Advances in Environmental Biology of laying hens

Advances in Environmental Biology, 7(13) November 2013, Pages: 4382-4389
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Advances in Environmental Biology
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Effect of dietary medicinal herbs on Performance, egg quality and immunity response
of laying hens
1
Jafari Navid, 2Mahdizadeh Mozaffar and 1Karimi Kazem
1
Department of Animal Science, Varamin-Pishva Branch, Islamic Azad University, Varamin, Iran.
Animal Science Research Institute (ASRI) karaj, Iran.
2
ARTICLE INFO
Article history:
Received 11 September 2013
Received in revised form 21 November
2013
Accepted 25 November 2013
Available online 29 December 2013
Keywords:
Medical herbs, Egg quality, Layer
hens,
Probiotic,
Performance,
Immunity.
ABSTRACT
This study was conducted to evaluate the effect of a dietary mixed medicinal herbs
powders (MHPs) (Thymus vulgaris, Pennyroyal, Cumin, Alhagi, Garlic and Eucalyptus
globules) on performance, egg quality and immunity response of laying hens. The
experiment was carried out under completely randomized design, including five
treatments (Treatment 1= basal diet without additive (control group), Treatment 2=
basal diet + probiotics at 0.1%, T3, T4 and T5=basal diet +1, 2 and 3% MHPs
respectively) with five replicates (12 birds each replicate). 300 Bovans laying birds (49
wks old) were distributed randomly in cages and allotted into one of experimental diets
for 84 days. The performance was recorded every week. In order to examine the
antibody against the sheep red blood cell (SRBC), 2 piece of hen from each replicate
was randomly chosen (1cc) SRBC (%0.5) was injected and ten days later blood samples
were collected. To examine the blood parameters (Serum Cholesterol, Red blood cells,
White blood cells, Lymphocyte, Packed cell volume), In the day 70th , from each
replicate 2 piece of hen blood samples was taken. Egg quality parameters (High unit,
specific gravity, shell weight and thickness and yolk cholesterol) at the end of the
experiment were evaluated. Diets supplemented with 1, 2, or 3% MHPs had no
significant effects on percentage of egg production, egg weight mass, and feed
conversion ratio (FCR). However, at the level of 2 and 3%, daily feed intake decrease
significantly (P< 0.05) compared to the control, probiotic and 1% MHPs. Probiotic and
different levels of MHPs had no significant effect on SRBC. Probiotic and 1 and 2%
MHPs significantly (P < 0.05) decrease IgG in compare to control and 3% MHPs.
Using probiotic in the diet significantly increase IgM in laying hens in compare to
control and different levels of MHPs. Probiotic and levels of MHPs had no significant
effect on egg weight, shell weight, shell thickness, specific gravity and haugh unit and
serum cholesterol. However, MHPs at the level of 1% increase yolk cholesterol in
compare to probitic, control, 2 and 3% MHPs significantly. In conclusion, using
medical herbs in mixed form may suppress positive effect of medical herbs in compare
to using individually.
© 2013 AENSI Publisher All rights reserved.
To Cite This Article: Jafari Navid, Mahdizadeh Mozaffar and Karimi Kazem., Effect of dietary medicinal herbs on Performance, egg
quality and immunity response of laying hens. Adv. Environ. Biol., 7(13), 4382-4389, 2013
INTRODUCTION
Antibiotic growth promoters have undoubtedly improved animal performance and health status. It is
apparent that antibiotics act by modifying the intestinal micro flora. The microbes can develop resistance to
these antibiotics and when transferred to human beings, may be problematic because of the resistance to these
antibiotics. This topic has been discussed many. Many antibiotics in livestock and poultry production as growth
promoters are banned in several countries and all antibiotics may soon be banned. Therefore, there is a demand
for alternatives to antibiotics in an increasingly health conscious market [9, 14, 17]. Nowadays, the searches
for alternative feed supplements have been increased extensively and considerable attention has been given to
the medicinal herbs as replacements for antibiotics [25].
