Effect of Rosemary (Rosmarinus Officinalis) Dietary Supplementation in Broiler Chickens Concerning immunity, Antioxidant Status, and Performance

Abstract

This study was conducted to evaluate the effect of rosemary leaves on the growth performance, blood parameters, and immune response of broiler chickens by determination of the serum immunoglobulins (IgA, IgM, and IgG), interferon-γ (INF-γ) and interleukin-10 (IL-10). Also, malondialdehyde (MDA), total superoxide dismutase (T.SOD), glutathione S-transferase (GST), and glutathione reduced (GSH) levels in the thigh and breast muscles were determined to evaluate the effect of rosemary in the broiler chicken’s muscles. To achieve this aim, 120 Cobb of one-day-old chicks were allocated into four equal groups as a control group that supplemented by the basal diet, while the other three groups were fed basal diet supplemented with 0.5, 1.0, and 1.5% of rosemary. The data of growth performance indicated that supplementation of broiler with rosemary had no growth-promoting effects. Feeding diet with rosemary leaves meal significantly increased the serum total protein and globulin, while significantly decreased total cholesterol and triacylglycerol levels. Rosemary significantly increased the IgG, IgM, INF-γ, IL-10, and muscle GSH levels and T. SOD and GST activities. Whereas, muscle MDA levels were significantly decreased, so rosemary could be considered as a natural antioxidant in broiler diet. Concomitantly, provide a healthy broiler’s meat with less MDA that favorable to human consumption

Full text

152
Alexandria Journal of Veterinary Sciences
www.alexjvs.com
AJVS. Vol. 55 (1):152-161. Oct. 2017
DOI: 10.5455/ajvs.275350
Effect of Rosemary (Rosmarinus Officinalis) Dietary Supplementation in Broiler Chickens
Concerning immunity, Antioxidant Status, and Performance
Safaa A. Ghozlan1*, Ali H. El-Far1, Kadry M. Sadek1, Abdelrahman A. Abourawash2, Mervat A. Abdel-
Latif3
1Department of Biochemistry, Faculty of Veterinary Medicine, Damanhour University, Damanhour, 22511, Egypt
2Department of Veterinary Pathology and Clinical Pathology, Faculty of Veterinary Medicine, Damanhour University, Damanhour
22511, Egypt
3Department of Nutrition and Veterinary Clinical Nutrition, Faculty of Veterinary Medicine, Damanhour University, Damanhour,
22511, Egypt
ABSTRACT
Key words:
Broilers,
Immunity,
Antioxidant,
Performance,
Feed additives,
Rosemary
This study was conducted to evaluate the effect of rosemary leaves on the growth
performance, blood parameters, and immune response of broiler chickens by
determination of the serum immunoglobulins (IgA, IgM, and IgG), interferon-γ (INF-γ)
and interleukin-10 (IL-10). Also, malondialdehyde (MDA), total superoxide dismutase
(T.SOD), glutathione S-transferase (GST), and glutathione reduced (GSH) levels in the
thigh and breast muscles were determined to evaluate the effect of rosemary in the
broiler chicken’s muscles. To achieve this aim, 120 Cobb of one-day-old chicks were
allocated into four equal groups as a control group that supplemented by the basal diet,
while the other three groups were fed basal diet supplemented with 0.5, 1.0, and 1.5% of
rosemary. The data of growth performance indicated that supplementation of broiler with
rosemary had no growth-promoting effects. Feeding diet with rosemary leaves meal
significantly increased the serum total protein and globulin, while significantly decreased
total cholesterol and triacylglycerol levels. Rosemary significantly increased the IgG,
IgM, INF-γ, IL-10, and muscle GSH levels and T. SOD and GST activities. Whereas,
muscle MDA levels were significantly decreased, so rosemary could be considered as a
natural antioxidant in broiler diet. Concomitantly, provide a healthy broiler’s meat with
less MDA that favorable to human consumption
Corresponde
nce to: *
safaa.gozlan1
@gmail.com
1. INTRODUCTION
The European Commission banned antibiotic
growth promoters in broiler nutrition. Therefore,
many research studies have been conducted to explore
the use of possible effective substitutes. One
possibility is the application of herbs or their essential
oils (Sarica et al., 2007). Numerous in vitro studies
have already confirmed the antibacterial actions of
these feed additives. Consequently, several in vivo
studies were performed to confirm their beneficial
qualities. Medicinal plants are resources of new drugs
and many of the modern medicines that improve the
health status of animals (El-Far et al., 2016b; El-Far et
al., 2017). The active principles of essential oils act as
a digestibility enhancer, balancing the gut microbiota
and stimulating the secretion of endogenous digestive
enzymes and thus improving the growth performance
in poultry (Cross et al., 2007; Ayoub et al., 2011;
Barakat et al., 2016; El-Far et al., 2016a).
