Received: 26 August 2019
Accepted for publication: 10 January 2020
Slov Vet Res 2020: 57 (4): 149–57
DOI 10.26873/SVR-954-2020
UDC  636.5.082.4:636.087.8:638.17:612.616.2
Original Research Article
Introduction
Although domestic roosters may live for more 
than 10 years, the fertility of commercial breeder 
strains declines during the last phase of their 
reproductive life, so they are kept for a much shorter 
period. Commercial broiler breeders have a relatively 
shorter reproductive life span, and their reproductive 
performance decreases, particularly after 45 weeks 
of age (1, 2). Therefore it is recommended to keep 
broiler breeder flocks in production until 61 to 64 
weeks of age. The decline of fertility in ageing roosters 
was accompanied by a reduction in spermatozoa 
ANTI-AGEING EFFECTS OF DIETARY BEE PRODUCTS AND 
CALORIE RESTRICTION ON SEMEN PRODUCTION AND 
OXIDATIVE DAMAGE IN OLDER BROILER BREEDER MALES
Çiğdem Şeremet Tuğalay*, Özge Altan
Department of Animal Science, Faculty of Agriculture, Ege University, 35040, Bornova, Izmir, Turkey
*Corresponding author, E-mail: cigdemseremet@gmail.com
Abstract: This study was conducted to investigate the effects of calorie restriction and dietary bee products (apilarnil plus 
royal jelly) supplementation on reproductive and oxidative responses and to determine the possibilities that these treatments 
may be used in retarding the reproductive ageing of broiler breeder males. At 52 weeks of age, broiler breeder males were 
assigned to four treatment groups. The control group was fed on restricted feed as recommended by the breeder company 
throughout the study; the ad libitum group was fed ad libitum for a four-week period; the bee products group was fed similar 
to the control group except that their diet was supplemented with apilarnil and royal jelly for a four-week period and in the last 
group calorie restriction (45 % of standard diet) was applied for a four-week period. After a four-week adaptation period, the 
experiment was continued for 18 weeks. The results obtained in the present study have demonstrated that the percentage of 
dead sperm was the most affected semen characteristic by reproductive ageing. Long-term moderate feed restriction could 
not prevent age-related declines in sperm production. Dietary bee products supplementation or calorie restriction for a four-
week period positively affected the semen characteristics, and these beneficial effects could be maintained to some extend 
up until 72 weeks of age. Calorie restriction enhanced antioxidant defence for the first four-week period; however, this benefi-
cial effect could not be sustained until the end of the experiment.
Key words: broiler breeder males; ageing; semen characteristics; oxidative stress; bee products; calorie restriction
production and by a decrease in motility and viability 
of spermatozoa (3, 4, 5). In contrast, the demand 
for spermatozoa increases with ageing because of 
the changes in the oviduct conditions, especially in 
sperm storage tubules (6, 7). Therefore, one of the 
most important goals in the poultry industry is to 
extend the reproductive life span and retard signs 
of reproductive senescence, both in terms of animal 
welfare and production costs.
Calorie restriction (CR) has been shown to 
extend the life span and retard the onset of many 
age-related disorders in a variety of animal models 
(8, 9, 10). Short-term calorie or food restriction 
has been routinely used by the poultry industry 
for many years to extend the reproductive life span 
and delay the maturation of laying hens (11, 12). 
Ç. Ş. Tuğalay, Ö. Altan150
However, it has been suggested that severe CR 
caused a significant decrease both in testis weight 
and in plasma testosterone levels and an increase 
in plasma corticosterone levels (13, 14).
The cumulative oxidative damage to macromol-
ecules caused by reactive oxygen species (ROS) is 
considered to be related to cellular senescence, life 
span, and fitness (15, 16). It is suggested that in 
avian cells the accumulation of ROS-induced dam-
age would be slower due to lower ROS production, 
better resistance to oxidative stress, enhanced an-
tioxidant capacity, and better DNA repair (16, 17, 
18). If ageing is caused by ROS as described by 
the free radical theory or oxidative damage theory 
of ageing, supplementation of exogenous antioxi-
dants may enhance the antioxidant defence capac-
ity, slow down ageing, and prolong the reproduc-
tive life span.
