Acta Physiol Scand 2001, 172, 3952

Brief intense exercise followed by passive recovery

modies the pattern of fuel use in humans during
subsequent sustained intermittent exercise

M . A . C H R I S T M A S S , 1 , 2 B . D A W S O N , 2 C . G O O D M A N 2 and P . G . A R T H U R 1
1 Department of Biochemistry, The University of Western Australia, Nedlands, Australia
2 Department of Human Movement and Exercise Science, The University of Western Australia, Nedlands, Australia

ABSTRACT
The role of work period duration as the principal factor influencing carbohydrate metabolism during
intermittent exercise has been investigated. Fuel oxidation rates and muscle glycogen and free
carnitine content were compared between two protocols of sustained intermittent intense exercise
with identical treadmill speed and total work duration. In the first experiment subjects (n 6)
completed 40 min of intermittent treadmill running involving a work : recovery cycle of 6 : 9 s or
24 : 36 s on separate days. With 24 : 36 s exercise a higher rate of carbohydrate oxidation
approached signicance (P 0.057), whilst fat oxidation rate was lower (P 0.01) and plasma lactate
concentration higher (P 0.01). Muscle glycogen was lower post-exercise with 24 : 36 s (P 0.05).
Muscle free carnitine decreased (P 0.05), but there was no difference between protocols. In the
second experiment a separate group of subjects (n 5) repeated the intermittent exercise protocols
with the addition of a 10-min bout of intense exercise, followed by 43 5 min passive recovery, prior
to sustained (40 min) intermittent exercise. For this experiment the difference in fuel use observed
previously between 6 : 9 s and 24 : 36 s was abolished. Carbohydrate and fat oxidation, plasma
lactate and muscle glycogen levels were similar in 6 : 9 s and 24 : 36 s. When compared with the
rst experiment, this result was because of reduced carbohydrate oxidation in 24 : 36 s (P 0.05).
There was no difference, and no change, in muscle free carnitine between protocols. A 10-min bout
of intense exercise, followed by 43 5 min of passive recovery, substantially modies fuel use
during subsequent intermittent intense exercise.

Keywords carbohydrate, carnitine, fat, intermittent exercise, metabolism, treadmill running.

Received 19 October 1999, accepted 19 December 2000

The concept that availability of fat regulates carbo-
hydrate metabolism (i.e. glucosefatty acid cycle) has
been the predominant theory of fuel selection in
skeletal muscle for many years (Randle et al. 1963).
However, results from several studies indicate that this
mechanism may have limited application in human
skeletal muscle during exercise (Sahlin 1990, Dyck
et al. 1993, Putman et al. 1993). Instead, under these
conditions the availability of carbohydrate may regu-
late fat oxidation in reverse of the glucosefatty acid
cycle (Sidossis et al. 1997). This proposal is supported
by recent ndings suggesting that increased carbohy-
drate metabolism (i.e. glycolytic ux) could directly
inhibit fat oxidation in human skeletal muscle by
limiting transport of long chain fatty acids (LCFA)
into mitochondria (Coyle et al. 1997, Sidossis et al.
1997).
During continuous exercise, glycolytic ux increases
as the work level increases from moderate- to high-
intensity and fat oxidation declines (Christensen &
Hansen 1939, Romijn et al. 1993). This outcome could
be a result of both limited availability of plasma non-
esteried fatty acid (NEFA) (Romijn et al. 1995) and
reduced transport of NEFA into mitochondria (Sidossis
et al. 1997). Mitochondrial LCFA transfer is dependent
on the carnitine palmitoyl-transferase I reaction for
which free carnitine is essential (Bremer 1983). At high
metabolic rates free carnitine also functions as an acetyl
group buffer (Alkonyi et al. 1975) preventing excess
accumulation of acetyl CoA (Foster & Harris 1987).
Depletion of skeletal muscle free carnitine levels, as a
consequence of rapid acetylation, represents a potential
mechanism limiting mitochondrial LCFA transport and
oxidation in the presence of accelerated glycolytic ux

Correspondence: M.A. Christmass, Department of Biochemistry, The University of Western Australia, Nedlands, 6907, Australia.

2001 Scandinavian Physiological Society 39

Fuel use in intermittent exercise
M A Christmass et al. Acta Physiol Scand 2001, 172, 3952

(Foster & Harris 1987). During continuous exercise indirect calorimetry to estimate whole body fuel oxi-
carbohydrate metabolism may, therefore, regulate fat dation rates during intermittent intense exercise. A
oxidation in accordance with the concept of the glucose second experiment repeated this comparison with the
fatty acid cycle reversed (Coyle et al. 1997, Sidossis et al. inclusion of a 10-min bout of intense exercise (i.e. brief
1997). Increased rates of carbohydrate metabolism with intense exercise), followed by approximately 45 min of
increases in exercise intensity during continuous exercise passive recovery (i.e. passive recovery) prior to LE and
are probably to be a consequence of several factors SE. On the basis of results from an earlier study
including increased cytosolic calcium release, catechol- (Christmass et al. 1999b) we proposed that increased
amine levels, phosphocreatine depletion and recruitment work period duration, resulting in lower muscle O2
of type II muscle bres as well as reduced mitochondrial availability, may be responsible for accelerated carbo-
O2 availability (Chastiosis 1983, Katz & Sahlin 1990). hydrate metabolism and hence, in turn, the inhibition of
Intermittent exercise consists of repeated periods of fat oxidation during intermittent intense exercise
intense work followed by lower intensity recovery. (Christmass et al. 1999b). The current study shows that
During sustained intermittent exercise, glycolytic ux any effect of muscle O2 availability on carbohydrate
increases with proportionate increases in work and metabolism, as a result of work period duration, is
recovery period duration and fat oxidation declines abolished when sustained intermittent exercise is
(Christmass et al. 1999b). Therefore, carbohydrate preceded by brief intense exercise and passive recovery.
metabolism may regulate fat oxidation during sustained
intermittent exercise according to the mechanisms
M AT E R I A L S A N D M E T H O D S
proposed for continuous exercise (i.e. glucosefatty
acid cycle reversed; Coyle et al. 1997, Sidossis et al. Subjects
1997). In this context, the factors inuencing glycolytic
The study comprised two experiments: (1) an initial
ux during sustained intermittent exercise are likely to
experiment compared fuel oxidation and capillary
be important for regulation of the carbohydratefat
plasma metabolite, blood gas and muscle metabolite
interaction under these conditions. Unlike continuous
concentrations during LE and SE with no prior exer-
exercise, glycolytic ux increased with increases in work
cise (fuel use during sustained intermittent intense
and recovery period duration although exercise inten-
exercise without prior brief intense exercise and passive
sity, represented as a proportion of peak aerobic power
recovery, Experiment 1). This experiment was repeated
(V_ O2peak), during the work period (i.e. work period
with the addition of brief intense exercise, followed by
intensity) was constant (Christmass et al. 1999b).
passive recovery, prior to SE and LE to examine work
During intermittent exercise more rapid rates of
period duration as the principal factor inuencing
glycolytic ux, at the same work period intensity, have
carbohydrate metabolism during intermittent exercise
been associated with proportionately longer work and
with the same work period intensity (effect of prior
recovery periods for almost 40 years (Astrand et al.
brief intense exercise and passive recovery on fuel use
1960b, Saltin & Essen 1971). With constant work period
during sustained intermittent intense exercise, Experi-
intensity, the increase in glycolytic ux could be a result
ment 2).
of reduced O2 availability because of depletion of
Separate groups of subjects were used for each
myoglobin stores as work duration increases (Astrand
experiment (Experiment 1, n 6; Experiment 2, n 5).
et al. 1960b, Saltin & Essen 1971). This is supported by
The (mean SE) age, height, body mass and V_ O2 for
the nding that the discrepancy between O2 supply and
the subjects were: 21 1 years; 184 1 cm;
demand increases with longer work period duration in
78.3 3.7 kg; 58.0 1.8 mL kg1 min1 (Experiment
intermittent exercise (Christmass et al. 1999b).
1) and 23 1 years; 180 2 cm; 77.4 3.9 kg;
The aim of the current experiment was to examine
57.5 1.9 mL kg1 min1 (Experiment 2). Subjects
the role of work period duration and, in turn, muscle
were healthy males participating in interval type running
O2 availability, as central factors affecting the rate of
exercise on a regular basis and provided informed
carbohydrate metabolism during intermittent exercise
consent for the studies. The experimental procedures
with the same work period intensity. For this purpose
were approved by the Human Rights Committee of The
an initial experiment replicated our earlier study
University of Western Australia in accordance with the
(Christmass et al. 1999b) to include a comparison of
Declaration of Helsinki.
skeletal muscle metabolism between intermittent exer-
For both experiments, the subjects' standing height
cise involving 24 s work : 36 s recovery (i.e. long
(0.1 cm; Holtain Stadiometer), body mass (0.05 kg;
interval exercise, LE) and 6 s work : 9 s recovery (i.e.
Sauter Multirange Electronic scales) and V_ O2peak were
short interval exercise, SE). A secondary aim was to
measured during a separate laboratory session as
examine changes in glycogen levels in muscle samples
described previously (Christmass et al. 1999a).
taken from the exercising leg as support for the use of