It is reported that supplementation of laying hens diet with Thyme (Thymus vulgaris) improve feed
conversion and egg production and reduce E. coli concentration in feces [6]. It is reported that black cumin at
level of 2 or 3% can positively influence egg production, egg weight, and shell quality and decrease the
concentration of cholesterol in the egg yolk [5]. Feeding powdered Nigella sativa L. seeds significantly reduce
yolk cholesterol content. The serum triglycerides, low-density lipoprotein, cholesterol and total cholesterol
levels were also reduced, while serum high-density lipoprotein cholesterol level was increased by
supplementing the commercial layer diet with Nigella sativa seeds [1]. Supplementation of commercial laying
Corresponding Author: Jafari Navid, Department of Animal Science, Varamin-Pishva Branch, Islamic Azad University,
Varamin, Iran.
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Advances in Environmental Biology, 7(13) November 2013, Pages: 4382-4389
hens diet with Eucalyptus improve productive performance and immunocompetence [16]. It has been showed
that an essential oil combination derived from selected herbs growing wild in Turkey may be considered as a
potential growth promoter in broiler production [2]. In addition, It has been demonstrated that garlic powder
would increase egg weight and would decrease egg yolk cholesterol concentration, serum triglyceride and
cholesterol concentrations without adverse effects on performance and egg traits [37]. Results of evaluation of
the effects of using probiotic, enzyme and Pennyroyal herbs on performance of laying hens has been showed
that combination of 1 and 2 % Pennyroyal individually, or in combination with probiotic, enzyme and both
could not provide an improved performance of laying hens. There is little evidence to show the effects of
addition of medicinal herbs to diet in mixed form on performance, egg quality characteristics and immunity
response of commercial laying hens. Therefore, we examine the effect of 6 medicinal herbs powders (Thymus
vulgaris, Pennyroyal, Cumin, Alhagi, Garlic and Eucalyptus globules) in mixed form in laying hens diet. There
is growing evidence have show positive effects of probiotics on animal health and performance [7, 12, 19]. We
also used a commercial probiotic (Protexin) in order to compare our treatments.
MATERIAL AND METHOD
This study was conducted in the research farm at the Animal Science Research Institute (ASRI) of Iran. In
this study, three hundred 49-wk-old laying hens (Bovans strain) were randomly assigned into 5 groups with 5
replicates of 12 birds each (60 laying hens per group) and fed diets supplemented with; Treatment 1= basal diet
without additive (control group), Treatment 2= basal diet + %0.1 probiotic (Protexin® Probiotic International
Limited, Uk), treatment 3, 4 and 5=basal diet +1, 2 and 3% MHPs respectively for 12 wk. Table 1 presents the
composition of the experimental diets.
Table 1: Composition of the diets and their nutrients.
Ingredients
Control (%)
Corn
Wheat
Soybean meal
NaCl
Concentrate
Dicalcium phosphate
Vegetable oil
Oyster shell
MHP
Metabolizable energy (Kcal/kg)
Crud protein (%)
Calcium (%)
Phosphorus (%)
Na (%)
Methionine (%)
Methionine +cystine (%)
Cystine (%)
Tryptophan (%)
1. Medicinal herbs powders
34
30
20.5
0.1
4.5
0.6
1.2
9.1
0.0
2773
15.73
4.15
0.41
0.18
0.42
0.69
0.87
0.2
Probiotic (%)
34
30
20.1
0.1
4.5
0.6
1.1
9.1
0.0
-
1
33
30
20.4
0.1
4.5
0.6
1.3
9.1
1.0
2762
15.82
4.17
0.41
0.18
0.42
0.69
0.88
0.2
MHPs1 (%)
2
33
29
20.3
0.1
4.5
0.6
1.4
9.1
2.0
2763
15.89
4.17
0.41
0.18
0.42
0.69
0.88
0.2
3
32
29
20.2
0.1
4.5
0.6
1.5
9.1
3.0
2761
15.89
4.18
0.41
0.18
0.42
0.69
0.88
0.2
Medical herbs powders in mixed form were obtained from the Sabzineh company (Mash had, Iran) and
used in the diets after mixing homogenously with Corn, Soybean meal and wheat. Water and feed were provided