Sperm membranes have a higher content of 
PUFAs (polyunsaturated fatty acids); therefore, 
they are more susceptible to oxidative damage. 
Oxidative stress may be reduced by antioxidant 
supplementation of broiler breeder diets (19).
Therefore, in this study, the natural bee products 
apilarnil and royal jelly have been used to retard 
the ageing process because of their antioxidant 
and antiaging activities.
Royal Jelly (RJ) is a functional food secreted 
by the hypopharyngeal and mandibular glands of 
worker bees. RJ has many properties, including 
antitumor, antibacterial, antioxidant, antiaging, 
hypotensive, growth-stimulating, and anti-
inflammatory activities (20). It is believed that 
RJ can prolong life span because it prolongs the 
longevity of queens comparing to worker bees. In 
a mice model study, it is suggested that a 16-week 
RJ supplementation prolonged the average life 
span protecting the DNA and lowering oxidative 
stress (21). 
Apilarnil is mainly a drone larvae extract that 
also contains small amounts of royal jelly, bee 
bread, honey, and propolis. Apilarnil has many 
pharmacological activities, such as anabolism 
stimulator, antiviral, immunomodulator, bio-
stimulator (22, 23). Additionally, because it 
comes from a “male-like” structure, apilarnil is 
very rich in male-like hormones, so it stimulates 
the spermatogenesis (22, 24) and regulates the 
human endocrine system (25, 26, 27). It also has 
high levels of 10-HDA (10-hydroxy-2-decenoic 
acid), and many vitamins and minerals (27, 28) 
affecting antioxidant capacity. (29) reported that 
both the total antioxidant potential and activity of 
capturing the free radicals of RJ and drone larvae 
are sufficiently high, and their combination leads 
to extremely valuable products.
The present study was conducted to determine 
the possibilities that calorie restriction or dietary 
apilarnil and royal jelly supplementation may be 
used in retarding the reproductive ageing of broiler 
breeder males. We evaluated the effects of those 
dietary manipulations on the aging of roosters by 
quantifying lipid peroxidation, some antioxidant 
activities, and semen quality parameters. Moreover, 
these age-related parameters were compared with 
those of the ad libitum feeding group.
Materials and methods
 
The ethical committee approval of Ege University 
(2011-092) was granted in order to conduct this 
study.
Experimental design and diets
A total of 160 broiler breeder males (Ross 
308) at 52 weeks of age were used in the present 
study. Males were identified with a leg tag and 
randomly assigned to 4 treatment groups with four 
replications. The treatments were 1) males were fed 
a restricted feed (135 g/day) as recommended by the 
breeder company (Control-CONT); 2) males were fed 
ad libitum for a four-week experimental period (Ad 
libitum-ADL), 3) males were fed similar to the CONT 
group except that diet was supplemented with bee 
products (apilarnil 5 g/d/male and royal jelly 200 
mg/d/male) (Bee products-BP); 4) calorie restriction 
was applied at about 45 % of the standard diet for a 
four-week experimental period (Calorie restriction-
CR). After a four-week adaptation period, the 
experiment was continued for 18 weeks. Males were 
placed in floor pens and fed a standard diet (Table 1) 
except for the four-week experimental period.
1: Allzyme SSF, Aspergillus niger (CBS 114.94) 
amylase, cellulase, phytase, xylanase, beta-
glucanase, pectinase, protease.
2: 2 kg mineral mixture, antioxidant, 125.000 
mg; copper, 10.000 mg; calcium D pantothenat, 
15.000 mg; zinc, 100.000 mg; D-Biotin, 250 mg; 
iron, 60.000 mg; folic acid, 2.000 mg; iodine, 
2.000 mg; cobalt, 500 mg; manganese, 80.000 mg; 
niacin, 55.000 mg; selenium, 250 mg.
3: 1 kg vitamin mixture, retinol-acetate, 13.000.000 
Ç. Ş. Tuğalay, Ö. Altan151
IU; thiamine, 3.000 mg; cyanocobolamin, 40 mg; 
riboflavin, 12.000 mg; pyridoxine, 4.500 mg; 
cholecalciferol, 3.000.000 IU; α-tocopherol acetate, 
100.000 mg; menadione, 5.000 mg.
