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Aerobic fitness determines whole-body fat


oxidation rate during exercise in the heat
Juan Del Coso, Nassim Hamouti, Juan Fernando Ortega, and
Ricardo Mora-Rodriguez

Abstract: The purpose of this study was to determine whole-body fat oxidation in endurance-trained (TR) and untrained
(UNTR) subjects exercising at different intensities in the heat. On 3 occasions, 10 TR cyclists and 10 UNTR healthy subjects (peak oxygen uptake = 60 6 vs. 44 3 mLkg1min1; p < 0.05) exercised at 40%, 60%, and 80% peak oxygen uptake in a hot, dry environment (36 8C; 25% relative humidity). To complete the same amount of work in all 3 trials,
exercise duration varied (107 4, 63 1, and 45 0 min for 40%, 60%, and 80% peak oxygen uptake, respectively).
Substrate oxidation was calculated using indirect calorimetry. Blood samples were collected at the end of exercise to determine plasma epinephrine ([EPI]plasma) and norepinephrine ([NEPI]plasma) concentrations. The maximal rate of fat oxidation
was achieved at 60% peak oxygen uptake for the TR group (0.41 0.01 gmin1) and at 40% peak oxygen uptake for the
UNTR group (0.28 0.01 gmin1). TR subjects oxidized absolutely (gmin1) and relatively (% of total energy expenditure) more fat than UNTR subjects at 60% and 80% peak oxygen uptake (p < 0.05). At these exercise intensities, TR subjects also had higher [NEPI]plasma concentrations than UNTR subjects (p < 0.05). In the heat, whole-body peak fat
oxidation occurs at higher relative exercise intensities in TR than in UNTR subjects (60% vs. 40% peak oxygen uptake).
Moreover, TR subjects oxidize more fat than UNTR subjects when exercising at moderate to high intensities (>60% peak
oxygen uptake).
Key words: catecholamines, carbohydrate oxidation, indirect calorimetry, cycling, body composition, endurance exercise.
Resume : Cette etude se propose danalyser le degre doxydation des gras dans tout lorganisme de sujets entranes en endurance (TR) et de sujets non entranes en endurance (UNTR) au cours dexercices accomplis a` differentes intensites dans
un environnement chaud. En trois occasions, 10 cyclistes TR et 10 sujets UNTR en bonne sante (consommation doxyge`ne
de pointe = 60 6 mLkg1min1 comparativement a` 44 3 mLkg1min1; p < 0,05) font un effort dans un environnement chaud (36 8C; 25 % HR), et ce, aux intensites suivantes : 40 %, 60 % et 80 % du consommation doxyge`ne de
pointe. Afin de maintenir constante la quantite de travail accompli, la duree des efforts varie comme suit : 107 4 min,
63 1 min et 45 0 min pour les intensites respectives de 40 %, 60 % et de 80 % du consommation doxyge`ne de
` la fin des seances dexercice, on prele`ve des
pointe. On mesure le degre doxydation des gras par calorimetrie indirecte. A
echantillons sanguins pour en determiner les concentrations plasmatiques depinephrine ([EPI]plasma) et de norepinephrine
([NEPI]plasma). On enregistre le plus haut taux doxydation des gras chez les TR a` 60 % du consommation doxyge`ne de
pointe (0,41 0,01 gmin1) et, chez les UNTR, a` 40 % du consommation doxyge`ne de pointe (0,28 0,01 gmin1). Les
sujets TR oxydent plus de gras que les sujets UNTR en valeur absolue (gmin1) et en valeur relative (% du total denergie
` ces intensites dexercice, la [NEPI]plasma
depensee) a` 60 % et 80 % du consommation doxyge`ne de pointe (p < 0,05). A
chez les sujets TR est plus elevee que chez les sujets UNTR (p < 0,05). Dans un environnement chaud, loxydation des
gras dans lorganisme se manifeste chez les sujets TR a` une plus haute intensite relative : 60 % comparativement a` 40 %
du consommation doxyge`ne de pointe chez les UNTR. De plus, les sujets TR oxydent plus de gras que les sujets UNTR
au cours defforts dintensite moderee a` elevee (>60 % du consommation doxyge`ne de pointe).
Mots-cles : catecholamines, oxydation des sucres, calorimetrie indirecte, cyclisme, composition corporelle,
exercice dendurance.
[Traduit par la Redaction]