Rosemary (Rosmarinus officinalis) has been
used as a medicinal and aromatic herb since ancient
Greek and Roman (al-Sereiti et al., 1999). In folk

Ghozlan et al. 2017. AJVS 55(1): 152-161
153
medicine, rosemary extract is a treatment for urinary
ailments, chronic weakness, nervous disorders, hair
loss, and peripheral vascular diseases. In addition,
rosemary is a traditional astringent, carminative, tonic,
rubefacient, antispasmodic, anti-inflammatory,
expectorant, emmenagogue, digestive, and diaphoretic
(Haloui et al., 2000). Rosemary is broadly used in the
food industry, and it is highly appreciated for its
several functional properties, such as aromatic
properties, antioxidant, and antimicrobial (Afonso et
al., 2013). Therefore, the current study was conducted
to evaluate the impact of rosemary feed
supplementation on broiler chicken’s health status
targeting the immunity, antioxidant potential, and
performance.
2. MATERIAL AND METHODS
2.1. Birds, accommodation, and
management
The present study is affirmed by the Ethics of
Animal Experiments Committee, Damanhour
University, Egypt. Whereas, one hundred and twenty
Cobb of one-day-old broiler chicks were incubated
and randomly allocated into four equal groups at the
first week of age. Each one was subdivided into three
replicates (10 birds per replicate). The housing of
chicks was done in a clean well-ventilated room. The
room temperature was adjusted according to age by
electric heaters. Furthermore, the birds were
vaccinated by Hitchner IB (7th day), Gumboro (14th
day), and Gumboro and Clone (21st day) by eye drop.
2.2. Diet and experimental design
The chicks were fed on the two phases feeding
programs from 1st to 21st days on the starter and from
22nd to 35th days on grower diets. The control diet
composition was represented in Table 1 and analyzed
according to AOAC (2005). The diet was formulated
to meet the requirements of NRC (1994). Rosemary
was obtained from a local market, washed, ground,
refined, and mixed with the ration at the concentration
of 0.5% in Rosemary I, 1% in Rosemary II and 1.5%
in Rosemary III groups, while control one was fed a
basal diet. The water was accessed ad libitum to all
birds.
Table 1. The starter and grower diet’s ingredients percentage and calculated composition (as fed basis)
Ingredients
Starter diet
Grower diet
Corn
52.87
60.47
SBM (CP 44%)
34.26
29.31
Corn gluten (CP 60%)
5.5
3.0
Corn oil
3.3
3.26
Limestone
1.35
1.53
Dicalcium phosphate
1.74
1.47
L-Lysine
0.11
0.13
Dl-methionine
0.17
0.13
Vitamins and minerals premix
0.3
0.3
NaCl
0.4
0.4
Total
100
100
Composition
ME (Kcal/Kg diet)
3061.2
3119.35
CP %
23.0
20.0
Calorie/protein ratio
133.1
155.97
Lysine %
1.3
1.16
Methionine %
0.58
0.48
Calcium %
1.0
0.9
Av. (P) %
0.45
0.40
NaCl
0.15
0.15
SBM= Soybean meal, ME = Metabolizable Energy, CP = crude protein, Av. (P) = Available phosphorous
*L-lysine 99% feed grade
**Dl-methionine 99% feed grade China
***Vitamin and mineral premix (Hero mix) produced by Heropharm and composed (per 3 kg) of vitamin A 12000000 IU, vitamin D3
2500000 IU, vitamin E 10000 mg, vitamin K3 2000 mg, vitamin B1 1000 mg, vitamin B2 5000 mg, vitamin B6 1500 mg, vitamin B12 10
mg, niacin 30000 mg, biotin 50 mg, folic acid 1000 mg, pantothenic acid 10000 mg, manganese 60000 mg, zinc 50000 mg, iron 30000 mg,
copper 4000 mg, iodine 300 mg, selenium 100 mg, and cobalt 100 mg.

