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Oxidative stress induced by corticosterone

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gallus domesticus) 1. Chronic exposure. Comp
Biochem Physiol B
ARTICLE in COMPARATIVE BIOCHEMISTRY AND PHYSIOLOGY PART B BIOCHEMISTRY AND MOLECULAR BIOLOGY
JANUARY 2005
Impact Factor: 1.55 DOI: 10.1016/j.cbpc.2004.09.013 Source: PubMed

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Comparative Biochemistry and Physiology, Part B 139 (2004) 737 744

www.elsevier.com/locate/cbpb

Oxidative stress induced by corticosterone administration

in broiler chickens (Gallus gallus domesticus)
1. Chronic exposure
H. Lina,b, E. Decuyperea, J. Buysea,*
a

Lab of Physiology and Immunology of Domestic Animals, Department of Animal Production, Catholic University Leuven,
Kasteelpark Arenberg 30, 3001 Leuven, Belgium
b
Department of Animal Science, Shandong Agricultural University, Taian, Shandong 271018, PR China
Received 24 May 2004; received in revised form 13 September 2004; accepted 14 September 2004

Abstract
The effects of long-term dietary administration of corticosterone (CORT) on the induction of oxidative injury in broiler chickens (Gallus
gallus domesticus) were evaluated. The experimental broiler chickens were fed with a diet supplemented with 30 mg CORT/kg diet for 2
weeks from 14 days of age onwards, while control chickens continued to consume the control diet. The growth performance parameters were
recorded weekly, and a blood sample was obtained from eight birds of both groups before CORT administration and at 3, 7 and 14 days after
treatment. The results showed that chronic CORT administration resulted in enhanced proteolysis and gluconeogenesis. Furthermore, CORT
administration may initially induce the formation of reactive oxygen species (ROS) as indirectly reflected by an increase in lipid
peroxidation. However, the significantly increased plasma uric acid (UA) and ceruloplasmin (CP) levels after 3 days of treatment indicates an
enhancement of the nonenzymatic antioxidant capacity during stress, and in this way, the development of a more severe oxidative injury is
alleviated. Broiler chickens seem to adapt to high circulating CORT levels in terms of their redox homeostasis after 3 days of treatment under
the present experimental conditions.
D 2004 Elsevier Inc. All rights reserved.
Keywords: Antioxidant; Chickens; Corticosterone; Oxidative injury; Stress; Superoxide dismutase; Uric acid

1. Introduction
Broiler chickens are continuously confronted by a
multitude of stressors that can last for a few hours (e.g.,
catching, crating and transport) or for nearly the entire
rearing period (e.g., heat stress, immune challenges).
Consequently, their internal homeostasis is constantly
challenged by intrinsic and extrinsic adverse forces or
stressors. The redox homeostasis is maintained by prooxidant/antioxidant balance, and the imbalance in favor of
the pro-oxidant system will result in oxidative stress (Sies,
1991). The maintenance of the redox balance has been

* Corresponding author. Tel.: +32 16 328525; fax: +32 16 321994.

E-mail address: Johan.Buyse@agr.kuleuven.ac.be (J. Buyse).
1096-4959/$ - see front matter D 2004 Elsevier Inc. All rights reserved.
doi:10.1016/j.cbpc.2004.09.013

found to be important for the health of broiler chickens

(Bottje et al., 1998; Iqbal et al., 2002).
Glucocorticoids, as the final effectors of the hypothalamicpituitaryadrenal (HPA) axis, participate in the control
of whole body homeostasis and the organisms response to
stress. In mammals, lipid peroxidation was induced while
the nonenzymatic antioxidant capacity was decreased, and
enzymatic antioxidant systems were suppressed in the liver
(Ohtsuka et al., 1998), erythrocytes (Orzechowski et al.,
2000), heart and spleen (Rajashree and Puvanakrishnan,
1998), skeletal muscle and lymphoid organs (Pereira et al.,
1999) after chronic administration of glucocorticoid hormones. In broiler chickens, there is evidence that the redox
balance is affected by heat stress (Lin et al., 2000; Altan et
al., 2003; Mahmoud and Edens, 2003). In our previous
work, a clear positive relationship between circulating

