V̇O2 Kinetics and Performance in Soccer

Players after Intense Training and Inactivity

PETER M. CHRISTENSEN1, PETER KRUSTRUP1, THOMAS P. GUNNARSSON1, KRISTIAN KIILERICH2,
LARS NYBO1, and JENS BANGSBO1
1
Copenhagen Muscle Research Centre, Department of Exercise and Sport Sciences, Section of Human Physiology, University
of Copenhagen, Copenhagen, DENMARK; and 2Copenhagen Muscle Research Centre, Department of Biology, University of
Copenhagen, Copenhagen, DENMARK
Downloaded from http://journals.lww.com/acsm-msse by BhDMf5ePHKav1zEoum1tQfN4a+kJLhEZgbsIHo4XMi0hCywCX1AWnYQp/IlQrHD3i3D0OdRyi7TvSFl4Cf3VC4/OAVpDDa8KKGKV0Ymy+78= on 03/08/2021

ABSTRACT
CHRISTENSEN, P. M., P. KRUSTRUP, T. P. GUNNARSSON, K. KIILERICH, L. NYBO, and J. BANGSBO. V̇O2 Kinetics and
Performance in Soccer Players after Intense Training and Inactivity. Med. Sci. Sports Exerc., Vol. 43, No. 9, pp. 1716–1724, 2011.
Purpose: The study’s purpose was to examine the effects of a short-term period with intensified training or training cessation of trained soccer
players on V̇O2 kinetics at 75% maximal aerobic speed, oxidative enzymes, and performance in repeated high-intensity exercise. Methods:
After the last match of the season, 18 elite soccer players were, for a 2-wk period, assigned to a high-intensity training group (n = 7)
performing 10 training sessions mainly consisting of aerobic high-intensity training (8
2 min) and speed endurance training (10–12
30-s
sprints) or a training cessation group (n = 11) that refrained from training. Results: For the training cessation group, V̇O2 kinetics became
slower (P G 0.05) with a larger time constant (T = 21.5 T 2.9 vs 23.8 T 3.2 s (mean T SD, before vs after)) and a larger mean response time
(time delay + T = 45.0 T 1.8 vs 46.8 T 2.2 s). The amount of muscle pyruvate dehydrogenase (17%, P G 0.01) and maximal activity of
citrate synthase (12%) and 3-hydroxyacyl-CoA (18%, P G 0.05) were lowered. In addition, the fraction of slow twitch fibers (56% T 18% vs
47% T 15%, P G 0.05), Yo-Yo intermittent recovery level 2 test (845 T 160 vs 654 T 99 m), and the repeated sprint performance
(33.41 T 0.96 vs 34.11 T 0.92 s, P G 0.01) were reduced. For the high-intensity training group, running economy was improved (P G 0.05),
and the amount of pyruvate dehydrogenase (17%) and repeated sprint performance (33.44 T 1.17 vs 32.81 T 1.01 s) were enhanced
(P G 0.05). Conclusions: Inactivity slows V̇O2 kinetics in association with a reduction of muscle oxidative capacity and repeated high-
intensity running performance. In addition, intensified training of already well-trained athletes can improve mechanical efficiency and
repeated sprint performance. Key Words: REPEATED SPRINTING, YO-YO INTERMITTENT RECOVERY TEST, RUNNING
ECONOMY, OXIDATIVE ENZYMES

R
epeated high-intensity work is performed in sports
such as soccer (36), basketball (35), and rugby (43)
and taxes both the aerobic and anaerobic energy
systems. The mean HR is around 85% of maximal HR in
soccer (5) and basketball (35), with average blood lactate
being 4–7 mM and peak values more than 10 mM (31,35).
Nearly 40 sprints have been reported to occur for a player
during a soccer game with the average distance being around
10–15 m (36). When comparing teams at different compet-
itive standards, it is evident that the teams at the highest
level carry out more high-intensity running during compe-
tition in soccer (36) and rugby (43). Furthermore, the players
in the high-level teams perform better than players in lower
APPLIED SCIENCES
ranked teams in the Yo-Yo intermittent recovery (Yo-Yo IR)
tests consisting of 2
20-m runs with increasing speed
interspersed by 10 s of recovery between work bouts until
exhaustion (6). Thus, the players’ ability to conduct repeated
high-intensity work is an important factor for team success.
The aerobic energy system makes a significant contribu-
tion to energy turnover during repeated high-intensity work
(8,12), and several studies have found a correlation between
maximal oxygen uptake (V̇O2max) and performance in re-
peated sprint tests (RST) (44) and the Yo-Yo IR tests (6,41).
In addition, recent studies have shown a correlation between
fast pulmonary V̇O2 kinetics at 60% maximal aerobic speed
(MAS) and performance in repeated 40-m sprints (19,42)

and the Yo-Yo IR levels 1 (IR1) and 2 (IR2) tests (41) in

trained soccer players. However, sprints in soccer have an
average duration of È2 s (15 m) (36), and the validity of the
Address for correspondence: Jens Bangsbo, Department of Exercise and
Sport Sciences, August Krogh Building, Universitetsparken 13, DK-2100
results for match play may be questioned.
Copenhagen K, Denmark; E-mail: jbangsbo@ifi.ku.dk. In untrained subjects, V̇O2 kinetics has been shown to be
Submitted for publication May 2010. improved by different types of training ranging from low-
Accepted for publication December 2010. intensity work at 60% V̇O2max in 6 wk (10) to high-intensity
0195-9131/11/4309-1716/0 interval training using 4–7
30-s all-out sprints with 4 min
MEDICINE & SCIENCE IN SPORTS & EXERCISEÒ of recovery for 2 wk (2), 20
1 min at 90% V̇O2max with
Copyright Ó 2011 by the American College of Sports Medicine 1-min recovery periods for 6 wk (10), and 15
1 min at
DOI: 10.1249/MSS.0b013e318211c01a 150% leg V̇O2max separated by 3 min of recovery for 7 wk