40 2001 Scandinavian Physiological Society

Acta Physiol Scand 2001, 172, 3952
Experimental protocols
Fuel use during sustained intermittent intense exercise without
prior brief intense exercise and passive recovery: Experiment 1.
This experiment involved two separate experimental
laboratory sessions. During each experimental session
subjects completed a warm-up consisting of 10 min
continuous running (10 km h1) on a motorized
treadmill and stretched for approximately 20 min.
Immediately following this warm-up and stretch
routine, subjects completed intermittent running
protocols (40 min) on a motorized treadmill requiring a
cycle of work : recovery of LE or SE (Fig. 1). The
alternate schedule of work and recovery duration was
completed during the second experimental session. The
two experimental sessions (i.e. one for LE and one for
SE) were separated by 7 days and the order of
completion was randomized and balanced. Treadmill
speed was determined for each subject on a separate
day during a laboratory familiarization session. This
procedure involved 510 min of intermittent running
(LE) during which treadmill speed was adjusted to elicit
a respiratory exchange ratio (R) between 0.960.99 as
determined by a computerized gas analysis system
(Christmass et al. 1999a). Treadmill speed for both LE
and SE was identical and was not changed during the
40 min exercise protocols (range, 19.021.5 km h1).
On arrival at the laboratory subjects were prepared
for needle biopsy of the vastus lateralis with two inci-
sions of the skin (approximately 1 cm length and 10 cm
apart) on one thigh to the level of the deep fascia, under
local anaesthesia (2.5 mL, 1% Xylocaine). A resting
muscle biopsy sample was removed from one incision
and both incisions were closed with Steristrips (3M
Health Care, St Paul, USA). A second muscle biopsy
was obtained from the alternate incision immediately
post-exercise. Biopsy samples were obtained with the
Figure 1 Experimental protocol.
Separate subject groups completed
intermittent exercise (40 min) on two
occasions with a work : recovery cycle
of either 24 : 36 s (LE) or 6 : 9 s (SE).
One subject group completed the
intermittent exercise protocols without
(a) and one group with (b), a 10 min
bout of intense exercise, followed by
approximately 45 min of passive
recovery prior to intermittent exercise.
Capillary blood and muscle biopsy
samples and respiratory gas exchange
were collected at the times indicated
with a downward arrow. Warm-up and
stretch (j); LE ( ); passive recovery
( ); intermittent exercise ( ).
2001 Scandinavian Physiological Society
M A Christmass et al.
Fuel use in intermittent exercise
subject supine on an examination couch. For determi-
nation of plasma metabolite concentration and for
measurement of blood gases and haematocrit, capillary
blood was collected at rest (i.e. immediately following
the muscle biopsy) and at approximately 10-min inter-
vals during exercise (i.e. 10, 21, 32 min). Respiratory gas
exchange was measured at 10 min intervals (i.e. 16, 27,
38 min) during exercise. Subjects breathed through the
spirometry system for 5 min at each sampling interval
and expired gases were collected for analysis across the
nal 1 min for each subject the order and timing of
procedures during the two experimental sessions were
identical.
Effect of prior brief intense exercise and passive recovery on
fuel use during sustained intermittent intense exercise: Experi-
ment 2. This experiment was identical with the excep-
tion that brief intense exercise, followed by passive
recovery, preceded the 40-min intermittent exercise
protocols. Brief intense exercise consisted of the
procedure for treadmill speed determination as
described for Experiment 1 with the exception that all
subjects completed 10 min of LE. Thus, during two
separate experimental laboratory sessions subjects
completed a warm-up consisting of 10 min of
continuous running (10 km h1) on a motorized
treadmill and stretched for approximately 20 min.
Immediately following this warm-up and stretch
routine, subjects completed a 10-min bout of LE
(brief intense exercise), followed by 43 5 min
supine rest (passive recovery). At the completion of
the passive recovery period, subjects performed a
second warm-up and stretch routine (15 2 min).
Immediately following this warm-up, subjects
completed 40 min of LE or SE as described for
Experiment 1 (Fig. 1). The two experimental sessions
involving each exercise protocol were separated by a
41
Fuel use in intermittent exercise
M A Christmass et al.
minimum of 14 days (range 1416 days) and the order
of completion of the two sessions was randomized
and balanced. Treadmill speed for both LE and SE
was identical and was not changed during the 40 min
exercise protocols (range 19.522.7 km h1).
Pre-exercise muscle biopsy samples were obtained
in the nal 10 min of the passive recovery period
(Fig. 1) and immediately post-exercise in the manner
described for Experiment 1. Capillary blood was
collected immediately following the pre-exercise
muscle biopsy (i.e. pre-exercise) and at approximately
10, 21, 32 min. All other measurements and proce-
dures were performed as described for Experiment 1
(Fig. 1).
Nutrient intake
For both experiments, subjects were requested to
maintain a similar dietary and exercise regimen in the
3 days prior to each experimental laboratory session.
All subjects completed a 3-day dietary record to assist
compliance with this request. Subjects reported to the
laboratory in the post-absorptive state for all experi-
mental sessions.
Respiratory gas exchange and indirect calorimetry
Respiratory gas exchange measurements were
performed according to methods previously described
(Christmass et al. 1999a, b). Briey, ventilation (V_ E)
was determined by collection of expired air into a
350 L Tissot spirometer (W.E. Collins Inc., USA). A
sample (2 L) of this gas was removed from the
spirometer for measurement of FECO2 and FEO2
using CO2 (CD3A analyser, Ametek, Paoli, PA, USA)
and O2 (S3A/1 analyser, Ametek) analysers. Aerobic
energy expenditure was calculated using caloric
equivalents of the non-protein respiratory quotient
(Lusk 1928) and measurements of V_ O2 and V_ CO2.
Rates of carbohydrate and fat oxidation were calcu-
lated from measurements of V_ O2 and V_ CO2