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H. Lin et al. / Comparative Biochemistry and Physiology, Part B 139 (2004) 737744

corticosterone (CORT) levels and plasma lipid peroxidation

was observed in broiler chickens exposed to a high ambient
temperature for a long-term period (Lin et al., 2004b).
Furthermore, the impaired integrity of muscular membranes,
as reflected in increased plasma creatine kinase (CK)
activity, is speculated to be associated with oxidative injury
elicited by dietary administration of CORT (Malheiros et al.,
2003).
Recently, it was reported that lipid peroxidation in the
liver and muscle of broiler chickens might be induced by
dietary supplementation of CORT (Eid et al., 2003).
Meanwhile, the glucocorticoid-induced oxidative injury is
alleviated by the supplementation of antioxidant substances,
such as tea polyphenols (Eid et al., 2003). Birds have a more
potent antioxidant system than mammals (Klandorf et al.,
1999; Simoyi et al., 2002). Uric acid (UA) is excreted as a
major end product of nitrogen metabolism by birds. The
antioxidant nature of UA has been recognized recently both
in mammals (Hellstein et al., 1997) and surely in poultry
(Simoyi et al., 2002, 2003; Machn et al., 2004). Therefore,
we hypothesize that the changes in antioxidant systems of
chickens upon stress may be different from that of
mammals.
Ceruloplasmin (CP), a blue copper glycoprotein with
ferroxidase and oxidase activities, is an important serum
antioxidant. Meanwhile, as an acute phase protein (APP)
exhibiting a moderate response in humans (Min et al., 1991)
and chickens (Koh et al., 1996), it can be stimulated by the
administration of corticosterone (Curtis and Butler, 1980)
and by oxidative stress in mammals (Sato and Bremner,
1993; Levy et al., 2001). Therefore, the response of CP, as
an antioxidant, to stress was also evaluated in the present
study.
The objective of the present study was to determine the
effects of long-term dietary administration of CORT on the
induction of oxidative injury (lipid peroxidation and CK
activities) and the response of antioxidant systems (enzymatic antioxidantsuperoxide dismutase [SOD]; nonenzymatic antioxidantstotal antioxidant capacity, CP and UA)
in broiler chickens. Furthermore, the impact of CORT on the
growing performance and circulating thyroid hormone and
glucose levels were also determined to establish possible
alterations in metabolic status and to see if this is related to
oxidative stress. In a companion paper, we describe the shortterm effects of CORT administration (Lin et al., 2004a).

2. Materials and methods

2.1. Chickens and diets
One hundred eighty broiler chicks (Cobb; Gallus gallus
domesticus) were obtained from a local hatchery at 1 day of
age and assigned randomly to nine pens, with 20 chicks per
pen, in an environmentally controlled room. The brooding
temperature was maintained at 35 8C for the first 2 days and