1716

Copyright © 2011 by the American College of Sports Medicine. Unauthorized reproduction of this article is prohibited.
(29). Limitations to oxygen uptake in the initial part of ex-
ercise is attributed either to local limitation in the muscles
(‘‘metabolic inertia’’) or in the oxygen delivery (40). In
the aforementioned training studies performed on untrained
subjects, it was unclear what caused the faster V̇O2 kinetics
because the improvement was accompanied by increases in
V̇O2max, capillary density, enzyme activity, and muscle blood
flow during work. In trained runners, a combination of mod-
erate continuous running and high-intensity interval training
at intensities close to V̇O2max during 8 wk also resulted in
faster V̇O2 kinetics, which seems to be caused by peripheral
adaptations because V̇O2max was unaltered (18). This as-
sumption is supported by results from another study per-
formed on physical education students in which V̇O2 kinetics
became faster and V̇O2max remained unchanged after 4 wk
of intense interval training (11). A few weeks (4–9 wk) of
intense intermittent and reduced amount of training have been
shown to improve performance during short-term intense
exercise in already trained runners (4,26). However, it is un-
clear whether a shorter period of intensified and reduced
training can improve performance during repeated high-
intensity work and V̇O2 kinetics of trained soccer players
accustomed to high-intensity intermittent work.
The effect of inactivity on V̇O2 kinetics has not been in-
vestigated to the same extent as training. Inactivity causes
among other things a decrease in oxidative enzymes (37),
which may affect V̇O2 kinetics. Convertino et al. (17) found
that V̇O2 kinetics became slower after 7 d of bed rest in
untrained subjects. However, the time resolution was low
(30-s intervals), and only one work period was used to
describe the V̇O2 kinetics (33). In two other studies with
untrained subjects, V̇O2 kinetics was not changed by 2 wk
of bed rest (16) or 3 wk of inactivity in one leg after harness
suspension (24), which may be because of the low training
status of the subjects before the studies. Therefore, the effect
of inactivity of trained subjects on V̇O2 kinetics and the
relation to oxidative enzymes need to be studied.
Thus, the aim of the present study was to examine the
effect of a 2-wk period of intensified training and cessation
of training of trained soccer players on V̇O2 kinetics and
how it is linked to muscle oxidative enzymes and the ability
to perform repeated high-intensity work. In addition, another
aim of the study was to evaluate whether V̇O2 kinetics was
related to performance during repeated high-intensity work,
using a higher intensity to describe V̇O2 kinetics and a
shorter sprint distance than that used previously to mimic the
work demands during intermittent sports such as soccer.
METHODS
Subjects
Eighteen male soccer players participated in the study. All
subjects were members of teams from the Danish second
division (third best league), with the exception of one player
coming from the first division. All subjects were healthy,
V̇O2 KINETICS AND PERFORMANCE IN SOCCER PLAYERS
Copyright © 2011 by the American College of Sports Medicine. Unauthorized reproduction of this article is prohibited.
and none was on medication. Age, height, weight, and
pulmonary V̇O2max were 23.4 T 3.5 (mean T SD) yr, 181.8 T
6.7 cm, 78.5 T 8.5 kg, and 55.0 T 3.1 mL O2Ikgj1Iminj1,
respectively. The subjects had been training and competing
on the elite level on a regular basis for at least 2 yr. Their
normal week schedule consisted of three to four training
sessions and one match. After receiving oral and written
information about any possible risks and discomforts asso-
ciated with the experimental procedures, all participants
gave their written informed consent to participate. The study
conformed to the Code of Ethics of the World Medical
Association (Declaration of Helsinki) and was approved by
the Ethics Committee of the Copenhagen and Frederiksberg
communities.
Experimental Protocol and Design
The present study was conducted as a randomized two-
group longitudinal experiment. After the last match of the
season, the subjects were either assigned to a high-intensity
training group (HI, n = 7) or to a training cessation group
(TC, n = 11) for a period of 2 wk. Before the intervention
period, the subjects completed an incremental running test
for determination of maximal oxygen uptake (V̇O2max),
MAS, and HRmax. Furthermore, both before and after the
intervention period, the subjects carried out approximately
six runs at 75% MAS for estimation of V̇O2 kinetics (see
below), the Yo-Yo IR2 test, and an RST. The tests were
performed during two separate days with three submaximal
exercise bouts and the Yo-Yo IR2 being performed on the
first day and three submaximal exercise running bouts and
the RST on the second day using the same test order before
and after the intervention period. All the testing took place at
least 36 h after the last practice or test session and more than
72 h after a match. In addition, TC performed the Yo-Yo
IR2 test 72 h into the inactivation period. A muscle biopsy
was obtained from the musculus Vastus lateralis with the
Bergström technique at rest during the first visit to the lab-
oratory both for HI and TC as well as 36–40 h after the last
training in the intervention period for HI and after 14 d with
training cessation for TC. The biopsies were split into two
parts. One part was immediately frozen in liquid nitrogen,
and the other part was embedded in Tissue-Tek (Sakura
Finetek, Zoeterwoude, The Netherlands) and frozen in iso-
pentane. The biopsies were stored at j80-C until analyzed.
APPLIED SCIENCES
Intervention Period
During the intervention period, HI had 10 training ses-
sions including five aerobic high-intensity (AHI) sessions
and five speed endurance training (SET) sessions. Each
training session included an approximately 25-min warm-up
program. The AHI sessions were performed as small-sided
soccer drills (four vs four and three vs three) on artificial
grass consisting of eight repetitions of 2 min of exercise
interspersed by 1 min of recovery. HR was recorded at 5-s
intervals (Polar Team System; Polar Electro Oy, Kempele,
Medicine & Science in Sports & Exercised 1717
 Finland) during every session and afterward downloaded to
a personal computer for further analysis. Mean HR during
the 2-min exercise period in AHI was 87.7% T 1.2% of
HRmax (determined during incremental running), and the
total duration of the exercise in AHI was 75.4 T 7.8 min. Four
of the SET sessions consisted of 10–12
25- to 30-s all-out
sprinting bouts including changes of directions and parts
with ball contacts. Peak HR was 88.4% T 1.9% of HRmax,
and the total duration of the exercise periods was 19.9 T
0.6 min. In one of the SET sessions, the players performed
16 exercise bouts lasting 40–60 s separated by a recovery
time between the work bouts of a similar duration. Mean HR
during the exercise periods was 84.4% T 1.7% of HRmax,
and the total duration of exercise periods was 14 min. SET
was performed the day after an AHI session, and no training
was performed the day after an SET session. Overall, the
players in HI reduced the total training duration during the
2-wk training intervention by È30% compared with their
normal training.
The subjects in TC did not perform any training during the
2-wk intervention period, and they maintained their normal
daily activities but refrained from intense physical activity.
Testing Procedures
On the days of testing, subjects reported to the laboratory
at least 2 h after consuming a light meal. Subjects refrained
from strenuous physical activity in the last 36 h before
testing and abstained from alcohol and caffeine consumption
24 h before the testing. During all testing sessions, the HR
was recorded during 5-s intervals (Polar Team System; Polar
Electro Oy).
Incremental running test. The subjects completed an
incremental running protocol on a motorized treadmill,
starting with 5 min at 10 kmIhj1 followed by 4 min at
14 kmIhj1, and then, the running speed was increased by
1 kmIhj1 every minute until exhaustion. Throughout the
test, pulmonary oxygen uptake was measured by a breath-
by-breath gas analyzing system (JAEGER MasterScreen
CPX; Viasys Healthcare GmbH, Hoechberg, Germany). The
analyzer was calibrated before each test with two gases of
known O2 and CO2 concentrations as well as by the use of
a 3-L syringe for the tube flow meter calibration. V̇O2max
was determined as the highest value achieved during a 20-s
period. A plateau in oxygen uptake, despite an increased
APPLIED SCIENCES
power output, and an RER 91.10 were used as criteria for
V̇O2max achievement. MAS was determined as the lowest
running speed when V̇O2max was achieved, and HRmax was
determined as the highest value observed during the test.
Submaximal running. The subjects performed approx-
imately six runs at 75% MAS (average speed = 14.1 kmIhj1)
corresponding to È85% V̇O2max on two separate days both
before and after the intervention period to describe V̇O2 ki-
netics. This intensity was chosen because it is well known
that high-intensity aerobic exercise is of great importance for
elite soccer players (30,36). The pulmonary V̇O2 was mea-
1718 Official Journal of the American College of Sports Medicine
Copyright © 2011 by the American College of Sports Medicine. Unauthorized reproduction of this article is prohibited.
sured breath by breath with the same equipment as in the
V̇O2max test. Each bout consisted of 1 min of rest standing on
the treadmill followed by 4 min of running at 75% MAS.
Because preceding work can influence V̇O2 kinetics (14), the
work bouts were separated by a 30-min rest period, and the
first run was not included in the calculations. Thus, V̇O2 ki-
netics was modeled from four transitions both before and
after HI and TC. To estimate V̇O2 kinetics, the average pul-
monary V̇O2 in 5-s intervals was calculated for each running
bout. The cardiopulmonary phase corresponding to 15–20 s
was removed before modeling of the V̇O2 responses for each
individual (9,15,45). The individual V̇O2 values for the four
transitions were modeled using monoexponential fitting
(Mathcad; Parametric Technology Corp., Needham, MA):
 
