Measurement of energy expenditure in humans by doubly labeled water method

The utility of the doubly labeled water method for the determination of energy expenditure and water output was investigated in humans. Approximately 10 g of 18O and 0.5 g of 2H as water was orally administered to four healthy adults. Total body water was determined from the isotope dilution, and the ensuing 18O and 2H disappearance rates from body water were determined for 13 days by mass spectrometric isotope ratio analysis of the urinary water. During this period, subjects were maintained on a measured diet to determine energy and water intake. The energy expenditure from the doubly labeled water method differed from dietary intake plus change in body composition by an average of 2%, with a coefficient of variation of 6%. The water outputs determined by the two methods differed by 1%, with a coefficient of variation of 7%. The doubly labeled water method is noninvasive, and the subjects could maintain their daily activities without restriction.     

Estimation of doubly labeled water energy expenditure with confidence intervals

Bivariate regression is used to estimate energy expenditure from doubly labeled water data. Two straight lines are fitted to the logarithms of the enrichments of oxygen-18 and deuterium simultaneously as a bivariate regression, so that the correlations between the oxygen and deuterium regression coefficients can be estimated. Maximum likelihood methods are used to extend bivariate regression to unbalanced situations caused by missing observations and to include replicate laboratory determination from the same urine samples, even if one of the replicates is missing. Use of maximum likelihood allows the determination of a confidence interval for the energy expenditure based on the log likelihood surface rather than use of the propagation of variance methods for nonlinear transformations. The model is extended to include the subject's deviations from the two lines as a bivariate continuous-time first-order autoregression to allow for serial correlation in the observations. The analysis of data from two subjects, one without apparent serial correlation and one with serial correlation, is presented.     

Use of the doubly labeled water method for measurement of energy expenditure, total body water, water intake, and metabolizable energy intake in humans and small animals

The basis of the doubly labeled water method is measurement of the differential rates of disappearance of two isotopes of water (H2 18O and either 2H2O or 3H2O, administered at the start of the study) from body water. Published studies indicate that, in its current forms, this technique can be used to provide accurate and reasonably precise information on carbon dioxide production, total body water, and water intake in free-living humans and many small animals. Total energy expenditure can be calculated from carbon dioxide production with little loss of precision. Metabolizable energy intake can also be predicted, as the sum of total energy expenditure plus an estimate for the change in body energy stores during the measurement, but this prediction is unlikely to be accurate and precise unless the subject is in approximate energy balance.     

Sensitivity of methods for calculating energy expenditure by use of doubly labeled water

Attempts to estimate human energy expenditure by use of doubly labeled water have produced three methods currently used for calculating carbon dioxide production from isotope disappearance data: 1) the two-point method, 2) the regression method, and 3) the integration method. An ideal data set was used to determine the error produced in the calculated energy expenditure for each method when specific variables were perturbed. The analysis indicates that some of the calculation methods are more susceptible to perturbations in certain variables than others. Results from an experiment on one adult human subject are used to illustrate the potential for error in actual data. Samples of second void urine, 24-h urine, and breath collected every other day for 21 days are used to calculate the average daily energy expenditure by three calculation methods. The difference between calculated energy expenditure and metabolizable energy on a weight-maintenance diet is used to estimate the error associated with the doubly labeled water method.     

Energy expenditure of elite female runners measured by respiratory chamber and doubly labeled water.

To determine whether female athletes have unusually low energy requirements as suggested by many food intake studies, energy expenditure (EE) and intake were assessed in nine elite distance runners [26 +/- 3 (SD) yr, 53 +/- 4 kg, 12 +/- 3% body fat, and 66 +/- 4 ml.kg-1.min-1 maximal O2 uptake]. Subjects were admitted to a metabolic ward for 40 h during which 24-h sedentary EE was measured in a respiratory chamber. Free-living EE was then assessed by the doubly labeled water method for the next 6 days while the women recorded all food intake, daily body weight, and training mileage (10 +/- 3 miles/day). Energy intakes estimated from free-living EE (2,826 +/- 312 kcal/day) and body weight changes (-84 +/- 71 g/day) averaged 221 +/- 550 kcal/day in excess of those calculated from food records (2,193 +/- 466 kcal/day). The energy cost of training (1,087 +/- 244 kcal/day) was calculated as the difference between free-living EE and 24-h EE in the respiratory chamber (1,681 +/- 84 kcal/day) corrected for the thermic effect of food of the extra energy intake. These data do not support the hypothesis that training as a distance runner results in metabolic adaptations that lower energy requirements in women.     

