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Vitamin E and organic Se enhances the performance, antioxidative

properties and immune response of Japanese quail under high ambient
temperature

T. A. Ebeid*, A. A. Kirrella, S. B. Abou El-Soud, M. M. Ragab, Y. Z. Eid and M. M. El-

Habbak

Dept. of Poultry Production, Fac. of Agric., Kafrelsheikh Univ., 33516 Kafr El-Sheikh,
EGYPT
* Corresponding author: tarek.ebeid@agr.kfs.edu.eg, tarkamin@gmail.com

Abstract

The objective of the present study was to examine the effects of supplemental dietary vitamin
E (vit. E), organic selenium (Se) or vit. E+Se on growth performance, lipid peroxidation,
antioxidative status and immune response in growing Japanese quail under high ambient
temperature. A total of 200 14-day-old Japanese quail chicks were randomly divided into 4
experimental treatments (50 each): (1) control (basal diet without any supplementation of vit.
E or Se); (2) vit. E (basal diet +250 mg α-tocopherol acetate/ kg diet); (3) Se (basal diet + 0.3
mg organic Se/ kg diet); and (4) vit. E+ Se (basal diet +250 mg α-tocopherol acetate/ kg diet +
0.3 mg organic Se/ kg diet). The birds were given the experimental diets from 14 to 42 d of
age. The average daily temperature ranged from 33 to 36 ºC and relative humidity from 60 to
70% during the experimental period. Supplemental dietary vit. E, organic Se and vit. E+ Se
increased the final body weight and daily weight gain, while, feed conversion ratio was
reduced in vit. E+ Se group. Interstingly, the inclusion of vit. E plus organic Se in the growing
quails diet markedly enhanced the serum glutathione peroxidase (GSH-Px) activity and total
antioxidant capacity significantly and, simultaneously, reduced the thiobarbituric acid reactive
substance (TBARS) concentration in plasma. It is noteworthy to mention that supplementing
diets with vit. E and/or organic Se increased antibody titre in Japanese quail under high
ambient temperature. In conclusion, supplemental dietary vit. E, Se, or vit. E+Se enhanced
growth performance, serum antioxidative status, lipid peroxidation and immune
responsiveness in growing Japanese quail under high ambient temperature.

Keywords: Quail; vitamin E; organic selenium; immunity; antioxidative status.

Introduction

Heat stress is one of the most important stressors negatively affecting poultry industry leading
to loss of millions of dollars each year. It has been noted that modern broiler breeds are more
susceptible to heat stress than earlier genotypes. High ambient temperature reduces feed
intake, live weight gain, feed efficiency and immune response of chicken broilers (Melesse et
al., 2011). Moreover, it was reported that hyperthermia can potentially promote reactive
oxygen species (ROS) formation. Therefore, a balance between ROS production and the
antioxidant system must be established to maintain immune function, health and productivity
(Suraï, 2002).

Vitamin E (vit. E) is essential for such body functions as growth, immune function

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enhancement, tissue integrity, reproduction, disease prevention, and antioxidant function in

biological systems (Rooke et al., 2004). Dietary vit. E may enhance immune competence and
increase antibody production in broiler chicks (Singh et al., 2006) and mammals (Rooke et al.
2004). Selenium (Se) is an essential element required for normal animal growth and
reproduction (Suraï, 2002). Se-enriched meat could be achieved through the fortification of
diets with organic Se (selenomethionine) as it is incorporated into proteins in place of
methionine (Rayman, 2004). It has been demonstrated that dietary Se supplementation
increases Se concentration in the meat of chickens (Wang and Xu, 2008), rabbits (Ebeid et al.,
2013), lambs (Liu et al., 2011) and calves (Skřivanová et al., 2007). Se plays an important role
in activating the antioxidative properties via its involvement in the active site of the enzyme
glutathione peroxidase (GSH-Px) in blood, liver and edible tissues which might be connected
with enhancing the immune response (Suraï, 2002).

There is a synergistic relationship between Se and vit. E to protect against cellular damage by
ROS and this synergism may enhance meat quality and antioxidative properties (Suraï, 2002;
Ebeid et al., 2013). The metabolic function of Se is closely linked to Se-containing enzyme
GSH-Px functions in the cytosol of the cell, while vit. E is an integral component of lipid
membranes (Suraï, 2002). Studies reported that the combined supplementation of vit. E and
Se significantly increased GSH-Px activity (Ebeid, 2012; Ebeid et al., 2013) and improved the
lipid stability of meat compared to the control diet in rabbits (Ebeid et al., 2013) and chickens
(Kim et al., 2010). Moreover, studies had shown that administration of both Se and vit. E may
result in synergistic enhancement of the immune response and disease resistance (Suraï, 2002;
Singh et al., 2006; Ebeid et al., 2013). However, scientific information available on
performance, meat quality and immune responsiveness in growing Japanese quail fed
combinations of vit. E and Se under heat stress condition are still limited. Therefore, the
objective of the present study was to examine the effects of supplemental dietary vit. E (α-
tocopherol acetate), organic Se (Sel-Plex®) or vit. E+Se on growth performance, lipid
peroxidation, antioxidative status and immune response in growing Japanese quail under heat
stress condition.