ad libitum during the study. The photoperiod was set at 16L: 8D throughout the study. Body weight, feed
consumption, egg production, Weight of egg mass, feed conversion ratio (FCR), were determined at the
beginning and every week of the study. The FCR was expressed as grams of feed consumed per gram of egg
produced. In order to examining egg quality (egg components and parameters) ten eggs per group (2 eggs/each
replicate) were collected at the end of 4th and 8th Week. Eggs were examined for interior (shell weights and Shell
thickness) and exterior (special egg weight and Haugh unit) quality. Mean eggs weight was determined for two
consecutive day every week. Egg shell thickness was determined by gauge thickness ultrasonic (Echometer
1062) [27] .Egg mass was calculated by method describe by North and Bell [28]. Albomine height measured
using micrometer and shaugh units calculated using formula described by Roush (1981) [33]. At the end of
study, 2 eggs from replicates of all groups (10 egg per group) were analyzed for egg cholesterol. The
cholesterol contents of the egg yolks were determined according to the commercial kit instructions. Briefly,
Yolks were separated from albumin, and at first step 1-g of yolks were weighed and mixed with 9 ml Nacl 2%
in a tube and shacked for 2 hr. In step2, 100 µl of homogenized solution mixed with 900 µl Nacl 2%. In step 3,
10 µl of step 2 solutions mixed with 100 µl NaCl 2% and 1ml Kit reagent and store in water bath at 37 °C for 10
min. In the final step, when color of the solution was pink, the yolk cholesterol concentration (mg of
cholesterol/g of egg yolk) was determined in samples by ultraviolet spectrophotometer at 540 nm wavelength.
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For measuring sheep red blood bell (SRBC) antibody titration, 0.2 ml sterile sheep red blood cell (0.8%)
suspension injected through limb vein of two hens per replicate. After 10 days blood samples collected and total
response titration (IgM+IgG) measured by hemoaglutination as described by [3], Isakov et al., 2005 [3, 21].
IgG and IgM measured by separating mercapto ethanol resistant (MER) antibody (IgG) and this amount is
deducted from the total antibody response sensitive to mercapto ethanol (IgM) [13]. Blood cells counted at day
70 of study after blood collecting (two hens per replicate) in venojects having EDTA. Counting of red blood
cells (RBC) and differentially white blood cells (WBC) was using Hemocytometer. Packed Cell Volume
(hematocrit) measured by centrifuging heparin coated capillary tubs for 15 min at 12000 rpm. For measuring
total cholesterol, at day 70 of study, after blood collecting (two hens per replicate), serum separated and total
cholesterol measured by spectrophotometer using commercial kits (Parsazmoon, Iran). Experimental design of
this study was completely random design (CRD). The following model was assumed in the analysis of all traits.
Yij = µ + Ai + eij. Where Yij = observed value for a particular character, µ=overall mean, Ai= effect of the ith
treatment and eij random error associated with the ijth recording. Data were analyzed by SPSS version 20.0 for
Windows (SPSS Inc., Chicago, IL). The differences between means were determined by ANOVA. Means were
compared by Duncan's multiple range tests with 95% confidence interval. In addition, Means were compared by
orthogonal contrasts.
Results:
Table 2: Effects of dietary feed additives on performance of laying hen at the termination of experimental period.
Performances
MHPs1 (%)
Control
Probiotic
1
2
3
Egg weight (g)
63.74a
63.24b
63.20b
63.22b
63.09b
Daily feed intake (g/hen)
110.00a
109.71a
109.61a
108.88b 108.22c
Egg production (%)
71.72
71.05
71.08
65.47
60.25
Egg weight mass (g)
45.72
44.94
44.95
41.39
38.01
FCR*
2.41
2.46
2.47
2.65
2.86
1. Medicinal herbs powders
*Feed conversion ratio.
Values in the same rows with different superscripts differ significantly (p<0.05).