then decreased gradually to 21 8C (45% RH) until 28 days

of age.
The broilers received a commercial starter diet (12.10 MJ
ME/kg and 22.2% CP) until 7 days of age, after which a
commercial grower diet (12.68 MJ/kg, 21.4% CP) was
provided until the end of the experiment. For a detailed
description of both diets, see Buyse et al. (2001). The light
regime was 23 L:1 D. The birds had free access to feed and
water during the entire rearing period. From 14 days of age
onwards, the chickens of experimental groups (six pens of
20 chickens) received an experimental diet supplemented
with 30 mg corticosterone /kg basal diet (Malheiros et al.,
2003), whereas control chickens (three pens of 20 chickens)
continued to consume the control diet. Feed intake (FI) and
body weight gain (BWG) were recorded weekly during the
2-week experimental period, and feed efficiency (feed/gain)
was calculated.
A blood sample was obtained from eight birds per
treatment before CORT administration and at 3, 7 and 14
days after dietary treatment. At 09:00 a.m. of each bleeding
day, the eight chickens of each treatment were selected at
random from the pens, and the chickens were bled on only
one occasion. Blood was drawn from a wing vein using a
heparinized syringe within 30 s and collected in iced tubes.
Plasma was obtained after centrifugation at 1200g for 10
min at 4 8C and stored at 20 8C.
2.2. Plasma parameters
Commercial colorimetric diagnostic kits were used to
measure plasma glucose (GLU; IL Testk kit, No. 18250800), uric acid (UA; IL Testk kit, No. 181685-00) and
creatine kinase (CK; IL Testk kit, No. 181605-90), using
the Monarchk 2000 Chemistry system Model 760 (Monarch Chemistry System, Instrumentation Laboratories, Belgium). Plasma superoxide dismutase (SOD) activity was
measured with a commercial kit (Dojindo), using a microplate reader (Titertek MultiskanR, MCC/340).
Corticosterone (CORT) was measured using a sensitive
and highly specific radioimmunoassay kit (IDS, Boldon,
UK), with a sensitivity of 0.39 ng/ml and low cross
reactivity with aldosterone (0.20%), cortisol (0.40%) and
deoxycorticosterone (3.30%). The intra-assay variability
was 3.8%. Before assay, plasma samples were heated at
80 8C for 10 min to inactivate corticosterone-binding
proteins.
Thyroid hormones were measured by radioimmunoassay
according to the method described by Darras et al. (1991).
Briefly, triiodothyronine (T3) measurements were performed
using a commercial available T3 antiserum (Byc-Sangtec
Diagnostica, Dietzenbach, Germany), in combination with a
specific tracer (Amersham International, Slough, England).
The intra-assay coefficient of variation was 4.5%. Thyroxine (T4) concentrations were assayed using tracer from
Amersham and a rabbit T4 antiserum (Mallinckrodt Diagnostica, Dietzenbach, Germany). This T4 antiserum had

H. Lin et al. / Comparative Biochemistry and Physiology, Part B 139 (2004) 737744

739

Table 1
Growing performance of broiler chickens subjected to dietary supplementation of corticosterone (CORT, 30 mg/kg diet) from 14 to 28 days of age
CORT

Body weight gain, g/d

First week
Second week
Feed consumption, g/d
First week
Second wk
Gain:feed, g/g
First week
Second week

NORM

29.3F2.5a
42.4F5.8a

50.5F4.4b
98.1F5.8b

69.5F4.4
116.8F14.6a

65.9F3.8
157.6F9.9b

0.42F0.02a
0.37F0.07a

P-value
Treatment

Time

Interaction

Pb0.0001

Pb0.0001

Pb0.0001

Pb0.0001

Pb0.002

Pb0.0005

Pb0.0001

Pb0.001

NS

0.77F0.04b
0.62F0.04b

CORTcorticosterone treatment; NORMcontrol group.

a,b
Values (meansFS.E.M., n=3 [control treatment] or n=6 [CORT treatment] pens of 20 chickens) within the same row with different superscript
significantly differ ( Pb0.05). NS: not significant (PN0.05).

0.16% cross-reactivity with T3. The intra-assay coefficient

of variation was 3.2%.
Plasma lipid peroxidation was estimated by spectrophotometric determination of thiobarbituric acid reacting substances (TBARS) with the method of Yagi (1984) modified
according to Jentzsch et al. (1996) and Lapenna et al.
(2001). In brief, 100 Al plasma was mixed with 2.0 ml 0.17
M H2SO4 and 0.3 ml of 10% phosphotungstic acid and
centrifuged (1200 g, 10 min). After standing at room
temperature for 5 min, the sediment was obtained and mixed
with 2.0 ml 0.17 M H2SO4 and 0.3 ml of 10% phosphotungstic acid and centrifuged again. The sediment was
suspended in 2.0 ml distilled water and mixed with 0.5 ml of
0.34% thiobarbituric acid (TBA, 50% acetic acid solution)
and 25 Al 0.5 M butylated hydroxytoluene (BHT, Sigma).
The mixture was heated at 95 8C for 60 min in a water bath.
After cooling with tap water, the chromogen was extracted
with 2.0 ml of a mixture of n-butanol and pyridine (1:15, v/
v). After centrifugation at 1400 g for 10 min, the organic
layer was extracted and read spectrophotometrically at 532
nm against a reaction blank. To correct for the background,
absorbance values at 572 nm were subtracted. TBARS were
expressed as nmol of malondialdehyde (MDA) per ml
plasma.
Total plasma antioxidant activity was determined by the
ferric reducing/antioxidant power (FRAP) assay, as
described by Benzie and Strain (1996, 1999). The measurement was conducted at room temperature and a 5-min time
window was used. Ceruloplasmin (CP) was measured with
the o-dianisidine dihydrochloride end-point method, as
described by Sugiyama et al. (2000).
All samples were analyzed in the same assay to avoid
inter-assay variability.
2.3. Statistical analysis
The main effects of CORT, supplementation time and
their interaction were analyzed by two-way ANOVA
(version 8e, SAS Institute, 1998). The differences between
time points within treatment and the difference between