Introduction
During exercise, factors such as exercise intensity and duration, environmental conditions, training status, adiposity,
and the pre-exercise meal influence fat oxidation (Jeu-

kendrup 2003). From light to moderate exercise intensities,


fat is the main energy substrate for muscle contraction, with
carbohydrate making a lower contribution (Jones et al.
1980). The highest rate of fat oxidation during exercise
(Fatmax) in a thermonetural environment has been found at

Received 8 April 2010. Accepted 12 August 2010. Published on the NRC Research Press Web site at apnm.nrc.ca on 5 November 2010.
J. Del Coso, N. Hamouti, J.F. Ortega, and R. Mora-Rodriguez.1 Exercise Physiology Lab at Toledo, Universidad de Castilla
La Mancha, Avda. Carlos III, s/n, Toledo, 45071, Spain.
1Corresponding

author (e-mail: Ricardo.Mora@uclm.es).

Appl. Physiol. Nutr. Metab. 35: 741748 (2010)

doi:10.1139/H10-068

Published by NRC Research Press

742

exercise intensities ranging from 33% to 65% peak oxygen


_ 2 peak) (Romijn et al. 1993, 2000; Bergman and
uptake (VO
Brooks 1999; Achten et al. 2002; Venables et al. 2005;
Nordby et al. 2006; Stisen et al. 2006; Riddell et al. 2008).
The rate of fat oxidation declines at exercise intensities
_ 2 peak, and it is almost negligible at
higher than 70% VO
_ 2 max) (Achten et al.
90% maximal oxygen uptake (VO
2002). As exercise progresses in duration, there is an increasing contribution of fat to energy production, while the
use of carbohydrate decreases over time (Romijn et al.
1993). Because of this, individuals undergoing weight loss
programs should target exercise intensity close to their
Fatmax while exercising for the longest possible duration.
However, this population is often unable to maintain exercise at Fatmax for a long time (Hulens et al. 2001). Exercise
practitioners may then consider reducing exercise intensity
to benefit from exercise duration. Data that address the best
combination of exercise intensity and duration that maximizes total fat oxidation are scarce.
One of the most prominent metabolic adaptations to endurance training is the higher reliance on fat oxidation during exercise. Several studies have shown that 1012 weeks
of endurance training substantially increase the rate of fat
_ 2 peak)
oxidation at a given relative workload (65% VO
(Coggan et al. 1990, 1993; Friedlander et al. 1998). Endurance-trained subjects oxidize relatively (as a percentage of
energy expenditure) and absolutely (gmin1) more fat than
their untrained counterparts when exercising at the same relative intensity (Bergman and Brooks 1999; Stisen et al.
2006). In addition, endurance-trained individuals achieve
Fatmax at higher relative exercise intensities than untrained
_ 2 peak) (Nordby et al. 2006).
subjects (50% vs. 43% VO
Thus, the exercise intensity prescribed to maximize fat oxidation may vary, depending on the aerobic capacity of the
individual.
Most studies focusing on fat oxidation during exercise
have been conducted in thermoneutral environments. However, exercise is frequently performed outdoors, where ambient temperature can be high. It has been found that
_ 2 max in a hot environment (40 8C) deexercise at 70% VO
creases the rate of fat oxidation in comparison to a thermoneutral environment (20 8C) (Febbraio et al. 1994).
However, it is unclear if the reduction in fat oxidation in
the heat is present at all exercise intensities, or if the heat
affects the intensity at which Fatmax is achieved. The first
purpose of this study was to determine the exercise intensity
that elicits Fatmax in trained and untrained subjects in a hot
environment in a fed state. The second purpose was to investigate the best combination of exercise intensity and duration to maximize total fat oxidation and energy
expenditure during exercise in the heat. We hypothesized
that endurance-trained individuals would maximize fat oxidation in the heat at a higher relative exercise intensity than
their untrained counterparts.