according to the stoichiometric equations and
assumptions outlined by Frayn (1983).
Blood sample collection and treatment
For determination of plasma metabolite concentra-
tions, capillary blood (60 lL) was collected from
the ear lobe and treated according to methods
previously described (Christmass et al. 1999a). For
lactate and glycerol determination, the plasma
obtained was immediately frozen in liquid nitrogen
and subsequently stored at 80 C. Plasma for
NEFA determination was acid denatured immediately
(Christmass et al. 1998), immersed in liquid N2 and
42
Acta Physiol Scand 2001, 172, 3952
subsequently stored at )80 C. Capillary whole blood
from the ear lobe was collected rapidly (<30 s) into
100 lL blood collection tubes (Chiron Diagnostics,
Medeld, MA, USA) and analysed for blood gases
according to methods outlined previously (Christmass
et al. 1999a). Haematocrit was measured in duplicate
using a microhaematocrit reader (Hawksley, Sussex,
UK).
Muscle sample collection and treatment
The percutaneous needle biopsy technique (Bergstrom
1962), with suction applied manually, was used to
obtain each muscle sample. Samples were frozen in
liquid N2 and removed from the biopsy needle under
liquid N2. The time between the end of exercise (i.e. the
nal work period) and needle penetration was 25
1.6 s (Experiment 1) and 33 2.9 s (Experiment 2),
and between needle penetration and immersion of the
needle/sample in liquid N2 was 16 1.9 s (Experi-
ment 1) and 21 3.2 s (Experiment 2). For both
experiments there was no difference between LE and
SE for the total time between the end of exercise and
immersion of the needle/sample in liquid N2. Samples
were freeze dried (40 C), dissected essentially free of
blood and connective tissue and stored dry at 80 C
for analysis.
Plasma metabolite analysis
Untreated plasma for lactate and glycerol determina-
tion was deproteinized and analysed according to
methods previously described (Christmass et al.
1999a). Briey, untreated plasma was deproteinized
with cooled (04 C) perchloric acid and neutralized
with a potassium hydroxide (1.2 M)/phosphate buffer
(0.32 M, pH 7) solution according to a modication of
the method of Arthur et al. (1989). Enzymatic assays
for lactate (Passonneau & Lowry 1993) and glycerol
(Bergmeyer et al. 1984) were modied by way of a
coupled bioluminescent assay (Arthur et al. 1989) for
small sample volumes (5 lL). In addition, the coupled
luminometric assays were modied according to
Thompson et al. (1997) for rapid sample throughput
using a Dynatech ML 2250 Microtiter Plate Lumi-
nometer (Dynatech Laboratories, Chantilly, VA,
USA). Acid denatured plasma for NEFA determina-
tion was analysed according to Christmass et al.
(1998). All samples were measured in triplicate and
were remeasured if two samples in a triplicate differed
by more than 10%. For each method interassay
variability was less than 10%. Recovery of known
concentrations of appropriate standards added to
plasma were 98 2% (lactate), 99 2% (glycerol)
and 100 5% (NEFA).
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Acta Physiol Scand 2001, 172, 3952 M A Christmass et al.
Fuel use in intermittent exercise

Paired t-tests were used to determine differences

Muscle metabolite analysis
A portion of the dried muscle sample was prepared for
determination of glycogen concentration according to
the method of Palmer et al. (1983). The glycogen
content was determined using an enzymatic assay for
glucose (Bergmeyer et al. 1984) modied in the manner
described for plasma lactate and glycerol determination
(Arthur et al. 1989, Thompson et al. 1997). The
remaining dried muscle was extracted in a solution of
0.6 M perchloric acid and 1 mM EDTA and neutralized
with 2.2 M KOH and 620 mM TEA. Lactate concen-
tration was determined as for plasma according to
Passonneau & Lowry (1993). Muscle free carnitine
concentration was determined using a microtiter plate
reader (SpectraMAX, Molecular Devices, Sunnyvale,
CA, USA) according to a modication (Foster & Harris
1987) of the method of Marquis & Fritz (1964).
Samples for lactate and glucose were measured in
triplicate and for carnitine in duplicate. All samples
were remeasured if two determinations differed by
more than 10%. For each method interassay variability
was less than 10%. Recovery of known concentrations
of appropriate standards added to deproteinized muscle
were 100 4% (glycogen), 107 4% (lactate) and
97 1% (carnitine).
Statistical analysis
For comparisons between LE and SE within the two
experiments respiratory gas exchange, plasma metabo-
lite and blood gas data were analysed using a two-way
analysis of variance with repeated measures for
comparisons across time. When a signicant interaction
between time and exercise protocol was revealed, the
Bonferroni post hoc test was used to compare means.
between mean for muscle metabolite concentrations.
For comparisons between Experiment 1 and Experi-
ment 2 respiratory gas exchange and plasma metabolite
data were analysed using a three-way analysis of vari-
ance for factors Time, Exercise Protocol (LE or SE)
and Experiment (1 and 2). Data analysis and statistical
calculations were performed using DATA DESK
(ANOVA; Data Description, Ithaca, NY, USA) or
STATVIEW SE (Paired t-test; Abacus Concepts,
Berkeley, CA, USA, 1989). Results were considered
signicant at P 0.05 and are presented as mean SE
unless stated otherwise.
RESULTS
Fuel use during sustained intermittent intense exercise
without prior brief intense exercise and passive recovery:
Experiment 1
Respiratory gas exchange and indirect calorimetry. From
16 min to 38 min of exercise, overall (i.e. work and
recovery) V_ E, V_ O2, V_ CO2, (Fig. 2), R and aerobic
energy expenditure remained constant (Fig. 3). Overall
V_ O2 (38.5 0.8 mL kg1 min1, LE; 42.2 0.9 mL
kg1 min1 SE; P 0.01; Fig. 2) and aerobic energy
expenditure (0.77 0.01 kJ min1 kg1, LE; 0.84
0.02 kJmin1.kg1, SE; P 0.05; Fig. 3) were both
lower in LE. Overall (i.e. work and recovery) exercise
intensity was 66.9 4.0% V_ O2peak during LE and
73.2 4.2% V_ O2peak during SE.
Although overall exercise intensity was lower in LE
compared with SE, the R was higher (0.96 0.01, LE;
0.91 0.01, SE; P 0.01). There was no difference
between LE and SE forV_ E (Fig. 2). A higher rate of
carbohydrate oxidation in LE (247 11 lmol kg1

