Advances in Physical Education
2013. Vol.3, No.1, 43-49
Published Online February 2013 in SciRes (
Copyright © 2013 SciRes. 43
Changes of Selected Haematological Parameters in a Female
Team during the 25th African Handball Winners’ Cup Played at
Cotonou (Benin)
Issiako Bio Nigan1*, Polycarpe Gouthon1, Mansourou Arèmou1, Jean-Marie Falola2,
Houndjovi Pierre Dansou3, Jean Koudouvo Houngbélagnon1, Basile Kocou Nouatin1,
Brigitte Affidéhomè Tonon1, Raïmath Yon-Taro Bio Nigan4
1Laboratory of “PSA and Motricity”, National Institute of Youth, Physical Education and Sport (INJEPS),
University of Abomey-Calavi, Cotonou, Benin
2Laboratory of Biomechanics and Performance (LABIOP), National Institute of Youth, Physical Education and
Sport (INJEPS), University of Abomey-Calavi, Cotonou, Benin
3Laboratory of Exercise Physiology, National Institute of Youth, Physical Education and Sport (INJEPS),
University of Abomey-Calavi, Cotonou, Benin
4Pediatrics Department of the Regional Hospital of Kouandé, Kouandé, Benin
Email: *
Received November 17th, 2012; revised December 20th, 2012; accepted January 4th, 2013
The performance in team sports partly depends on the oxygen transport capacity that is also associated
with the erythrocyte values, which should be regularly assessed throughout the sports season. This con-
trolled observational study aims at measuring the haematologic modifications induced by the physical
load associated with a series of matches in a Benin Division 1 female handball team. It was carried out at
Cotonou during the 25th African Handball Winners’ Cup, with 22 female handball players of the Be-
ninese champion team, divided into an experimental group (EG: n = 16) and a control one (CG: n = 6).
The red blood cell count (RBC), the rates of Hb (Hb) and haematocrit (Ht), the Wintrobe’s constants and
four iron parameters were assessed prior to and after a series of four matches played in six days. At the
end of the competition, RBC increased significantly (p < 0.05), but the increases in Hb and Ht were
non-significant (p > 0.05). Serum iron, serum transferrin and total iron binding capacity decreased sig-
nificantly (p < 0.05). The results suggest an improvement in the players’ diet by highly protein enriched
foods and erythropoiesis raw materials. They also suggest that the girls may adopt adequate hydric prac-
tices, in order to compensate for water losses during matches and training sessions.
Keywords: Erythrocytes; Iron; Competition; Handball; Tropical Climate
Sports performance strongly depends on the oxygen trans-
portation capacity to supply exercising muscles. This capacity
is associated with the erythrocyte values, which may thus be
regularly assessed throughout the sports season (Fallon, 2004;
Lesesve et al., 2000) to allow trainers and medical staff mem-
bers to collect useful fitness and health related information on
players. Many factors (environment, seasons, physiological
state, nutritional profile, alcoholism, etc.) tend to modify hae-
matological parameters, which sometimes leads to anaemia
(Malcovati et al., 2003; Rietejen, 2002). In a sportsman prac-
ticing an intense long duration activity, water losses are ac-
companied by a decrease in iron store. In case this situation
persists, it may lead to anaemia, as the sport considered requires
important energy expenditure. More than one quarter of the
male and the three quarters of the female long distance race
specialists would be affected by iron deficiency (Clement &
Sawchuck, 1984). The decrease in iron stores is indeed deeper
in sporting women, because of the periodic menstruation which
represents a source of blood loss. A decrease in Hb and haema-
tocrit rates and red blood cell count was also recorded among
runners and swimmers, in spite of a sufficient iron dietary in-
take (Pizza et al., 1997). The deficiencies may also appear in an
environment like that of Benin where the prevalence of malnu-
trition and enteric parasites is high and malaria endemic (IN-
SAE/Benin, 2012; Ministère de la Santé Publique du Bénin,
2009; Kindé-Gasard et al., 2000). On the first hand, the hae-
molysis and enteric bleedings that these last affections are
likely to cause may induce a decrease in the Hb rate (Dillon,
2000). On the other hand, the higher an individual’s Hb rate,
the stronger his/her oxygen transportation capacities (Gledhill
et al., 1999). Many studies reported the effects of a training and
competition period on the erythrocyte values and the iron status
in long distance runners, swimmers, cyclists and soccer players
(Santhiago et al., 2009; Silva et al., 2008; Zapico et al., 2007).