Materials and methods


Subjects
Ten endurance-trained cyclists (TR) and 10 untrained
(UNTR) healthy subjects volunteered to participate in this
study. A detailed summary of subjects physical characteris-

Appl. Physiol. Nutr. Metab. Vol. 35, 2010

tics is shown in Table 1. Each group was composed of


7 males and 3 females. TR subjects were licensed cyclists
who routinely trained at least 2 h a day, 47 days a week,
at the time of the study and during the previous 4 years.
UNTR subjects were university students who regularly
performed <2 h of physical activity per week during the previous year. All subjects were fully informed of any risks and
discomforts associated with the experiment before giving
their informed written consent to participate. Once the participants fulfilled the inclusion criteria and agreed to participate in the study, they underwent a physical examination
(including rest and exercise electrocardiogram) to ensure
that they were in good health. The study was approved by
the local Hospital Research Ethics Committee of the Hospital Virgon de la Salud, Spain, and conformed to the Declaration of Helsinki.
Preliminary testing
TR and UNTR subjects were tested using a continuous incremental test to volitional exhaustion to determine their
_ 2 peak. After a standardized 10-min warm-up, the initial
VO
power output was set at 75 W for UNTR and 125 W for
TR subjects, and it increased 25 Wmin1 until voluntary
cessation (or when pedaling cadence dropped below
_ 2) was measured using an
60 rmin1). Oxygen uptake (VO
automated breath-by-breath system (Quark b2, Cosmed,
Rome, Italy), and the data were averaged each 15 s. The
_ 2 value during the in_ 2 peak was defined as the highest VO
VO
cremental test (Balady et al. 2000). The test was carried out
1 week before the onset of the experimental trials; it was
performed on a cycle ergometer (Ergoline 200, Cosmed).
Subjects withdrew from all dietary sources of caffeine 48 h
_ 2 peak test. The body composition of the subbefore the VO
jects was assessed using skinfold measurements (Jackson
and Pollock 1978; Jackson et al. 1980).
Experimental design
On 3 occasions, each subject pedaled a cycle ergometer at
_ 2 peak in a hot, dry ventilated
40%, 60%, or 80% of their VO
environment (36 0.5 8C; 25% 5% relative humidity;
_ 2 peak test data, a VO
_ 22.5 ms1 wind speed). Using the VO
workload best-fit equation was generated for each subject to
determine individual workloads corresponding to the desired
_ 2 peak test was performed
_ 2 peak. However, since the VO
%VO
under temperate conditions (25 8C; 40% relative humidity),
exercise intensity for experimental protocols might be
slightly overestimated in both groups (Arngrmsson et al.
2003). To complete the same amount of work in each trial
(658 17 kJ for TR and 508 11 kJ for UNTR subjects;
p < 0.05), exercise duration was set at 107 4, 63 1, and
_ 2 peak, respectively
45 0 min for 40%, 60%, and 80% VO
(see Table 2 for details). Total amount of work was set to
_ 2 peak, since pilot data revealed that
equal 45 min at 80% VO
UNTR subjects could barely finish that trial in our hot environment. The exercise durations for the remaining trials (i.e.,
_ 2 peak) were calculated by dividing the total
40% and 60% VO
amount of work by the individual workload assigned for
these trials. Experimental trials were performed in a
randomized order, and were separated by at least 4 days to
ensure proper recovery and glycogen repletion. Data on carPublished by NRC Research Press

Del Coso et al.

743
Table 1. Morphological characteristics of endurance-trained cyclists (TR) and untrained healthy subjects (UNTR).
Subjects
TR
UNTR

Age (y)
22.96.6
24.14.4

Height
(cm)
17410
17613

Weight
(kg)
67.011.9*
76.815.5

Body mass index


(kgm2)
21.81.9*
24.52.6

Body fat
(%)
9.05.2*
18.46.4

VO2 peak
(mLkg1min1)
59.65.7*
43.63.2

Note: VO2 peak, peak oxygen uptake.


*Different from UNTR (p < 0.05).

Table 2. Power output, exercise times, and total work


during the 3 experimental trials at 40%, 60%, and
80% peak oxygen uptake for trained and untrained individuals.
Trial VO2 peak
(%)
TR
40
60
80
UNTR
40
60
80

Intensity
(W)

Time
(min)

1099*
17614*
24420*

1014
621
450

778
13210
18813

1125
641
450

Total work
(kJ)
65817*

50811

Note: VO2 peak, peak oxygen uptake.