SEM
0.07
0.15
1.57
1.01
0.06
P.value
0.03
0.00
0.74
0.06
0.11
Table 2 represents the effect of different levels of medical herbs powders (Thymus vulgaris,
Pennyroyal, Cumin, Alhagi, Garlic and Eucalyptus globules) in mixed form on the egg weight, daily feed
intake (g/d per hen), FCR, egg production, and egg weight mass in laying hens. Diets supplemented with 1, 2,
or 3% medical herbs powders had no significant effects on percentage of egg production, egg weight mass, and
FCR. Inclusion of medical herbs powders at the level of 2% and 3% in the diet significantly (P < 0.05) decrease
daily feed intake ccompared with the control, probiotic and 1% medical herbs powders. Additionally, egg
weight was lower in probiotic, 1, 2 and 3% medical herbs powders in compare to control. Also, orthogonal
comparisons of performance did not show significant differences between groups (Table 3).
Table 3: Comparing effects of dietary feed additives on performance at the termination of experimental period.
Performances
Control Probiotic MHPs1 SEM
(%)
Egg weight (g)
63.74a
63.24b
63.17b
0.07
Daily feed intake (g/hen)
110.00a
109.71a
108.90b 0.15
Egg production (%)
71.72
71.05
65.60
1.57
Egg weight mass (g)
45.72
44.94
41.45
1.01
Feed conversion ratio FCR
2.42
2.42
2.66
0.06
1. Medicinal herbs powders
Values in the same rows with different superscripts differ significantly (p<0.05).
P.value
0.00
0.00
0.20
0.17
0.20
Table 4 represents the effect of different levels of medical herbs powders on antibody production against
SRBC, IgG and IgM. Probiotic and different levels of medical herbs powders had no significant effect on
SRBC. Inclusion of probiotic and medical herbs powders at the level of 1 and 2% significantly (P < 0.05)
decrease IgG in compare tControl and 3% medical herbs powders. Using probiotic in the diet significantly
increases IgM in laying hens in compare to control and different levels of medical herbs powders. In table 5
Orthogonal comparisons confirm results of Table 4.
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Table 4. Effects of dietary supplementation of feed additives on immune response of laying hens
MHPs1 (%)
2
3
7.60
7.60
3.40c
4.80ab
4.20b
2.80b
Control
Probiotic
1
SEM
Immune response
SRBC
8.20
9
7.25
2.81
IgG
6a
2d
4.50bc
0.33
IgM
3.20b
7.00a
2.75b
0.40
1.Medicinal herbs powders
Values in the same rows with different superscripts differ significantly (p<0.05).
P.value
0.38
0.00
0.001
Table 5: Orthogonal comparisons of dietary supplementation of feed additives on immune response of laying hen.
Immune response
Control
Probiotic
MHPs1
SRBC
8.20
9.00
7.50
IgG
6.00a
2.00c
4.21b
IgM
3.20b
7.00a
3.29b
SEM
0.28
0.33
0.40
P.value
0.12
0.00
0.00
Table 6. Effect of different levels of dietary feed additives on blood biochemistry of laying hen
MHPs* (%)
Blood biochem.
Control
Probiotic
W.B.C (×103)1
R.B.C (×106)2
PCV (%)3
LYP (%)4
MCV5
29.26
2.56a
30.67
67.00
30.96
28.61
2.55a
29.60
67.90
28.64
1
26.56
2.09b
27.83
69.50
25.80
2
27.80
2.31ab
30.25
65.25
28.72
3
25.27
2.28ab
28.50
68.75
26.65
SEM
0.48
0.05
0.60
0.58
0.57
P.value
0.07
0.03
0.24
0.25
0.16
*. Medicinal herbs powders
1. WBC, 2. RBC, 3. Packet cell volume, 4. Lymphocytes, 5. Mean corpuscular volume
Values in the same rows with different superscripts differ significantly (p<0.05).
Table 6. Orthogonal comparisons of effect of dietary feed additives on blood biochemistry of laying hen
Control
Probiotic
MHPs* (%)
SEM
P.value
W.B.C (×10 )
28.74
26.85
29.26
0.48
0.09
R.B.C (×106)2
2.57a
2.23b
2.56a
PCV (%)3
28.89
28.95
30.67
0.05
0.60
0.00
0.55
LYP (%)4
64.67
67.58
67.00
0.57
0.92
MCV5
28.81
27.27
30.96
0.57
0.05
Blood biochem.
3 1
* Medicinal herbs powders
1. White blood cells 2. Red blood cells 3. Packet cell volume 4. Lymphocytes 5. Mean corpuscular volume Values
in the same rows with different superscripts differ significantly (p<0.05).