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154
2.3. Gas chromatography–mass
spectrometry (GC-MS) analysis
The fine powder of rosemary was extracted by
n-hexane by a dilution of 1: 3 (w: v). 10 µl of thyme
n-hexane extract was injected in Trace GC Ultra-ISQ
mass spectrometer with a direct capillary column TG–
5MS (30 m×0.25 mm×0.25 µm). Helium carrier gas
was used with a flow rate of 1ml/min. The oven
temperature program was initiated at 50°C for 2 min,
the rate of 4°C/min up to 160°C for 5 min, the rate of
8°C/min up to 220°C for 2 min, the rate of 15°C/min
up to 280°C for 5 min. Injector and flame ionization
detector temperatures were 250°C and 290°C,
respectively. 1 μl of each extract was injected with a
split ratio of 1:200 (Hay et al. 2015). The mass spectra
of the identified components were determined by
comparison to Wiley Registry mass spectral database
of 8th edition.
2.4. Serum parameters
The blood samples were collected from wing
vein at 3rd and 5th weeks. Each blood sample was left
to coagulate at room temperature and centrifuged at
3000 rpm for 5 min. The clear sera were subjected to
determination of total protein, albumin, alanine
aminotransferase (ALT, EC 2.6.1.2), creatinine, total
cholesterol, and TAG following the instructions
enclosed in the manufactured kits produced by
Biodiagnostic Company, Egypt. Also, serum globulin
levels were calculated by subtraction of albumin value
from the total protein value of the same sample
(Coles, 1986).
2.5. ELISA assays
The serum levels of immunoglobulin A (IgA),
immunoglobulin G (IgG), immunoglobulin M (IgM),
Interferon-γ (INF-γ), and interleukin-10 (IL-10) were
determined by ELISA kits manufactured by
Elabscience Co.
2.6. Preparation of muscle tissue
homogenate
At the end of the experimental period, the birds
of control and rosemary-treated groups (n= 10) were
sacrificed under anesthesia with an intramuscular
injection of sodium pentobarbital (50 mg/kg BW), and
then muscle samples from left breast and left thigh of
each bird were immediately dissected and soaked in
ice-cold saline 0.9%. They were homogenized using a
motor-driven Teflon and glass Potter-Elvehjem
homogenizer in 0.1 M Tris-HCl buffer of pH 7.4
containing 5 mM β-mercaptoethanol (1:4 w/v). The
homogenates were centrifuged at 105,000 ×g for 60
min at 4°C; the supernatants were divided into
aliquots then stored at -20°C for further evaluation of
oxidative stress and antioxidant parameters.
2.7. Determination of oxidative stress
parameters
The frozen aliquots of muscle homogenates
were utilized for the colorimetric assessment of MDA
and GSH contents, as well the T.SOD and GST
activities.
2.7.1. Determination of lipid peroxidation
Malondialdehyde is the main aldehyde by-
product of lipid peroxidation in biological systems. It
was analyzed after the incubation of supernatants with
thiobarbituric acid at 95°C for 30 min (pH 3.6) to
form thiobarbituric acid-reactive substances
(TBARS), a pink colored compound. MDA levels
were measured at 532 nm and expressed as nmol
MDA /mg protein (Ohkawa et al., 1979).