Figure 2 Expired volume (d),

volume of O2 consumed (s) and
volume of CO2 produced (j) during
intermittent exercise with a
work : recovery cycle of either
24 : 36 s (solid lines) or 6 : 9 s (dashed
lines). One subject group completed
the intermittent exercise protocols
without (a), one group with (b), a
10 min bout of intense exercise,
followed by approximately 45 min of
passive recovery prior to intermittent
exercise. Values are mean SEM.
Signicant difference between
exercise protocols.

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Fuel use in intermittent exercise
M A Christmass et al.
min1, LE; 222 8 lmol kg1 min1, SE) approached
signicance (P 0.057). Fat oxidation rate was 2.2-fold
lower in LE (10 2 lmol kg1 min1, LE; 22
2 lmol kg1 min1, SE; P 0.01). In both LE and SE,
rates of carbohydrate and fat oxidation remained
constant from 16 to 38 min of exercise (Fig. 3).
Plasma metabolite and blood gas concentrations. There was
no difference between LE and SE for resting plasma
lactate concentration. In both exercise protocols the
plasma lactate level increased from rest to 10 min
(P 0.001) and remained constant from 10 to 32 min.
During exercise, plasma lactate was higher in LE
compared with SE for all measurements (P 0.01;
Fig. 4). Following an initial increase from rest to the
rst measurement (P 0.001), plasma glycerol
concentration remained constant between 10 and
32 min (Fig. 4). There was no difference in the plasma
glycerol response between the two exercise protocols.
Plasma NEFA concentration decreased from rest to
10 min (P 0.05), returned to the resting level and
subsequently remained constant from 10 to 32 min in
LE and SE (Fig. 4). There was no difference in the
plasma NEFA response to exercise for the two
protocols despite an apparently lower total body fat
oxidation during LE.
44
Acta Physiol Scand 2001, 172, 3952
Figure 3 (a) Energy expenditure,
(b) rate of carbohydrate oxidation and
(c) rate of fat oxidation during
intermittent exercise with a
work : recovery cycle of either
24 : 36 s (solid lines) or 6 : 9 s (dashed
lines). One subject group completed
the intermittent exercise protocols
without (d), and one group with (e), a
10 min bout of intense exercise,
followed by approximately 45 min of
passive recovery prior to intermittent
exercise. Values are mean SEM.
Signicant difference between
exercise protocols; Signicant
difference between experiments.
A protocol * time interaction effect indicated that
the pattern of response across time for capillary blood
pH was different between the exercise protocols
(P 0.001). In both exercise protocols, the level of
blood pH was constant from 10 to 32 min (Fig. 4).
However, blood pH was lower at all measurements
for LE compared with SE (P 0.001) suggesting that
the difference in response across time may have been
because of differences in the magnitude of decline in
blood pH from rest to the rst measurement. This is
supported by the nding that capillary blood pH
decreased from rest to 10 min in LE (P 0.001),
whereas in SE the value at 10 min was not different
to the resting level. For both exercise protocols blood
partial pressure of CO2 (pCO2) declined signicantly
from the resting value (P 0.01) and subsequently
remained constant between 10 and 32 min. A proto-
col * time interaction effect indicated that the
response for blood bicarbonate (HCO3 ) concentra-
tion during exercise was different for LE and SE
(P 0.05). In a similar pattern to that for pH, the
HCO3 concentration was constant from 10 to 32 min
of exercise in both protocols, and was lower at all
measurements for LE compared with SE (P 0.05).
Blood HCO3 decreased from rest to 10 min in LE
(P 0.001), although in SE there was no difference
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Acta Physiol Scand 2001, 172, 3952 M A Christmass et al.
Fuel use in intermittent exercise

Figure 4 (a) Capillary blood pH,

(b) plasma glycerol and non-esteried
fatty acid (NEFA) and (c) plasma
lactate concentration during
intermittent exercise with a
work : recovery cycle of either
24 : 36 s (solid lines) or 6 : 9 s (dashed
lines). Glycerol (closed symbols), NEFA
(open symbols). One subject group
completed the intermittent exercise
protocols without (d), and one group
with (e), a 10-min bout of intense
exercise, followed by approximately
45 min of passive recovery prior to
intermittent exercise. Values are
mean SEM. Signicant difference
between exercise protocols.

between the resting HCO3 concentration and the

value at 10 min.
Muscle metabolite concentrations. Resting muscle glycogen
concentrations were not different in LE and SE
(Fig. 5). There was a decline in muscle glycogen
content between pre- and post-exercise measurements
for LE (118 mmol kg1 dw; P 0.05) but not for SE
(54 mmol kg1 dw). As a result, the post-exercise
muscle glycogen concentration in LE was lower than
the value observed for SE (P 0.05; Fig. 5) which
supports the ndings for carbohydrate oxidation rates
estimated from indirect calorimetry. Muscle lactate
concentrations at rest were not different in LE and
SE. Exercise resulted in an approximately two-fold
increase in muscle lactate in both LE and SE
(P 0.05) and consequently there was no difference
in post-exercise muscle lactate levels (Fig. 5). The
muscle free carnitine content decreased approximately
1.3-fold and 1.5-fold in LE (P 0.05) and SE
(P 0.01), respectively (Fig. 5). There was no differ-
ence in either resting or post-exercise muscle free
carnitine content between the two exercise protocols
(Fig. 5).
Effect of prior brief intense exercise and passive recovery
on fuel use during sustained intermittent intense exercise:
Experiment 2
Respiratory gas exchange and indirect calorimetry. Overall R
and V_ E were constant from 16 min to the nal
measurement during exercise (38 min), and from 27 to
38 min overall V_ O2, V_ CO2 (Fig. 2) and aerobic energy
expenditure were constant (Fig. 3). In a similar response
to Experiment 1, the overall V_ O2 (39.0 0.9 mL kg1
min1, LE; 42.3 0.8 mL kg1 min1, SE; P 0.01;
Fig. 2) and aerobic energy expenditure
(0.81 0.02 kJ min1 kg1, LE; 0.88 0.02 kJ min1
kg1, SE; P 0.01; Fig. 3) were lower in LE compared
with SE. Overall exercise intensity was 68.0 2.7%
V_ O2peak in LE and 73.8 2.8% V_ O2peak in SE.
There was no difference in overall exercise intensity,
although aerobic energy expenditure was slightly lower
(5%, P 0.05), in Experiment 1 compared with
Experiment 2.
There was no difference between the exercise
protocols for R when brief intense exercise and passive
recovery preceded LE and SE (0.93 0.01, LE and
SE) in contrast to the results for Experiment 1. There