Table 8 represents the effect of different levels of medical herbs powders on egg weight, shell weight, shell
thickness, specific gravity and haugh unit. Results of this study showed that probiotic and medical herbs
powders at the level of 1, 2 and 3% in the diet had no significant effect on egg weight, shell weight, shell
thickness, specific gravity and haugh unit. Orthogonal comparisons did not showed significant difference
between groups (Table 9).
Table 10 represents the effect of different levels of medical herbs powders on serum and yolk cholesterol
concentrations. Results showed that inclusion of probiotic and medical herbs powders at the level of 1, 2 and 3%
in laying hens diet did not affect serum cholesterol. But medical herbs powders at the level of 1% significantly
increase yolk cholesterol in compare to probitic, 2 and 3% medical herbs powders and control. Orthogonal
comparisons showed that addition of medical herbs powders in to the laying hen’s diet may lead to higher serum
cholesterol (Table 11).
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Table 7. Effect of dietary feed additives on egg qualities of laying hen
Egg qualities
Egg weight (g)
Shell weight (g/egg)
Shell thickness (µm)
Specific gravity (g/cm3)
Haugh unit
Control
67.89
7.06
428.3
1.08
82.23
Probiotic
66.53
6.89
425.1
1.08
87.83
MHPs1 (%)
2
64.09
6.62
404.3
1.08
89.02
1
64.66
6.81
425.6
1.08
79.53
3
64.01
6.72
413.6
1.08
80.30
SEM
0.63
0.07
0.38
1.00
1.36
P.value
0.26
0.52
0.25
0.62
0.27
1. Medicinal herbs powders
Values in the same rows with different superscripts differ significantly (p<0.05).
Table 8. Orthogonal comparisons of effect of feed additives on egg qualities of laying hen
Egg qualities
Control
Probiotic
MHPs1 (%)
SEM
P.value
Egg weight (g)
Shell weight (g/egg)
Shell thickness (µm)
Specific gravity (g/cm3)
Haugh unit
1. Medicinal herbs powders
66.20
6.88
425.3
1.08
76.74
64.47
6.72
414.8
1.08
83.42
67.89
7.06
428.3
1.08
82.23
0.63
0.07
0.38
0.00
1.63
0.09
0.23
0.30
0.70
0.31
Values in the same rows with different superscripts differ significantly (p<0.05).
Table 9. Effect of feed additives on blood metabolites of laying hen
Blood metabolites
Serum cholesterol (mg/dl)
Yolk cholesterol (mg/g Yolk)
Control Probiotic
215.2
130.50
15.52b 17.44ab
1
171.7
18.25a
MHPs1 (%)
2
3
182.40 157.4
b
15.93
17.53ab
SEM
11.06
0.37
P.value
0.16
0.02
1. Medicinal herbs powders
Values in the same rows with different superscripts differ significantly (p<0.05).
Table 10. Orthogonal comparisons of dietary feed additives on blood metabolites of laying hen
Blood metabolites
Serum cholesterol (mg/dl)
Yolk cholesterol (mg/g Yolk)
Control
133.56
15.52
Probiotic
168.32
17.44
MHPs1 (%)
215.20
17.40
SEM
11.06
0.37
P.value
0.06
0.13
1. Medicinal herbs powders
Values in the same rows with different superscripts differ significantly (p<0.05).
Discussion:
This is the first study showed the effect of addition of 6 medical herbs powders (Thymus vulgaris,
Pennyroyal, Cumin, Alhagi, Garlic and Eucalyptus globules) in mixed form on performance, egg quality
characteristics and immunity response of commercial laying hens. There are so many evidence shows the effects
of medical herbs individually on performance, egg quality characteristics and immunity response of commercial
laying hens, but synergistic or antagonistic effects of mixed form of different medical herbs powders at different
levels in laying hens diet did not studied.
Reddy et al., (1991) reported that egg production, egg mass, feed intake, and feed efficiency were not
affected through 8 wk feeding of 0.02% garlic oil [32]. Chowdhury et al., (2002) also observed that there were
no effect of dietary garlic on egg production rate, egg weight and feed consumption [10]. It has been
demonstrated that garlic powder addition to the laying hens diet had no adverse effects on performance [37].