*Different from UNTR (p < 0.05).

diovascular and thermoregulatory variables of this experimental design have been published elsewhere (MoraRodriguez et al. 2010).
Experimental protocol
Experimental trials were carried out from February to June
to avoid the possible effects of heat acclimation on substrate
utilization (Kirwan et al. 1987). Two days before the experimental trials, subjects withdrew from all dietary sources of
alcohol and caffeine. The day before, subjects were instructed to consume at least 7 gkg1 body weight of carbohydrate and to perform light exercise, if any. Food and training
diaries were collected to aid in the replication of diet and exercise before each trial. Each subject performed all trials at
the same time of day and in the postabsorptive state 4 h after
a standardized breakfast (~3 g carbohydratekg1 body
weight). Subjects also ingested 500 mL of tap water 2 h
before the onset of the experimental trials to ensure euhydration,
which was confirmed by urine specific gravity (pre-exercise
urine sample <1.020). On arrival, a 22-gauge Teflon catheter was inserted into an antecubital vein of each subject for
blood sampling. The catheter was frequently flushed with 3
to 4 mL of 0.9% sterile saline to ensure patency. Afterward, subjects entered the climatic chamber and sat quietly
on the cycle ergometer for 5 min while pre-exercise variables were recorded. After that, a venous blood sample was
withdrawn. Then, subjects pedaled in steady-state for the
duration and at the exercise intensity selected for the trial;
no fluid was provided during the trial.
Indirect calorimetry measurement and calculations
_
_ 2 and carbon dioxide production (VCO
VO
2) rates were
measured during the trials for 6-min periods, starting after

10 and 39 min, and at the end of trials. For these measurements, subjects wore a mouth piece and a noseclip while
samples of the expired air were collected breath by breath
to determine gas concentrations (Quark b2, Cosmed). The
volume of inspired and expired air was also measured with
a digital turbine flowmeter. Certified calibration gases
(16.0% O2; 5.1% CO2) and a 3-L syringe were used to calibrate the gas analyser and turbine flowmeter before each
trial. During the last 2 min of each collection period, the
ventilatory parameters were averaged to achieve a represen_ 2 and VCO
_
tative value for each period. From VO
2, energy
expenditure and substrate oxidation (fat and carbohydrate)
were calculated using the nonprotein respiratory quotient,
with the assumption that the urinary nitrogen excretion rate
was negligible (Brouwer 1957; Frayn 1983):
Energy expenditure rate (kJmin1) was calculated as
_ 2) + 91.195  (VCO
_
[(3.869  VO
2)]  4.186.
Fat oxidation rate (gmin1) was calculated as (1.67 
_
_ 2) (1.67  VCO
VO
2).
Carbohydrate oxidation rate (gmin1) was calculated as
_
_
_
_
(4.55  VCO
2) (3.21  VO2), where VO2 and VCO2 are
in Lmin1.
Total energy expenditure and total fat oxidation were calculated using the rate of energy expenditure or fat oxidation
multiplied by exercise duration.
Blood analysis
Venous blood samples (5 mL) were obtained before exercise and at the end of each trial. A portion of whole blood
was introduced in a clinical blood gas analyzer (ABL 520,
Radiometer, Copenhagen, Denmark) to determine H+ concentration in the plasma portion of the blood. The remaining
blood (i.e., 3 mL) was added to a tube containing 0.3 mL of
reduced glutathione (4.5 mg), sodium heparin (50 IU), and
20 mL of 0.24 molL1 EGTA. Tubes were then centrifuged
at 2000g to separate the plasma. At a later date, plasma epinephrine ([EPI]plasma) and norepinephrine ([NEPI]plasma) concentrations were measured using high performance liquid
chromatography with electrochemical detection (Hjemdahl
1984). Plasma catecholamine concentrations were not corrected for changes in plasma volume so that its real bioavailability in blood was presented. A limitation of this study
is that [EPI]plasma and [NEPI]plasma concentrations were only
measured at the end of each trial.
Statistics
Unpaired Students t tests were used to compare subjects
physical and morphological characteristics. One-way repeated-measures analysis of variance (ANOVA) was used
to compare the total energy expenditure and total carbohyPublished by NRC Research Press

744

drate and fat oxidations among trials within the same group
of subjects. Factorial ANOVA was used to compare energy
expenditure and substrate oxidation rates between TR and
UNTR subjects at the different exercise intensities. After a
significant F test (GeisserGreenhouse correction for the assumption of sphericity), differences between means were
identified using Tukeys HSD post hoc procedure. The significance level was set at p < 0.05. All data are presented
as means standard error of the measurement.