4: Toxin binder, probiyotic, vitamin D3
Table 1: Nutrient composition and analysis of standard 
and calorie restriction diets (g/kg)
Sample collection and analysis
Blood samples for biochemical analysis were 
collected from 10 males of each treatment group 
at the 4th, 8th, and 18th weeks of the trial. Lipid 
peroxidation (LPO) was ascertained by the 
formation of malondialdehyde (MDA), which was 
estimated using the thiobarbituric acid (TBARS) 
method (30). Superoxide dismutase (SOD) 
activity was determined using the commercially 
available enzyme kit (Ransod, RANDOX/SD-125). 
Glutathione peroxidase (GSH-Px) activity was 
determined using a Ransel kit (RANDOX/RS-504). 
The Randox Uric Acid (UA) Enzymatic Colorimetric 
method kit was used for uric acid analysis. Total 
antioxidant capacity (TAC) was measured using an 
Abbott Architect Analyzer commercial kit (Abbott 
Lab. Illinois, USA).
Semen collection and evaluation
Semen samples were collected using the abdominal 
massage technique (at 2nd, 6th, 10th, and 16th weeks 
of trial) and evaluated in 20 minutes using Sperm 
Vision System (Minitüb Abfüll und Labortechnik 
Gmbh&Co.KG). Semen volume, motility (%), dead 
sperm (%), progressive motility (%) were determined 
in each semen sample.
Statistical analysis 
The data were analysed using a one-way analysis 
of variance (ANOVA) with the General Linear Models 
(GLM) procedure of the SAS software (31). Significant 
differences between groups were determined by 
Student’s t-test. Differences were considered to be 
significant at P<0.05 and the results were presented 
as the mean and standard error of the mean (SEM).
Results and discussion
The effects of dietary manipulations on semen 
quality parameters are presented in Table 2. As 
seen prior to treatments, there were no significant 
differences among groups for semen quality 
parameters. Dietary treatments caused dramatic 
changes in all semen parameters throughout 
the experimental period. At the 2nd week, BP 
supplementation positively affected semen quality. 
These males had the highest progressive sperm 
motility and semen volume. However, CR treatment 
adversely affected semen quality. Lower semen 
volume, lower sperm motility and higher dead 
sperm were observed in CR males compared to 
the other groups. After a four-week CR period, the 
detrimental effects of CR on semen production were 
gradually alleviated by increasing feed allocation.
Ingredients Standard diet (g/kg)
Corn 748.49
Full fat soybean   40.00
Soybean cake   12.42
Sunflower cake 148.76
Acid oil   10.00
Limestone   23.53
MCP-22.7     4.28
Sodium sulphate     1.21
Salt     2.40
DL-Methionine     0.89
Vitamin C     0.20
L-Lysine sulphate     0.32
Colin chloride liquid     0.50
Enzyme mixture1     0.20
Trace Mineral mixture2     0.50
Vitamin mixture3     2.00
Other additives4     4.80
Analysed composition            (g/kg)
Dry matter 875.70
Crude protein 133.40
Crude fat   50.00
Crude fibre   50.70
Crude ash   56.60
ME (MJ kg-1)   12.01
Ç. Ş. Tuğalay, Ö. Altan152
Table 2: Semen quality characteristics in broiler breeder males during the experimental period (±SEM)
Pr
et
re
at
m
en
t 
2 
w
k
6 
w
k
10
 w
k
16
 w
k
Pr
e 
vs
 
Po
st
-t
re
at
m
en
t
G
R
O
U
P
Semen volume (ml)
Motility (%)
Progressive sperm motility (%)
Dead sperm (%)
Semen volume (ml)
Motility (%)
Progressive sperm motility (%)
Dead sperm (%)
Semen volume (ml)
Motility (%)
Progressive sperm motility (%)
Dead sperm (%)
Semen volume (ml)
Motility (%)
Progressive sperm motility (%)
Dead sperm (%)
Semen volume (ml)
Motility (%)
Progressive sperm motility (%)
Dead sperm (%)
Semen volume
Motility
Progressive sperm motility
 Dead sperm
C
O
N
TR
O
L
0.
23 ±
0.
02
98
.0
6
±
0.