2.7.2. Determination of reduced glutathione
levels
Reduced glutathione assay was based on the
reductive cleavage of 5, 5′-dithiobis 2-nitrobenzoic
acid (DTNB) by compounds containing sulfhydryl
groups and development of a yellow color (Sedlak
and Lindsay, 1968). The quantity of reduced
chromogen is directly proportional to the GSH
content. The absorbance was recorded at 412 nm and
expressed as µmol GSH/mg protein.
2.7.3. Determination of the Total superoxide
dismutase activity
The reduction of nitro blue tetrazolium with
NADH-mediated by phenazine methosulfate (PMS)
under aerobic conditions was inhibited upon addition
of superoxide dismutase. This observation indicated
the involvement of superoxide anion radical in the
reduction of nitro blue tetrazolium, the radical being
generated in the reoxidation of reduced PMS. This
assay was determined at 560 nm and represented as
U/mg protein (Nishikimi et al., 1972).
2.7.4. Determination of the glutathione S-
transferase activity
The activity of GST was measured according to
the method of Vessey and Boyer (1984). This assay
was based on monitoring the rate of enzyme–
catalyzed conjugation of the 1-chloro-2,4-
dinitrobenzene (CDNB)with GSH. GST activity was
measured as the increase in absorbance at 340 nm and
represented as U/mg protein.
2.7.5. Determination of tissue protein
Protein concentrations in muscle homogenates
were determined using bovine serum albumin as the
standard according to the method of Bradford (1976).

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2.8. Performance parameters
The basal diets of both starter and grower
phases were formulated according to the
recommendation of National Research Council
Nutrient Requirements for Broiler Chickens (NRC,
1994). Performance parameters include the final body
weight, feed intake, feed conversion ratio (FCR)
(Lambert et al., 1936) and protein efficiency ratio
(McDonald et al., 1987) were determined throughout
the experimental period.
2.9. Statistical Analyses
The obtained data were analyzed by one-way
analysis of variance (ANOVA), with Duncan's
multiple range tests for significance between means
using SPSS software package v.20. The data of
ELISA assays and antioxidant status were analyzed by
One-way ANOVA, Tukey’s multiple range tests by
GraphPad Prism 5. All declarations of significance
depended on p <0.05.
3. RESULTS
The data illustrated in Table 2 and presented in
Fig.1 revealed the chemical composition of one
sample was carried out using the GC–MS analysis led
to the identification of eleven different
components;1,8-cineole (23.76%), camphor (3.87%),
α-terpineol (1.32%) eugenol (1,59) caryophyllene
oxide (0.17%) oleic acid (5,16%), and ethyl iso-
allocholate (3.15%).
The data illustrated in Table 2 represented that
at the 3rd and 5th weeks, the serum levels of total
protein and globulin were significantly increased in
Rosemary I, Rosemary II, and Rosemary III in
relation to control group, while serum albumin levels
were non-significantly increased. Also, no significant
changes were recognized in serum ALT activities and
creatinine levels when the rosemary-treated groups
were compared with control one. Furthermore, the
data shown in Table 2 revelated significant decreases
in the levels of total cholesterol and TAG at the 3rd
and 5th weeks in comparison to control.
The data illustrated in Fig. 2(A) represented the
effects of rosemary dietary supplementation on serum
immunoglobulins where IgA levels non-significantly
differed (p>0.05) at the 3rd and 5th weeks compared to
control. Serum IgG levels were significant increase in
Rosemary I (p<0.05) and Rosemary III (p<0.01) in
relation to control at the 3rd week, while at the 5th
week the IgG levels in rosemary-treated groups were
significantly increased in Rosemary I (p<0.001),
Rosemary II (p<0.01), and Rosemary III (p<0.01)
compared to control (Figure 2B).The data illustrated in
Figure 2.C represented the effects of rosemary dietary
supplementation on serum IgM and revealed that at the
3rd and 5th weeks, the IgM levels were significantly
increased (p<0.001) in Rosemary I, Rosemary II, and
Rosemary III compared to control.