In most of these studies, RBC, Hb and serum transferrin in-
crease at the end of a high-level intensity exercise (Ashenden et
al., 1999; Cordova & Escanero, 1992). The impact of the phy-
sical load associated with a tournament in team sports like
handball on the haematological parameters is not well known,
mainly in an environment like that of Benin, where it is likely
to find a high prevalence of anaemia. When they exist, studies
concern more male sportsmen (Santhiago et al., 2009; Silva et
*Corresponding author.
al., 2008; Wilkinson et al., 2002) or other sports than handball
(Douglas, 1989). In Benin, the modifications of the erythrocyte
values during the periods of competition and periods of break
(Gouthon et al., 2007) or during the first phase of the Division
1 championship (Gouthon et al., 2011), were assessed only in
soccer players. The modifications of these parameters induced
by a competition in clusters of female handball players during a
continent-level championship were never measured. However,
these players are likely to be a population at risk of anaemia,
due to their status of non-menopausal women, and to their daily
environment that favours the development of malaria and other
transmissible diseases (Fallon, 2004). The risk of physiological
disturbance is all the higher as the competition schedule does
not allow enough recovery between the matches and the diet
proposed by the restaurateurs does not often correspond to the
food habits of the players. Under these conditions, some players
feed badly throughout African competitions, in spite of the
costly meals offered by hotels (Bio Nigan et al., 2011). The
objective of this study is thus to measure the haematological
modifications induced by the repeated physical load related to a
series of matches played by the Benin Division 1 female hand-
ball team during the 25th African Handball Winners’ Cup.
Materials and Methods
Nature of the Study an d Se tt ings
It was a controlled observational study, carried out in May
2009 in Cotonou, Southern Benin (West Africa), during the
25th African Handball Winners’ Cup. The matches were played
indoor, at a mean temperature of 30˚C ± 2˚C with a relative
humidity of 70% ± 5%.
Study Sample
The study sample was composed of the 22 female players of
the Association Sportive Omnisports (ASO) Baobab which
represented the Republic of Benin in this continental level
competition. The players were divided into two groups: an ex-
perimental group (EG) composed of the 16 players who played
at least a match and a control group (CG) constituted with the 6
girls who took part in only two training sessions scheduled for
the team during break time.
Seventy two hours before the beginning of the competition,
the players produced a written informed consent to participate in
the study and gave information related to their oestrian status.
The morning before the first match scheduled for 9 a.m., the first
faecal samples were collected in individual test tubes to seek the
presence of enteric worms by a coprology test. Height (near to a
cm) and weight (near to 100 g) were then measured, using re-
spectively a stadiometer and a digital bio-impedancemeter BG
22 (Beurer, Germany). A multifunction station (Meteo-Star) ins-
talled near the playing ground made it possible to record the
ambient temperature and the relative humidity during the mat-
ches and training sessions. Resting heart rate (HRr) was taken by
the Physician of the team on an electronic monitor UA-767 plus
30 (AND A&D Medical, Japan) in the players, seated for 10 min
at least in a quiet room. A blood sample of 5 mL was drawn out
after a 12 hour-fasting by venepuncture at the fold of the left
elbow, to assess erythrocyte values and evaluate their malarial
and iron status.
The erythrocyte values (red blood cell count or RBC, Hb and
haematocrit rates, Wintrobe’ constants) were assessed using an
automat counter M-Series (Medonic, Sweden). The iron para-
meters i.e. serum iron (Fe), serum transferrin (TR), ferritin
(FERI), the rate of transferrin saturation (RTS) and the total
iron binding capacity (TBC), protidemia (PTD) and plasma
albumin (ALB) were assessed by the enzymatic and colorimet-
ric method, using a spectrophotometer RT-9200 (Rayto, Ger-
many). The same operations were repeated under the same con-
ditions 36 hours after the last match (Figure 1).
During the matches and the two training sessions, the energy
expenditure was evaluated with pedometers F700 (Geonaute,
China). The meals consumed were weighed and the samples
analysed in bomb calorimeters to determine the daily energy,
vitamin C and iron consumption in each player. The water
drunk was measured using individual bottles of 1.5 L given to
each player at the beginning of each match and training session.