2001 Scandinavian Physiological Society 45

Fuel use in intermittent exercise
M A Christmass et al.
was also no difference for V_ E between LE and SE
(Fig. 2). Consequently, although overall exercise
intensity, work period intensity (114% V_ O2peak,
Experiment 1; 111% V_ O2peak, Experiment 2) and LE
and SE were effectively identical to Experiment 1, there
was no difference between the exercise protocols for
rates of carbohydrate (219 10 lmol kg1 min1, LE;
238 11 lmol kg1 min1, SE) or fat oxidation (17
2 lmol kg1 min1, LE; 18 2 lmol kg1 min1, SE).
In both protocols, carbohydrate and fat oxidation
rate remained constant from 16 to 38 min (Fig. 3).
Comparison of fuel oxidation rates between the two
experiments showed that when brief intense exercise
and passive recovery was performed prior to LE,
carbohydrate oxidation was lower (P 0.05) and fat
oxidation was higher (P 0.01) compared with
Experiment 1 (no prior brief intense exercise and
passive recovery; Fig. 3). In contrast, the pattern of fuel
oxidation during SE was apparently similar between the
two experiments.
Plasma metabolite and blood gas concentrations. In both
LE and SE pre-exercise plasma lactate concentrations
were similar and there was an increase to the rst
measurement (P 0.001). Plasma lactate levels
remained constant from 10 to 32 min (Fig. 4).
46
Acta Physiol Scand 2001, 172, 3952
Figure 5 Skeletal muscle, (a) carnitine,
(b) lactate and (c) glycogen
concentration before (j) and after (h)
intermittent exercise with a
work : recovery cycle of either
24 : 36 s or 6 : 9 s. One subject group
completed the 24 : 36 s (d) 6 : 9 s (e)
protocols without, and one subject
group completed the 24 : 36 s (f) and
6 : 9 s (g) protocols with, a 10-min
bout of intense exercise, followed by
approximately 45 min of passive
recovery prior to intermittent exercise.
Values are mean SEM. Signicant
difference between exercise
protocols; *Signicant difference
before, compared to after, intermittent
exercise.
However, when brief intense exercise and passive
recovery were performed prior to intermittent exercise,
there was no difference in plasma lactate between LE
and SE. The contrast between the two experiments for
plasma lactate appeared to be because of a reduced
plasma lactate response when LE was preceded by brief
intense exercise and passive recovery (Fig. 4). The
protocol * study interaction term for plasma lactate did
not achieve signicance (P 0.10), although this
outcome may have been because of the variability in
plasma lactate levels between subjects. Pre-exercise
plasma glycerol levels were not different and increased
to the rst measurement (P 0.05). Plasma glycerol
levels were constant from 10 to 32 min and, similar to
the nding for Experiment 1, were not different
between LE and SE (Fig. 4). Plasma NEFA concen-
trations were also not different between exercise
protocols (Fig. 4). There was a signicant effect of time
(P 0.05) and plasma NEFA concentration with the
value at 32 min higher compared with the pre-exercise
level (P 0.05). There was no difference between the
two experiments for plasma glycerol or NEFA
concentrations.
Capillary blood pH was lower at 10 min compared
with the pre-exercise level (P 0.01) and with the value
at 32 min (P 0.05). Despite this decrease, blood pH
2001 Scandinavian Physiological Society
Acta Physiol Scand 2001, 172, 3952
remained constant between 21 and 32 min and during
this period was not different from the pre-exercise level
(Fig. 4). Blood pCO2 was lower throughout exercise
compared with the pre-exercise value (P 0.05). For
both LE and SE, following this initial decline there was
no further change in pCO2 between 10 and 32 min
Capillary blood HCO3 decreased from the pre-exercise
value to 10 and 21 min of exercise (P 0.01).
However, HCO3 concentration was constant between
21 and 32 min and was not different to the pre-exercise
level at this time (i.e. 32 min). There was no difference
between LE and SE for pH, HCO3 and pCO2. In
addition, there was no difference between the two
experiments for pH, HCO3 and pCO2.
In both experiments, the haematocrit level was not
different between LE and SE and there was no
difference for haematocrit between the two experi-
ments. Therefore, it is probable that observed differ-
ences between LE and SE for plasma metabolites in
both experiments are a consequence of differences in
the metabolic response to exercise rather than differ-
ences in the degree of haemoconcentration.
Muscle metabolite concentrations. There was no difference in
pre-exercise muscle glycogen concentrations between
the two exercise protocols (Fig. 5). In contrast to the
results for Experiment 1, there was no difference in
post-exercise muscle glycogen concentration. Pre- and
post-exercise muscle lactate concentrations were not
different in LE and SE (Fig. 5). Muscle free carnitine
content did not change across the exercise protocol for
either LE or SE which also contrasts with the results
for Experiment 1 (Fig. 5). There was no difference
between LE and SE for pre- or post-exercise muscle
free carnitine concentration.
DISCUSSION
Our results clearly show that work period duration
cannot be considered as the overriding inuence on the
pattern of fuel use during sustained intermittent exer-
cise with constant work period intensity. Under these
conditions, we previously demonstrated an effect of
longer work period duration to reduce muscle O2
availability. On the basis of this nding we proposed
that lower muscle O2 availability is responsible for the
higher rate of carbohydrate metabolism, and in turn the
decline in fat oxidation during LE compared with SE
(Christmass et al. 1999b). This pattern of fuel use was
replicated, and supported by measurement of muscle
glycogen concentrations, during Experiment 1. Without
prior brief intense exercise and passive recovery
(Experiment 1), a 1.1-fold higher rate of carbohydrate
oxidation approached signicance and muscle glycogen
depletion was two-fold greater, fat oxidation 2.2-fold
2001 Scandinavian Physiological Society
M A Christmass et al.
Fuel use in intermittent exercise
lower and plasma lactate levels 1.5-fold higher when
work and recovery period durations were proportion-
ately longer (Figs 35).
This difference in carbohydrate metabolism was
subsequently abolished when intermittent exercise was
preceded by a single 10 min bout of intense work,
followed by approximately 45 min of passive recovery
(Experiment 2). Under these conditions, carbohydrate
oxidation, muscle glycogen depletion, fat oxidation and
plasma lactate concentration were almost identical in
LE and SE (Figs 35). In contrast to our proposal
(Christmass et al. 1999b), factors other than work
period duration, and in particular the effect of muscle
O2 availability, are likely to be responsible for the rate
of carbohydrate metabolism during sustained intermit-
tent exercise with constant work period intensity.
The effect of a single work (30 s) and recovery
(240 s) cycle to slow carbohydrate metabolism in a
subsequent work bout during intermittent exercise is