treatments within each time point were analyzed by oneway ANOVA, followed by the Scheffe test when appropriate. Means were considered significantly different when
Pb0.05.

3. Results
The growing performance of broiler chickens was
impaired ( Pb0.0001) by dietary administration of CORT
(Table 1). Compared to control chickens, feed intake, BWG
and feed/gain ratio were significantly lower ( Pb0.0001) in
CORT chickens, except for the feed consumption during the
first week of supplementation. There were significant
interactions between treatment and time for BWG
( Pb0.0001) and feed consumption ( Pb0.0005) but not for
feed/gain ratio.
The CORT treatment had a significant ( Pb0.05) overall
effect on plasma CORT, CK, GLU, UA, T3, T4, T3/T4,
FRAP, CP and TBARS but not on SOD ( PN0.05) (Table 2).
There was a significant time effect for CORT, FRAP, CP,
UA, CK and T4 but not for SOD, TBARS, GLU, T3 and T3/
T4 ratio (Table 2). Significant interactions ( Pb0.05)
between CORT treatment and time were only observed for
Table 2
Analysis of variance for the effect of dietary supplementation of corticosterone (CORT, 30 mg/kg diet) and time of treatment on plasma parameters
of broiler chickens (n=8 birds per treatment group)
CORT, ng/ml
TBARS, nmol/ml
SOD, U/ml
CP, U/ml
FRAP, Amol/l
UA, mg/dl
GLU, mg/dl
CK, IU/l
T3, ng/ml
T4, ng/ml
T3/T4

Treatment

Time

Treatmenttime

b0.0001
b0.0001
NS
b0.0001
b0.0001
b0.0001
0.0097
0.0459
0.0029
b0.0001
0.0084

0.0049
NS
NS
0.0178
0.0372
0.0174
NS
b0.0001
NS
0.0276
NS

0.0353
0.0425
0.0093
0.0502
NS
NS
NS
0.0002
0.0310
0.0470
NS

NS: not significant (PN0.05).

740

H. Lin et al. / Comparative Biochemistry and Physiology, Part B 139 (2004) 737744

Fig. 1. Plasma levels of corticosterone (CORT) and thiobarbituric acid reacting substances (TBARS) in broiler chickens subjected to dietary supplementation of
corticosterone (CORT, 30 mg/kg diet). CORTcorticosterone treatment (o), NORMcontrol group (x); abmeans within the CORT treatment with
different superscript differ significantly ( Pb0.05); xzmeans within NORM treatment with different superscript differ significantly ( Pb0.05); *means within the
same time point significantly differ between treatments ( Pb0.05).