Statistical Analysis
The data were processed with the software Statistica of Stat
Soft Inc. (version 5.5). The normality of the distribution of the
variables was checked using the test of Kolmogorov Smirnov.
The mean value (M) and the standard error of the mean (SEM)
were calculated for each variable. The distribution of the vari-
ables not being normal, the Wilcoxon test was used for com-
paring measures within the same group, and the Mann-Whitney
U test, for inter-group comparisons. Otherwise, the parametric
tests were used. The level of significance of the statistical tests
was settled at p < 0.05.
Biometric Characteristics and the Results of the
There was no significant difference between the EG and CG
72 H before 1st match
- Contact with the subjects
- Written informed consent
- Oestrian status
2 H before 1st match
- Cardiovascular parameters
- Anthropometric measures
- Blood and faecal sampling
sang et selles
Every day
- Data of match
- Survey on dietary intake
- Ambient temperature and
relative humidity
36 H after last match
- Cardiovascular parameters
- Anthropometric measures
- Blood and faecal sampling
Prior to competition During competition After competition
H: hour
Figure 1.
Schematic presentation of the study design.
Copyright © 2013 SciRes.
groups (p > 0.05), so far as the biometric characteristics were
concerned (Table 1). The ASO Baobab team was eliminated
after having lost their three matches of the preliminary round
against Héritage of Congo (23 versus 25 goals), TKC of Came-
roun (24 versus 30 goals), and Electro of Angola (15 versus 29
goals), as well as their last match against Rombo of Côte d’i-
voire (30 versus 31 goals).
General Data of the Matches, Dietary Patterns,
Oestrian and Parasitic Status
As it is shown in Table 2, the EG and CG groups were dif-
ferent with regard to the data collected during the matches
and/or training sessions, i.e. the mean distance covered, the
mean energy expenditure and the average quantities of water
consumption were all higher in EG than in CG (p < 0.05). No
significant difference (p > 0.05) was observed between the two
groups regarding the daily energy, iron intake and C vitamin
intake (Table 3).
At the beginning of the competition, 14 players of EG were
in the follicular phase of their menstrual cycle versus two in the
luteal phase. In CG, they were four in the follicular phase ver-
sus two in the luteal phase. Protidemia was reduced by 8.7 ±
8.5 g/L (87.6 ± 7.4 g/L versus 78.8 ± 5.2 g/L, p = 0.001) in EG
and by 2.0 ± 5.7 g/L (81.0 ± 2.9 g/L versus 79.0 ± 2.9 g/L, p =
0.500) in CG. Plasma albumin was reduced by 4.8 g/L (48.1 ±
4.0 g/L versus 43.3 ± 2.8 g/L, p = 0.001) in EG against 1.1 g/L
in CG (44.5 ± 1.6 g/L versus 43.4 ± 1.6 g/L, p = 0.500).
Though opposite in both EG and CG (respectively 0.8% ±
3.3% versus +0.5% ± 3.0%), the variation in plasma volume
does not differ significantly between them (p = 0.396). In EG,
three positive cases of enteric parasites and five of infestation
by plasmodium falciparum were found versus respectively zero
and four cases of enteric parasites, then of infestation by plas-
modium in CG.
Modifications of the Erythrocyte Values during the
As shown by Table 4, only RBC increased in EG (p < 0.05)
at the end of the competition, the other recorded modifications
being non-significant (p > 0.05). Before and after the competi-
tion, all the girls presented normal values of the erythrocyte pa-
Modifications of the Iron Parameters during the
In the group GE, As indicated by Figures 2-4, the following
variables in the EG group presented significant reductions: Fe
(0.71 ± 0.17 mg/L versus 0.69 ± 0.17 mg/L, p = 0.005), TRF
(1.65 ± 0.12 mg/L versus 1.63 ± 0.17 mg/L, p = 0.008), CTF
(3.2718 ± 2.650 mg/L versus 3.2712 ± 2.950 mg/L, p = 0.045),
while serum iron (Figure 5) and RTS (Figure 6) did not vary
significantly (p > 0.05). In CG, all the recorded modifications
n the iron parameters were non-significant (p > 0.05). i
Table 1.
Biometric characteristics of the studied female handball players, prior to and after competition.
