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Effect of myrtle (Myrtus communis L.) oil on performance, egg quality, some
biochemical values and hatchability in laying quails

Article  in  Revue de Médecine Vétérinaire · January 2014

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280
Effect of myrtle (Myrtus communis L.) oil on
performance, egg quality, some biochemical values
and hatchability in laying quails
T. BULBUL1*, D. YESILBAG2, E. ULUTAS3, H. BIRICIK2, S. S. GEZEN2, A. BULBUL3
1
Department of Animal Nutrition and Nutritional Diseases, Faculty of Veterinary Medicine, Afyon Kocatepe University, 03200, Afyonkarahisar, TURKEY.
2
Department of Animal Nutrition and Nutritional Diseases, Faculty of Veterinary Medicine, 16059, Uludag University, Bursa, TURKEY.
3
Department of Physiology, Faculty of Veterinary Medicine, 03200, Afyon Kocatepe University, Afyonkarahisar, TURKEY.
*Corresponding author: tbulbul@aku.edu.tr
SUMMARY
The aim of this study was to evaluate the effect of myrtle (Mytrus communis
L.) oil when added to the diet of laying quails on performance, egg quality,
some biochemical values and hatchability. A total of 375 quails (250
females and 125 males; Coturnix coturnix japonica), aged eight weeks
old, were randomly allocated to five dietary treatments (five replicates of
ten females and five males). The birds were fed either a basal diet or the
basal diet supplemented with myrtle oil at a dose of 500, 1000, 2000 or
5000 mg/kg feed. The experiment was conducted for eight weeks. At the
end of the experiment, no significant differences were found among the
groups in terms of initial live weight, feed consumption, egg weight, egg
quality (fracture strength, haugh unit, shape index, yolk index, albumen
index), serum biochemical values, hatchability, early embryonic death, late
embryonic death and submembranous death. After 8 weeks, egg production
was decreased (p< 0.01), whereas feed conversion rate (FCR) was increased
(p< 0.05) in birds with a diet supplementation with myrtle oil doses of 5000
mg/kg. Eggshell thickness was decreased (p< 0.05) of groups receiving myrtle
oil doses of 2000 and 5000 mg/kg. Yolk color index was affected positively
by addition of myrtle oil (p< 0.001). Hatch performance was highest of the
groups with diet supplemented with mytrle oil doses of 1000 mg/kg, whereas
it was lowest of the groups received 5000 mg/kg mytrle oil (p< 0.01). The
addition of myrtle oil to the diets caused significantly decrease in serum
total cholesterol (p< 0.01), Ca (p< 0.01) and Malondialdehyde (MDA; p<
0.001) while determined significantly increase in blood urea (p< 0.001) and
serum β-carotene (p< 0.001) concentrations. Serum creatine kinase-MB
(CK-MB) was increased (p< 0.001) a dose of 5000 mg/kg, whereas albumin
concentration decreased (p< 0.01). Egg yolk MDA concentration was
decreased (p< 0.01) in all groups that received myrtle oil supplementation in
their diets on days 15th and 30rd of storage (p< 0.001). In conclusion, mytrle
oil supplementation was changed laying performance and biochemical
values in laying quails depending on supplemented quantity and duration.
It is recommended to supplement diets with 1000 mg/kg mytrle oil as egg
production, egg quality, yolk MDA concentrations and hatching parameters
were taken into consideration.
Keywords: Myrtle, performance, hatchability, lipid
peroxidation, quails.
Introduction
Adding aromatic plants or their extracts, separately or as a
mixture, to animal diets has antibacterial [13, 44], antifungal
[6, 47], anticoccidial [25], antioxidant [12] and antilipidemic
BULBUL (T.) AND COLLABORATORS
RÉSUMÉ
Effets de l’huile essentielle de myrte (Myrtus communis L.) sur les
performances, la qualité de l’œuf, certaines valeurs biochimiques et
l’éclosion chez les cailles de ponte
Le but de cette étude était d’évaluer l’effet de l’huile essentielle de myrte
(Mytrus communis L.) sur les performances, la qualité de l’œuf, certaines
valeurs biochimiques et l’éclosion lorsqu’elle est ajoutée à l’alimentation des
cailles de ponte. Un total de 375 cailles (250 femelles et 125 males: Coturnix
coturnix japonica), âgées de huit semaines, ont été réparties au hasard en
cinq groupes homogènes (cinq réplicas de dix femelles et cinq mâles). Les
animaux ont reçu une alimentation de base complétée par l’huile essentielle
de myrte à des doses de 0, 500, 1000, 2000 et 5000 mg/kg d’aliments.
L’expérience a été menée pendant huit semaines. A la fin de l’expérience,
aucune différence significative n’a été perçue entre les groupes en termes de
poids vif initial, de consommation d’aliments, du poids et la qualité de l’œuf,
les valeurs biochimiques du sérum, l’éclosion, la mort précoce ou tardive
de l’embryon et la mort sub-membraneuse. Après 8 semaines, la production
d’œufs avait diminué (p< 0.01) alors que l’indice de consommation avait
augmenté (p< 0.05) chez les cailles ayant reçu une alimentation complétée
par la dose de 5000 mg/kg d’huile essentielle de myrte. L’épaisseur des
coquilles avait diminué (p< 0.05) chez les groupes recevant des doses d’huile
essentielle de myrte de 2000 et 5000 mg/kg. L’indice de la couleur jaune a
été affecté positivement par l’ajout de l’huile essentielle de myrte (p< 0.001).
La performance de l’éclosion était plus élevée chez les groupes ayant reçu
une alimentation complétée avec des doses d’huile essentielle de myrte de
1000 mg/kg alors qu’elle était plus basse chez les groupes ayant reçu 5000
mg/kg (p< 0.01). L’ajout de l’huile essentielle de myrte à l’alimentation a
significativement diminué le cholestérol (p< 0.01), Calcium (p< 0.01) et
malondialdéhyde (MDA; p< 0.001) sérique et augmenté significativement
les concentrations d’azote uréique (p< 0.001) et de β-carotène (p< 0.001).
La créatine kinase sérique (CK-MB) a augmenté (p< 0.001) à la dose de
5000 mg/kg alors que la concentration d’albumine a diminué (p< 0.01).
La concentration MDA du jaune d’œuf a diminué (p< 0.01) dans tous les
groupes qui ont reçu un complément d’huile essentielle de myrte à leur
alimentation aux 15ème et 30ème jours de conservation (p< 0.001). En
conclusion, la complémentation par de l’huile essentielle de myrte a changé
la performance de ponte et les valeurs biochimiques chez les cailles de ponte
selon la quantité et la période d’apport. La prise en compte des paramètres de
production et qualité des œufs, de concentration en MDA dans le jaune d’œuf
et le taux d’éclosion conduisent à recommander de compléter l’alimentation
des cailles pondeuse à la teneur de 1000 mg/kg d’huile essentielle de myrte.
Mots-clés : Myrte, performance, éclosion, peroxydation
lipidique, cailles.
[19] effects. There are also growth promoting effects that are
due to the antimicrobial properties and positive influence
on the microflora of the digestive tract [32]. Thus, aromatic
plants and their extracts are accepted as natural and safe
substances for birds.
Revue Méd. Vét., 2014, 165, 9-10, 280-288
USE OF MYRTLE OIL IN LAYING QUAILS DIET 281