A rapid analytical technique for the determination of energy expenditure by the doubly labelled water method

The doubly labelled water method involves the administration of water enriched in 2H and 18O followed by determination of the turnover rates of these isotopes. Since 18O is eliminated from the body as both CO2 and water, while 2H leaves only as water, the difference between the two turnover rates provides a measure of CO2 production and hence energy expenditure. Isotopic analysis by conventional stable isotope ratio analysis (SIRA) is labour intensive and time consuming, as it requires off-line conversion of water samples to gases (H2 and CO2) followed by sequential analysis for each of the two isotopes using the mass spectrometer. Lack of suitable automated instrumentation with the ability to process large numbers of samples has prevented routine application of the method. We describe here an automated technique in which body water samples (urine, saliva, breath water or milk) are analysed simultaneously for 2H and 18O. The single bench system comprises two mass spectrometer analysers, one for measuring 2H from H2 gas, the other for measuring 18O from the water vapour (masses 18, 20). Both analysers share a common heated inlet system into which microlitre quantities of the body fluids are injected from an autosampler (102 samples). The water vapour flows both directly to one analyser for 18O measurement and into a uranium reduction furnace for conversion to H2, prior to 2H measurement by the second analyser. Both analysers also share vacuum and electronic components, enabling savings in both space and cost. In this paper we present results illustrating performance characteristics and procedures for routine application to human subjects.(ABSTRACT TRUNCATED AT 250 WORDS)     

Comparison of different approaches for the calculation of energy expenditure using doubly labeled water in a small mammal

The doubly labeled water (DLW) method is an isotope-based technique for the estimation of the CO(2) production, and hence energy expenditure, of free-living animals and humans. Several methods are available for the calculation of CO(2) production from the isotope fluxes, depending on different assumptions about the behavior of isotopes during the elimination process. We used the DLW method to estimate the daily energy expenditures (DEE) of 55 field voles (Microtus agrestis) held in a captive facility at 8 degrees C. We calculated DEE using both plateau and intercept approaches for estimating the sizes of the isotope dilution spaces, three different assumptions about fractionation processes, and two ways of treating the different dilution spaces of the oxygen and hydrogen isotopes. We compared the resultant DEE estimates with metabolizable energy intake (MEI) measured during a 3-d feeding trial immediately before the DLW measurements, during which the animals were in energy balance. By making different assumptions about the apparent energy absorption efficiency, we generated a range of direct estimates of MEI. When we compared DEE and MEI, we found that the two-pool model formulations consistently underestimated energy demands by up to 29.8%, depending on the assumptions made in the reference calculation. However, while our data suggest that some correction for fractionation is necessary, with the present data we were unable to separate the two most common treatments of fractionation. These data strongly support the previous suggestion that for small mammals single-pool models provide more accurate estimates of energy demands than two-pool formulation of the DLW method.     

Evaluation of dietary assessment instruments against doubly labeled water, a biomarker of habitual energy intake

Epidemiological studies of diet and disease rely on the accurate determination of dietary intake and subsequent estimates of nutrient exposure. Although methodically developed and tested, the instruments most often used to collect self-reported intake data are subject to error. It had been assumed that this error was only random in nature; however, an increasing body of literature suggests that systematic error in the reporting of true dietary intake exists as well. Here, we review studies in which dietary intake by self report was determined while energy expenditure was simultaneously measured using the doubly labeled water (DLW) method. In seeking to establish the relative accuracy of each instrument to capture true habitual energy intake, we conclude that none of the self-reported intake instruments demonstrates greater accuracy against DLW. Instead, it is evident that the physical and psychological characteristics of study participants play a significant role in the underreporting bias observed in these studies. Further research is needed to identify underreporters and to determine how to account for this bias in studies of diet and health.     