Materials and methods

A total of 200 14-day-old Japanese quail (Coturnix coturnix japonica) chicks were randomly
divided into 4 experimental treatments (50 each): (1) control (basal diet without any
supplementation of vit. E or Se); (2) vit. E (basal diet +250 mg α-tocopherol acetate/ kg diet);
(3) Se (basal diet + 0.3 mg organic Se/ kg diet); and (4) vit. E+ Se (basal diet +250 mg α-
tocopherol acetate/ kg diet + 0.3 mg organic Se/ kg diet). The birds were given the
experimental diets from 14 to 42 d of age. Birds were placed in separate temperature
controlled brooder floor pens with 24 h of light in the Poultry Research Farm, Faculty of
Agriculture, Kafrelsheikh University, Egypt. The average daily temperature ranged from 33 to
36 ºC and relative humidity from 60 to 70% during the experimental period. The diets are
formulated to meet the recommendations of the National Research Council for Japanese quail
(NRC, 1994).

Feed intake and body weight were measured weekly and feed conversion ratio (feed/gain) was
calculated. Lipid peroxidation in the blood plasma was measured in the form of thiobarbituric
acid reactive substance (TBARS) as described by Richard et al. (1992). The activity of the
antioxidative enzyme GSH-Px and total antioxidant capacity in the blood plasma were
measured according to Paglia and Valentine (1967) and Koracevic et al. (2001), respectively
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using kits produced by Biodiagnostic, Egypt. Birds were vaccinated with Newcastle disease
“La-Sota” vaccine (Intervet International B.V., Boxmeer, Holland) at 32 days of age for
immunological examination. Blood samples (10 samples from each treatment) were collected
7 days after the vaccination and antibody titer against Newcastle disease virus (NDV) was
detected by hemagglutination-inhibition test according to Alexander et al. (1983). The
differences among treatments were statistically analyzed with a one-way ANOVA test in a
completely randomized design using Statistical Packages for the Social Sciences (SPSS®,
2001) with the type of diet as the main source of variation. The significant differences among
means were compared using Duncan's new multiple-range test.

Results and discussion

The effect of dietary vit. E, Se, or vit. E+Se supplementation on growth performance of
growing Japanese quail is presented in Table 1. Supplemental dietary vit. E, organic Se and
vit. E+ Se increased the final body weight and daily weight gain, while, feed conversion ratio
was reduced in vit. E+ Se group (P≤ 0.05). Similar results were observed by Ebeid et al.
(2013) who demonstrated that vit. E addition resulted in higher body weight in rabbits.
Likewise, dietary organic Se improved body weight and feed conversion ratio of broiler
chickens (Wang and Xu, 2008). The possible reasons are that natural antioxidants can protect
intestinal mucosa against oxidative damage and pathogens and limit peristaltic activity in
digestive disorders preventing diarrhea (Kermauner and Laurenčič, 2008) as well as to the
immunomodulating properties of Se and vit. E (Suraï, 2002; Rooke et al., 2004). Therefore, it
could be suggested that dietary supplementation of vit. E, Se, or vit. E+Se may be useful to
improve health and overall growth performance in growing Japanese quail under heat stress
conditions.

Table 1: Effect of dietary vitamin E and organic Se on growth performance, lipid

peroxidation, antioxidative status and immune response in growing Japanese quail
under high ambient temperature (means ±SE).