CORT, SOD, TBARS, CP, CK, T3 and T4 indicating the

different developing trends of these parameters in the two
treatments (Table 2). Indeed, all the measured plasma
parameters significantly ( Pb0.05) changed with time in
CORT treatment, whereas the age-related changes ( Pb0.05)
were only observed for CK, GLU, UA, FRAP and TBARS
in control chickens.
Dietary supplementation of CORT resulted in significantly ( Pb0.0001) elevated plasma levels of CORT in the
experimental group compared with the control chickens

(Fig. 1A). Plasma concentrations of TBARS were significantly ( Pb0.05) increased by CORT treatment at day 3 and
maintained as such thereafter, while it dropped with time
( Pb0.05) in control chickens (Fig. 1B). In CORT chickens,
the significantly higher SOD activity was only observed at
day 3 when compared to the basal value and that of control
chickens. Thereafter, no significant differences in plasma
SOD activity were observed (Fig. 2A). There was a
significant and continuous increase in plasma level of CP
in CORT chickens after day 3 until the end of the

Fig. 2. Plasma levels of superoxide dismutase (SOD) activity, ceruloplasmin (CP), ferric reducing/antioxidant power (FRAP) and uric acid (UA) in broiler
chickens subjected to dietary supplementation of corticosterone (CORT, 30 mg/kg diet). CORTcorticosterone treatment (o); NORMcontrol group (x);
ac
means within the CORT treatment with different superscript differ significantly ( Pb0.05); xymeans within NORM treatment with different superscript differ
significantly ( Pb0.05); *means within the same time point with different superscript differ significantly between treatments ( Pb0.05).

H. Lin et al. / Comparative Biochemistry and Physiology, Part B 139 (2004) 737744

741

Fig. 3. Plasma concentration of glucose and activities of creatine kinase (CK) activity in broiler chickens subjected to dietary supplementation of corticosterone
(CORT, 30 mg/kg diet). CORTcorticosterone treatment (o), NORMcontrol group (x); abmeans within the CORT treatment with different superscript
differ significantly ( Pb0.05); xymeans within NORM treatment with different superscript differ significantly ( Pb0.05); * means within the same time point
differ significantly between treatments ( Pb0.05).

experiment, and values were significantly higher than those

in control chickens (Fig. 2B). After 3 days of administration,
FRAP and UA values of CORT chickens were significantly
higher than their basal values and those of control chickens,
while a significant elevation was only observed at day 14 in
control chickens (Fig. 2C, D).
The plasma levels of GLU were significantly higher than
the basal values after 3 days of administration, while a timedependent elevation was only observed at day 14 in control
chickens (Fig. 3A). The significant change in plasma CK
activity was observed in CORT chickens at day 3 and
thereafter, whereas this increase was only found at day 14 in
the control group (Fig. 3B).
Compared with the absence of changes with time in the
control group, plasma concentrations of T3 and T4 were both
significantly decreased by CORT administration from day 3
onwards, and values were significantly lower than those of
control broilers after day 3 with regard to T4 but not for T3
(Fig. 4A, B). In CORT chickens, significantly higher T3/ T4
ratios were only observed at day 3, compared to the basal
values and those of control chickens (Fig. 4C).

4. Discussion
4.1. The catabolic effect induced by CORT administration

Fig. 4. Plasma concentrations of triiodothyronine (T3) and thyroxine (T4),

and T3/T4 ratio in broiler chickens subjected to dietary supplementation of
corticosterone (CORT, 30 mg/kg diet). CORTcorticosterone treatment
(o), NORMcontrol group (x); abmeans within the CORT treatment
with different superscript differ significantly ( Pb0.05); *means within the
same time point differ significantly between treatments ( Pb0.05).

The significantly elevated circulating CORT levels of

broiler chickens with dietary CORT supplementation and
the significant interaction between treatment and time
illustrate the successfully induced hyperglucocorticoid
status in the treated chickens within reasonable physiological limits (Freeman, 1983). The significantly increased
plasma levels of GLU and UA after chronic CORT
administration compared to the basal levels and that of
control chickens indicate the enhanced glycogenolysis/
gluconeogenesis and protein catabolism in CORT chickens,
respectively. This stimulated protein catabolism was further reflected in the significantly suppressed BWG and