Appl. Physiol. Nutr. Metab. Vol. 35, 2010


Fig. 1. Average fat oxidation rate in endurance-trained (TR) and
untrained (UNTR) individuals at 40%, 60%, and 80% peak oxygen
_ 2 peak). Data are means standard error of measurement.
uptake (VO
_ 2 peak,
*, Different from UNTR, p < 0.05; {, different from 60% VO
_
p < 0.05; {, different from 40% VO2 peak, p < 0.05.

Results
Energy expenditure
As expected, the rate of energy expenditure increased
along with the exercise intensity (i.e., 40% < 60% < 80%
_ 2 peak) in both TR (39 3 < 55 4 < 76 5 kJmin1;
VO
p < 0.05) and UNTR (33 3 < 46 3 < 60 4 kJmin1;
p < 0.05) subjects. Total energy expenditure was higher
_ 2 peak for a long time (107
when exercising at 40% VO
4 min) than when the same amount of work was performed
_ 2 peak in TR (3946 250 > 3445
at 60% and 80% VO
226 & 3431 241 kJ; respectively; p < 0.05) and UNTR
(3458 223 > 2955 170 > 2717 172 kJ; p < 0.05) subjects. Total energy expenditure (kJ) and rate (kJmin1) were
higher for TR than UNTR subjects at all the relative exercise intensities tested (p < 0.05).
Fat oxidation
Figure 1 depicts the fat oxidation rate during each trial in
TR and UNTR subjects. While TR subjects achieved Fatmax
_ 2 peak (0.41 0.01 gmin1), UNTR subjects
at 60% VO
_ 2 peak (0.28 0.01 gmin1). The
achieved Fatmax at 40% VO
rate of fat oxidation was higher for TR than for UNTR subjects, although it only reached statistical difference at 60%
_ 2 peak (p < 0.05; Fig. 1). The total fat oxidation
and 80% VO
for the set amount of work was higher at 40% than at 60%
_ 2 peak for TR (36 5 > 26 4 > 11 2 g;
and at 80% VO
p < 0.05) and UNTR (29 3 > 14 3 > 2 1 g; p < 0.05)
subjects; it was higher for TR than UNTR subjects only at
_ 2 peak (p < 0.05).
60% and 80% VO

Fig. 2. Relative fat (FAT) and carbohydrate (CHO) oxidation for


endurance-trained and untrained individuals at 40%, 60%, and 80%
_ 2 peak. Data are means standard error of measurement. *, DifVO
_ 2 peak, p <
ferent from UNTR, p < 0.05; {, different from 60% VO
_ 2 peak, p < 0.05.
0.05; {, different from 40% VO

Carbohydrate oxidation
The rate of carbohydrate oxidation increased along with
_ 2 peak) in
the exercise intensity (i.e., 40% < 60% < 80% VO
both TR (1.6 0.2 < 2.5 0.2 < 4.2 0.3 gmin1; p <
0.05) and UNTR (1.4 0.2 < 2.4 0.1 < 3.6 0.3 gmin1;
p < 0.05) subjects. Despite the differences in the energy expenditure rates, the differences in the carbohydrate oxidation
rates between TR and UNTR subjects did not reach statistical significance.

_ 2 peak (28% 4% vs. 17% 3%, p < 0.05) and 80%


VO
_ 2 peak (11% 2% vs. 4% 1%, p < 0.05), but not at
VO
_ 2 peak (35% 5% & 33% 4%).
40% VO

Substrate contribution to total energy expenditure


Figure 2 depicts the relative contribution of carbohydrate
and fat to total energy expenditure in TR and UNTR subjects. Carbohydrate was the predominant substrate oxidized
during exercise in the heat at all exercise intensities for
both groups of subjects. Furthermore, the relative contribution of carbohydrate to total energy expenditure increased
with exercise intensity (p < 0.05). However, TR individuals
oxidized relatively more fat than UNTR subjects at 60%

Blood responses and respiratory exchange ratio


From similar resting values (7.42 0.01 for TR and
7.41 0.01 for UNTR), blood pH was well maintained during exercise at all exercise intensities in both TR (7.44
0.01, 7.43 0.01, and 7.41 0.01 at 40%, 60%, and 80%
_ 2 peak, respectively) and UNTR (7.44 0.01, 7.44 0.01,
VO
_ 2 peak,
and 7.41 0.01 for UNTR, at 40%, 60%, and 80% VO
respectively) subjects. At the end of exercise, [EPI]plasma concentrations increased along with the increases in exercise inPublished by NRC Research Press

Del Coso et al.