Treatment1
Sig.
Control Vit. E Se Vit. E+Se
Final body weight (g) 165.40±0.22b 172.47±0.29a 174.35±0.28a 175.12±0.29a *
Body weight gain (g) 110.2±0.33 b 116.75±0.56 b 119.57±0.24 b 124.33±0.32 a *
Feed intake (g) 400.29±18.53 500.57±15.69 503.51±13.56 488.22±18.36 *
b a a ab
c a ab
Feed conversion ratio (g 3.78±0.06 4.34±0.05 4.23±0.06 3.95±0.6 abc *
feed/g gain)
Plasma total antioxidant 0.34±0.01 d 0.50±0.02bc 0.45±0.02c 0.59±0.01 a *
capacity (mM/l)
Plasma glutathione 148.0±27.56 d 248.02±29.36 346.36±32.21 400.45±29.54 *
c b a
peroxidase activity (U/g
protein)
Plasma TBARS (nmol/l) 7.22±0.07 a 2.42±0.08 b 1.65±0.07 c 1.51±0.06 c *
Antibody titers (Log2) 3.54±0.30 b 4.33±0.23 a 5.00±0.35 a 4.66±0.28 a *
1
Experimental treatments include control (basal diet without any supplementation of Vit. E or Se); Vit. E (basal
diet +250 mg α-tocopherol acetate/ kg diet); Se (basal diet + 0.3 mg organic Se/ kg diet); and Vit. E+ Se (basal
diet +250 mg α-tocopherol acetate /kg diet + 0.3 mg organic Se/kg diet).
a,b,c,d
Means in the same row followed by unlike superscripts differ (P≤0.05).

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Dietary vit. E or Se resulted in a significant increase of blood serum antioxidative properties

as measured by GSH-Px activity and total antioxidant capacity as well as a decrease in blood
serum lipid peroxidation (i.e., TBARS) when compared with the control (P≤ 0.05, Table 1).
The inclusion of vit. E plus organic Se in the quail's diet markedly enhanced the serum GSH-
Px activity to more than two times the corresponding value of the controls. The vit. E and Se
combination reduced the TBARS concentration in blood serum to about 20% of the controls.
These results confirm that supplementing quail diets with vit. E has a positive effect on the
antioxidative properties, as it is a very efficient scavenger of free radicals (Ebeid., 2012; Ebeid
et al., 2013). Se plays an important role in the body's antioxidant defense system via activation
of GSH-Px and minimizing lipid peroxidation (Surai, 2002). Similarly, Saito et al. (2003) and
Ebeid (2012) reported that there is a synergistic relationship between Se and vit. E to protect
and maintain cell membrane integrity. The metabolic function of Se is closely linked to vit. E.
The Se-containing enzyme GSH-Px functions in the cytosol of the cell, while vit. E is an
integral component of lipid membranes (Suraï, 2002). Therefore, dietary vit. E in combination
with organic Se has a synergistic effect in minimizing lipid peroxidation in blood plasma and
maxmizing the antioxidant defense system, which probably translates into enhanced growth
performance, immunity and disease resistance in growing Japanese quail under high ambient
temperatutre. Getting such positive results is vital, especially under commercial production
conditions.

As shown in Table 1, dietary vit. E or Se caused an improvement in the antibody titers in

growing Japanese quail reared in high ambient temperature. Birds fed vit. E, Se, or vit. E+Se
had the highest antibody titers (4.33, 5.00 and 4.66 Log2, respectively) compared with the
control (3.66 Log2). These results suggested that vit. E and Se might have synergistic effects
on immune responses and these results support those concerning the antioxidative parameters
discussed above. The results from the current trial are in agreement with Singh et al. (2006)
and Ebeid et al. (2013). Singh et al. (2006) postulated that vit. E and Se act synergistically in
enhancing the humoral immune response in broiler chickens. Se intake appears to confer
additional health benefits on the immune system and reduction of inflammation (Rayman,
2004). Also, dietary vit. E plays an important role in enhancing the immune competence and
increases antibody production in broiler chicks (Singh et al., 2006). Rooke et al. (2004)
reported that both vit. E and Se are involved in multiple functions, at cellular and molecular
levels, such as regulation of cell signaling molecules (nuclear factor kappa B), reduction of
glucocorticoids (which are known to be immunosuppressive), minimizing rates and duration
of infections and regulation the function of neutrophils, lymphocytes and natural killer cells
via activation of interleukin-2 which finally results in enhancing cell-mediated and humoral
immune responses (Rayman, 2004). Since the effects of dietary Se and vit. E on the immune
response and synergistic effects between Se and vit. E are documented, it could be assumed
that the addition of natural antioxidants are beneficial to birds’ health especially under stress
conditions. Furthermore, by taking into account our results in lipid peroxidation and
antioxidative properties, it could be concluded that dietary vit. E, Se, or vit. E+Se may
enhance the immune responsiveness in growing Japanese quail under heat stress condition.

Conclusions

Based on the data presented above and taking into account antioxidant synergisms, it is
possible to suggest that supplemental dietary vit. E or Se might be involved in enhancing
growth performance, serum antioxidative status and immune responsiveness in growing
Japanese quail under heat stress condition.
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