SCVMJ, XVII (2) 2012

129

Effects of vitamin E and selenium complex on heat-stressed

rabbits
AL-Zafry, S. R. and Medan, M. S. *
Department of Animal Production, Faculty of Agriculture, Omar AL
Mukhtar University, AL Baida, Libya
* Department of Theriogenology, Faculty of Veterinary Medicine, Suez
Canal University, Ismailia, Egypt.
Abstract
The effects of vitamin E & selenium were evaluated on 27 heatstressed New Zealand white rabbits. After induction of heat stress,
rabbits were randomly divided into 3 groups (9 rabbits of each). The
first group (control group) did not receive any supplementation of
vitamin E or selenium. The second group was injected
subcutaneously with 0.2 ml (5.05 mg) vitamin E & selenium per
animal weekly. The third group was injected subcutaneously with 0.4
ml (10.10 mg) vitamin E & selenium per animal weekly. Heat stress
was induced through exposure of all groups to a temperature of 35 C
daily for 6 hours and continued for 35 days. Rectal and skin
temperatures were recorded 1 hour before and at the end of heat
stress. Blood samples were collected from the ear vein into
heparinized vacutainer tubes and plasma was separated for estimation
of glucose, urea, cholesterol, calcium, phosphorus, potassium,
sodium, albumin and globulin. The rectal and skin temperatures
increased significantly (P<0.05) after induction of heat stress. The
results showed that supplementation of vitamin E & selenium
complex decreased both rectal and skin temperatures in heat-stressed
rabbits. Plasma glucose and cholesterol were significantly higher in
all treated groups compared to control group. Regarding the effect of
vitamin E & selenium dose, there was no significant difference
between the two treated groups. From these results we can conclude
that vitamin E & selenium complex could be used to alleviate the
adverse effects of heat stress in rabbits.
Keywords: Vitamin E & selenium; Heat Stress; Rabbits
Introduction
High environmental temperature
challenges the animal's ability to
maintain energy, thermal, water,

hormonal and mineral balance. Heat

stress
stimulates
excessive
production of oxidative free radicals
[1] and these free radicals can in

130
turn produce harmful effects if they
are not eliminated. Heat production
is directly controlled by the nervous
and endocrine systems through
modifications of appetite, digestive
process
and,
indirectly,
by
alterations of protein synthesis.
The most obvious limitation to
rabbit production in the regions with
a hot climate is the susceptibility of
rabbits to heat stress. Heat stress is
defined as a stress inflicted by a
wide range of environmental
conditions that induce a state of
physiological strain within the
animals body, and means that
animals are not able to regulate their
heat homeostasis passively. It
mainly occurs when animals are
exposed
to
high
ambient
temperatures, high humidity, low
wind speed, and direct and indirect
high solar radiation [2].
Rabbits, as a homoeothermic
animal, can regulate the heat input
and output of their bodies using
physical,
morphological,
biochemical
and
behavioural
processes to maintain a constant
body temperature [3]. The thermoneutral zone of temperature in
rabbits is around 1821 C [3].
Thus, when rabbits are exposed to
elevated ambient temperatures,
imbalances are induced in their
body temperature [4,5], which
adversely affect their growth and
reproductive
traits
[6-8].
Furthermore, disturbances in feed
intake, feed utilization, water
metabolism, blood parameters,
enzymatic reactions, hormonal
secretions, in addition protein,

AL-Zafry and Medan

energy and mineral imbalances had
been also reported to be disrupted in
heat-stressed rabbits [8-10,11].
Studies in large animals showed that
a reduction in plasma vitamin C
concentration was reported in calves
stressed by housing conditions [12]
and in heat- stressed cows [13].
Ayo et al. [14] and Ghanem et al
[15] provided vitamin C to goats
and Awassi sheep, by dissolving
ascorbic acid in water, they found
that supplemented vitamin C was
effective in alleviating stress. AbouZeina et al [16] found that
supplementation of antioxidants
improved fertility in heat-stressed
buffaloes.
A major strategy to reduce the effect
of heat stress on animals is to alter
the environment through the use of
sheds, fans or evaporative cooling
[17]. Such practices are sometimes
not possible in desert areas. This
necessitates trying other strategies
to counteract the adverse effects of
heat stress such as supplementation
of antioxidants such as vitamin E
and selenium as free radical
scavengers. In rabbits, the literature
did not reveal sufficient information
regarding the role of antioxidants
during heat stress. Therefore, the
present study was conducted to
assess whether supplementation of
vitamin E and selenium has a
beneficial effect on heat stressed
rabbit.
Materials and methods
27 apparently healthy, New Zealand
white rabbits, 7 weeks of age, were
used in this experiment. The body