As an aromatic plant, Myrtus communis L. belongs to Material and Methods

the Myrtaceae family. It is widely used in medicine and the
pharmaceutical industry, because it contains volatile fatty
acids (VFAs) and other compounds. Volatile fatty acids of M.
communis L. include myrtenol, myrtenyl acetate, limonene,
linalool, α-pinene, 1,8-cineole, β-caryophyllenein, p-cymene,
geraniol, nerol, phenylpropanoid and methyleugenol [38]. It
has been reported that derivates of some myrtus extracts, such
as beta-triketones [30], tannens, myricetin, gallic acid and
ellgic acid, have antibacterial effects and the plant has strong
antioxidant activity due to its galloyl derivates [43]. Also,
it has antifungal [23], hypoglycaemic [46], anticonvulsant
[18], anticarcinogenic [15], anti-inflammatory [20] and
antimutagenic [28] effects.
Studies involving M. communis L. have focused mostly
on its volatile components and the composition of phenolic
compounds in leaves and fruit. However, no available
scientific data were found about the utilisation of the plant’s
versatile benefits in the diets of laying quails. The aim of this
research was to investigate the effects on performance, egg
quality, some biochemical values and hatchability of laying
quails that received varying concentrations of myrtle oil
added to their diets.
ANIMALS, DIETS AND EXPERIMENTAL DESIGN
This experimental study was carried out in The Avian
Research Farm at the Animal Research Center of Afyon
Kocatepe University, Turkey, following the ethical committee
approval. A total of 375 quails (250 females and 125 males;
Coturnix coturnix Japonica), aged eight weeks old, were
randomly allocated to receive one of five dietary treatments.
The birds were fed either a basal diet or the basal diet with
supplemented with myrtle oil doses of 500, 1000, 2000 or
5000 mg/kg feed. Each treatment consisted of five replicates
of ten females and five males. The birds were housed in
cages kept inside a windowed poultry house with a light
regimen of 16 hours light and 8 hours dark. Feed and water
were provided ad libitum. The experiment was conducted
for duration of eight weeks. The content of the basal diet is
shown in Table I. The basal diet was formulated to meet or
slightly exceed the nutrient requirements recommended by
the NATIONAL RESEARCH COUNCIL [37], and contained
12.15 MJ/kg Metabolisable Energy (ME) and 21.68% crude
protein. The diet was free of antibiotics, coccidiostats and
growth promoters. The nutrient composition of the basal
diet, including moisture, crude protein, crude fat, crude
fiber, crude ash, calcium and phosphorus contents was