well established (McCartney et al. 1986, Spriet et al.
1989, Putman et al. 1995). The current results extend
these ndings concerning individual work bouts to
include the pattern of net fuel use across a sustained
period of intermittent intense exercise. Thus, brief
intense exercise (10 min), followed by passive recovery
(approximately 45 min), substantially modies the net
metabolic response to subsequent whole body,
sustained (40 min) intermittent intense exercise. Our
ndings bring to light the potential for a novel
approach to the preservation of carbohydrate stores in
prolonged exercise by modifying pre-exercise warm-up
strategies.
A secondary objective was to monitor changes in
muscle glycogen concentration as indirect support for
estimates of carbohydrate oxidation rate based on indi-
rect calorimetry. At present there are no data comparing
muscle substrate levels during LE and SE treadmill
running. In the current study, and in our previous work
(Christmass et al. 1999a, b), rates of fuel oxidation were
estimated from measurements of respiratory gas
exchange using indirect calorimetry (Frayn 1983).
Appropriate conditions for interpretation of respiratory
gas exchange and application of indirect calorimetry
during intermittent exercise have been outlined previ-
ously (Christmass et al. 1999a). Briey, respiratory gas
exchange data are collected across work and recovery and
interpreted as an overall measurement. The validity of
indirect calorimetry during LE and SE has been
supported on the basis of a stable plasma lactate and
HCO3 pool (Christmass et al. 1999b).
Following an initial increase, plasma lactate
concentration was always constant between 10 and
32 min During Experiment 1, capillary HCO3, pCO2
and pH were stable during this period. During Experi-
ment 2 capillary pCO2 was also constant between 10
47
Fuel use in intermittent exercise
M A Christmass et al.
and 32 min and pH and HCO3 were stable between 21
and 32 min (Fig. 4). These results conrm our previous
ndings (Christmass et al. 1999b) and combined with
constant values for V_ E (Fig. 2) indicate the presence of
stable plasma lactate and blood HCO3 pools.
Measured changes in muscle glycogen concentra-
tions closely reected the results for estimated rates of
carbohydrate oxidation (Figs 3 and 5). Changes in
muscle glycogen concentration cannot be related
directly with whole body carbohydrate oxidation
because of uncertainty concerning the volume of active
muscle, the contribution of the sampled muscle to the
work output (Coyle 1996) and the extent of glycogen
resynthesis during recovery (Essen 1978). Nevertheless,
current results for muscle glycogen provide further
indirect support for estimates of fuel oxidation rates
during LE and SE based on indirect calorimetry
(Christmass et al. 1999b).
Carbohydrate metabolism is reduced during intermittent
exercise and with repeated intense exercise
The nding that carbohydrate metabolism was not
different when LE and SE were preceded by brief
intense exercise and passive recovery was apparently
the result of the slowing of carbohydrate metabolism in
response to LE. The metabolic response to SE was
similar during both experiments. The extent to which
carbohydrate metabolism was slowed is emphasised by
the nding that carbohydrate oxidation, plasma lactate
and muscle glycogen depletion in LE following brief
intense exercise and passive recovery were almost
identical to the values observed during SE for both
Experiment 1 and Experiment 2 (Figs 35).
It is well established that carbohydrate use decreases
considerably when the same intensity of heavy exercise
is performed intermittently (Essen 1978). Furthermore,
a reduction in glycogen depletion, glycolytic ux and
lactate production across successive individual work
and recovery cycles during brief intermittent maximal
and intense exercise has been repeatedly demonstrated
(McCartney et al. 1986, Spriet et al. 1989, Bangsbo et al.
1992a, Gaitanos et al. 1993, Putman et al. 1995, Trump
et al. 1996). The effect of a single, individual work and
recovery cycle to reduce lactate output in the successive
work bout persists across as much as 60 min of
recovery (Bangsbo et al. 1992b). Bangsbo (1996) has
reviewed several proposed mechanisms for the effect of
repeated exercise to slow glycolysis.
Briey, an effect of reduced pH and/or changes in
the level of adenine nucleotides following the brief
intense exercise is not likely to have inhibited glycolytic
ux (Bosca et al. 1985, Gaitanos et al. 1993) during
subsequent LE. Bangsbo et al. (1992b) found a decline
in lactate production across two bouts of exhaustive
48
Acta Physiol Scand 2001, 172, 3952
knee extensor exercise despite a separation of 60 min
during which muscle pH and lactate had returned to the
resting level. Although muscle ATP, ADP and AMP
levels were similar during the two exercise bouts
changes in the bound and unbound fraction of these
effectors could have inuenced glycolytic ux (Bangsbo
et al. 1992b). However, estimated lactate production
decreased with repeated isometric calf contractions
despite a progressive increase in free ADP determined
using nuclear magnetic resonance (Bangsbo 1994). The
pre-exercise muscle glycogen level is unlikely to inu-
ence the glycogenolytic rate during short-term high-
intensity exercise (Bangsbo et al. 1992b, Hargreaves
et al. 1997) but may have an effect in prolonged
submaximal exercise (Hargreaves et al. 1995). Never-
theless, pre-exercise glycogen levels were not different
for LE in Experiment 1 and Experiment 2 and,
therefore, could not have inuenced the extent of
glycogen depletion in the subsequent exercise.
The rate of carbohydrate metabolism during LE and
SE in Experiment 2 may have been inuenced either
directly or indirectly by hormonal responses during the
43 5 min of passive recovery. For example, cate-
cholamines may contribute to the increase in energy
metabolism observed during exercise recovery
(i.e. excess post-exercise oxygen consumption) through
an increase in both triglyceride/fatty acid cycling and
fat oxidation (see Borsheim et al. 1998). Increased
oxidation of NEFA and reduced citric acid cycle
activity during the passive recovery following brief
intense exercise (Fig. 1) could have impeded subse-
quent glycolytic ux during LE and SE through
increases in acetyl CoA and citrate (Randle et al. 1964).
Muscle citrate content is higher during the rest periods
of intermittent exercise (15 s work : 15 s recovery).
The increased citrate has been implicated in the slowing
of carbohydrate metabolism during intermittent
compared with continuous exercise (Essen 1978).
However, increased acetyl CoA production from fatty
acid oxidation during passive recovery would be
expected to correspond to increased acetylcarnitine
(Constantin-Teodosiu et al. 1991, Putman et al. 1993)
and in turn, decreased free carnitine pre-exercise
(Harris et al. 1987). High pre-exercise muscle free
carnitine levels (Fig. 5) following brief intense exercise
and passive recovery (Experiment 2) indicate minimal
formation of acetylcarnitine. Furthermore, pre-exercise