V̇O2 ðtÞ ¼ V̇O2 baseline þ A¶ 1  eððtTdÞ=TÞ
where t = time, V̇O2 baseline = average V̇O2 at standing rest,
A¶ = asymptote for the exponential rise in V̇O2, Td = time
delay (s), and T = time constant (s)—63% of time to reach the
final V̇O2 value. In addition, the mean response time (MRT)
was calculated as time delay + T (9). An iterative process
was used to minimize the sum of the squared errors between
the fitted function and the observed values. The 4-min work
period was chosen to avoid fatiguing the subjects before the
RST and the Yo-Yo IR2 test. This procedure allowed for a
calculation of the overall response by the monoexponential
fitting procedure described previously but did not allow a
prober two-component exponential fitting and determina-
tion of the slow component. Instead, the rise in pulmonary
V̇O2 in the last phase of exercise was evaluated as the dif-
ference in V̇O2 from 120–150 to 210–240 s. Running
economy (RE) (mL O2Ikgj1Ikmj1), RER (V̇CO2/V̇O2), and
HR (%HRmax) were calculated during the final 30 s of each
4-min running bout.
RST. The RST was performed indoor on a wooden floor.
A standardized 15-min warm-up procedure was performed
with the last part consisting of two 20-m sprints separated by
1 min of recovery. The RST was then performed 2 min after
the last sprint. The subjects carried out ten 20-m sprints in-
terspersed by 15 s of active recovery. All sprints started from
a standing position. In the recovery period, the subjects ran
slowly back to the starting point before the next sprint. Time
to cover the 20-m sprint was measured with ports of light
sensors (Newtest Powertimers; Newtest Oy, Oulu, Finland).
Fastest sprint time (FST) and total time for all 10 sprints
(TST) were determined. If the fastest sprint was obtained
during the warm-up, this was used as FST. In addition,
a sprint fatigue index (SFI) was calculated as SFI = (1 j
FST
10
TTj1)100%. Furthermore, the reliability of
RST was determined in six subjects, who performed the
RST twice separated by 4 d. The coefficients of variance for
TST, FST, and SFI were 0.7% T 0.2%, 0.6% T 0.2%, and
7.7% T 3.0%, respectively.
Yo-Yo IR2 test. The Yo-Yo IR2 test was performed in-
door on a wooden floor after a 15-min standardized warm-up
http://www.acsm-msse.org
procedure. The Yo-Yo IR2 test consists of 2
20-m shuttle
runs at increasing speeds, interspersed with 10 s of active
recovery controlled by audio signals from a CD (30). The test
was terminated when the subject was no longer able to
maintain the required speed. The total distance covered re-
presented the test result.
Muscle Analysis
Oxidative enzymes. To determine the maximal activity
of citrate synthase (CS) and 3-hydroxyacyl-CoA (HAD),
muscle samples were freeze-dried, and all connective tissue,
visible fat, and blood were removed under a stereomicro-
scope in a room with a temperature of 18-C and a relative
humidity below 30%. About 2 mg dry weight of muscle
tissue was homogenized (1:400) in a 0.3-M phosphate buffer
adjusted to pH 7.7 containing 0.5 mgImLj1 of bovine serum
albumin. Maximal activities of CS and HAD were deter-
mined by using fluorometric methods with nicotinamide
adenine dinucleotide–coupled reactions (Fluoroskan Ascent;
Thermo Scientific, Waltham, MA) (34).
The amount of pyruvate dehydrogenase (PDH) was deter-
mined by western blotting. Approximately 3 mg dry weight
of muscle tissue taken at rest was homogenized in a fresh
batch of buffer (10% glycerol, 20 mM of Na pyrophosphate,
150 mM of NaCl, 50 mM of HEPES, 1% NP-40, 20 mM of
A-glycerophosphate, 10 mM of NaF, 2 mM of phenyl-
methylsulfonyl fluoride, and 1 mM of EDTA and EGTA,
aprotinine, leupeptine, and benzamidine) two times of 30 s
(QIAGEN TissueLyser II; Retsch GmbH, Haan, Germany).
After rotation end over end for 1 h, the samples were
centrifuged for 30 min at 17,500g at 4-C, and the lysate
was collected as the supernatant. Protein concentrations
were determined in the lysates using bovine serum albumin
standards (Pierce Reagents, Chicago, IL). The lysates were
diluted to appropriate protein concentrations in a 6X sam-
ple buffer (0.5 M of Tris base, dithiothreitol (DTT), sodium
dodecyl sulfate, glycerol, and bromphenol blue), and an
equal amount of total protein was loaded for each sample in
different wells on precasted gels (Bio-Rad Laboratories,
Copenhagen, Denmark). For comparisons, samples from
the same subject were always loaded on the same gel. After
gel electrophoresis, the proteins were blotted to a poly-
vinylidene fluoride (PVDF) membrane, which was incubated
with È10 mL of a primary antibody overnight and then
washed for 5 min in Tris-buffered saline–Tween 20 (TBST)
before incubation with a secondary antibody for 1 h. PDH-
E1> protein expression was measured in muscle samples by
sodium dodecyl sulfate polyacrylamide gel electrophoresis
(Tris–HCl 10% gel; Bio-Rad Laboratories) and western blot-
ting using the PVDF membrane and semidry transfer. After
the transfer, the PVDF membrane was blocked overnight at
4-C (TBST + 2% skim milk). The following day, the mem-
brane was incubated with the primary antibody (39) in TBST
+ 2% skim milk for 2 h at room temperature and thereafter
washed in TBST and incubated with a horseradish peroxi-
V̇O2 KINETICS AND PERFORMANCE IN SOCCER PLAYERS
Copyright © 2011 by the American College of Sports Medicine. Unauthorized reproduction of this article is prohibited.
dase–conjugated secondary antibody (Dako, Glostrup, Den-
mark) for 1 h at room temperature (TBST + 2% skim milk).
Immobilon Western (Millipore Corp., Billerica, MA) was
used as a detection system. Bands were visualized using an
Eastman Kodak Co. Image Station 2000MM. Bands were
quantified using the Kodak Molecular Imaging Software
v. 4.0.5 (Eastman Kodak Co., Rochester, NY), and protein
content was expressed in units relative to control samples
loaded on each gel. The changes in PDH are expressed as an
arbitrary unit and relative to the levels before the interven-
tions. On the basis of our experience with variation in results
from western blotting, observations were removed if the delta
ratio (before vs after) differed more than the average delta
ratio T 2 SD for the two groups separately. This resulted in
removal of data from one subject from TC.
Muscle fiber types and size. Ten-micrometer-thick
sections of the muscle samples embedded in Tissue-Tek
were cut at j20-C and incubated for myofibrillar ATPase
reactions at pH 9.4, after preincubation at pH 4.3, 4.6, and
10.3 (13). On the basis of the myofibrillar ATP staining,
slow-twitch (ST) and fast-twitch a, x, and c (FTa, FTx, and
FTc) fiber types were defined under light microscopy. The
number and size of fibers were determined using a computer
program (TEMA 1995; CheckVision, Hadsund, Denmark).
On average, 170 muscle fibers were included in each of the
four analyses before and after HI and TC.
Statistics
Differences between the two groups HI and TC before the
intervention period were examined by an unpaired t-test.
Changes in V̇O2 kinetics, RE, RER, enzyme activity, and
muscle fiber distribution and size were examined by a paired
Student’s t-test separately for HI and TC. Changes in TST
and SFI in RST and Yo-Yo IR2 performance were tested by
a two-way ANOVA for repeated measurements (one-factor
repetition) with group (HI vs TC) and time (before vs after)
as factors. Changes in Yo-Yo IR2 performance for TC were
examined by a one-way ANOVA for repeated measure-
ments (before, 72 h, after). When a significant main effect
was detected in the one- or two-way ANOVA repetition
maximum, a Student–Newman–Keuls post hoc analysis was
performed for pairwise multiple comparison. The Pearson
correlation coefficient was calculated to test the relation-
ship between two variables. All the statistical analyses
APPLIED SCIENCES
were made in SigmaPlot version 11.0 (Systat Software, Inc.,
Chicago, IL), and the level of statistical significance was
set at P G 0.05. Data are expressed as means T SD.
RESULTS
V̇O2 kinetics. V̇O2 kinetics was unaffected in HI
(Fig. 1), but in the last 30 s of the 4-min exercise bout, the
oxygen uptake was lower (P G 0.05), and the RE was im-
proved after the intensified training period (197.8 T 10.2
(before) vs 192.8 T 7.2 (after) mL O2Ikgj1Ikmj1, P G 0.05)
(Table 1). After TC, V̇O2 kinetics became slower (P G 0.05)
Medicine & Science in Sports & Exercised 1719
 uptake during the last 2 min was not changed after the inter-
vention period either in HI (72 T 48 vs 63 T 33 mL O2Iminj1)
or in TC (101 T 51 vs 91 T 44 mL O2Iminj1).
HR response and RER during submaximal run-
ning. HR and RER during submaximal running were not
changed in HI. In TC, HR was elevated (P G 0.05) throughout
exercise, and RER was higher (P G 0.05) from 80 s of exer-
cise after the inactivity period (Table 1).
RST and Yo-Yo IR2 test performance. Performance
in the RST is displayed in Figure 2. In HI, the fastest sprint
(3.15 T 0.08 and 3.16 T 0.13 s) and fatigue index (5.8% T
2.8% and 3.9% T 1.6%) were not changed, but performance
was improved (P G 0.05) in the fourth (3.35 T 0.13 vs 3.26 T
0.14 s), sixth (3.43 T 0.21 vs 3.32 T 0.10 s), and tenth (3.39 T
0.20 vs 3.29 T 0.12 s) sprints. In addition, total sprint time
was reduced (P G 0.05) after the intervention period (33.44 T
1.17 vs 32.81 T 1.01 s). In TC, the fastest sprint (3.14 T 0.08
and 3.15 T 0.14 s) and fatigue index (5.9% T 2.3% and
7.6% T 2.8%) were the same. Performance was reduced
(P G 0.05) in the fifth (3.38 T 0.11 vs 3.46 T 0.14 s), sixth
(3.41 T 0.15 vs 3.48 T 0.11 s), seventh (3.41 T 0.13 vs
3.50 T 0.11 s), eighth (3.42 T 0.13 vs 3.50 T 0.12 s), ninth
(3.42 T 0.10 vs 3.56 T 0.09 s), and tenth (3.35 T 0.12 vs
3.46 T 0.09 s) sprints. Total sprint time was also longer
(P G 0.01) after the inactivity period (33.41 T 0.96 vs
34.11 T 0.92 s). In HI, no change in performance of the
Yo-Yo IR2 test was observed (937 T 56 and 994 T
72 m), whereas it was reduced (P G 0.01) in TC from