Ingredients %
Corn 47.23
Full fat soybean 25.00
Sunflower meal 9.32
Soybean meal 6.20
Corn gluten 0.60
Meat and bone meal 4.00
Vegetable oil 1.00
CaCO3 4.60
Dicalcium phosphate 0.40
Salt 0.25
NaHCO3 0.25
L-Lysine 0.10
DL-Methionine 0.70
Vitamin mineral premixa 0.35
Chemical composition
Dry matter % 92.16
Crude protein % 21.68
Crude fat % 8.06
Crude fiber % 4.29
Crude ash % 6.32
Nitrogen free extract % 51.81
Calcium % 2.55
Phosphorus % 0.40
Metabolisable energyb (MJ/kg) 12.15
a
Added per kg of diet: retinol acetate, 4.1 mg; cholecalciferol, 0.075 mg; DL-α-tocopherylacetate, 11 mg; menadione, 1 mg; thiamin, 2.8 mg; riboflavin sodium
phosphate, 5.8 mg; niacin, 44 mg, pyridoxine, 4 mg; cyanocobalamin, 0.026 mg; Ca-D-pantothenate, 8.8 mg; folic acid, 1 mg; choline, 220; biotin, 0.11 mg; Cu,
6 mg; I, 1.1 mg; Fe, 30 mg; Se, 0.1 mg; Mn, 60 mg; Zn, 50 mg b Metabolisable energy content of diets was estimated using the equation devised by Carpenter and
Clegg [33]