The data illustrated in Fig. 3(A) represented the
effects of rosemary dietary supplementation on INF-γ
and revealed that at the 3rd and 5th weeks, its levels
were significantly increased (p<0.001) in the
Rosemary I, Rosemary II, and Rosemary III compared
to control. Results in Fig. 3(B) represented significant
increases (p<0.05) in the levels of IL-10 in Rosemary
II and Rosemary III compared to control at the 3rd
week. Similarly, at the 5th week, the IL-10 levels
showed significant increases in the Rosemary I
(p<0.05), Rosemary II (p<0.01), and Rosemary III
(p<0.01) compared to control.
Table 2. GC-MS analysis of rosemary n-hexane extract (antioxidant constituents)
Compound Name
RT (minutes)
Area %
Molecular Formula
1
1,8-Cineole
12.43
23.67
C10H18O
2
(-)-camphor
18.01
3.87
C10H16O
3
α-Terpineol
18.91
1.32
C10H18O
4
Eugenol
24.58
1.59
C10H12O2
5
Caryophyllene oxide
25.35
0.17
C15H24O
6
Oleic acid
38.88
5.16
C18H34O2
7
Ethyl iso-allocholate
39.58
3.15
C26H44O5

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156
Table 3. Effect of dietary Rosemary supplementation on serum total protein, albumin, globulin, ALT,
creatinine, total cholesterol, and triacylglycerol
Item
Control
Rosemary supplementation
Rosemary I
Rosemary II
Rosemary III
3rd week
Total protein (g/dl)
4.45±0.05b
5.12±0.21a
5.28±0.06a
5.25±0.05a
Albumin (g/dl)
3.48±0.15a
3.56±0.09a
3.61±0.08a
3.57±0.11a
Globulin (g/dl)
0.97±0.13b
1.56±0.18a
1.66±0.08a
1.69±0.06a
ALT(U/l)
17.5±0.5a
17.25±0.48ab
16±0.41ab
15.75±0.63b
Creatinine (mg/dl)
0.33±0.05a
0.33±0.02a
0.39±0.03a
0.39±0.02a
Cholesterol (mg/dl)
144.11±3.56a
126.76±1.59b
125.14±2.31b
127.76±4.02b
TAG (mg/dl)
153.37±5.69a
131.97±3.37b
127.96±3.89b
130.32±1.94b
5th week
Total protein (g/dl)
5.14±0.05b
5.62±0.1a
5.62±0.04a
5.84±0.12a
Albumin (g/dl)
3.53±0.05a
3.62±0.09a
3.57±0.02a
3.45±0.09a
Globulin (g/dl)
1.6±0.08c
2±0.17b
2.05±0.05ab
2.39±0.12a
ALT (U/l)
16.25±0.48a
16.5±0.65a
15±1.08a
14.25±0.63a
Creatinine (mg/dl)
0.47±0.02ab
0.43±0.02b
0.53±0.03a
0.47±0.03ab
Cholesterol (mg/dl)
150.38±2.49a
127.59±2.63b
126.63±3.32b
126.05±3.73b
TAG (mg/dl)
158.53±1.98a
132.23±2.94b
122.58±0.84c
130.62±2.57b
Mean values with different letters in the same row differ significantly at P<0.05
Table 4. Effect of dietary turmeric supplementation on growth performance of broiler chickens
Rosemary supplementation
Control
Rosemary III
Rosemary II
Rosemary I
157.39±1.72a
157.08±2.13a
155.4±2.12a
160±1.54a
IBW (g)
1578.04±23.05a
1595.21±23.68a
1646.2±21.56a
1608.25±24.59a
FBW (g)
1420.65±21.45b
1438.13±21.66ab
1490.8±19.48a
1448.25±23.1ab
BWG (g)
2901.76±25.35 a
2916.89±30.23 a
2907.5±23.65 a
2884±16.44a
FI (g)
2.05±0.03a
2.04±0.03ab
1.96±0.03b
2.00±0.03ab
FCR
2.56±0.03a
2.56±0.04a
2.66±0.03a
2.58±0.04a
PER
Mean values with different letters in the same row differ significantly at P<0.05
IBW = initial body weight; FBW = final body weight; BWG = body weight gain; FI= Feed intake; FCR = feed conversion ratio; PER =
protein efficiency ratio
Fig. 1. GC-MS analysis chromatogram of rosemary n-hexane extract

Ghozlan et al. 2017. AJVS 55(1): 152-161
157
The data illustrated in Fig. 4 represented the
effects of rosemary dietary supplementation on MDA
and antioxidant status in breast and thigh muscles. In
Fig. 4(A) the breast muscle MDA levels were
significantly decreased (p<0.05) in rosemary-treated
groups compared to control. Also, the thigh muscles
MDA levels were significantly decreased (p<0.01) in
rosemary-treated groups. The data illustrated in Fig.
