of true ileal digestible (TID) lysine intake, body weight (BW), and growth potential on lysine catabolism in growing pigs. Starting at 15 kg BW, 16 female Yorkshire pigs were offered a purified diet providing all nutrients in excess of requirements for maximum protein deposition (PDmax). At approximately 25 kg BW, the pigs' PDmax was determined using the N-balance method. Thereafter, 4 pigs were allocated to each of 4 diets, first-limiting in lysine, providing lysine intakes corresponding to 60, 70, 80, and 90% of estimated requirements for PDmax. The pigs were surgically fitted with catheters in the jugular and femoral veins. Lysine catabolism was determined at 2 BW (40-45 kg, low; 70-75 kg, high) either directly (oxidation) using a primed, constant infusion of l-[1-(14)C]-lysine or indirectly (disappearance) using the N-balance method. There was no effect of BW on the rate (g/d) or fraction of TID lysine intake catabolized. Lysine catabolism decreased with increasing growth potential. Lysine disappearance and lysine oxidation (% of TID lysine intake) were independent of lysine intake, except for the lowest lysine intake level, where they were lower. When lysine catabolism was independent of intake, lysine oxidation based on plasma free lysine specific radioactivity (SRA) was lower (9.9% of TID intake) than lysine disappearance (17.4% of TID intake) or lysine oxidation based on liver free lysine SRA (13.4% of TID intake).

Pattern of carbon dioxide production and retention is similar in adult pigs when fed hourly, but not when fed a single meal.

BACKGROUND: The understanding of bicarbonate kinetics and CO2 retention in the body is necessary to conduct amino acid tracer oxidation studies in both humans and laboratory animals. Significant metabolic activity is associated with eating which can affect bicarbonate steady state kinetics. A study was conducted to assess the impact of feeding regimen on the recovery of labelled bicarbonate and energy expenditure in adult female pigs (sows). Five catheterized sows (235 +/- 5 kg) were fed semi-synthetic diets as: a single meal 2 h into the infusion after an overnight fast, or in eight hourly meals starting 2 h before the infusion. Oxygen consumption, CO2 production and 14CO2 recovery (ie fraction not retained) were determined during primed, constant intravenous infusions of NaH14CO3. RESULTS: The 14CO2 recovery (%) after fasting (58.1 +/- 4.8) was lower than that after single meal feeding (78.8 +/- 5.9) or hourly meal feeding (81.0 +/- 2.6, P = 0.03). CO2 production correlated with 14CO2 recovery during hourly feeding (r = 0.40, P = 0.01); this relationship was not significant after single meal feeding (P = 0.30), probably due to physical activity-associated CO2 production. CONCLUSIONS: The correlation of CO2 retention factors with CO2 production during hourly feeding suggests that this regimen should be preferred for future amino acid kinetics studies.

Indicator amino acid oxidation responds rapidly to changes in lysine or protein intake in growing and adult pigs.

There is disagreement about the adaptation time required when using the indicator amino acid oxidation (IAAO) technique. Our objective was to establish the adaptation time required to obtain a plateau in indicator (L-[1-(14)C]-phenylalanine) oxidation in response to a test diet using growing and adult pigs. Four barrows (20 kg) and 4 sows (240 kg) were surgically implanted with venous catheters for isotope infusion. Growing Pigs: After 7 d of adaptation to an adequate lysine intake of 8.8 g/d, phenylalanine oxidation in growing pigs was 9.38 +/- 1.25% of the infused dose. At 2, 3, 4, or 6 d after reducing lysine intake to 3.8 g/d, and then increasing it back to 8.8 g/d, phenylalanine oxidation was 16.94 +/- 0.84% (P < 0.05) and 9.70 +/- 0.80% (P < 0.05), respectively, with no significant effect of days of adaptation to diet. Adult Pigs: After 14 d of adaptation to an intake of 200% of the amino acid maintenance requirement, phenylalanine oxidation in sows was 4.23 +/- 0.45% of dose. Changing the intake to 100 and 50% of the maintenance requirement, increased (P < 0.05) phenylalanine oxidation to 5.95 +/- 0.26 and 7.90 +/- 0.26%, respectively, with no significant effect of time (1, 2, 5, 6, 9, and 10 d) after diet change. The CV for repeated phenylalanine oxidation measurements within pigs and diets was 13.5% for growing and 8.8% for adult pigs. This demonstrates that the IAAO requires <2 d of adaptation regardless of age, dietary challenge (individual amino acid or total protein) or direction (increase or decrease) of change, and that the measured oxidation rate (% of dose) is highly repeatable.

Lysine requirements of pre-lay broiler breeder pullets: determination by indicator amino acid oxidation.

The indicator amino acid oxidation (IAAO) method allows the determination of amino acid requirements under conditions of low growth rate as found in pre-laying broiler breeder pullets. Cobb 500 breeder pullets (20 wk old; 2290 +/- 280 g, n = 4) were adapted (6 d) to a pelleted, purified control diet containing all nutrients at >or=110% of NRC recommendations. After recovery from surgery for implantation of a jugular catheter, each bird was fed, in random order, test diets containing one of nine levels of lysine (0.48, 0.96, 1.92, 2.88, 3.84, 4.80, 7.68, 9.60 and 14.40 g/kg of diet). Indicator oxidation was determined during 4-h primed (74 kBq/kg body), constant infusions (44 kBq x h(-1). kg body(-1)) of L-[1-(14)C]phenylalanine. Using the breakpoint of a one-slope broken-line model, the lysine requirement was determined to be 4.88 +/- 0.96 g/kg of diet or 366 +/- 72 mg x hen(-1) x d(-1) with an upper 95% CI of 6.40 g/kg of diet or 480 mg x hen(-1) x d(-1). IAAO allows determination of individual bird amino acid requirements for specific ages and types of birds over short periods of time and enables more accurate broiler breeder pullet diet formulation.