Table I: Ingredients and chemical composition of the basal diet

Revue Méd. Vét., 2014, 165, 9-10, 280-288

282
determined according to the AOAC [4]. Metabolisable
energy of each diet was estimated using the following
equation devised by Carpenter and Clegg [33]: ME, kcal/
kg = 53 + 38 [(crude protein, %) + (2.25 x crude fat, %) +
(1.1 x starch, %) + (sugar, %)]. Group feeding was applied for
all treatment groups. Moreover, specific gravity values were
considered when calculating the amount of myrtle oil to add
to the diets. The specific gravity value of myrtle oil (0.8897
g/mL) was determined by the suppliers (NBT Company,
Alanya, Turkey). The essential oils were extracted by
hydrodistillation. Hydrodistillation is a simple form of steam
distillation. High pressure steam is forced through crushed
plant, picking up the oil. The vapor containing the oil is then
condensed, producing a liquid containing a mixture of water
and the plant oil. The oil is then separated from the water
[7]. Then the composition of myrtle oil was determined by
GS-MS. Mytrle oil was added to a small portion of the diet
and then sprayed on to the total diet. All diets were prepared
freshly every week.
LAYING PERFORMANCE
Quails were weighed individually at the beginning and
end of the study. Eggs were collected daily and egg production
was calculated on % production for egg numbers. Egg weight
was recorded daily for each replicate. Feed consumption was
recorded every two weeks. The feed conversion rate (FCR)
was calculated as kilograms of feed per kilogram of eggs.
EGG QUALITY TRAITS
Twenty eggs from each group (five eggs from each
replicate) were collected on the second, fourth, sixth and
eighth weeks of the experiment to determine the quality of
the interior and exterior of the eggs at monthly intervals. Egg
quality traits were measured within 24 hours of collection.
The eggs were examined for weight (g), length (mm), width
(mm), egg shell thickness (mm), fracture strength, albumen
index (%), yolk index (%), Haugh Unit (HU), egg shape index
and yolk color index. Egg weights were measured to the
nearest 0.1 g using a digital scale. Egg width, egg length, yolk
width, albumen length and albumen width were measured by
digital compass to the nearest 0.01 mm. Measurements of the
shell thickness of dried shells with the membrane still intact
were obtained from two sides in the equatorial region, as well
as on the blunt and the pointed edges with a micrometer to
the nearest 0.01 mm. Also, yolk and albumen heights were
measured by micrometer to the nearest 0.01 mm. The egg
yolk visual color score was determined by matching the
yolk with one of the 15 bands of the “1961, Roch Improved
Yolk Color Fan”. Egg quality traits were calculated using the
following formulas [27,54]:
Shape index (%) = [egg width (mm)/egg length (mm)]
x 100
Yolk index (%) = [yolk height (mm)/yolk width (mm)]
x 100
Haugh Unit = 100 log [albumen height (mm) + 7.57 - 1.7
0.37
x egg weight (g)]
BULBUL (T.) AND COLLABORATORS
Albumen index (%) = albumen height (mm)/[(average
albumen length (mm) + width (mm) /2) x 100]
DETERMINATION OF SERUM BIOCHEMICAL VALUES
At the end of experimental period , blood specimens
collected from two animals per replicate, from a total of
10 animals, into dry tubes were centrifuged 1500 g for 15
minutes at 4°C. Then, the sera were stored at -18 ˚C for further
analysis. The serum concentrations of calcium (Ca; limit of
quantitation, 0.50 mmol/L; intra- and interassay coefficients
of variation, 0.7% and 2.1%), alkaline phosphatase (ALP;
sensitivity of assay, 1 U/L; intra- and interassay coefficients
of variation, 0.8% and 2.9%), alanine aminotransferase (ALT;
sensitivity of assay, 1 U/L; intra- and interassay coefficients of
variation, 0.8% and 1.1%), aspartate aminotransferase (AST;
sensitivity of assay, 1 U/L; intra- and interassay coefficients
of variation, 0.7% and 2.9%), creatinine (sensitivity of assay,
1.52 µmol/L; intra- and interassay coefficients of variation,
1.1% and 1.6%), urea (sensitivity of assay, 0.357 mmol/L;
intra- and interassay coefficients of variation, 1% and 1.6%),
creatine kinase - MB (CK-MB; sensitivity of assay, 10 U/L;
intra- and interassay coefficients of variation, 1.9% and 3.6%),
albumin (sensitivity of assay, 0.2 g/L; intra- and interassay
coefficients of variation, 1.5% and 3.6%, total cholesterol
(sensitivity of assay, 0.0259 mmol/L; intra- and interassay
coefficients of variation, 1.6% and 2.6%), high-density
lipoprotein cholesterol (HDL-C; sensitivity of assay, 0.0259
mmol/L; intra- and interassay coefficients of variation, 1.7%
and 2.2%) and low-density lipoprotein cholesterol (LDL-C;
sensitivity of assay, 0.0259 mmol/L; intra- and interassay
coefficients of variation, 1.5% and 2.2%) (Cormay, Poland)
concentrations using autoanalyser (Tokyo Boeki Prestige
24i, Japan) [29,34,45]. Serum MDA [17] and β-carotene
concentrations were determined using the method of [51]
were determined with enzyme-linked immunosorbent
assays and a spectrophotometric reader (MWGt Lambda
Scan 200, Bio-Tek Instruments, USA). Precision of the assay
was assured by use of a quality control sample, which was
included on each plate.
ANALYZING EGGS MALONDIALDEHYDE (MDA)
CONCENTRATIONS
Malondialdehyde was measured as a secondary oxidation
product according to the TBA method described by [31]
using spectrophotometry with some modifications. At the
end of the experimental period, 100 egg yolk specimens (20
egg yolk specimens from each group) were tested. The lipid
oxidation value of raw egg yolk specimens stored at +4oC was
determined at 1., 7., 15., and 30. days of storage. A modified
2-thiobarbituric acid method was used, and the results were
expressed as the amount of 2-thiobarbituric acid reactive
substances (mg MDA). This method is based on observing
the red color that is created by the oxidation of unsaturated
fatty acids with thiobarbituric acid (TBA) after heating MDA.
For the analyses, 10 g of the specimen was homogenized with
distilled water in a blender and then transferred to a Kjeldahl
Revue Méd. Vét., 2014, 165, 9-10, 280-288
USE OF MYRTLE OIL IN LAYING QUAILS DIET
flask, where the specimen was distilled to aggregation by
adding 2.5 mL of 4 N HCl (Merck, Germany) and 1 mL of
Antifoam A. The reactant, 5 mL of TBA (Merck, Germany),
was added to 5 mL distillate and incubated in a water bath for
up to 30 min. The final solution and a blank were measured
in a spectrophotometer at 538 nm. The final value was
expressed as mg MDA/kg specimen (yolk).
DETERMINATION OF HATCHABILITY
A total of 160 eggs were collected per group (32 eggs from
each replicate) at weeks four and eight of the study. The eggs
that had suitable characteristics for brooding were placed in
a brooder on a group basis. The number of hatched chicks
was recorded for three days after the 17 days of brooding.
Then, the remaining non-hatched eggs were cracked, and the
number of fertile and unfertile eggs and number of embryonic
deaths were recorded. The hatchability characteristics were
calculated using the following formulas:
Fertility rate (%) = (fertile egg count / hatched egg count)
x 100
Hatching performance (%) = (number of eggs hatched
out / hatched egg count) x 100
Hatchability (%) = (number of eggs hatched out / hatched
fertile egg count) x 100
Fertility checks were performed during the early (less
than six days) and late (7 to 15 days) periods. Embryonic and
submembranous deaths at 16 to 17 days plus those that died
after egg perforation were determined by breaking the egg
shell of unhatched eggs at the end of the hatching period.
STATISTICAL ANALYSIS
The effect of different treatment doses of myrtle oil on
laying performance, egg quality traits, serum biochemical
values and egg yolk MDA concentration were subjected
to ANOVA procedures appropriate for a completely
randomized design. All replicates were the experimental unit
for all analysis. When differences (p< 0.05) among means
were found, means were separated using Tukey test. A Chi
square test was used to evaluate hatching properties. The
values were given in median and range.
Components
α-pinene
1,8-cineole
Limonene
Linalool
α-terpineol/ α-terpinyl acetate
Linalyl acetate
Geraniol
β-caryophylle
Table II: Chemical composition (%) of the volatile oil of Myrtus communis L.
Revue Méd. Vét., 2014, 165, 9-10, 280-288
283
Results
The ingredients and chemical composition of the diets are
presented in Table I. The volatile oil composition of myrtle
oil is summarized in Table II. The main active components
of myrtle oil are α-pinene (31.2%), 1,8-cineole (24.2%) and
limonene (13.8%).
It was observed that addition of 500 to 2000 mg/kg of
M. communis L. oil to the diets had no effect on final LW;
however, supplementation of 5000 mg/kg reduced the final
LW in both male and female quails (p< 0.05). M. communis
L. oil added diets did not altered feed consumption of the
groups. Supplementation of 5000 mg/kg M. communis L.
oil reduced egg production in whole study except second
week (p< 0.001). However, supplementation of 1000 mg/kg
M. communis L. oil to the diets increased egg production in
entire study. Supplementation of 5000 mg/kg M. communis
L. oil to the diets had negative effects on FCR during whole
study although there was no difference in egg weight between
groups (Table III).
During eighth week of experiment, it was found that
eggshell thickness was 35,23 mm in control group, whereas
the groups supplemented with 500, 1000, 2000 and 5000
mg/kg/day myrtle oil had 34.55, 33,78, 31.30 and 32.21 mm
thickness, respectively. Furthermore, yolk color index in
control and experimental groups (500, 1000, 2000 and 5000
mg/kg/day) were 6.1 and 7.80, 8.40, 7.80, 8.20, respectively.
However, diet supplemented with myrtle oil doses of 2000
and 5000 mg/kg were decreased eggshell thickness (p< 0.05).
But addition of myrtle oil to laying quail diets increased yolk
color index at eight weeks. Other egg quality traits (fracture
strength, haugh unit, shape index, yolk index and albumin
index) were not affected by myrtle oil addition to the diets.
Biochemical results obtained during the study are shown
in Table IV and egg yolk MDA concentrations are shown
in Table V. As presented, the addition of myrtle oil to the
diets decreased serum Ca concentrations (p< 0.01), whereas
it increased urea (p< 0.001) and β-carotene (p< 0.001)
concentrations in all groups. Supplementation with 5000
mg/kg myrtle oil was increased CK-MB activity (p< 0.001)
and decreased albumin concentration (p< 0.05); during all
doses (500-5000 mg/kg) of myrtle oil supplementation,
%
31.2
24.2
13.8
8.8
4.9
3.9
1.4
1.0
 284