4(B) revealed that the breast muscle levels of GSH
were a significant increase (p<0.001) in rosemary-
treated groups compared to control. In comparison to
Rosemary I the GSH levels were significantly
increased (p<0.01) in Rosemary II and Rosemary III.
The GSH levels in thigh muscles were significantly
increased (p<0.05) in Rosemary II and Rosemary III
compared to control. Also, it showed significant
increases (p<0.05) in Rosemary III compared to
Rosemary I.
The data illustrated in Fig. 4(C) represented that
the T. SOD activities in breast muscles were
significantly increased (p<0.01) in rosemary-treated
groups compared to control, while in thigh muscles its
activities were significant increase in Rosemary I
(p<0.05), Rosemary II (p<0.05), and Rosemary III
(p<0.01) compared to control.
Results in Fig. 4(D) revealed significant
increases (p<0.01) in the breast muscles GST activities
in the rosemary-treated groups compared to control.
Fig. 2. Represents the effects of rosemary on
serum (A) IgA, (B) IgG, and (C) IgM. *p <
0.05, **p < 0.01, and ***p < 0.001 vs. control.
Fig. 3. Represents the effects of rosemary
on (A) INF-γ and (B) IL-10. *p < 0.05, **p
< 0.01, and ***p < 0.001 vs. control.

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158
But in the thigh muscle, its activities revealed non-
significant differences in rosemary-treated groups.
The data presented in Table 4 showed non-
significant improvement (p≥0.05) in growth
performance measurements in 0.5% rosemary-treated
group in comparison to control while inclusion of
high levels of rosemary deteriorates the growth
performance parameters.
4. DISCUSSION
Continuous use of antibiotics in poultry diets has
evoked numerous problems including the cross-
resistance and environmental pollution. So that the
search for alternative substances to replace classical
antibiotics should be continued (Dickens et al. 2000).
Therefore, vegetables, herbs, spices and edible plants
were suggested as feed additives in animal nutrition
(Abaza 2001).
The GC-MS analysis of rosemary stated the
cineole content of rosemary was 23,67% area in the
chromatogram. The essential oil isolated from
rosemary was characterized by its greater content of
1,8-cineole as stated by Mathlouthi et al. (2011). The
main active components were camphor (11-16%),
alpha-pinene (15-20%) and cineole (30-35%) which
has a high degree of inhibition against many bacteria
and fungi (Ali and Ghazalah, 2008). The same
compounds have the antioxidant potential as studied
by Rašković et al. (2014).
The obtained data showed an immunostimulant
effect of rosemary through significant increases in the
serum levels of IgM, IgG, INF-γ, and IL-10 in
rosemary-treated groups at the 3rd and 5th weeks, while
the serum IgA level was significantly increased in the
Rosemary II at the 3rd week and was significantly
increased in the Rosemary III at the 5th week in
comparison to control. These results come in
accordance with that obtained in the study of
ELnaggar et al. (2016) who studied the addition of
rosemary to the basal diet at the concentrations of
0.25, 0.5, 0.75, and 1.0 % had significantly improved
the serum immunoglobulins (IgY, IgM, and IgA),
IFN-γ, and IL-10.