_ 2 peak), but it was similar


tensity (i.e., 40% < 60% < 80% VO
between groups (Fig. 3A). [NEPI]plasma concentrations also
increased with exercise intensity, but it was higher in TR
than in UNTR subjects at the end of the 60% and 80%
_ 2 peak trials (Fig. 3B; p < 0.05). Finally, respiratory exVO
change ratios were 0.89 0.01 vs. 0.90 0.01, 0.91 0.01
vs. 0.94 0.01 (p < 0.05), and 0.96 0.01 vs. 0.99 0.01
(p < 0.05) for TR vs. UNTR subjects at 40%, 60%, and 80%
_ 2 peak, respectively.
VO

745
Fig. 3. (A) Plasma epinephrine and (B) plasma norepinephrine concentrations in trained and untrained individuals at the end of exer_ 2 peak. Data are means standard
cise at 40%, 60%, and 80% VO
error of measurement. *, Different from UNTR, p < 0.05; {, differ_ 2 peak, p < 0.05; {, different from 40% VO
_ 2 peak,
ent from 60% VO
p < 0.05.

Discussion
Several studies have been geared to elucidate the exercise
intensity that elicits Fatmax in a thermoneutral environment
(Romijn et al. 1993, 2000; Bergman and Brooks 1999;
Achten et al. 2002; Venables et al. 2005; Nordby et al.
2006; Stisen et al. 2006; Riddell et al. 2008). The practical
finding is that the exercise intensity that produces Fatmax
_ 2 peak. This ample range of
may range from 33% to 65% VO
exercise intensities for Fatmax can be explained by the different nature of the subjects participating in the cited studies
(i.e., different aerobic capacities). When comparing data
from Achten et al. (2002), who used TR subjects, and Venables and colleagues (2005), who used UNTR subjects, TR
subjects obtained Fatmax at a higher relative exercise inten_ 2 max). Similarly,
sity than UNTR subjects (64% vs. 48% VO
Nordby et al. (2006) found that Fatmax was attained at a
higher relative exercise intensity in TR than in UNTR sub_ 2 max). Likewise, in a thermoneutral
jects (50% vs. 43% VO
environment, we found in this study that Fatmax in the heat
occurred at a higher relative intensity in TR than in UNTR
_ 2 peak). In addition, during exersubjects (60% vs. 40% VO
cise in the heat, the curve depicting fat oxidation rate as a
function of relative exercise workload was displaced upward
and rightward in TR, compared with UNTR, individuals
(Fig. 1).
Exercise in the heat (40 8C) increases muscle glycogen
oxidation and reduces whole-body fat oxidation (Febbraio et
al. 1994), in comparison to the same exercise intensity performed at 20 8C. In agreement with data from Febbraio et
al. (1994), Fatmax levels found in this study (0.41 and
0.28 gmin1 for TR and UNTR, respectively) were substantially lower than those found in temperate environments. For
example, Achten and coworkers (2002) found that TR subjects oxidized fat at a rate as high as ~0.6 gmin1, whereas
Venables and colleagues (2005) reported fat oxidation rates
as high as ~0.4 gmin1 in UNTR subjects. In addition, fat
_ 2 peak represented 35% 5% for TR
oxidation at 40% VO
and 33% 4% for UNTR subjects of total energy expenditure (Fig. 2), whereas Bergman and Brooks (1999) reported
a contribution of 40% of total energy derived from fat when
_ 2 peak in a thermoneutral environment.
exercising at 40% VO
It is then possible that our hot environmental conditions induced to the reported lower rates of fat oxidation, in comparison to those studies.
To increase the applicability of our study, subjects ingested a standardized breakfast prior to each trial, containing
3 g of carbohydrate per kilogram of body weight. The ingestion of a similar breakfast has been shown to decrease exercise fat oxidation rates in both trained and untrained subjects
(Bergman and Brooks 1999). However, in the Bergman and