Dietary myrtle oil supplementation (mg/kg/day)

0 500 1000 2000 5000 p<
Initial live weights, g
Female 188.4 (181.1-195.6) 187.3 (180.2-193.4) 187.1 (179.8-193.2) 188.3 (182.5-194.5) 188.7 (183.1-196.7) NS
Male 175.3 (165.9-178.5) 172.1 (168.1-176.3) 172.5 (163.4-178.1) 168.12 (165.9-174.5) 172.77 (163.4-178.3) NS
Final live weights, g
Female 210.4 (203.9-211.0)a 209.9 (204.7-216.4)a 207.5 (198.6-214.3)ab 204.8 (202.1-213.2)ab 199.3 (193.8-205.02)b 0.05
Male 190.2 (183.3-197.8)a 193.36 (185.5-193.3)a 190.4(185.1-193.3)a 186.7 (183.0-192.0)ab 184.8 (173.1-189)b 0.05
Feed consumption, g/day 31,44 (27.44-33.12) 31.32 (28,34-32,91) 33.04 (29,88-35.78) 31.93 (27.11-33.78) 30.78 (28.11-33.34) NS
Egg production, % 83.14 (80.12-86.91)c 84.57(77.12-89.14)ab 89.03 (84.62-93.56)a 84.21 (81.45-85.88)bc 76.70(70.84-80.56)d 0.01
FCR, 2.78 (2.70-2.89)b 2.72 (2.62-2.80)b 2.78 (2.64-2.86)b 2.81 (2.70-2.84)ab 3.02 (2.89-3.10)a 0.05
Egg weight, g 13.60 (12.80-13.87) 13.68 (13.06-13.78) 13.32 (13.04-13,82) 13.46 (13.10-13.98) 13.48 (13.23-14.12) NS
FCR: Food conversion ratio (feed consumed, (g)/egg production (g).
Letters (a, b, c, d) in the same line indicate significant differences between different letters. NS = not significant

Table III: Effects of Myrtus communis L. oil diets on laying performance

Dietary myrtle oil supplementation (mg/kg/day)