71
87
.6
2
±
0.
75
2.
13 ±
0.
34
0.
27 ±
0.
01
c
95
.2
1
±
0.
47
b
88
.2
1
±
0.
52
b
9.
56 ±
0.
92
a
0.
25 ±
0.
01
b
88
.3
9
±
0.
70
b
84
.6
9
±
0.
92
b
11
.2
7
±
1.
07
a
0.
18 ±
0.
02
b
85
.3
6
±
0.
86
b
78
.2
5
±
0.
97
c
16
.3
2
±
1.
83
b
0.
18 ±
0.
01
c
84
.2
6
±
0.
88
b
80
.3
9
±
0.
93
c
16
.9
2
±
1.
48
a
N
S
**
*
**
**
*
A
D
L
0.
31 ±
0.
04
98
.1
2
±
0.
55
84
.9
7
±
1.
89
2.
07 ±
0.
68
0.
39 ±
0.
02
b
98
.5
6
±
0.
53
a
87
.2
1
±
0.
59
b
2.
06 ±
0.
62
b
0.
21 ±
0.
02
bc
82
.1
1
±
1.
39
c
69
.3
2
±
1.
03
d
8.
66 ±
1.
55
a
0.
25 ±
0.
04
b
78
.2
1
±
0.
83
c
60
.1
0
±
1.
06
d
24
.3
4
±
1.
49
a
0.
33 ±
0.
04
b
80
.9
5
±
2.
21
b
72
.6
9
±
2.
03
d
20
.4
8
±
2.
43
a
N
S
**
*
**
**
*
B
P
0.
30 ±
0.
07
97
.5
1
±
0.
73
84
.4
1
±
2.
31
1.
73 ±
0.
36
0.
46 ±
0.
03
a
95
.3
1
±
0.
54
b
91
.2
7
±
1.
31
a
2.
33 ±
0.
50
b
0.
72 ±
0.
10
a
98
.9
6
±
0.
16
a
94
.3
2
±
0.
49
a
0.
41 ±
0.
11
b
0.
61 ±
0.
02
a
99
.2
6
±
0.
11
a
93
.8
5
±
0.
30
a
0.
79 ±
0.
13
d
0.
43 ±
0.
01
a
97
.8
9
±
0.
12
a
93
.0
0
±
0.
35
a
1.
51 ±
0.
21
b
N
S
N
S
**
N
S
C
R
0.
36 ±
0.
05
97
.6
4
±
0.
74
87
.2
3
±
2.
32
2.
45 ±
0.
29
0.
11 ±
0.
01
d
88
.2
1
±
0.
72
c
81
.5
1
±
0.
73
c
7.
89 ±
0.
42
a
0.
07 ±
0.
01
c
81
.5
8
±
0.
43
c
80
.6
3
±
1.
31
c
9.
23 ±
0.
73
a
0.
27 ±
0.
02
b
87
.1
5
±
0.
68
b
84
.5
3
±
0.
45
b
5.
51 ±
0.
48
c
0.
39 ±
0.
01
ab
96
.2
5
±
0.
66
a
89
.3
3
±
0.
91
b
4.
62 ±
0.
94
b
N
S
N
S
N
S
*
X
±S
E
M
0.
30 ±
0.
02
97
.8
3
±
0.
32
86
.0
6
±
0.
94
2.
09 ±
0.
21
0.
31 ±
0.
03
94
.3
2
±
0.
90
87
.0
5
±
0.
89
5.
46 ±
0.
81
0.
31 ±
0.
06
87
.7
6
±
1.
64
82
.2
4
±
2.
10
7.
39 ±
1.
05
0.
33 ±
0.
04
87
.5
0
±
1.
76
79
.1
8
±
2.
85
11
.7
4
±
2.
18
0.
33 ±
0.
02
89
.8
4
±
1.
77
83
.8
5
±
1.
89
10
.8
8
±
1.