FIGURE 1—Mean values T SD and individual values of the fast com-

ponent of oxygen uptake kinetics during running at 75% of MAS for
trained soccer players before (pre) and after (post) 2 wk of high-intensity
training (HI, left) or training cessation (TC, right). A. The time constant
(T) describing the time to reach 63% of the final V̇O2 value after deletion
of the initial time delay. B. MRT (MRT = time delay + T). *P G 0.05, post
significantly different from pre.

reflected as an increase in the time constant from 21.5 T 2.9 to

23.8 T 3.2 s and an increase in the MRT from 45.0 T 1.8 to
46.8 T 2.2 s (Fig. 1 and Table 1). The increase in the oxygen

TABLE 1. V̇O2 kinetics, pulmonary V̇O2, RE, RER, and HR in trained soccer players
running for 4 min at 75 % MAS before and after 2 wk of high-intensity training group
(HI, n = 7) and training cessation group (TC, n = 11).
APPLIED SCIENCES

HI TC
Pre Post Pre Post
Time delay (s) 21.9 T 3.5 24.5 T 3.3 23.6 T 3.0 23.0 T 2.8
T (s) 21.7 T 2.5 18.8 T 4.0 21.5 T 2.9 23.8 T 3.2*
A¶ (mL O2Iminj1) 3556 T 324 3484 T 353** 3583 T 294 3566 T 289
pV̇O2 at 4 min 3596 T 353 3523 T 351** 3643 T 280 3608 T 289
(mL O2Iminj1)
RE at 4 min 197.8 T 10.2 192.8 T 7.2* 196.6 T 7.0 195.2 T 7.2
(mL O2Ikgj1Ikmj1)
RER at 4 min 0.94 T 0.03 0.95 T 0.03 0.93 T 0.03 0.97 T 0.03**
HR at 4 min (%HRmax) 83.2 T 1.6 82.3 T 1.9 84.0 T 3.7 86.7 T 3.6***
Values are means T SD. Values at 4 min are calculated as the average during the last FIGURE 2—Mean sprint times T SD during 10
20-m sprinting with
30 s of each interval. 15 s of recovery before and after 2 wk of high-intensity training (HI, A) or
* P G 0.05 and ** P G 0.01, *** P G 0.001, post significantly different from pre. training cessation (TC, B). *P G 0.05, post significantly different from pre.

1720 Official Journal of the American College of Sports Medicine http://www.acsm-msse.org

Copyright © 2011 by the American College of Sports Medicine. Unauthorized reproduction of this article is prohibited.
845 T 160 m before the intervention to 801 T 162 and
654 T 99 m after 3 and 14 d of inactivity, respectively.
V̇O2 kinetics and performance during repeated
intense work. Before the interventions, there was no cor-
relation between V̇O2 kinetics (expressed as T and MRT,
respectively) and total sprint time (r2 = 0.03 and 0.001, NS),
fatigue index (r2 = 0.02 and 0.02, NS), and Yo-Yo IR2
performance (r2 = 0.002 and 0.003, NS). A correlation
(P G 0.05) was observed between the changes in V̇O2 ki-
netics and in fatigue index (r2 = 0.43 for T and 0.27 for
MRT) and total sprint time and T (r2 = 0.25) but not MRT
(r2 = 0.21, NS). No correlation was found between the
changes in T and MRT and the change in Yo-Yo IR2 per-
formance (r2 = 0.18 and 0.03, NS).
Muscle enzymes and fiber type distribution. In HI,
FIGURE 4—Mean values T SD and individual values of the change in the
the maximal activity of CS (30.8 T 2.8 and 32.7 T 3.6 Kmol amount of PDH in trained soccer players after 2 wk of high-intensity
per dry weight per minute) and HAD (23.7 T 2.1 and 22.5 T training group (HI, left, n = 7) or training cessation group (TC, right,
2 Kmol per dry weight per minute) did not change (Fig. 3), n = 8). *P G 0.05, **P G 0.01, post significantly different from pre. AU,
arbitrary units.
whereas the amount of PDH was elevated by 17% T 14%
from before level (P G 0.05) after the intervention period
(Fig. 4). In TC, CS (32.7 T 2.9 and 28.8 T 4.8 Kmol per (P G 0.01) by 12% and 18% after the inactivation period, and
dry weight per minute) and HAD (25.3 T 3.6 and 20.8 T the amount of PDH decreased by 17% T 11% (P G 0.01)
3.1 Kmol per dry weight per minute, P G 0.05) were lowered from before level (Figs. 3 and 4).
In HI, no changes in muscle fiber distribution and size
were observed (Table 2). In TC, the relative number (56% T
18% and 47% T 15%) and area (51% T 18% and 39% T
16%) of ST fibers were lowered (P G 0.05) after the inacti-
vation period, whereas fiber size was unaltered (Table 2).