SCVMJ, XVII (2) 2012

weight ranged from 400 to 600 gm.
The rabbits were maintained under
uniform
management
and
husbandry conditions. The animals
were fed commercial rabbit diet ad
libitum and were provided with
clean drinking water ad libitum.
Animals were randomly divided
into 3 groups, 9 animals of each.
The group I (control group) did not
receive any treatment of vitamin E
or selenium, while the second group
(treat1) was subcutaneously injected
with 0.2 ml (5.05 mg) vitamin E and
selenium per animal weekly
throughout the stress period. The
third
group
(treat2)
was
subcutaneously injected with 0.4 ml
(10.10 mg) vitamin E and selenium
per animal weekly throughout the
stress period.
All groups were exposed to a
temperature of 35C for 6 hours per
day starting from 12 AM until 6 PM
for 35 days. Body temperature was
recorded by inserting a digital
clinical thermometer into the rectum
for one minute. Also skin
temperature was recorded. Blood
samples of all groups were collected
from ear vein into heparinized
vacutainer tubes at the end of the
experimental
period.
Plasma
samples
were
separated
by
centrifugation at 3,000 rpm for 15
min and were frozen and stored at 20C until analysis.
Plasma glucose concentration was
determined by glucose analyzer and
plasma urea was determined by
BUN analyzer as provided by
Beckman Company. Total protein
and cholesterol were determined by

131
kits provided by Human Company,
Germany using Semi-Automated
Clinical
Chemistry
Analyzer,
Germany. Moreover, plasma levels
of Albumin, calcium, phosphorus
and sodium were determined by
spectrophotometric method (kits
provided by Jenway Company). The
globulin level was calculated
through the following equation:
Globulin = total protein - albumin
Statistical data for all measured
parameters were performed using
the general linear model (GLM)
produced by the Statistical Analysis
Systems Institute [18]. Significant
differences among means were
evaluated using t-test (temperatures
before and after heat stress) or
Duncan multiple range test in other
parameters (P 0.05).
Results
Rectal and skin temperatures in
rabbits exposed to heat stress (35
C) for 35 days:
Rectal and skin temperatures before
and after heat stress in rabbits are
shown in Fig. 1. After induction of
heat stress, rectal temperature
significantly (P < 0.05) increased
from 39.03 C to 39.91C and also
skin temperature significantly (P <
0.05) increased from 34.96 C to
36.16C.
The effect of vitamin E and selenium
treatment on rabbits exposed to heat
stress (35 C) for 35 day:
Rectal and skin temperatures in the
control and treated groups are
shown in Fig. 2. Rectal temperature
in the treated groups (39.35 and
39.24C, respectively) was lower

132

AL-Zafry and Medan

than in the control group (39.52C).

Similarly, skin temperature, was
also significantly (P < 0.05)
decreased in the treat2 group
compared with the control group.
Effect of vitamin E and selenium
complex
on
some
plasma
biochemical parameters in rabbits
exposed to heat stress (35 C) for 35
day:
Table 1 shows plasma levels of
glucose, urea, sodium, potassium,
calcium, phosphorus, cholesterol,
total protein, albumin and globulin
in the control (heat-stressed, non
treated) and treated New Zealand
white rabbit.

As shown in table 1, plasma glucose

level was significantly (P < 0.05)
higher in the heat stressed-treated
groups (91.00 3.39 mg/dl and
88.17 3.67 mg/dl, respectively)
compared with control non-treated
group (74.17 4.33 mg/dl). In
addition, plasma level of cholesterol
was significantly (P < 0.05) higher
in the heat stressed-treated groups
(65.67 1.85 mg/dl and 66.67
2.95 mg/dl, respectively) compared
with control non-treated group
(55.67 1.33 mg/dl). However, the
effect of vitamin E and selenium
complex on the other parameters
was not significant.

Table 1: Biochemical parameters in plasma of New Zealand white rabbits

exposed to heat stress (35 C) for 35 days.
Parameters
Glucose
(mg/dl)
Urea
(mg/dl)
Sodium
(mmol/L)
Potassium
(mmol/L)
Calcium
(mg/dl)
Phosphorus
(mg/dl)
Cholesterol
(mg/dl)
Total protein
(g/dl)
Albumin
(g/dl)
Globulin
(g/dl)
a,,b

Control (n = 9)

Groups
Treat1 (n = 9 )

Treat 2 (n = 9 )

74.17 a 4.33

91.00 b 3.39

88.17 b 3.67

30.00 1.48

30.00 2.92

31.00 1.89

147.67 0.88

146.00 0.68

145.5 1.26

5.87 0.14

5.72 0.11

5.80 0.32

12.42 0.71

11.77 0.61

11.58 0.65

8.47 0.26

8.62 0.21

8.00 0.31

55.67 a 1.33

65.67 b 1.85

66.67 b 2.95

6.21 0.14

5.86 0.15

5.85 0.11

4.58 0.11

4.43 0.09

4.47 0.05

1.63 0.10

1.43 0.14

1.38 0.13

Means within rows with different superscripts are significantly different

(P < 0.05).