0 500 1000 2000 5000 p<
b b
CK-MB (U/L) 2667 (2123-3212)b 3060 (2164-3412) 3412 (2345-3821) 2711 (2043-3567)b 5413 (4213-6798)a 0.001
b a
Urea (mmol/L) 16.30 (12.18-21.13)c 33.40 (24.00-42.11) 47.16 (23.44-61.23) 45.10 (33.21-58.12)a 54.50 (32.45-65.72)a 0.001
a a
Albumin (g/L) 19.3 (16.5-19.8)a 18.8 (16.4-19.7) 18.6 (17.2-20.2) 17.1 (16.0-18.3)ab 15.0 (14.0-15.8)b 0.01
b b
Total cholesterol (mmol//L) 10.62 (9.30-14.97)a 9.87 (9.30-10.26) 10.11 (8.50-10.16) 8.32 (7.39-10.08)b 9.38 (7.44-10.65)b 0.01
HDL-C (mmol/L) 1.39 (0.46-2.07)c 2.25 (1.86-3.57)cb 2.44 (1.69-3.22)b 3.76 (2.64-4.91)a 3.10 (2.53-4.08)b 0.001
b ab
LDL-C (mmol//L) 1.86 (1.53-2.09)b 1.79 (1.53-1.91) 2.45 (2.15-4.11) 3.13 (1.62-3.48)a 2.93 (2.56-3.31)a 0.01
b b
Ca (mmol/L) 7.67 (6.80-8.52)a 5.55 (4.82-6.53) 6.00 (4.80-7.20) 4.70 (4.10-7.35)b 5.28 (4.94-5.57)b 0.01
a a
β-carotene (µmol/L) 0.245 (0.207-0.262)b 0.313 (0.268-0.387) 0.295 (0.281-0.299) 0.351 (0.261-0.387)a 0.369 (0.294-0.396)a 0.001
MDA (µmol/L) 3.56 (3.21-4.95)a 2.87 (2.64- 3.36)b 2.54 (2.24-2.93)bc 2.56 (2.21-2.96)bc 2.11 (1.88-2.82)c 0.001
CK-MB: creatine kinase-MB, HDL-C: High-density lipoprotein cholesterol,
LDL-C: Low-density lipoprotein cholesterol, MDA: Malondialdehyde.
Letters (a, b,c) in the same line indicate significant differences between different letters, NS = not significant

Table IV: Effects of Myrtus communis L. oil diets on some serum biochemical values