A review of seabird energetics using the doubly labeled water method

The doubly labeled water (DLW) method has been essential for understanding animal energetics of free-ranging individuals. The first published studies on free-ranging seabirds were conducted on penguins in the early 1980s. Since then, nearly 50 seabird species with representatives from each major taxonomic order have been studied using DLW. Although the basic methodology has not changed, there are at least nine different equations, varying with respect to assumptions on fractionation and the total body water pool, to estimate field metabolic rate (FMR) from isotopic water turnover. In this review, I show that FMR can vary by as much as 45% depending on the equation used to calculate CO₂ production in five albatross species. Energy budgets derived from DLW measurements are critical tools for understanding patterns of energy use and allocation in seabirds. However, they depend on accurate and representative measurements of FMR, so analyses that include greater partitioning of activity specific FMR yield more realistic cost estimates. I also show how the combined use of DLW and biologging methods can 1) provide greater clarity for explaining observed variation in FMR measurements within a species and 2) allow FMRs to be viewed in a wider physiological, behavioral, or ecological context. Finally, I update existing allometric equations with new FMR data. These updates reaffirm that albatrosses have the lowest at-sea FMRs per equivalent body mass and that individuals of other seabird orders have FMRs ranging between 1.39 and 2.24 times higher than albatrosses.     

Adaptation of the doubly labeled water method for subjects consuming isotopically enriched water

Gretebeck, Randall J., Dale A. Schoeller, Rick A. Socki,Janis Davis-Street, Everett K. Gibson, Leslie O. Schulz, and Helen W. Lane. Adaptation of the doubly labeled water methodfor subjects consuming isotopically enriched water. J. Appl. Physiol. 82(2): 563-570, 1997.The use ofdoubly labeled water (DLW) to measure energy expenditure is subject toerror if the background abundance of the oxygen and hydrogen isotopetracers changes during the test period. This study evaluated theaccuracy and precision of different methods by which such backgroundisotope changes can be corrected, including a modified method thatallows prediction of the baseline that would be achieved if subjectswere to consume water from a given source indefinitely. Subjects inthis study were eight women (4 test subjects and 4 control subjects)who consumed for 28 days water enriched to resemble drinking water aboard the United States space shuttle. Test subjects and control subjects were given a DLW dose on days1 and 15,respectively. The change to an enriched water source produced a bias inexpenditure calculations that exceeded 2.9 MJ/day (35%), relative tocalculations from intake-balance. The proposed correction based on thepredicted final abundance of ¹⁸Oand deuterium after equilibration to the new water source eliminated this bias, as did the traditional use of a control group. This newmodified correction method is advantageous under field conditions whensubject numbers are limited.

     

Doubly labeled water measurement of human energy expenditure during strenuous exercise

The energy expenditures (EE) of 23 adult male Marines were measured during a strenuous 11-day cold-weather field exercise at 2,200- to 2,550-m elevation by both doubly labeled water (2H2 18O, DLW) and intake balance methods. The DLW EE calculations were corrected for changes in baseline isotopic abundances in a control group that did not receive 2H2 18O. Intake balance EE was estimated from the change in body energy stores and food intake. Body energy-store changes were calculated from anthropometric [-1,574 +/- 144 (SE) kcal/day] and isotope dilution (-1,872 +/- 293 kcal/day) measurements made before and after the field exercise. The subjects kept daily logbook records of ration consumption (3,132 +/- 165 kcal/day). Mean DLW EE (4,919 +/- 190 kcal/day) did not differ significantly from intake balance EE estimated from food intake and either anthropometric (4,705 +/- 181 kcal/day) or isotope dilution (5,004 +/- 240 kcal/day) estimates of the change in body energy stores. The DLW method can be used with at least the same degree of confidence as the intake balance method to measure the EE of active free-living humans.     

Inaccuracies in self-reported intake identified by comparison with the doubly labelled water method

To test the accuracy of self-reported energy intake, reported intake was compared with measured energy expenditure. Results from nine studies were reviewed in which intake data were obtained by recall or weighed record for at least 7 days. Expenditure was measured for 7 days or more by the doubly labelled water method. Individual differences between reported intake and expenditure were large (range +25 to -76%). Group mean differences were smaller. Lean, nonathletic groups living in industrialized countries demonstrated the smallest mean difference between self-reported energy intakes and expenditure (0 to -20%). Obese populations demonstrated the largest mean differences (-35 and -50%), but women living in the Gambia and elite athletes also demonstrated large mean differences. Most of the difference appears to be due to under-reporting, but some subjects lost weight during the reporting period indicating that some of the difference was due to undereating. Because the greatest bias was observed in obese subjects, current methods for self-reported energy intake are not recommended for use in obesity research.     