SCVMJ, XVII (2) 2012

133

Fig. 1. Rectal and skin

temperatures of New
Zealand white rabbits
before and after exposure
to heat stress (35 C).

Fig. 2: Effects of vitamin E

and selenium injection on
rectal and skin temperatures
in New Zealand white heatstressed rabbits.

Discussion
Antioxidants are free radical
scavengers which protect the body
systems
against
excessively
produced free radicals and stabilize
health status of the animal. Vitamin
E is a free radical scavenger on the
cell membrane and metabolic
functions of selenium are closely
linked to vitamin E which not only
protects biological membranes from
oxidative degeneration but also

makes up an integral part of the

enzyme glutathione peroxidase of
the antioxidant system [19]. The
body temperature is recognized as
important measures of physiological
status [20] as well as ideal
indicators for assessment of stress
in animals. Hence, in the present
study, the increased rectal and skin
temperature after exposing rabbits
to heat stress suggested that the

134
rabbits were under a stressful
environment.
In the present study, it was found
that
heat
stress
increased
significantly rectal and skin
temperatures. Plasma glucose level
was higher in heat-stressed rabbit
treated with vitamin E and selenium
compared with that non-treated
rabbits. Ondruska et al [21] found
that the overall mean of plasma
glucose levels was significantly
lowered by elevated ambient
temperature in rabbits compared to
their counterparts in the control
group. The authors stated that the
decrease in glucose levels in the
heat-stressed rabbits could be due to
increases in glucose utilization
during
muscular
movements
required for high respiratory
activity [8,22], or due to increases
in corticosteroid concentrations
[23].
Nevertheless,
other
researchers have demonstrated that
decreases in energy metabolism
(gluconeogenesis
and
glycogenolysis)
during
heat
exposure correlated with decreases
in plasma insulin and thyroxin
concentrations [11, 24]. In a similar
study in goats [25], there was an
increase in rectal temperature
during the periods of heat stress. In
hot environments, animals attempt
to maintain heat balance by
increasing their respiratory activity,
thereby losing more heat by
evaporation from the respiratory
tract
than
under
normal
circumstances. The decreased rectal
temperature in the present study in

AL-Zafry and Medan

treatment groups indicated that
injection of vitamin E and selenium
complex ameliorated the heat stress
in rabbits. Similar decrease in rectal
temperature by vitamin E and C
supplementation was reported in
goats [25] and pigs [26] under heat
stressed conditions. Vitamin E and
C directly alters thermal set point
by decreasing prostaglandin output,
especially of prostaglandin E series,
whose turnover increases during
stress [27] which has a direct effect
on
the
hypothalamic
thermoregulatory
zone
[28].
Therefore, these vitamins may have
an ameliorating effect upon heat
stressed animals by affecting the
prostaglandin output.
It is generally recognized that
environmental factors may cause
stress and might alter animals
resistance to disease or affect
metabolism.
Amici et al [29]
reported that the short term
exposure of rabbits to heat stress
(one hour at 42 C) significantly
decreased the level of glucose to
(115 mg/dl) compared with the
control (126 mg/dl) after 54 hours.
Also, he found a significant
decrease of urea from 30 mg/dl to
22 mg/dl.
The overall mean of plasma glucose
levels in the present study was
significantly lower in heat-stressed
non treated rabbits compared with
heat-stressed rabbits treated with
vitamin E and selenium. The
decrease in glucose levels in the
heat-stressed adult rabbits could be
due to increases in glucose

SCVMJ, XVII (2) 2012

utilization
during
muscular
movements required for high
respiratory activity [8], or due to
increases
in
corticosteroid
concentrations [23]. Nevertheless,
other researchers have demonstrated
that decreases in energy metabolism
(gluconeogenesis
and
glycogenolysis)
during
heat
exposure correlated with decreases
in plasma insulin and thyroxin
concentrations [11, 24].
An explanation of this response
may be derived from a previous
observation on functional and
metabolic changes in heat-stressed
rabbits [30]. Rabbits stressed at
33.5C strongly reduced their feed
intake on the first day of treatment
(13.6 vs 161.6 g/d) but presented a
gradual increase toward the end of
the trial (98.8 vs 177.3 g/d).
However, a significant increase of
the
rectal
temperatures
in
comparison to rabbits under
thermoneutral
conditions
was
observed throughout the entire
period of exposure. However,
changes
in
a
variety
of
compensatory
physiological
processes
(e.g.,
antioxidant
systems) could also play a role in
the ability of rabbits to recover from
stress. This hypothesis is supported
by the results of Amici et al [30],
which reported a significant
increase in the plasma level of
vitamin E and a reduction in the
plasma concentration of sulphydrylgroups and in the total radical
trapping antioxidant parameter in
heat stressed rabbits.

135
The results of the present study
suggest that vitamin E and selenium
have an ameliorative effect on
physiological parameters during
heat stress. Hence, vitamin E and
selenium could be used as anti-heat
stressors in rabbits.
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