DISCUSSION
The main findings of the present study performed on
trained soccer players were impaired V̇O2 kinetics after 2 wk
of training cessation, which was associated with a lowered

TABLE 2. Muscle fiber distribution (%), percent area, and muscle fiber size in trained
soccer players before and after 2 wk of high-intensity training group (HI, n = 6) and
training cessation group (TC, n = 9).
HI
Distribution (%) Distribution (% area) Fiber Size (Km)
ST Pre 65 T 18 60 T 20 3958 T 421
Post 61 T 14 55 T 16 4285 T 1131
FTa Pre 32 T 17 36 T 18 5169 T 879
Post 35 T 11 41 T 13 5600 T 657
FTx Pre 3 T 4 3 T 4 4833 T 1531 APPLIED SCIENCES
Post 3 T 3 3 T 2 4572 T 575
FTc Pre 2 T 3 1 T 1 3373 T 247
Post 1 T 1 1 T 1 3430 T 1284
TC
Distribution (%) Distribution (% area) Fiber size (mm)
ST Pre 56 T 18 51 T 18 4556 T 776
Post 47 T 15* 39 T 16* 4285 T 776
FTa Pre 33 T 16 39 T 16 6054 T 1273
Post 37 T 19 45 T 12 6087 T 960
FTx Pre 8T8 9T9 5771 T 1419
Post 13 T 7 14 T 8 5160 T 840
FIGURE 3—Mean values T SD and individual values of maximal activity FTc Pre 2T4 2T4 4993 T 1706
Post 2T4 2T3 3603 T 567
of HAD (A) and CS (B) for trained soccer players before and after 2 wk
of high-intensity training group (HI, left, n = 6) or training cessation Values are means T SD.
group (TC, right, n = 7). *P G 0.05, post significantly different from pre. * P G 0.05, post significantly different from pre.

V̇O2 KINETICS AND PERFORMANCE IN SOCCER PLAYERS Medicine & Science in Sports & Exercised 1721