EG (n = 16) CG (n = 6)
Prior to competition
After competition
M ± SEM Prior to competition
M ± SEM After competition
Age (years) 23.81 ± 4.02 - 22.83 ± 3.26 -
Height (cm)‘ 167.31 ± 4.93 169.67 ± 4.03
Weight (kg) 60.56 ± 4.60 60.76 ± 5.12 0.19 ± 2.03 74.08 ± 8.73 74.28 ± 7.56 0.20 ± 1.68
BMI (kg·m2) 21.59 ± 1.50 21.87 ± 1.51 0.28 ± 1.11 25.82 ± 3.35 25.90 ± 3.09 0.8 ± 0.58
Resting HR (bpm) 54 ± 1 52 ± 2 1 ± 1 54 ± 1 55 ± 0 2 ± 1
Note: n: sample size; EG: experimental group; CG: control group; M: mean value; SEM: standard error of the mean; BMI: body mass index; Resting HR: resting heart rate;
bpm: beats per minute. There is no significant difference between EG and CG groups for these values.
Table 2.
General data collected during the matches and the training sessions.
EG (n = 16) CG (n = 6)
Mean distance covered
during the matches and the
training sessions (m)
3062.7 ± 746.3 1763.0 ± 311.4*
Mean energy expenditure
during the matches and the
training sessions (kcal)
439.17 ± 142.81 174.78 ± 46.96*
Amount of water drunk
during the matches and the
training sessions (L) 1.38 ± 0.64 0.76 ± 0.38*
Note: The data are reported by player; n: sample size; M: mean value; SEM:
standard error of the mean; EG: experimental group; CG: control group;
*difference with the experimental group significant at p < 0.05.
Table 3.
Dietary intake data of the female handball players surveyed during the
competition period.
EG (n = 16) CG (n = 6)
Daily energy supply
(kcal) 2185.95 ± 126.92 2176.55 ± 189.09
Dietary iron (mg/day) 65.89 ± 1.64 65.99 ± 1.21
Dietary C vitamin
(mg/day) 42.98 ± 1.90 43.60 ± 1.92
Note: n: sample size; EG: experimental group; CG: control group; M: mean value;
SEM: standard error of the mean.
Copyright © 2013 SciRes. 45
Table 4.
Erythrocyte values in the studied female handball players, prior to and after the competition.
EG (n = 16) CG (n = 6) RV
Prior to competition
After competition
M ± SEM Prior to competition
M ± SEM After competition
RBC (1012·L1) 4.79 ± 0.44 4.85 ± 0.44 0.06 ± 0.12* 4.53 ± 0.43 4.65 ± 0.44 0.12 ± 0.44 4.17
Hb (g·dL1) 13.00 ± 0.88 1.03 ± 1.01 0.03 ± 0.28 12.35 ± 1.15 12.30 ± 1.29 0.05 ± 0.3511.6
Ht (%) 40.44 ± 3.18 40.48 ± 3.18 0.44 ± 1.03 38.67 ± 3.07 38.67 ± 3.26 0.0 ± 0.63 37
MCV (fL) 83.69 ± 2.77 83.25 ± 2.35 0.44 ± 6.7185.67 ± 6.50 82.67 ± 2.73 3.00 ± 7.3488
MCHC (g·dL1) 32.44 ± 1.09 32.06 ± 0.92 0.38 ± 0.8032.17 ± 1.72 32.00 ± 1.09 0.17 ± 0.7532
MCH (pg) 27.00 ± 1.26 26.63 ± 1.02 0.38 ± 0.8027.33 ± 2.65 26.17 ± 0.75 1.17 ± 2.4829
Note: n: sample size; EG: experimental group; CG: control group; RV: reference values for female adults, used in the National Laboratory in the Republic of Benin; M:
mean value; SEM: standard error of the mean; Δ (EG): difference between the data collected prior to and after competition in EG; Δ (CG): difference between the data
collected prior to and after competition in CG; RBC: red blood count; Ht: haematocrit; Hb: haemoglobin; MCV: mean corpuscular volume; MCH: mean corpuscular hae-
moglobin; MCHC: mean corpuscular haemoglobin concentration; *p < 0.05.
Figure 2.