The effects of fermented garlic powder on production performance, egg quality, blood profiles and fatty acid
composition of egg yolk in laying hens were studied in a 35-d experimental period. Results showed that addition
of fermented garlic powder reduced plasma cholesterol concentration and did not cause adverse effects on
production performance. Moreover, addition of 3.0% garlic powder decreased saturated fatty acids (SFA) but
increased polyunsaturated fatty acids (PUFA) and PUFA:SFA ratio in egg yolk [4]. It is reported that laying
hens dietary supplementation with 0.1 , 0.5 and 1% Thyme (Thymus vulgaris) improve feed conversion and egg
productions and reduce E. coli concentration in feces at the level of 0.1 and 0.5% but not %1 [6]. It has been
showed that black cumin at the level of 2 or 3% would positively influence egg production, egg weight, and
shell quality and decrease the concentration of cholesterol in the egg yolk [5].
Using mixed medicinal herbs powders at the 1 and 2% level did not affect egg weight, feed intake, and egg
production. But at 3% level, egg weight and other parameters showed decreasing trend. Mixed medicinal herbs
powders failed to increase feed intake and egg production. Presence of high crud fiber or antagonistic effects of
active elements in used herbs powders with together or with nutrients of the diet, may be main cause of this
failure.
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The effect of probiotic (Lactobacillus sporogenes) feeding on egg production and quality, yolk cholesterol
and humoral immune response of White Leghorn (WL) layer breeders was studied. peobiotic at 100 mg (6 × 108
spore) kg−1 diet significantly enhanced the egg production, shell quality and humoral immune response [29].
Probiotics can inhibit pathogens by competition for a colonization sites or nutritional sources and production of
toxic compounds, or stimulation of the immune system. In order to enhance the efficacy of probiotics, it is
necessary to obtain additional knowledge on their mode of action. The efficacy of probiotics may be
potentate by several methods: The selection of more efficient strains; gene manipulation; the combination of
several strains and the combination of probiotics and synergistically acting components. The adoption of logical
criteria for the in vitro selection of probiotic bacteria can result in the isolation of strains capable of performing
effectively in the gastrointestinal tract [26] .It has been reported that supplementation of probiotic in layer hen's
diets did not appear to cause any adverse effects on egg production, egg quality and immunity. Moreover, the
dietary supplementation of probiotic may lead to the development of low-cholesterol eggs as demanded by
health-conscious consumers [24].Therefore, results of this study were in argument with previous studies. Diets
supplemented with Eucalyptus improved performance and immunocompetence of laying hens [16]. Hmamouchi
et al., (1992) indicated that oil Eucalyptus oil extract (globules) has antibacterial activities against 9
microorganisms, including salmonella type, Klebsiella spp, Streptococcus A, Proteus spp. Staphylococcus
aureus [20]. In Eucalyptus globules, the major monoterpenoid component is eucalyptol (1,8-cineole),
constituting the 60–90% [22], that has been reported to inhibit the production and synthesis of tumor necrosis
factor-α (TNF-α), interleukin-1β (IL-1β), leukotriene B4,and thromboxane B2 in human blood monocytes [22,
23]. Medical herbs powders of this study did not affect antibody titration against SRBC, IgG and IgM in laying
hens. It is possible that medicinal herbs affect immunity response by different mechanism instead of SRBC, IgM
or IgG. For example, little is known about the influence of Eucalyptus oil extract on the monocytic/macrophagic
system, one of the primary cellular effectors of the immune response against pathogen attacks. It is
demonstrated that Eucalyptus oil extract is able to implement the innate cell-mediated immune response [35]. It
has been reported that antibody production in response to the inoculation of sheep red blood cells and the
cetaceous basophilic hypersensitivity (CBH) responses to inoculation with phytohaemagglutinin in white
leghorn layers from 25 to 72 weeks of age did not differ significantly at either 24 or 40 weeks of age. The
antibody titration was significantly higher in the groups supplemented with probiotic at 64 weeks of age. The
addition of 100 mg/kg of probiotic in diet significantly increased the CBH response at 64 weeks of age [30].