Copyright © 2011 by the American College of Sports Medicine. Unauthorized reproduction of this article is prohibited.
 activity and amount of oxidative enzymes as well as a re-
duced performance in repeated sprint work and the Yo-Yo
IR2 test. In addition, 2 wk of intensified training improved
RE as well as performance in an RST and content of PDH.
To our knowledge, this is the first controlled study, using
several exercise bouts to describe V̇O2 kinetics for elite
athletes, showing slower V̇O2 kinetics after a period of in-
activity. Convertino et al. (17) found that V̇O2 kinetics was
slower after 7 d of bed rest in untrained subjects with a
V̇O2max of 38 mL O2Ikgj1Iminj1 before the bed rest period,
although it should be mentioned that only one work bout
was used to estimate V̇O2 kinetics, which may reduce the
validity of the calculations (33). In contrast to the present
finding, several studies did not find any effect of inactivity
on V̇O2 kinetics (16,24). A possible explanation is the low
training status of the subjects, indicating that they had lim-
ited activities already before the intervention period. Al-
though the fitting procedure used in the present study does
not allow for a clear distinction between fast and slow
components of the V̇O2 response, the finding that the rise in
V̇O2 during the last 2 min of exercise was unchanged after
the inactivity period suggests that the slower V̇O2 kinetics
after inactivity was primarily related to the early phase of
exercise. There is good evidence to support that a limited
extraction of oxygen by the contracting muscle cells causes
the delay in oxygen utilization in the initial phase of exercise
(7,20,29,40), often termed ‘‘metabolic inertia.’’ The slower
V̇O2 kinetics in the present study was associated with a 17%
decrease in the amount of PDH and a lowering of the max-
imal activity of CS and HAD by 12% and 18%, respectively.
In agreement with this, several studies using short periods of
detraining on well-trained subjects have shown reductions in
oxidative enzymes (37). The reduction in PDH and CS may
have slowed the V̇O2 kinetics, but it is questionable whether
these enzymes are limiting for muscle oxygen uptake per se.
In several studies where the PDH activity was increased by
ingestion of dichloroacetate, no effect was seen on muscle
and pulmonary V̇O2 (3,27). In addition, when correlating the
changes in T with the alterations in PDH and CS in the in-
activity group, no correlations were observed, and in the HI,
PDH was elevated by 17% after the 2-wk training period
without V̇O2 kinetics being changed. Furthermore, after 5 d
of training V̇O2 kinetics was improved in untrained subjects
despite no changes in CS (38). The lowering of the HAD
activity probably also had little influence on the slower V̇O2
APPLIED SCIENCES
kinetics because glucose metabolism dominates while run-
ning at 75% MAS (È85% V̇O2max) (1), which is supported
by the high RER values of 0.93–0.97 at the end of the run-
ning bouts (Table 1). Nevertheless, further studies also in-
cluding measurements of oxygen delivery are needed to
clarify the effect of inactivity on V̇O2 kinetics. After TC, the
distribution of ST fibers was lowered. This is in contrast to
observations in some other studies with detraining of trained
subjects where fiber type distribution is reported to be
unaltered (37) but in agreement with observations after
11 d of space flight in astronauts (46), after 3 wk of knee
1722 Official Journal of the American College of Sports Medicine
Copyright © 2011 by the American College of Sports Medicine. Unauthorized reproduction of this article is prohibited.
immobilization in recreationally active subjects (23), and
6 wk after knee surgery in trained athletes (21). Neverthe-
less, it may be speculated that a larger recruitment of FT
fibers during exercise may have slowed V̇O2 kinetics be-
cause support for slower V̇O2 kinetics in FT fibers can be
found in the literature. Thus, an increase in T during mod-
erate work after inactivation of ST fibers by cisatracurium
has been observed (32), and T is often reported higher when
the relative workload is increased (28,29), which may be
explained by a higher activation of the FT fibers.
An interesting finding was that the RE was improved after
just 2 wk with 10 high-intensity training sessions substitut-
ing the normal training of the soccer players. Similarly, two
recent studies with trained runners performing speed en-
durance production training consisting of runs at 95% of
maximal speed for 30 s either for 4 (25) or 6–9 wk in
combination with AHI training (4) found an improved RE. It
may be related to a specific adaptation of the high-order
motor units (FT fibers), e.g., an improved P:O ratio, because
these fibers were likely maximally taxed during the sprints
used in the training. In support, Henriksson and Reitman
(22) have demonstrated a training-specific fiber adaptation
in the aerobic enzyme succinate dehydrogenase (SDH) after
an 8-wk training period. However, this issue warrants more
investigation. The high-intensity training was not sufficient
to improve the V̇O2 kinetics in the soccer players in the pre-
sent study, which may have been because of the high initial
fitness level and the short intervention period. Many studies
involving training of untrained subjects have shown im-
provement within 2 wk (2,38), and intervention periods last-
ing 4–8 wk have shown faster kinetics in moderately (11,29)
and highly trained subjects (18).
Previous soccer studies have demonstrated a correlation
between T at 60% MAS and performance in both Yo-Yo IR2