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H. Lin et al. / Comparative Biochemistry and Physiology, Part B 139 (2004) 737744

decreased feed efficiency in broiler chickens consuming the

CORT diet, in line with previous reports in broiler chickens
(Eid et al., 2003; Malheiros et al., 2003) and in rats
(Ohtsuka et al., 1998; Orzechowski et al., 2002). Furthermore, the absence of further changes in GLU and UA since
day 3 after CORT exposure in experimental group shows the
adaptation of chickens to chronic stress. In the present study,
no significant interaction between treatment and time for
GLU and UA was detected, indicating the effect of age in
both treatments. However, the significant lower values in
control chickens at each time point after day 3 of CORT
administration imply a different metabolic status in control
chickens compared to CORT chickens.
There are controversial reports on the effect of CORT
administration on feed intake. A significantly elevated feed
consumption after CORT treatment was observed in some
studies (Bartov et al., 1980a,b; Nasir et al., 1999) but not in
others (Buyse et al., 1987; Malheiros et al., 2003), and it
seems to be related to the duration, dose and the way of
CORT administration. In the present study, the absolute feed
intake (expressed as g/chicken) of CORT chickens was not
significantly affected during the first week but was
significantly decreased in the second week after CORT
treatment. However, if feed consumption was expressed as
feed consumed per kilogram metabolic body weight
(BWkg0.75), then the CORT chickens ate significantly more
( Pb0.05) feed than control chickens (Fig. 5). This indicates
that the reduced absolute feed intake per chicken of the
CORT chickens compared to control chickens is due to their
suppressed body weight and in turn the smaller volume of
gastrointestinal tract. The negative feedback of high
circulating levels of CORT on the anorexigenic, hypothalamic corticotrophin-releasing hormone is likely to be the
underlying causal mechanism of the relative high feed
consumption per unit of metabolic BW in CORT chickens
(Denbow et al., 1999). The high feed consumption relative

Fig. 5. Feed intake on the basis of metabolic body weight (BWkg0.75) in

broiler chickens subjected to dietary supplementation of corticosterone
(CORT, 30 mg/kg diet). CORTcorticosterone treatment (o), NORM
control group (x); abmeans within the CORT treatment with different
superscript differ significantly ( Pb0.05); xymeans within NORM treatment
with different superscript differ significantly ( Pb0.05); *means within the
same time point with different superscript differ significantly between
treatments ( Pb0.05).

to metabolic body weight and the poorer feed efficiency of

CORT chickens suggest increased energy expenditure or fat
deposition or both.
The significantly decreased plasma T3 and T4 levels after
3 days of CORT exposure and the significant interaction
between treatment and time illustrate the persistent suppression of CORT on the secretion of thyroid hormones, as
found previously (Decuypere et al., 1983; Buyse et al.,
1987; Darras et al., 1996).
4.2. Lipid peroxidation induced by chronic CORT
administration
In accordance with the results of Eid et al. (2003), the
significantly increased plasma concentrations of TBARS in
CORT chickens indicates the enhanced lipid peroxidation
induced by chronic CORT treatment. This result is also in
line with the findings in mammals (Rajashree and
Puvanakrishnan, 1998; Ohtsuka et al., 1998; Pereira et
al., 1999; Orzechowski et al., 2000) and suggests that the
oxidative stress-induced effect of CORT exists also in
chickens. This conclusion is supported further by the
significantly increased SOD activity after 3 days of dietary
CORT administration, which may be considered as a
reaction of the enzymatic scavenger system to oxidative
stress. The presumably or likely increased energy expenditure triggered by high circulating CORT levels might be
responsible, at least partially, for the augmented formation
of reactive oxygen species (ROS) as reflected by increased
lipid peroxidation. High concentrations of glucocorticoids
are capable of inducing a high degree of oxidative stress in
mammals; both the enzymatic and nonenzymatic antioxidant levels are suppressed by long-term glucocorticoid
administration in most studied tissues, such as the whole
blood, soleus muscle and spleen (Orzechowski et al.,
2002), heart (Rajashree and Puvanakrishnan, 1998) and
skeletal muscle (Ohtsuka et al., 1998). The transient upregulation in plasma SOD activity coincides with the
simultaneously increased nonenzymatic antioxidants
(FRAP, UA, CP), suggesting that the nonenzymatic
antioxidants may act as the first defense line to quench
the increased formation of free radicals. This speculation is
supported by the observation that there was no further
significant increase in TBARS after 3 days of CORT
treatment. The significant interaction between treatment
and time for TBARS showed the different trend with time
in CORT and control treatments. The decreased plasma
concentrations of TBARS with time in control chickens
indicate a decreased lipid peroxidation, which might be due
to the simultaneous rise in UA and FRAP, suggesting the
increased antioxidant capacity with age.
CORT treatment induced muscle proteolysis resulting in
high UA levels in the circulation. The significant positive
correlation between plasma UA and FRAP (r=0.967;
Pb0.001) suggests an important contribution of UA to the
increased total antioxidant capacity after CORT adminis-