Results of this study were in agreement. Hematological traits (WBC, RBC, PCV, Lyp and MCV) were not
affected by dietary supplementation by mixed herbs powders, But RBC in probiotic group had decreasing and
MCV in mixed herbs powders groups showed increasing tendency. It has been reported that addition of 2
medicinal herbs including garlic (Allium sativum) and Thyme (Thymus vulgaris) to diet of laying hens, white
blood cell counts (% of total) were not affected, except for lymphocyte that was higher in hens fed diets
included 2% medical herbs comparing to the two other experimental diets [18]. Also, their experiment showed
that the addition of garlic powder had no significant effect on the egg shell thickness, egg albumen index, egg
yolk index and egg Haugh unit. Lim et al. (2006) reported that egg shell thickness was not affected by the
dietary supplementation of garlic powder. Results of this study were in agreement with the results of Yalçın et
al., 2006 and Lim et al. 2006. Additionally, It is reported that egg quality characteristics yolk index, Haugh
unit and egg shell thickness were not significantly affected by laying hens dietary supplementation with
medicinal herbs Including Garlic (Allium sativum) and Thyme (Thymus vulgaris); however, diets including 2%
medicinal herbs, increase egg yolk color and decreased shell weight [18].
Serum total cholesterol concentration did not responded with increase levels of mixed herbs powders and
probiotic, but yolk cholesterol concentration increase significantly with 1% mixed herbs powders in compare to
probiotic and control groups. However, in orthogonal comparisons medical mixed herbs powders inclusion in
laying hen’s diet showed decreasing tendency for serum cholesterol. Sklan et al. (1992) reported that hepatic
cholesterol concentrations in chickens were reduced when 2% garlic was fed [36]. The activity of hepatic 3hydroxy-3-methylglutaryl coenzyme A reductase was decreased by 50-69% in 12-week-old broilers and by 7283% in 12-week-old Leghorn pullets with concomitant decreases in serum cholesterol of 7-25 and 20-25%,
respectively, by the feeding of diets containing garlic paste at 3.8% or solvent fractions of garlic oil equivalent
to 3.8% garlic paste [31]. Chowdhury et al. (2002) observed there were no effects on egg production but
significantly decreased serum cholesterol by the addition of garlic [10]. However, Birrenkott et al [8]
demonstrated that the levels of cholesterol in egg yolk and serum were not decreased significantly by 3% garlic
powder after feeding of several months [8]. It has been reported that when a few common spices of cumin
(Cuminum cyminum), cinnamon (Cinnamomum zeylanicum), ginger (Zingiber officinale), mustard (Brassica
nigra) and tamarind (Tamarindus indica) was added to normal and hypercholesterolemia inducing diets of rats,
these spices did not show any cholesterol lowering effect at about 5-fold the normal human intake level [34].
However, black cumin seeds are of interest as potential egg-yolk cholesterol-lowering agents [15]. Cumin active
compound act as inhibitors to active enzyme hepatic 3-hydroxy-3-ethylglutaryl coenzyme A (HMG-CoA) that
synthesized cholesterol [11].
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Advances in Environmental Biology, 7(13) November 2013, Pages: 4382-4389
Conclusion:
Using different concentrations of different feed additives such as (mixed medicinal herbs powder,
probiotics and antibiotics), did not affect adversely performance, egg mass during 84-days of experimental
period. However, the herbs powders at the level of 2 and 3% decrease feed intake and egg weight as compare to
control and probiotic groups. None of egg quality parameters (egg weight, shell weight, shell thickness, huagh
unit, specific gravity), antibody titrations against SRBC, Lymphocyte, packed cell volume, mean corpuscular
volume and serum cholesterol did not show significantly different, but RBC, IgG, IgM and yolk cholesterol was
affected. Using herbs powders in mixed form may suppress positive effect of herbs in compare to using
individually.
ACKNOWLEDGMENT
We gratefully acknowledge the efforts of the staff of the department of Animal Science, Azad Islamic
University, Varamin, Iran and Animal Science Research Institute (ASRI) in karaj, Iran.
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