(41) and repeated 40-m sprints (19,42). In the present study,
no association was observed before the intervention between
the V̇O2 kinetics and performance in repeated intense exer-
cise, which may be related to the protocol used to determine
V̇O2 kinetics and intermittent exercise performance. We
used four repetitions at 75% MAS in comparison with two
repetitions at 60% MAS (18,41) and a 20-m sprint distance
making the aerobic contribution to the total energy turnover
less pronounced compared with the 40-m sprints applied in
the previous studies (19,42). Considering that the time
constants become greater with increasing work intensity
(28,29), the kinetics was fast in the present study with time
constants of È22 s before the interventions in comparison
with values of 23 and 27 s in the other studies using 60%
MAS (19,41,42). Interestingly, the observed changes in T
and MRT after the two interventions were correlated with
the changes in RST performance, and Yo-Yo IR2 was
markedly reduced after inactivity in association with slower
V̇O2 kinetics, confirming a link between V̇O2 kinetics and
intermittent exercise capacity.
In summary, 2 wk of training cessation of elite soccer
players led to slower V̇O2 kinetics, which was associated
http://www.acsm-msse.org
with a reduced content and activity of oxidative enzymes
and a lower fraction of ST fibers. In addition, performances
in the Yo-Yo IR2 test and the RST were also impaired. A 2-wk
training period with 10 high-intensity training sessions sub-
stituting the normal training improved work efficiency during
submaximal running and performance during repeated
sprinting. No association between fast V̇O2 kinetics and per-
formance during repeated high-intensity work was observed,
REFERENCES
1. Achten J, Gleeson M, Jeukendrup AE. Determination of the ex-
ercise intensity that elicits maximal fat oxidation. Med Sci Sports
Exerc. 2002;34(1):92–7.
2. Bailey SJ, Wilkerson DP, Dimenna FJ, Jones AM. Influence of re-
peated sprint training on pulmonary O2 uptake and muscle deoxy-
genation kinetics in humans. J Appl Physiol. 2009;106:1875–87.
3. Bangsbo J, Gibala MJ, Krustrup P, González-Alonso J, Saltin B.
Enhanced pyruvate dehydrogenase activity does not affect muscle
O2 uptake at onset of intense exercise in humans. Am J Physiol
Regul Integr Comp Physiol. 2002;282:R273–80.
4. Bangsbo J, Gunnarsson TP, Wendell J, Nybo L, Thomassen M.
Reduced volume and increased training intensity elevate muscle
Na+-K+ pump alpha2-subunit expression as well as short- and
long-term work capacity in humans. J Appl Physiol. 2009;107:
1771–80.
5. Bangsbo J, Iaia FM, Krustrup P. Metabolic response and fatigue in
soccer. Int J Sports Physiol Perform. 2007;2:111–27.
6. Bangsbo J, Iaia FM, Krustrup P. The Yo-Yo intermittent recovery
test: a useful tool for evaluation of physical performance in inter-
mittent sports. Sports Med. 2008;38:37–51.
7. Bangsbo J, Krustrup P, González-Alonso J, Boushel R, Saltin B.
Muscle oxygen kinetics at onset of intense dynamic exercise in
humans. Am J Physiol Regul Integr Comp Physiol. 2000;279:
R899–906.
8. Bangsbo J, Krustrup P, González-Alonso J, Saltin B. ATP pro-
duction and efficiency of human skeletal muscle during intense
exercise: effect of previous exercise. Am J Physiol Endocrinol
Metab. 2001;280:E956–64.
9. Bearden SE, Moffatt RJ. V̇O2 and heart rate kinetics in cycling:
transitions from an elevated baseline. J Appl Physiol. 2001;90:
2081–7.
10. Berger NJ, Tolfrey K, Williams AG, Jones AM. Influence of
continuous and interval training on oxygen uptake on-kinetics.
Med Sci Sports Exerc. 2006;38(3):504–12.
11. Billat VL, Mille-Hamard L, Demarle A, Koralsztein JP. Effect of
training in humans on off- and on-transient oxygen uptake kinetics
after severe exhausting intensity runs. Eur J Appl Physiol. 2002;
87:496–505.
12. Bogdanis GC, Nevill ME, Boobis LH, Lakomy HK. Contribution
of phosphocreatine and aerobic metabolism to energy supply dur-
ing repeated sprint exercise. J Appl Physiol. 1996;80:876–84.
13. Brooke MH, Kaiser KK. Three ‘‘myosin adenosine triphosphatase’’
systems: the nature of their pH lability and sulfhydryl dependence.
J Histochem Cytochem. 1970;18:670–2.
14. Burnley M, Koppo K, Jones AM. ‘‘Priming exercise’’ and V̇O2
kinetics. In: Jones AM, Poole DC, editors. Oxygen Uptake Kinetics
in Sport, Exercise and Medicine. Abingdon (UK) Routledge,
Taylor & Francis Group; 2005. p. 230–60.
15. Burnley M, Roberts CL, Thatcher R, Doust JH, Jones AM. Influ-
ence of blood donation on O2 uptake on-kinetics, peak O2 uptake
and time to exhaustion during severe-intensity cycle exercise in
humans. Exp Physiol. 2006;91:499–509.
16. Capelli C, Adami A, Antonutto G, Cautero M, Tam E. Oxygen
deficits and oxygen delivery kinetics during submaximal intensity
V̇O2 KINETICS AND PERFORMANCE IN SOCCER PLAYERS
Copyright © 2011 by the American College of Sports Medicine. Unauthorized reproduction of this article is prohibited.
but the changes found after the interventions indicate that
they may be related.
This work was supported by the Danish Ministry of Culture
(Kulturministeriets Udvalg for Idr&tsforskning).
There are no conflicts of interest.
The authors thank Jens Jung Nielsen, Rikke Leihof, and Benjamin
Leisvig for excellent technical assistance.
The results of the present study do not constitute endorsement
by the American College of Sports Medicine.
exercise in humans after 14 days of head-down tilt-bed rest. Eur
J Appl Physiol. 2009;107:51–9.
17. Convertino VA, Goldwater DJ, Sandler H. V̇O2 kinetics of con-
stant-load exercise following bed-rest-induced deconditioning.
J Appl Physiol. 1984;57:1545–50.