H. Lin et al. / Comparative Biochemistry and Physiology, Part B 139 (2004) 737744

tration. High circulating levels of UA have been suggested

as one of the reasons of greater longevity of birds than
mammals of similar size (Klandorf et al., 1999).
In the present study, the significant increase in plasma
concentrations of CP in CORT chickens is in accordance
with previous work (Curtis and Butler, 1980). Furthermore,
the steady increase in plasma CP, while CORT leveled off
and even tended to decrease at day 14, suggests the
potential induction effect of CORT in the production of CP
in chickens. CP exhibits a two-edged property: antioxidative as well as pro-oxidant activities in biological systems.
It can protect polyunsaturated fatty acids in cellular
membranes from ROS (Kim et al., 1998; Park et al.,
1999), and on the other hand, there is evidence suggesting
its pro-oxidant activity in the oxidation of low-density
lipoprotein and in turn the development of cardiovascular
disease (Fox et al., 2000). The collateral effects of the
significantly increased CP in stressed chickens should be
further investigated.
A significant increase in plasma CK with age was
observed in control chickens, suggesting its relation with
body weight. This observation is supported by the results
of Wilson et al. (1990) and Hocking et al. (1998), who
reported that plasma CK activities were higher in rapidly
growing turkey lines compared to slower growing lines. In
control chickens, the significant rise in plasma CK activity
at 28 days of age (day 14) coincides with a substantial
increase in body weight and breast muscle at this stage
(Scheuermann et al., 2003). Therefore, the higher body
weight and growing rate of control chickens (1.45F0.07
vs. 0.92F0.03 kg) compared to CORT chickens should be
responsible, at least partially, for the significantly higher
plasma CK levels at the same age. In a previous study, it
was hypothesized that the impaired integrity of muscular
membranes is associated with oxidative injury elicited by
dietary administration of CORT (Malheiros et al., 2003). In
accordance with this hypothesis, the plasma CK activities
were significantly increased from day 3 onwards in CORT
chickens compared to basal level, while plasma TBARS
concentration were concurrently increased in the present
experiment. The absence of a difference in CK levels
between CORT and control treatments at day 3 and day 7
is not in contradiction with the hypothesis of Malheiros et
al. (2003) because of the lower BW of the CORT chickens.
The significant interaction between treatment and time also
indicates a differential age-related CK pattern between
control and CORT chickens. The result suggests that the
response of CK values is the combined effect of stress and
decreased BWG, and CK may not be a good parameter to
reflect long-term stress.
In conclusion, chronic dietary CORT administration to
broiler chickens resulted in a stimulated proteolysis and
gluconeogenesis. Furthermore, after 3 days of CORT
administration, lipid peroxidation was significantly elevated, which may reflect an augmented formation of ROS.
However, this oxidative injury induced by chronic CORT

743

treatment was not additionally aggravated along with the

treatment duration. The significantly increased plasma UA
and CP levels may enhance the nonenzymatic antioxidant
capacity during stress elicited by high circulating CORT
levels and prevent the development of more severe
oxidative injury. The significance of the steady elevation
in plasma CP levels during the whole experimental period
needs to be identified. Broiler chickens seem to adapt to
chronic stress induced by CORT administration after 3 days
of treatment under the present experimental conditions.

Acknowledgement
The diligent technical assistance of C. Borgers and G.
Nackaerts is greatly appreciated. Hai Lin is supported by
Grants F/00/89 from the Research Fund, K. U. Leuven and
Grant 2004CB117507 from National Basic Research Program of China.

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