plasma NEFA levels were similar in Experiment 1 and
Experiment 2.
During intense dynamic knee extensor exercise,
blood ow to the working muscle increases 30-fold and
remains above resting levels for 2030 min of recovery
(see Bangsbo & Hellsten 1998). The persistence of
hyperaemia in exercise recovery varies with the type,
intensity and duration of the prior exercise and is
2001 Scandinavian Physiological Society
Acta Physiol Scand 2001, 172, 3952
thought to be inuenced by a number of systemic and
local factors (see Bangsbo & Hellsten 1998). Hyper-
aemia during the passive recovery following brief
intense exercise in Experiment 2 could have resulted in
a higher pre-exercise muscle O2 availability compared
with Experiment 1. However, it is unlikely that this
hyperaemia substantially inuenced muscle O2 avail-
ability throughout the subsequent 40 min intermittent
intense exercise.
The current ndings do not permit a denitive
explanation for the slowed carbohydrate metabolism
when LE is preceded by brief intense exercise and
passive recovery. Further research is needed concerning
the effect of brief intense exercise followed by passive
recovery on metabolic and hormonal regulation of
glycolytic rate both prior to, and during, subsequent
sustained heavy exercise.
Implications of reduced carbohydrate metabolism for fat
oxidation during intermittent exercise
Essen et al. (1977) suggested that the slowing of glyc-
olysis during work periods of intermittent exercise
could promote the oxidation of fat. In the current
study, the decline in carbohydrate metabolism during
LE in Experiment 2 compared with Experiment 1 was
accompanied by a 1.7-fold higher rate of fat oxidation.
The difference in fat oxidation rate is unlikely to
be the result of a limitation in plasma NEFA avail-
ability. There was no difference between the two
experiments for plasma glycerol concentration.
Furthermore, the plasma NEFA concentrations in
Experiment 1 and Experiment 2 were not different and
were higher (Fig. 4) than the levels suggested to impair
fat oxidation (Romijn et al. 1995).
With similar work period intensity (114% V_ O2peak,
Experiment 1; 111% V_ O2peak, Experiment 2) and
identical total work duration (16 min), the difference in
whole body fat oxidation during LE performed without
(Experiment 1), compared to with (Experiment 2),
prior brief intense exercise and passive recovery could
be because of a metabolic factor/s within skeletal
muscle (see Christmass et al. 1999b). Sidossis et al.
(1997) proposed that the availability of carbohydrate
regulates fat oxidation in skeletal muscle in reverse of
the classic glucosefatty acid cycle (Randle et al. 1963).
This concept is supported by recent evidence from
continuous exercise suggesting that increased glycolytic
ux, as a result of either pre-exercise glucose feeding
(constant exercise intensity; Coyle et al. 1997) or
increased exercise intensity (Sidossis et al. 1997), could
reduce fat oxidation in part by direct inhibition of
mitochondrial LCFA transport.
Increased glycolytic ux could reduce LCFA trans-
port by inhibition of carnitine palmitoyl-transferase I
2001 Scandinavian Physiological Society
M A Christmass et al.
Fuel use in intermittent exercise
(Sidossis et al. 1996). Carnitine palmitoyl-transferase I is
essential for transfer of LCFA across the mitochondrial
membrane and is inhibited by malonyl CoA (McGarry
1995). Sidossis et al. (1997) proposed that increased
carbon ow through acetyl CoA carboxylase could
inhibit carnitine palmitoyl-transferase I by increasing
the malonyl CoA concentration. The decline in malonyl
CoA observed in response to submaximal treadmill
exercise in rats has been suggested as important for the
increase in fatty acid oxidation during prolonged exer-
cise (Winder et al. 1989). However, the malonyl CoA
content in human skeletal muscle at rest was lower
compared with the rat and was unchanged during
prolonged (60 min) moderate-intensity (6570%
V_ O2max) exercise (Odland et al. 1996). Furthermore,
the skeletal muscle malonyl CoA concentration in
humans following 10 min of cycling exercise at 35, 65,
90% V_ O2max was not different to the resting level and
was not related to the fat oxidation rate (Odland et al.
1998). These ndings suggest that reduced transfer of
LCFA into mitochondria in the presence of accelerated
glycolytic ux during exercise in humans is unlikely to
involve inhibition of carnitine palmitoyl-transferase I by
malonyl CoA.
During intense exercise (Constantin-Teodosiu et al.
1991) free carnitine serves as an acetyl group buffer
(Alkonyi et al. 1975) preventing excess accumulation of
acetyl CoA and thereby maintaining viable CoA levels
(Foster & Harris 1987). Free carnitine is also essential
for the carnitine palmitoyl-transferase I reaction and
hence for mitochondrial LCFA transfer and oxidation
(Bremer 1983). Rapid formation of acetylcarnitine with
increased glycolytic ux could deplete free carnitine
levels thereby providing a mechanism for the inhibition
of fat oxidation when carbohydrate metabolism is
accelerated (Foster & Harris 1987).
In this context, skeletal muscle free carnitine
concentration remains at the resting level during low-
intensity (40% V_ O2max) exercise but may decrease
three-fold at high-intensity (75% V_ O2max) exercise
(Sahlin 1990). Corresponding to these changes in free
carnitine availability, the percentage of [113C]oleate
uptake that is oxidised during low-intensity (40%
V_ O2max) exercise is reduced at high-intensity (80%
V_ O2max) exercise. The percentage of [114C]octanoate
uptake oxidised is not inuenced by exercise intensity.
Oleate transfer is dependent on carnitine palmitoyl-
transferase I whilst octanoate diffuses freely across the
inner mitochondrial membrane. Thus, a limitation in
mitochondrial LCFA transfer could be responsible for
the decline in fat oxidation (Sidossis et al. 1997) which
parallels the fall in free carnitine availability during
intense exercise.
According to the proposed mechanism reduced
glycolytic ux during LE, following brief intense exer-
49
Fuel use in intermittent exercise
M A Christmass et al.
cise and passive recovery (Experiment 2), could facili-
tate an increase in fat oxidation by attenuating the
inhibition of mitochondrial LCFA transfer. On this
basis we measured skeletal muscle free carnitine
content following LE and SE in Experiment 1 and
Experiment 2 to examine free carnitine availability as a
factor responsible for the increase in fat oxidation with
decreased carbohydrate metabolism during intermittent
intense exercise.
A 1.3-fold higher post-exercise muscle free carnitine
content for LE in Experiment 2 compared with
Experiment 1 approached signicance (P 0.07;
Fig. 5). This outcome occurred in the presence of
reduced carbohydrate metabolism and increased fat
oxidation during LE in Experiment 2 compared with
Experiment 1. In contrast, a 1.7-fold higher post-
exercise free carnitine level for SE in Experiment 2
relative to Experiment 1 (P 0.05) was observed
despite similar rates of fat oxidation (Figs 3 and 5).
Muscle free carnitine content was also compared