96
Pr
ob
ab
ili
ty
N
S
N
S
N
S
N
S
**
*
**
*
**
*
**
*
**
*
**
*
**
*
**
*
**
*
**
*
**
*
**
*
**
*
**
*
**
*
**
*
ab
c d
M
ea
n
 v
al
u
es
 w
it
h
in
 t
h
e 
sa
m
e 
co
lu
m
n
 s
h
ar
in
g 
a 
co
m
m
on
 s
u
pe
rs
cr
ip
t 
le
tt
er
 a
re
 n
ot
 s
ta
ti
st
ic
al
ly
 d
iff
er
en
t 
at
 P
<0
.0
5.
*:
 P
<0
.0
5;
 *
*:
 P
<0
.0
1;
 *
**
: *
: 
P<
0.
05
; *
*:
 P
<0
.0
1;
 *
**
: P
<0
.0
00
1;
 N
S
: N
ot
 S
ig
n
ifi
ca
n
t 
(P
>0
.0
5)
. A
D
L:
 A
d 
lib
it
u
m
; B
P:
 B
ee
 P
ro
du
ct
s;
 C
R
: C
al
or
ie
 R
es
tr
ic
ti
on
.
Ç. Ş. Tuğalay, Ö. Altan153
At the end of the experiment (16th week), all 
semen quality parameters of CR and BP males were 
significantly better than those of CONT and ADL. 
Semen volume was higher in ADL males than that 
of CONT, and there were significant differences 
in sperm motility and dead sperm between these 
groups.
The semen characteristics significantly changed 
throughout the experimental period (Table 2). The 
percentage dead sperm of ADL males markedly 
increased from 2.07 % at the beginning to 
20.46 % on the 16th week of the experiment. 
Similarly, in CONT males, the percentage of dead 
sperm increased from 2.13 % to 16.92 % during the 
experimental period. At the end of the experiment, 
a 13-fold increase in percentage dead sperm of ADL 
males occurred in comparison with the BP males. 
It is noteworthy that dead sperm (%) was one of 
the semen characteristics most affected by age and 
dietary treatments.
At the end of the experiment, the ADL, BP, and 
CR males maintained semen volume similar to 
those observed at the beginning of the experiment 
while long-term restricted males (CONT) produced 
lower semen volume than the other groups did.
The results of the present study showed that 
sperm motility and progressive sperm motility 
significantly decreased while dead sperm 
dramatically increased in CONT males during 
the experimental period. These data imply that 
long-term feed restriction (135 g/d) recommended 
by the breeder company for males could not 
prevent age-related declines in semen production. 
Although during CR treatment period, all semen 
quality parameters were adversely affected, calorie 
restriction at 45 % of the standard diet for weeks 
retarded onset of age-related decline in semen 
quality. The effect of CR treatment observed in 
the present study was in agreement with previous 
studies reporting inadequate ME intake could 
be detrimental to semen production (4, 32, 33) 
and these detrimental effects could be revised by 
increasing male feed allocation (34, 35).
(36) suggested that there were no significant 
differences in sperm production, semen volume 
or sperm concentration between full-fed and feed 
restricted males. Moreover, they reported that while 
overfeeding might assist a male in semen production 
for the short-term, the long-term effects of being 
over-weight are negative ones. In agreement with 
these findings, we obtained no significant differences 
in sperm motility and dead sperm between ADL and 
CONT males, but semen volume was higher in ADL 
males.
In the BP group, progressive motile sperm 
significantly increased while there were no negative 
changes in the other semen characteristics at the 
end of the experiment compared to the pretreatment 
period, suggesting that age-related declines in 
semen production could be alleviated with dietary 
BP supplementation. Congruent with these results, 
it was reported that RJ administration caused an 
increase in sperm production, sperm motility, and 
higher testosterone levels in lab animals (37, 38, 
39). (40) suggested that the androgenic effect of 
apilarnil on chickens was higher than its anabolic 
effect. Supporting this report, (41) obtained that 
apilarnil administration at an early age increased 
testicular weights and testosterone production and 
stimulated comb growth in broiler males. (42) also 
reported that RJ administration has a positive effect 
on libido, semen quality, sperm output, testosterone 
level, and fertility of heat-stressed male rabbits. 
Controversially, (43) suggested that high dose RJ 
(800 mg/kg) administration for four weeks adversely 
affected the reproductive system of pubescent male 
rats, but these detrimental effects are alleviated to 
some extend by the cessation of RJ.