Modifications of serum iron in female handball players of Benin during
the competition period (n = 22). *Difference between measures taken
prior to and after competition in the experimental group, significant at p
< 0.05.
Figure 3.
Modifications of serum transferrin in female handball players of Benin,
during the competition period (n = 22). *Difference between measures
taken prior to and after competition in the experimental group, signifi-
cant at p < 0.05.
Figure 4.
Modifications of total iron binding capacity in female handball players
of Benin during the competition period (n = 22). *Difference between
measurements taken prior to and after competition in the experimental
group, significant at p < 0.005.
Figure 5.
Modifications of serum ferritin in female handball players of Benin,
during the competition period (n = 22). Differences between measures
taken prior to and after competition in both groups were all non-sig-
nificant (p > 0.05).
Copyright © 2013 SciRes.
Figure 6.
Modifications of transferrin saturation rate in female handball players
of Benin, during the competition period (n = 22). Differences between
measures taken prior to and after competition in both groups were all
non-significant (p > 0.05).
The main concern at the origin of this study was to measure
the impact of a group form competition on the haematological
parameters whose monitoring is necessary in athletes during the
different periods of the season (Désidéri-Vaillant et al., 2003).
This monitoring is all the more indicated as the players practise
in a risky environment, i.e. an environment marked by strong
heat, malnutrition and a high prevalence of transmissible dis-
eases. As a hole, the characteristics of their daily environment
contribute to increasing the risk of anaemia in the players (Ben
Rayana et al., 2002). Therefore, in case the dietary survey re-
vealed deficiencies, it was likely to expect an observation of
modifications in the haematological parameters at the end of
the competition.
This study was carried out with the players of the female
champion handball team of Benin which took part in a con-
tinental-level competition. The studied team appeared relatively
homogeneous regarding anthropometric parameters, since the
differences between the experimental and control groups are
not significant. It is consequently reasonable to think that the
trainer’s choice to use only the 16 players of the experimental
group during the matches, rather took into account their effec-
tiveness during the preparatory phase.
The Erythrocyte Values
Compared with the literature data (Aboussaleh et al., 2004;
Moulessehoul et al., 2004), the mean erythrocyte data collected
at the beginning of the competition in our female handball
players are all within normal values. These results confirm
those reported in French athletes practising various sports and
whose biochemical and biological parameters were all within
the normal limits (Désidéri-Vaillant et al., 2003). Similar re-
sults were previously recorded in female hockey and football
players (Douglas, 1989).
The erythrocyte values do not differ considering the two
groups of female handball players involved in the study sample.
They were all trained players. Many studies revealed differ-
ences between trained athletes in various sports and untrained
subjects (Boyadjiev & Taralov, 2000; Biancotti, 1992). The
stability of the erythrocyte values observed in the study samples
differs from data reported in Division 1 soccer players after five
weeks of competition (Gouthon et al., 2011). Indeed, Gouthon
and his contributors recorded an increase of RBC and Hb be-
tween test and retest in their study sample.
At the end of the competition, the erythrocyte values did not
vary and remained within the reference ranges for female sub-
jects, except for the RBC, which increased to a significant de-
gree in the experimental group. Under these conditions, the lack
of variation of the erythrocyte values in our series is probably
due to the relatively short duration of the study, since repeated
exercises at high intensity, are known to stimulate erythropoi-
esis (Szygula, 1990). The relatively low-level practice of the
studied players could also explain the stability recorded in the
erythrocyte values. A priori, one can also assert that the play-
ers’ dietary intake was deficient with regard to the nutrients
constituting the raw materials for haematopoiesis, i.e. dietary
iron, C and B12 vitamins. Since the erythrocyte parameters did
not decrease and that C vitamin and iron intake are below the
reference ranges and recommended values for athletes, i.e. res-
pectively 110 mg per day and 16 mg per day (Martin, 2001;
Agence Française de Sécurité Sanitaire des Aliments, 2000),
one can suppose that the exercise-stimulating effect on the hae-
matopoiesis intervened to compensate for the light nutritional
deficiency (Szygula, 1990; Schumacher et al., 2002; Schobers-
berger et al., 1990).