Lipid peroxidation leads to the formation of
various products as the MDA. Therefore, blood MDA
level is often determined in some studies as an
indicator of lipid peroxidation in the body (Droge,
2002). In a biological system, many oxidation
reactions are essential for our survival. Sometimes,
inside the normal cells, oxidation reactions release
uncontrolled reactions and produce unstable oxygen
molecules “free radicals”. These produced compounds
will react with many different important molecules in
vital organs like lipid, protein, and DNA forming a
new compound that damage DNA (Ercegovac et al.,
2010). Antioxidants are the first line of defense
against free radical damage and are critical for
maintaining optimum health (Lobo et al., 2010). The
data of the present study revealed that the
supplementation of rosemary to the diet of broiler
chickens reduced the muscle MDA levels, which
indicates a decreased lipid peroxidation. In contrast,
T.SOD and GST activities in them were significantly
increased in comparison to control, while GSH was
significantly increased in Rosemary III and non-
significantly increased in Rosemary I and Rosemary
II. These results came in accordance with that
obtained in the study of Polat et al. (2011) where they
recognized an enhancement in serum SOD activity in
rosemary fed birds. The meat from broilers fed on the
diet containing 500 mg/kg rosemary and sage extracts
had smaller concentrations of total cholesterol
oxidation products than meat from the control group
(Lopez-Bote et al., 1998).
Inspection of the blood data in Table 2 showed
that values of serum total protein and globulin were
significantly increased with the addition of rosemary
compared to control, while no significant differences
were observed in blood albumin. This reflects the
ability of chicks to store reserve protein even after the
body has reached its maximum capacity for depositing
protein to tissues. In addition, the increase in the
globulin indicates the effective role of rosemary in
increasing immunity due to its role in developing and
protecting cells and inhibiting non-enzymatic
oxidation (Houghton et al., 2007). These results came
in accordance with that obtained in the study of Ali
and Ghazalah (2008). The non-significant alterations
in ALT activities and creatinine levels indicated the
safe use of rosemary as a feed additive in broiler
chickens diet on liver and kidney functions.
Moreover, the serum total cholesterol and
triacylglycerol levels were significantly reduced due
to rosemary feed supplementation compared to
control.

Ghozlan et al. 2017. AJVS 55(1): 152-161
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The average body weight, average body weight
gain, feed conversion, and average protein efficiency
were statistically non-significantly differed in the
rosemary-treated groups compared to control. These
findings are confirmed by the study done by Abd El-
Latif et al. (2013) who investigated the effects of
dietary inclusion of different levels of rosemary and
garlic essential oils on broilers performance and
concluded that the data of growth performance
indicated that supplementation of broiler diets with
rosemary oil had no growth-promoting effects.
Similarly, Hernandez et al. (2004) found no
significant differences in body weight gain of broilers
when a blend of extracts of sage, thyme, and rosemary
was added to diets. Also, dietary rosemary and yarrow
herb powders had no significant differences in the
final body weight (Norouzi et al., 2015). Also, these
results were supported by ELnaggar et al. (2016) who
found that low levels (0.25%) of rosemary give better
growth performance compared to higher one. On the
other hand, Al-Kassie et al. (2008) stated that 0.5 and
1.0% rosemary herb supplementation in the diet
clearly improved broiler growth performance at the
42nd days of age, compared with the control treatment
(Ali and Ghazalah, 2008) also found that 0.5%
rosemary herb supplementation in the diet gave better
results than the control treatment at the 49th days of
age.
5. CONCLUSION
From the obtained results, it could be concluded
that supplementing rosemary into broiler diet show no
positive effect on growth performance especially
higher levels. Remarkably, it, in a concentration-
dependent manner, increased immunity and
antioxidant activity in the broiler chickens producing
a meat with fewer quantities of MDA.
6. COMPETING INTERESTS
The authors have no conflict of interest.
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