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BULBUL (T.) AND COLLABORATORS
USE OF MYRTLE OIL IN LAYING QUAILS DIET
total cholesterol concentration decreased, while HDL-C
and LDL-C increased. However, it was observed that the
supplementation of myrtle oil did not affect the ALP, ALT,
AST and creatinine concentrations. In this study it was
found that the addition of myrtle oil decreased serum MDA
concentrations as compared with the control group, but the
effect was dependent on the dose (p< 0.001). Egg yolk MDA
concentrations were varied depending on storage time. The
lowest MDA concentration (p< 0.01) was observed on days
15th and 30th in the group received a daily diet supplemented
with 2000 mg/kg of myrtle oil.
No differences were found in fertility rate, hatchability,
early embrionic death, late embrionic death and sub-
membraneous death at 4th and 8th weeks. During eighth week
of the experiment, hatch performance was 65.8%, 70.0%,
78.3%, 64.2% and 54.3% in control and experimental groups
(500, 1000, 2000 ve 5000 mg/kg myrtle oil), respectively. It
was seen that the most increased (p< 0.01) hatch performance
was in the groups consumed a diet supplemented by 1000
mg/kg myrtle oil at week 8th.
Discussion
The aim of this study was to determine the effects
of a diet supplemented with M. communis L. oil in the
performance, egg quality, some serum biochemical values
and lipid peroxidation of serum and egg, and hatchability
in laying quails. The dosage range selected was between 500
and 5000 mg/kg on the basis of other essential oil studies
[7,11,14,40] because there was no available data about myrtle
oil supplementation of diets. M. communis L. oil used in
this research, extracted from M. communis L. leaves which
is rich in volatile oil as analysed composition showing high
proportion of α-pinene (31.2%) and 1,8-cineole (24.2%) and
moderate proportion of limonene (13.8%), linalool (8.8%)
lesser proportion of α-terpineol / α-terpinyl acetate (4.9%),
linalyl acetate (3.9%) and geraniol (1.4%). This content
complies with the former reports [21,35,41]. The specific
gravity of the myrtle oil added to the diets in this study was
0.8897 g/ml. The specific gravity value of myrtle oil has been
reported to range between 0.940 and 0.980 g/ml at 25oC and
between 0.875 and 0.893 g/ml at 20oC (US/EU/EUROPA/
FDA/JECFA Information). These reported values are in the
normal ranges found in our study. In this study, the specific
gravity value of the essential oil was taken into consideration
when calculating the essential oil dosage added to the diets.
In this study, it was found that the addition of 500 to 2000
mg/kg myrtle oil to the diet has no effect on the final LW of
female and male quails. It was reported that the addition of
eucalyptus at a dose of up to 3 g to layer diets, which is rich
in 1,8-cineol and α-pinene and is similar to myrtle oil, has
no effect on live weight [1]; however, essential oils increase
the secretion of pancreatic digestive enzymes, i.e., amylase,
lipase, trypsin, and chymotrypsin, therefore increasing the
bioavailability of digestives [32]. Similarly, it is reported that
Revue Méd. Vét., 2014, 165, 9-10, 280-288
285
the addition of essential oils from some plants such as thymol
to chicken rations increases feed intake and may induce
growth [32]. YESILBAG et al. [55] noted that, rosemary
volatile oil supplementation at 100 mg/kg has a positive effect
on performance variables. In the present study, it was found
that the addition of 5000 mg/kg myrtle oil decreased final LW.
Although the composition of thyme and myrtle are different,
ALI et al. [3] reported that the addition of 0.25% thyme leaf
to layer diets decreased LW. Although the authors concluded
that the difference between the results was because of the
difference in essential oil sources and quantity, it was shown
that myrtle oil has quantity dependent effects on LW.
In the present study, there was no difference in feed
consumption between the groups. The results of the study
showed supplementation of 24 mg/kg essential oil mixture
(oregano oil, laurel leaf oil, sage leaf oil, myrtle leaf oil, fennel
seed oil, citrus peel oil) [14], and also supplementation of
50 and 100 mg/kg oregano to the diets did not altered feed
intake which complies with the relevant former reports [22].
However, BOLUKBASI et al. [8] reported that bergamot
oil depresses feed intake depending on the quantity added.
Difference in feed intake during study may be due to different
taste and odor of essential oils.
In this research, we have observed that myrtle oil did not
affect egg weight. However, it was observed that myrtle oil
supplementation altered egg production and FCR depending
on added quantity and consumption period. Other reports
show that supplementation of rosemary, oregano, saffron
[11] or oregano essential oil [22] to the diets did not alter
egg production and FCR. On the contrary, it was reported
that supplementation of 24 mg/kg essential oil mixture has
improved FCR [14]. Supplementation of 5000 mg/kg myrtle
oil to the diet has worsen FCR in whole study, whereas
other quantities did not alter it. Egg production increased in
whole study by supplementation of 500 and 1000 mg/kg of
myrtle oil, although 5000 mg/kg supplementation of myrtle
oil suppressed it significantly at weeks 4th, 6th, 8th and during
entire study. Thus, low FCR is related to low egg production
of the same group.
In the current study, no difference was found between
groups in the internal quality parameters except yolk color
index; yolk color index increased in groups receiving myrtle
oil at week eight (Table IV). Similarly, it was reported that the
addition of 20 mg/kg saffron [11] or 1% curcuma longa [40] to
the diets improved yolk color. However, it was shown that the
addition of 5000 mg/kg Rosmarinus officinalis or 5000 mg/
kg Origanum vulgare to the diets did not change yolk color
[11]. Yolk color depends on the deposition of carotenoids in
the yolk [26]. There is a relation between plasma carotene
concentrations and yolk color [10]. Higher plasma carotene
concentrations in all groups in the present study as compared
with the control were thought to be due to the effect of myrtle
oil, which prevents the carotene in feed from degrading or
maintains plasma carotene concentrations by its antioxidant