Brooks (1999) study, the ingestion of pre-exercise carbohydrates did not affect the exercise intensity that produced
Fatmax. Thus, the lower Fatmax found in our study, in comparison to other studies, might be due to a combination of
the high heat load and the effect of the pre-exercise ingestion of carbohydrates. A limitation of our study is that we
cannot distinguish between the effects of heat load and those
of carbohydrate ingestion in reducing the fat oxidation rate.
Some studies have used a single progressive exercise test
to determine the exercise intensity that elicits Fatmax (Achten
et al. 2002; Venables et al. 2005; Stisen et al. 2006). A different approach is to perform bouts of exercise at diverse intensities on different days, and then to compare the fat
oxidation rate obtained at each exercise intensity (Romijn et
al. 1993; Bergman and Brooks 1999; Meyer et al. 2007). For
our study, we chose the latter approach (3 trials separated by
4 days) to avoid the influence of progressive fatigue and
substrate depletion on fat oxidation during a progressive exercise test. Besides, this approach allowed us to calculate total energy expenditure and total fat oxidation required to
complete the same amount of work despite exercising at difPublished by NRC Research Press

746

ferent intensities. Interestingly, both groups of subjects expended more energy and oxidized significantly more fat
_ 2 peak) for a
when exercising at light intensity (i.e., 40% VO
long time (107 min), despite their differences in the relative
exercise intensity for Fatmax (Fig. 1). In the case of TR sub_ 2 peak (0.41 gmin1),
jects, they achieved Fatmax at 60% VO
but they needed only 62 min to complete the assigned work
at this intensity. In contrast, the fat oxidation rate at 40%
_ 2 peak was high (0.36 gmin1), but subjects were required
VO
to exercise for more than 100 min to complete the trial. This
information may be relevant for trained and healthy untrained subjects aiming to maximize energy cost and fat oxidation during training. Our data suggest that prolonged
exercise at a low intensity is the best choice to increase total energy expenditure and fat oxidation during exercise in the heat.
Figure 2 depicts the relative contribution of fat and carbohydrate oxidations to the total energy expended in each trial.
During exercise in the heat, TR subjects relied more on fat
oxidation than their UNTR counterparts when exercising at
_ 2 peak; while there were no differences be60% and 80% VO
_ 2 peak. Our data are in agreement
tween groups at 40% VO
with previous authors, who found a higher relative contribution of fat oxidation to the energy expenditure at moderate
_ 2 peak (Stisen et al. 2006)) and high intensities
(45%60% VO
_ 2 peak (Coggan et al. 2000)) in TR than in
(75%80% VO
UNTR subjects when exercising in thermoneutral environments. In the case of light exercise intensities (20%40%
_ 2 peak), Bergman and Brooks (1999) found a higher conVO
tribution of fat oxidation in TR subjects, whereas Stisen and
coworkers (2006) failed to find differences between groups,
as we did in this study. Our data confirm that TR subjects
had a superior reliance on fat as fuel during exercise in the
heat, compared with UNTR subjects, at least at moderate to
high intensities. In addition, we speculate that TR subjects
did not oxidize significantly more fat than UNTR subjects
_ 2 peak, because UNTR subjects achieved their
at 40% VO
Fatmax at this exercise intensity, whereas TR subjects did so
_ 2 peak.
when exercising at 60% VO
The mechanisms for a higher fat oxidation in TR than
UNTR subjects may be related to different metabolic adaptations derived from aerobic training. During exercise,
[EPI]plasma increases lipolysis (Mora-Rodriguez et al. 2001),
although high levels of [EPI]plasma induce a higher rate of
glycogen utilization, which in turn decreases fatty acid oxidation during exercise (Mora-Rodriguez and Coyle 2000). In
our study, [EPI]plasma concentrations rose with the exercise
intensity, but the increase was similar in TR and UNTR subjects (Fig. 3A). On the other hand, [NEPI]plasma concentrations were higher in TR than UNTR subjects at 60% and
_ 2 peak (Fig. 3B), in agreement with data from a train80% VO
ing study (Greiwe et al. 1999) and during exercise in an uncompensable hot environment (Wright et al. 2010).
Interestingly, these precise exercise intensities (60% and
_ 2 peak) were the ones at which TR subjects had
80% VO
higher absolute and relative fat oxidation than UNTR subjects (Fig. 2). It has been found that a higher [NEPI]plasma
concentration increases the rate of free fatty acid release
from adipose tissue (Hodgetts et al. 1991), thus increasing
the availability of free fatty acids and, potentially, fat oxidation during exercise.