Few subjects (3 out of 22) were infested by the malaria para-
site (plasmodium falciparum) and enteric worms were present
in only 40% of the girls. These data contrast with that of the
overall Beninese population within which the prevalence of the
enteric parasites is higher (INSAE, 2012). They are certainly
the expression of the good quality of the counsels and the
medical follow-up from which these female players of ASO
Baobab profited and that make them a privileged group. In spite
of the fact that an important number of the EG players were
menstruating during the study period, the RBC increased. So,
blood losses do not seem to have had a negative impact on
blood volume in these players, but it would have been interest-
ing to collect information relating to the duration of the men-
struation. Physical exercise may indeed cause menstrual disor-
ders in athletes, i.e. a reduction in the duration of the blood
flow during the menses (Wilmore & Costill, 2006; Portal et al.,
The reduction in plasma volume allows suspecting a lack of
compensation of the hydric losses caused by match-related
efforts, although the players of the experimental group drank an
average of 1.38 L of water per match or training session. How-
ever, the low values of plasma albumin recorded before and
after the competition, do not permit to retain this assumption.
Indeed, only high values of this blood protein, i.e. those higher
than 52 g/L (Janssens, 2009) are associated with water loss-
related dehydration. It is why the weight gain (0.31% of the
initial weight) at the end of the competition in EG players
should be associated with an increased protein synthesis during
recovery periods (Poortmans, 1984; Guezennec, 1989).
Iron Parameters
In this study, serum ferritin remained stable from the begin-
ning to the end of the competition, thus testifying the fact that
body iron stores were not depleted during this period. It was the
same for the rate of transferrin saturation which constitutes one
Copyright © 2013 SciRes. 47
of the iron bioavailability markers. TBC was on the other hand
reduced, as well as RBC suggesting a decrease in the oxygen
transportation capacity in the players of the experimental group.
These data suggest the following two observations.
The first one is about the dietary intake related to the raw
materials for erythropoiesis in these female players. The data
collected reveal weak iron and C vitamin intakes during the
competition period. Under these conditions, a reduction of the
erythropoiesis in the handball players could explain the de-
crease in TBC recorded at the end of the competition. Since
physical exercise is regarded as a trigger of erythropoiesis
(Szygula, 1990; Schobersberger et al., 1990; Schumacher et al.,
2002), other factors may be taken into account to explain this
result. A more important red cell destruction than red cell pro-
duction which requires at least one week may be an explanatory
hypothesis. Ultimately, the factor most likely to explain the
reduction in TBC at the end of the competition remains the
decrease in serum transferrin, which is one of the iron carrier
The second observation relates to the stability of iron stores
in the players’ bodies, contemporary to the decrease in serum
iron during this period. If there was a concomitant reduction of
the bioavailable iron (RTS and TBC), (Haute Autorité de Santé,
2011) there would be the suspicion of an inflammatory process
in the players as a result of the high intensity of the match-
related physical loads. Since Hb has not decreased below the
normal value (12 g/100 mL), such a hypothesis is given up.
Dietary deficit needs to be pronounced and chronic beyond a
few days, to induce the depletion of iron stores in the body.
Though their average daily 2185 kcals intake near the mini-
mum of 2200 kcals recommended for female athletes (Martin,
2001), these girls’ diet has to be improved qualitatively as well
as quantitatively during competition periods.
This study belongs to the few ones addressing the modifica-
tions of the haematological parameters during a handball con-
tinental-level competition, played in a tropical environment.
The four matches played in six days did not significantly mod-
ify the players’ erythrocyte values. Serum iron, TBC and serum
transferrin were however reduced at the end of the matches of
the preliminary round, without any case of anaemia. Thus, in
the Beninese environment characterized by a high prevalence of
transmissible diseases, it is possible to reduce the risk of anae-
mia in athletes, as long as the medical follow-up makes it pos-
sible to avoid parasites. The data of this study suggest associat-
ing good medical follow-up in these handball players, to an
improvement of their diet with highly protein-enriched food
and erythropoiesis raw materials such as iron, C and B 12 vita-
mins. The data also suggest that these girls adopt adequate hy-
dration practices, in order to compensate for the effort-induced
water losses during matches and training sessions.
The authors thank the President and Members of the ASO
Baobab handball club of Lokossa (Republic of Benin) who
agreed and helped for carrying out this research. We are also
grateful to all the female players who accepted to take part in
the study and to the Members of the Fitness Centre VITA
FORM of Porto-Novo, for their technical assistance.
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