286
effect. In the current study, it was concluded that improved
yolk color was due to the effect of elevated plasma β-carotene
concentrations in the myrtle oil supplemented group.
It was observed that egg shell thickness was decreased in
addition of 2000 and 5000 mg/kg of myrtle oil to the diets in
eight weeks (p< 0.05, Table IV). BOLUKBASI et al. [8] reports
that the addition of bergamot oil to the diets decreases shell
index. However, some reports indicate that the addition of an
essential oil to the diets increases egg shell thickness [5,36]
or does not change it [22,24]. In the present study, it was
found that the addition of myrtle oil to the diet significantly
decreased serum Ca concentrations (p< 0.05, Table V). A
decrease in egg shell thickness may be due to a decrease in
serum Ca concentrations. The major components of myrtle
oil are 1,8-cineole, myrtenyl acetate, α-pinene, myrtenol,
limonenes and monoterpenes. It stops in vivo and in vitro
bone resorption reversibly in rats [16]. It was reported that
plasma Ca concentrations of hens were decreased depending
on the added quantities of monoterpenes to the diet. Also,
it has been reported that monoterpenes directly effect
osteoclastic formation in homopoietic cells and blocks bone
resorption [16]. The addition of myrtle oil related to low
Ca concentrations in the current study, may be due to the
blocking of intestinal absorption of Ca, accelerated kidney
clearance and/or blocking bone resorption.
Serum total cholesterol concentrations (p< 0.05)
were decreased, whereas HDL-C (p< 0.01) and LDL-C
concentrations (p< 0.05), which are included in the
cholesterol profile, were increased depending on added
quantity of myrtle oil to the diet (Table V). Similar to the
current results, essential oils were shown to decrease serum
cholesterol concentrations in previous studies [3,40]. It
is reported that 3 hydroxy-3 methylglutaryl coenzyme
A (HMG-CoA) reductase plays a key role in cholesterol
synthesis and essential oils down regulate HMG-CoA and
therefore shows a hypocholesteromic effect [32]. Similarly, it
was observed that the addition of 25 to 100 mg/kg limonene
to the diet inhibits hepatic HMG-CoA and reduces serum
cholesterol concentrations [39]. However, BOLUKBASI
et al. [9] report that the addition of bergamot oil does not
reduce cholesterol concentrations.
It has been reported that AST, ALT and ALP are
remarkable values for the detection of liver damage in birds
[29,33,45]. There were no statistically significant (p> 0.05)
Dietary myrtle oil supplementation (mg/kg/day)
0 500
1st day 0.08 (0.05-0.14)a 0.04 (0.031-0.09)b
7th day 0.13 (0.09-0.23)a 0.13 (0.08-0.19)a
15th day 0.24 (0.14-0.32)a 0.14 (0.09-0.28)b
30th day 0.61 (0.30-0.78)a 0.21 (0.16-0.27)b
Letters (a, b) in the same line indicate significant differences between different letters
Table V: Effects of Myrtus communis L. oil diets on egg yolk MDA concentration (mg MDA /kg specimen) in raw egg specimens at different storage times
(at +4°C)
BULBUL (T.) AND COLLABORATORS
differences in serum indicators originating from the liver
(ALP, ALT, AST) in this study. Enzyme CK has high activity in
skeletal muscles and in the myocardium. The isoenzyme CK-
MB is present in many tissues, mostly in the myocardium.
CK is more active in skeletal muscles than in myocardium.
An increase in serum CK and CK-MB activities is generally
linked to defects in skeletal muscles and myocardium [29,52].
Elevated concentrations of CK-MB and urea indicate the
presence of muscle tissue degeneration as well [29]. In the
present study, CK-MB (p< 0.001), urea (p< 0.01) activities
were raised, whereas albumin concentration decreased (p<
0.05, Table IV) in the groups fed a diet supplemented with
5000 mg/kg myrtle oil showing that myrtle oil may be cause
muscle degeneration. Also, decreased LW in these groups
was in accordance with biochemical results.
In the present study, it was observed that yolk MDA
concentrations were decreased on days 15th and 30rd in all
myrtle groups (p< 0.001). It was reported that the antioxidant
effects of aromatic plants are due to their phenolic compounds
[48]. Phenolic compounds act as antioxidants by trapping
free radicals, which reduces singlet oxygen formation by
making compounds with metallic ions [42]. Also, 1.8-cineole
α-pinen and β-pinenin, which are some major components
of myrtle oil, are known to have antioxidant activity [53].
Similarly, antioxidant characteristic of myrtle oil was also
reported [2]. Due to the presence of phenolic OH groups;
these groups act as hydrogen donors to the peroxy radicals
produced in the first step in lipid oxidation, thus retarding
the formation of hydroxyl peroxide.
In the present study, it was observed that the addition of
1000 mg/kg M. communis L. oil to the diet increased the hatch
performance statistically at eight weeks (p< 0.01). ALI et al.
[3] reported that individually added 1% thyme, 1% rosemary,
1% oregano and 0.5 to 1% curcuma longa to chicken diets
increased fertility and hatchability. Oxidative metabolism
increased, especially in the final few days before hatching,
as a normal result of embryonic growth. It is reported that
over increment lipid peroxidation may lead to tissue damage
[49], whereas diets with added antioxidants may protect the
embryo and therefore increase survival rate [50].
As a conclusion, this study suggested that myrtle oil
supplementation, especially at a concentration of 500 and
1000 mg/kg may be considered a potential natural growth
promoter. However, more studies are needed to define the
1000 2000 5000 p<
0.07 (0.06-0.14)a 0.08 (0.07-0.12)a 0.08 (0.03-0.13)a NS
0.07 (0.05-0.12)b 0.11 (0.06-0.16)a 0.05 (0.09-0.15)b 0.01
0.10 (0.07-0.16) c
0.08 (0.05-0.13)c 0.09 (0.05-0.15)c 0.01
0.21 (0.13-0.28)b
0.11 (0.06-0.30)b 0.15 (0.09-0.26)b 0.001
Revue Méd. Vét., 2014, 165, 9-10, 280-288
USE OF MYRTLE OIL IN LAYING QUAILS DIET
effect of myrtle oil supplementation on the performance of
poultry with regard to environmental conditions, effective
dosage, active oil substances, dietary ingredients and nutrient
density. Furthermore, this study indicated that the use of
myrtle oil as a natural antioxidant may improve poultry egg
quality and may extend the shelf life of poultry egg products.
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