PSXI-34 Evaluating the lysine requirements of adult and senior Labrador Retrievers using low and high crude protein inclusion levels

Despite consumer interest in senior pet care, little is known about the effect of changing crude protein (CP) levels on the lysine (Lys) requirements in young and senior dogs. Feeding crude protein in excess can lead to long-term health problems. Moreover, changing the CP content of the diet can lead differences in Lys requirements. Two experiments were conducted to determine the Lys requirements in adult and senior Labrador Retrievers through the indicator amino acid oxidation technique. Six young (< 1.5 yrs) and six old (>8 yrs) dogs were allocated to two trials (6 dogs/exp) to determine the effects of feeding 13 and 18% CP on the Lys requirement. All dogs were supplied with constant dietary Phe across diets. The control diet was fed for two days, followed by a day in which the test diet was fed, a tracer amino acid was supplied, and breath samples were collected. On test day, a priming dose of L-[1-13C]phenylalanine (Cambridge Isotope Laboratories, Inc.) based on the subject’s body weight was supplied, followed by [1-13C]Phe doses every 30 minutes, spanning a four hour period. A respiration mask was placed on each subject every 30 minutes (Oxymax, Columbus Instruments), 13CO2 was collected, and enrichment was determined by isotope ratio mass spectrometry (IRMS). Results for IRMS were converted to atom percent excess (APE) and analyzed using a broken-line model of best fit (JMP® Pro 15). Oxidation results showed that for a 13% CP diet, the Lys mean and population safe requirements (mean±2SD) were 1.56±0.25 and 0.81±0.37 g/1000kcal for adult and senior dogs, respectively. For the 18% CP diet, adult and senior dog mean and population safe requirements were 1.36±0.18 and 0.86±0.07 g/1000kcal accordingly. The results suggest that as dogs age, the lowered lean mass requires less AA nitrogen and Lys to maintain body tissues.

Rapid and accurate13CO2isotopic measurement in whole blood: comparison with expired gas

Determination of13CO2enrichment on the CO2 released from blood by acid has been used in situations in which breath sampling is difficult. This method can be improved by measuring this enrichment on the CO2 spontaneously released from blood. Therefore, simultaneous comparisons of13CO2content between breath and arterialized blood added with or without acid were performed in 51 samples from human studies, using the statistical method of Bland and Altman (J. M. Bland and D. G. Altman. Lancet 1: 307–310, 1986). Strong relationships exist between the methods ( r > 0.99) expressed in atom percent excess (APE). Compared with breath, the acid method overestimates the13CO2enrichment (0.318 ± 0.632 APE × 1,000, P < 0.001). The acid-free method shows similar enrichments to breath (0.003 ± 0.522 APE × 1,000, P = 0.97) with good precision and degree of agreement (95% confidence interval 0.15 APE × 1,000). The analysis can be performed up to 5 days after sampling with a good reproducibility. In conclusion, measuring13CO2enrichments on the CO2spontaneously released from blood is feasible, gives identical results to the breath method, and lessens operator manipulations. It allows study of situations in which the breath sampling method is not feasible.

Recovery of 13C in breath from NaH13CO3 infused by gut and vein: effect of feeding

Estimates of substrate oxidation obtained from appearance of 13C or 14C from tracers in breath must be corrected for retention of labeled carbon in the body. We aimed to determine the effect of a defined experimental diet and metabolic status on recovery of infused Na [13C]bicarbonate in breath. Six healthy male subjects consumed an experimental diet for 7 days before receiving a continuous infusion of formula without tracer on day 8 and received either an intragastric (ig) or intravenous (iv) infusion of Na [13C]bicarbonate on day 9 or 11 during a 4-h postabsorptive (PA), 4-h continuously fed period. A trend toward increasing PA breath enrichment during the first 7 diet days approached statistical significance (P = 0.051), whereas breath enrichments measured 3 h postbreakfast were consistently higher than PA values throughout and did not change over the 7-day period. Breath enrichments during a 4-h continuous ig infusion of formula without tracer on day 8 rose 2.0 +/- 0.5 atom percent excess (APE).10(-3) above base line (P less than 0.001, ANOVA). In the tracer studies, breath enrichments were similar for the ig and iv routes of tracer infusion. For the ig infusion the fraction of infused Na [13C]bicarbonate recovered in breath as 13CO2 was 0.74 +/- 0.02 for the PA period and 0.79 +/- 0.02 for the fed period. For the iv infusion the fraction recovered was 0.70 +/- 0.04 for the PA period and 0.82 +/- 0.03 for the fed period. Fractional recoveries were not significantly different for ig and iv routes of administration but were different for PA and fed periods (P less than 0.0001, 2-way ANOVA). The fractional recoveries for the fed period obtained here were similar to the value 0.81 reported in a number of other studies. Recovery of tracer in breath increased linearly with O2 uptake and CO2 production, suggesting that factors affecting respiratory gas exchange may alter recovery. We conclude that the primary factor determining label recovery is the immediate and recent nutritional status of the host.

Energy expenditure by doubly labeled water: validation in humans and proposed calculation

To further validate the doubly labeled water method for measurement of CO2 production and energy expenditure in humans, we compared it with near-continuous respiratory gas exchange in nine healthy young adult males. Subjects were housed in a respiratory chamber for 4 days. Each received 2H2(18)O at either a low (n = 6) or a moderate (n = 3) isotope dose. Low and moderate doses produced initial 2H enrichments of 5 and 10 X 10(-3) atom percent excess, respectively, and initial 18O enrichments of 2 and 2.5 X 10(-2) atom percent excess, respectively. Total body water was calculated from isotope dilution in saliva collected at 4 and 5 h after the dose. CO2 production was calculated by the two-point method using the isotopic enrichments of urines collected just before each subject entered and left the chamber. Isotope enrichments relative to predose samples were measured by isotope ratio mass spectrometry. At low isotope dose, doubly labeled water overestimated average daily energy expenditure by 8 +/- 9% (SD) (range -7 to 22%). At moderate dose the difference was reduced to +4 +/- 5% (range 0-9%). The isotope elimination curves for 2H and 18O from serial urines collected from one of the subjects showed expected diurnal variations but were otherwise quite smooth. The overestimate may be due to approximations in the corrections for isotope fractionation and isotope dilution. An alternative approach to the corrections is presented that reduces the overestimate to 1%.