Measurement of CO2 production by the doubly labeled water technique

Recent applications of the doubly labeled water technique to the study of human metabolism have employed multiple sampling of body water over protracted periods, rather than the more traditional method of taking only an initial and final sample for isotopic analysis. In addition fractional turnovers of the body pools have been estimated by fitting curves to the sequential log-converted isotope enrichment against time. By manipulation of data collected in the field in a study of metabolism of vespertilionid bats, it is shown the curve-fitting technique results in an accurate estimate of CO2 production only when the rate of CO2 production is constant. Biologically realistic nonsteady-state conditions result in errors in estimates of CO2 production of up to 30%. In conditions where CO2 production is known to be temporally variable, the more traditional two-sample method may provide a more accurate estimate of CO2 production.     

Milk intake and carbon dioxide production of piglets determined with the doubly labelled water technique

The present study was undertaken to study different methodological aspects of quantifying CO2 production and milk intake of suckling piglets using the doubly labelled water (DLW) technique. In total, 37 piglets were enriched intraperitoneally with DLW to study equilibration time of 18O (n = 3), to validate the estimation of milk intake and CO2 production (n = 10) of piglets fed milk replacer and to quantify milk intake and CO2 production of piglets nursed ordinarily by sows (n = 24). Enrichment of 18O in expired air was analysed without any sample preparation, whereas enrichment of 18O in serum was analysed after a minimum step of sample preparation, which included pipetting of the sample, blowing gaseous CO2 into the vial for 3 s and equilibrating for 24 h. The 18O enrichment of CO2 in expired air was constant within 30-40 min of intraperitoneal injection, suggesting that DLW was equilibrated within the body water by that time. For piglets fed milk replacer, the estimation of the daily CO2 production by the DLW method (64.0 ± 2.7 l CO2/day) was in agreement with that obtained by respiration trials (64.7 ± 1.8 l CO2/day). Furthermore, the intake of milk replacer (891 ± 63 g/day) determined by deuterium oxide (D2O) dilution was similar in magnitude to that found by weighing the milk disappearance (910 ± 58 g/day). The milk intake of piglets fed milk replacer was comparable with that of sucking piglets, but sucking piglets had a remarkably higher CO2 production than artificially reared piglets, which likely was caused by a higher intake of milk solids and a higher activity level. For sucking piglets, the daily CO2 production increased curvilinearly with increasing live weight (LW) in kg: piglet CO2 production (l/day) = 25.75 × LW - 1.01 × LW2. In conclusion, 18O equilibrates fast within the body water pool when administered intraperitoneally, and the accuracy of assessing milk intake and rate of CO2 production using the DLW technique is promising. Assessment of excess enrichment of 18O in serum proved to be robust. Finally, the CO2 production of piglets fed milk replacer differs considerably from that of sucking piglets.     

Estimating the energy expenditure of free‐ranging polar bears using tri‐axial accelerometers: A validation with doubly labeled water

Measures of energy expenditure can be used to inform animal conservation and management, but methods for measuring the energy expenditure of free‐ranging animals have a variety of limitations. Advancements in biologging technologies have enabled the use of dynamic body acceleration derived from accelerometers as a proxy for energy expenditure. Although dynamic body acceleration has been shown to strongly correlate with oxygen consumption in captive animals, it has been validated in only a few studies on free‐ranging animals. Here, we use relationships between oxygen consumption and overall dynamic body acceleration in resting and walking polar bears Ursus maritimus and published values for the costs of swimming in polar bears to estimate the total energy expenditure of 6 free‐ranging polar bears that were primarily using the sea ice of the Beaufort Sea. Energetic models based on accelerometry were compared to models of energy expenditure on the same individuals derived from doubly labeled water methods. Accelerometer‐based estimates of energy expenditure on average predicted total energy expenditure to be 30% less than estimates derived from doubly labeled water. Nevertheless, accelerometer‐based measures of energy expenditure strongly correlated (r² = 0.70) with measures derived from doubly labeled water. Our findings highlight the strengths and limitations in dynamic body acceleration as a measure of total energy expenditure while also further supporting its use as a proxy for instantaneous, detailed energy expenditure in free‐ranging animals.