The effects of dietary manipulation on some 
antioxidant activities and MDA levels are presented 
in Table 3. On the 4th week, dietary treatment 
significantly affected TAC and UA levels, but no 
significant effects were observed in MDA, SOD, 
and GSH-Px levels. The TAC and UA levels were 
significantly higher in the CR group than in 
others. At the end of the experiment, there were no 
significant differences in antioxidant capacity and 
MDA levels among the treatment groups.
Ç. Ş. Tuğalay, Ö. Altan154
4 wk
GROUP UA(mg/dL)
TAC
(mmol/L)
SOD
(U/L)
GSH-Px
(U/L) MDA (umol/L)
CONTROL 10.18±1.09b 0.76±0.00b 1.27±0.26 22983.17±1710.69 1.92±0.10
ADL 7.22±1.02b 0.78±0.14b 1.59±0.18 23553.67±2851.07 1.87±0.19
BP 10.28±1.24b 1.04±0.14b 1.62±0.24 28788.20±3675.19 1.72±0.24
CR 25.38±1.58a 1.62±0.07a 1.42±0.20 20808.80±3371.93 2.02±0.37
Probability *** ** NS NS NS
8 wk
CONTROL 9.80±0.99 0.78±0.05 1.70±0.08 24845.67±2707.96ab 2.58±0.83
ADL 7.94±1.12 0.96±0.10 1.57±0.16 30443.83±2994.62a 2.62±0.49
BP 8.56±1.18 1.08±0.26 1.25±0.17 19736.00±910.62b 1.58±0.19
CR 8.98±0.99 0.97±0.12 1.41±0.21 18596.00±1869.43b 1.22±0.16
Probability NS NS NS ** NS
18 wk
CONTROL 7.30±0.50 0.74±0.07 1.16±0.38 28829.17±4625.01 1.88±0.17
ADL 8.14±0.84 0.89±0.11 1.64±0.21 28227.33±1525.50 2.37±0.65
BP 8.10±0.87 0.77±0.00 1.54±0.17 24122.00±2406.23 3.40±1.21
CR 8.33±1.08 0.81±0.08 1.66±0.15 29366.00±6130.38 1.14±0.21
Probability NS NS NS NS NS
Table 3: Some biochemical parameters of broiler breeder males during the experimental period (±SEM)
abMeansvalues within the same column sharing a common superscript letter are not statistically different at P<0.05.
*: P<0.05; **: P<0.01; ***: P<0.0001; NS: Not Significant (P>0.05). ADL: Ad libitum; BP: Bee Products; CR: Calorie Restriction. UA: Uric Acid; TAC: 
Total Antioxidant Capacity; SOD: Superoxide Dismutase; GSH-Px: Glutathione Peroxidase; MDA: Malondialdehyde. 
Overexpression of antioxidant enzymes has 
been considered to be a protective response to 
oxidative stress (44, 45, 46). Therefore, both 
increased antioxidant activity and no significant 
increase in MDA level of CR males may be 
interpreted as a protective response to LPO 
damage. Supporting our results, previous studies 
showed that in calorie-restricted animals, free 
radical generation and LPO decreased (47, 48) 
antioxidant defence capacity was enhanced (49), 
DNA damage decreased (50), and the rate of 
the ageing process was modulated by retarding 
many age-related physiological declines (51). (52) 
suggested that a-10 day fasting treatment can be 
used to decrease oxidative stress-mediated injury 
in aged hens without affecting the welfare of hens 
from previous fasting experiments.
After a four-week treatment period, it was 
observed no significant difference among 
treatment groups in MDA levels and antioxidant 
capacity except GSH-Px activity on the 8th week, 
suggesting that all groups had similar LPO 
responses and that there were no significant 
differences in possible age-related oxidative stress. 
It is reported that RJ has high antioxidant 
activity and scavenging ability against free radical 
(20, 29, 39). However, in the present study, BP 
supplementation (royal jelly plus apilarnil) for a 
four-week period could not enhance the antioxidant 
capacity, indicating that the antioxidant 
properties of these products were probably not 
adequate to affect the oxidative status of males. 