between LE and SE within each experiment. Without
prior brief intense exercise and passive recovery
(Experiment 1), the differences in carbohydrate and fat
oxidation between LE and SE are comparable with the
differences observed between LE in Experiment 1 and
Experiment 2. Under these conditions (Experiment 1),
the pre- to post-exercise decrease in skeletal muscle free
carnitine content was almost identical in both exercise
protocols (Fig. 5) despite the more than two-fold lower
rate of fat oxidation during LE (Fig. 3). This result is in
accordance with the apparent dissociation between
rates of whole body fat oxidation and muscle free
carnitine levels for comparisons between the two
experiments. Our ndings do not support the proposal
that rapid glycolytic ux reduces the availability of free
carnitine and thereby inhibits the oxidation of fat
(Foster & Harris 1987) under conditions of intermittent
exercise.
There was no difference in pre- and post-exercise
skeletal muscle free carnitine content between LE and
SE during Experiment 2 in a similar response to that
for Experiment 1. At rest, 96% of human skeletal
muscle carnitine content exists in either the free or the
acetylated form (Harris et al. 1987). During exercise, a
decline in free carnitine corresponds to formation of
acetylcarnitine such that the sum concentration of these
two metabolites remains constant (Harris et al. 1987,
Constantin-Teodosiu et al. 1992). Furthermore, the
concentration of acetylcarnitine and acetyl CoA is
highly correlated in human skeletal muscle at rest and
during exercise (Constantin-Teodosiu et al. 1991).
Therefore, the decrease in free carnitine in LE and SE
without prior brief intense exercise and passive
recovery (Experiment 1; Fig. 5) provides indirect evi-
dence to suggest a rate of acetyl CoA formation higher
50
Acta Physiol Scand 2001, 172, 3952
than the rate of oxidation within the citric acid cycle.
This is likely to have occurred in the initial 10 min of
the experiment when plasma lactate levels were
increasing and could be because of limitations at the
level of citric acid cycle enzyme activities or substrate
and cofactor availability for the cycle (Constantin-
Teodosiu et al. 1991). In contrast, following brief
intense exercise and passive recovery (Experiment 2)
the muscle free carnitine content did not change across
the exercise protocol for LE and SE (Fig. 5) suggesting
that formation and oxidation of acetyl CoA is balanced
under these conditions (Constantin-Teodosiu et al.
1991). The current results indicate that the coordination
of acetyl CoA formation and oxidation during inter-
mittent exercise is likely to be similar for LE and SE but
may be inuenced by prior brief intense exercise and
passive recovery (Fig. 5).
Without prior brief intense exercise and passive
recovery (Experiment 1), there was no difference in pre-
and post-exercise muscle lactate concentration between
LE and SE despite the higher plasma lactate in LE
(Figs 4 and 5). Because an increase in circulating lactate
accumulation occurs as a result of increases in muscle
lactate content, a higher muscle lactate concentration in
LE might be anticipated. However, blood and muscle
lactate concentrations in continuous exercise are not
always correlated (Tesch et al. 1982) and may not be
directly related under conditions of intermittent exer-
cise. Alternatively, small differences in muscle lactate
content between LE and SE in relation to the difference
in plasma lactate could have been obscured by variations
in muscle lactate inherent to mixed muscle bre
samples. In this context, the range for muscle lactate
(7.618.1 mM) was approximately twice that for plasma
lactate (3.38.9 mM) during LE in Experiment 1.
Our results suggest that muscle free carnitine avail-
ability is unlikely to inuence the rates of fat oxidation
during LE or SE. Further research is needed to den-
itively exclude the presence of metabolic factor/s
within skeletal muscle responsible for the decline in fat
oxidation during LE. For example, determination of the
carnitine content in relation to bre type in human
skeletal muscle during LE and SE could be of benet
on the basis of the difference in oxidative capacity
amongst type I, type IIa and type IIb bres. In addition,
Hultman (1995) has suggested that increases in the
acetyl CoA : CoA ratio could limit LCFA transport by
reducing the availability of mitochondrial CoA.
In summary, the current results do not support the
concept that differences in muscle O2 availability, as a
result of differences in work period duration, are the
principal inuence on carbohydrate metabolism during
sustained intermittent intense exercise with constant
work period duration (Christmass et al. 1999b). In LE
compared with SE during Experiment 1, a higher rate of
2001 Scandinavian Physiological Society
Acta Physiol Scand 2001, 172, 3952
carbohydrate oxidation approached signicance and
was accompanied by lower rates of fat oxidation,
elevated plasma lactate levels and lower post-exercise
muscle glycogen. These results conrm our previous
ndings for sustained intermittent intense exercise
(Christmass et al. 1999b). Results for muscle glycogen
provide support for the use of indirect calorimetry to
estimate fuel oxidation rates under these conditions.
The current ndings also suggest that a decrease in free
carnitine availability is unlikely to be responsible for the
inhibition of fat oxidation during LE. Most importantly
however, the difference in fuel use between LE and SE
was abolished when intermittent exercise was preceded
by brief intense exercise and passive recovery. This
effect appeared to be because of the slowing of carbo-
hydrate metabolism in LE. An important implication of
these ndings is the potential for even brief intense
exercise followed by passive recovery to substantially
modify the pattern of fuel use in subsequent whole body
intermittent exercise. This presents the possibility for a
novel approach to the problem of sparing carbohydrate
fuel stores through relatively simple modications to
preparation strategies for prolonged exercise.
The authors are indebted to Professor Peter Hartmann of the
Department of Biochemistry, The University of Western Australia for
his helpful advice and support throughout these experiments. We
especially thank Dr Paul Fournier of the Department of Human
Movement and Exercise Science, The University of Western Australia
for valuable discussions and advice on the development of the manu-
script. We acknowledge and thank Professor Roger Harris of the
Department of Sports Sciences, University College of Chichester, UK
for assistance with assay procedures for determination of muscle free
carnitine concentration. The authors also thank Dr Arjun Rao for
performing several muscle biopsy procedures, Mr David Preen and Ms
Christine Wakeford for assistance during experimental testing and the
staff of the Western Australian Institute of Sport for the use of their
exercise physiology laboratory facilities.
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2001 Scandinavian Physiological Society