Appl. Physiol. Nutr. Metab. Vol. 35, 2010

Muscle pH reductions inhibit the transport of free fatty


acids into the mitochondria by inhibiting CPT1 activity
(Starritt et al. 2000). In our study, blood pH (a surrogate of
muscle acidity (Juel et al. 2004)) was maintained to a similar degree between groups at the end of exercise. On the
other hand, b-oxidative muscle enzyme activity (e.g., b-hydroxyacyl-CoA-dehydrogenase activity) is higher in TR
than in UNTR subjects (Nordby et al. 2006; Stisen et al.
2006), which in turn facilitates higher fat oxidation at moderate and high exercise intensities in the TR population
(Stisen et al. 2006). Thus, the higher [NEPI]plasma concentration and increased b-oxidative muscle enzyme activity induced by aerobic training may be responsible, in part, for
the higher fat oxidation in TR than in UNTR subjects in
our study.
Since, for a given relative exercise intensity, TR subjects
exercised at a higher absolute workload than UNTR subjects, their sweat rate during exercise was higher (table 3, in
Mora-Rodriguez et al. 2010), which likely allowed them to
remain at core temperatures similar to their UNTR counter_ 2 peak). The increased sweat rate
parts (at least up to 80% VO
induced higher dehydration in TR than in UNTR subjects
(p < 0.05). It could then be argued that the larger dehydration in the TR group may have influenced metabolism
through an effect on muscle glycogen utilization (Hargreaves et al. 1996). Although the differences in dehydration
between TR and UNTR subjects were statistically significant, the metabolic effect of the small absolute differences
(~0.6% dehydration) is dubious. In addition, it has been
found that a reduction in body-water content does not affect
whole-body fat oxidation during exercise (Roy et al. 2000).
One limitation of this study was the utilization of indirect
calorimetry to determine fat and carbohydrate oxidation dur_ 2 reliably reflects tissue O2 uptake,
ing exercise. While VO
_
VCO2 is a good estimation of tissue CO2 production up to
moderate exercise intensities, where H+ production is minimal and the bicarbonate pool is maintained stable (Del
Coso et al. 2009). At high exercise intensities, the increase
in H+ production is buffered by bicarbonate, producing nonmetabolic CO2, which in turn will produce an overestimation of carbohydrate oxidation and an underestimation of fat
oxidation. In our study, the acidbase balance was well
maintained in both groups at all exercise intensities, and respiratory exchange ratio was always maintained at less than
1.0, indicating minimal production of nonmetabolic CO2. In
addition, it has been reported that indirect calorimetry accurately determines substrate oxidation (compared with a 13C_ 2 max,
to-12C ratio technique) during exercise up to 85% VO
at least in trained subjects (Romijn et al. 1992). For all these
reasons, we consider that the substrate oxidation rates calculated in this study were minimally affected by nonmetabolic
CO2 production.

Conclusion
When exercising in hot environments, whole-body peak
fat oxidation occurs at higher relative exercise intensity in
aerobically trained individuals than untrained counterparts
_ 2 peak). In addition, trained individuals oxi(60% vs. 40% VO
dize more fat than their untrained counterparts when exercis_ 2 peak). This
ing at moderate to high intensities (>60% VO
Published by NRC Research Press

Del Coso et al.

elevated reliance on fat as a fuel during exercise in the heat


could be related to the higher plasma norepinephrine concentration observed in the aerobically trained individuals.

Acknowledgements
The authors thank the subjects for their invaluable contribution to the study. Juan Del Coso and Nassim Hamouti
were supported by a predoctoral fellowship from the
CastillaLa Mancha Government in Spain. Juan F. Ortega
received funds from the Gatorade Sports Science Institute.

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