Supporting this result, (53) reported that RJ and 
its bioactive component 10-HDA, did not scavenge 
any ROS; dietary RJ might have protective effects 
against tissue damage through other mechanisms 
other than ROS scavenging. There was no report 
focusing on the antioxidant effect of apilarnil in 
poultry; therefore, the present results could not 
be compared with other studies in the literature.
As a result, it was obtained that the ageing 
effect was most pronounced in the percentage 
of dead sperm, especially in males fed ad 
libitum and restricted as recommended by the 
breeder companies. Royal jelly plus apilarnil 
supplementation or calorie restriction for a 
Ç. Ş. Tuğalay, Ö. Altan155
four-week period positively affected the semen 
characteristics of broiler breeder males at 56 
weeks of age, and these beneficial effects could 
be maintained to some extent until 72 weeks of 
age. Supporting these results, (54) determined 
that antioxidant diet supplementation resulted in 
a higher percentage of normal sperm cells in male 
broiler breeders older than 50 weeks.
Calorie restriction has been enhanced 
antioxidant defence for the first four-week period, 
indicating a protective mechanism against 
oxidative stress. However, after the calorie 
restriction period, this positive effect could not be 
sustained. A four-week calorie restriction period 
may not be long enough to induce a long-lasting 
oxidative response; a longer time may be required 
to obtain long term antiaging and antioxidative 
effects.
In conclusion, it seems to be possible that 
dietary bee products supplementation or calorie 
restriction can be used to slow down the rate of 
the ageing process and extended reproductive life 
span of broiler breeder males by retarding an age-
related decline in semen production. However, the 
long-term moderate feed restriction recommended 
by the breeder companies could not prevent age-
related decline in semen quality parameters.
Acknowledgements 
The study was supported by The Scientific 
& Technological Research Council of Turkey 
(TUBITAK), Project No: 111O272 and Scientific 
Research Projects Committee at Ege University, 
Project No: 2011ZRF068.
The authors declare that they have no conflict 
of interest.
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VPLIV ČEBELJIH PRIDELKOV IN OMEJEVANJA KALORIJ NA PROIZVODNJO SEMENA 
IN OKSIDATIVNI STRES PRI STAREJŠIH SAMCIH PLEMENSKIH BROJLERJEV
Ç. Ş. Tuğalay, Ö. Altan
Povzetek: Študija je bila izvedena z namenom raziskovanja učinkov omejevanja kalorij in dodajanja prehranskih čebeljih 
pridelkov (apilarnil in matični mleček) na reprodukcijske in oksidativne odzive ter ugotoviti možnosti uporabe prehranskih 
dodatkov za zaviranje reproduktivnega staranja samcev plemenskih brojlerjev. Pri starosti 52 tednov so bili samci plemenskih 
brojlerjev razporejeni v štiri skupine. Kontrolna skupina je bila ves čas študije krmljena z restrikcijsko krmo po priporočilih 
podjetja, ki se ukvarja z gojenjem plemenskih broilerjev; skupina ad libitum je bila štiri tedne hranjena ad libitum; skupina, pri 
kateri so bili dodani čebelji pridelki je bila krmljena podobno kot kontrolna skupina, le da je bila njihova prehrana štiri tedne 
dopolnjevana z apilarnilom in matičnim mlečkom, zadnja skupina pa je štiri tedne dobivala kalorično omejeno hrano (45 % 
običajne prehrane). Po štiritedenskem prilagoditvenem obdobju se je poskus nadaljeval še 18 tednov. Rezultati, pridobljeni v 
tej študiji, so pokazali, da je bila najbolj prizadeta značilnost staranja povišan odstotek mrtvih semenčic v ejakulatu. Dolgoročna 
zmerna omejitev krme ni preprečila starostnega zmanjšanja proizvodnje smenčic. Dodatek prehranskih čebeljih pridelkov ali 
omejevanje kalorij v obdobju štirih tednov je pozitivno vplival na značilnosti semena. Ti blagodejni učinki so se ohranili vse do 
starosti do 72 tednov. Omejitev kalorij je okrepila tudi antioksidativno obrambo v prvih štirih tednih raziskave; vendar pa se je ta 
ugodni učinek kasneje izgubil.
Ključne besede: samci plemenskih brojlerjev; staranje; značilnosti semena; oksidativni stres; čebelji proizvodi; omejitev kalorij