18. Demarle AP, Slawinski JJ, Laffite LP, Bocquet VG, Koralsztein
JP, Billat VL. Decrease of O2 deficit is a potential factor in in-
creased time to exhaustion after specific endurance training. J Appl
Physiol. 2001;90:947–53.
19. Dupont G, Millet GP, Guinhouya C, Berthoin S. Relationship be-
tween oxygen uptake kinetics and performance in repeated running
sprints. Eur J Appl Physiol. 2005;95:27–34.
20. Fukuba Y, Ohe Y, Miura A, et al. Dissociation between the time
courses of femoral artery blood flow and pulmonary V̇O2 during
repeated bouts of heavy knee extension exercise in humans. Exp
Physiol. 2004;89:243–53.
21. Häggmark T, Eriksson E, Jansson E. Muscle fiber type changes in
human skeletal muscle after injuries and immobilization. Ortho-
pedics. 1986;9:181–5.
22. Henriksson J, Reitman JS. Quantitative measures of enzyme activi-
ties in type I and type II muscle fibres of man after training. Acta
Physiol Scand. 1976;97:392–7.
23. Hortobágyi T, Dempsey L, Fraser D, et al. Changes in muscle
strength, muscle fibre size and myofibrillar gene expression after
immobilization and retraining in humans. J Physiol. 2000;524(pt 1):
293–304.
24. Hotta N, Sato K, Katayama K, et al. Oxygen uptake kinetics fol-
lowing 20 days of unilateral lower limb suspension. J Physiol Sci.
2006;56:347–53.
25. Iaia FM, Hellsten Y, Nielsen JJ, Fernström M, Sahlin K, Bangsbo J.
Four weeks of speed endurance training reduces energy expendi-
ture during exercise and maintains muscle oxidative capacity
despite a reduction in training volume. J Appl Physiol. 2009;106:
73–80.
26. Iaia FM, Thomassen M, Kolding H, et al. Reduced volume but
increased training intensity elevates muscle Na+-K+ pump alpha1-
subunit and NHE1 expression as well as short-term work capacity
in humans. Am J Physiol Regul Integr Comp Physiol. 2008;294:
R966–74.
27. Jones AM, Koppo K, Wilkerson DP, Wilmshurst S, Campbell IT. APPLIED SCIENCES
Dichloroacetate does not speed phase-II pulmonary V̇O2 kinetics
following the onset of heavy intensity cycle exercise. Pflugers
Arch. 2004;447:867–74.
28. Koppo K, Bouckaert J, Jones AM. Effects of training status and
exercise intensity on phase II V̇O2 kinetics. Med Sci Sports Exerc.
2004;36(2):225–32.
29. Krustrup P, Hellsten Y, Bangsbo J. Intense interval training
enhances human skeletal muscle oxygen uptake in the initial phase
of dynamic exercise at high but not at low intensities. J Physiol.
2004;559:335–45.
30. Krustrup P, Mohr M, Nybo L, Jensen JM, Nielsen JJ, Bangsbo
J. The Yo-Yo IR2 test: physiological response, reliability, and
application to elite soccer. Med Sci Sports Exerc. 2006;38(9):
1666–73.
Medicine & Science in Sports & Exercised 1723
 31. Krustrup P, Mohr M, Steensberg A, Bencke J, Kjaer M, Bangsbo J.
Muscle and blood metabolites during a soccer game: implications
for sprint performance. Med Sci Sports Exerc. 2006;38(6):1165–74.
32. Krustrup P, Secher NH, Relu MU, Hellsten Y, Söderlund K,
Bangsbo J. Neuromuscular blockade of slow twitch muscle fibres
elevates muscle oxygen uptake and energy turnover during sub-
maximal exercise in humans. J Physiol. 2008;586:6037–48.
33. Lamarra N, Whipp BJ, Ward SA, Wasserman K. Effect of in-
terbreath fluctuations on characterizing exercise gas exchange ki-
netics. J Appl Physiol. 1987;62:2003–12.
34. Lowry OH, Passonneau JV. A Flexible System of Enzymatic
Analysis. New York (NY): Academic Press; 1972. p. 237–49.
35. McInnes SE, Carlson JS, Jones CJ, McKenna MJ. The physio-
logical load imposed on basketball players during competition.
J Sports Sci. 1995;13:387–97.
36. Mohr M, Krustrup P, Bangsbo J. Match performance of high-
standard soccer players with special reference to development of
fatigue. J Sports Sci. 2003;21:519–28.
37. Mujika I, Padilla S. Detraining: loss of training-induced physio-
logical and performance adaptations. Part I: short term insufficient
training stimulus. Sports Med. 2000;30:79–87.
38. Phillips SM, Green HJ, MacDonald MJ, Hughson RL. Progressive
effect of endurance training on V̇O2 kinetics at the onset of sub-
maximal exercise. J Appl Physiol. 1995;79:1914–20.
APPLIED SCIENCES
1724 Official Journal of the American College of Sports Medicine
Copyright © 2011 by the American College of Sports Medicine. Unauthorized reproduction of this article is prohibited.
39. Pilegaard H, Birk JB, Sacchetti M, et al. PDH-E1alpha dephos-
phorylation and activation in human skeletal muscle during exer-
cise: effect of intralipid infusion. Diabetes. 2006;55:3020–7.
40. Poole DC, Barstow TJ, McDonough P, Jones AM. Control of
oxygen uptake during exercise. Med Sci Sports Exerc. 2008;40(3):
462–74.
41. Rampinini E, Sassi A, Azzalin A, et al. Physiological determinants
of Yo-Yo intermittent recovery tests in male soccer players. Eur J
Appl Physiol. 2010;108:401–9.
42. Rampinini E, Sassi A, Morelli A, Mazzoni S, Fanchini M, Coutts
AJ. Repeated-sprint ability in professional and amateur soccer
players. Appl Physiol Nutr Metab. 2009;34:1048–54.
43. Sirotic AC, Coutts AJ, Knowles H, Catterick C. A comparison of
match demands between elite and semi-elite rugby league com-
petition. J Sports Sci. 2009;27:203–11.
44. Tomlin DL, Wenger HA. The relationships between aerobic fit-
ness, power maintenance and oxygen consumption during intense
intermittent exercise. J Sci Med Sport. 2002;5:194–203.
45. Whipp BJ, Ward SA, Lamarra N, Davis JA, Wasserman K.
Parameters of ventilatory and gas exchange dynamics during ex-
ercise. J Appl Physiol. 1982;52:1506–13.
46. Zhou MY, Klitgaard H, Saltin B, Roy RR, Edgerton VR, Gollnick
PD. Myosin heavy chain isoforms of human muscle after short-
term spaceflight. J Appl Physiol. 1995;78:1740–4.
http://www.acsm-msse.org