https://doi.org/10.52973/rcfcv-e34465
Received: 05/06/2024 Accepted: 06/08/2024 Published: 05/12/2024
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Revista Científica, FCV-LUZ / Vol. XXXIV, rcfcv-e34465
ABSTRACT
In cattle metabolism, fatty acids are basic biological components that
meet the body’s energy needs and are used in important metabolic
processes. In this study, sweat, urine and blood samples were taken
from cows and the fatty acids of the samples were determined by
gas chromatography. Sweat samples contained fewer fatty acids
than blood and urine (14 in sweat, 25 in blood and 19 in urine). In the
correlation analysis, there was a moderately positive, statistically
significant (P<0.01) relationship between sweat fatty acids and
P
correlation was found between blood and urine fatty acids. Regression
blood fatty acids, and sweat and urine fatty acids could explain 81%
moderate correlation in urine fatty acids and that it could explain 79%
of the changes in sweat fatty acids. It was determined that the changes
in blood fatty acids were due to the changes in sweat and urine fatty
acids. Therefore, it was concluded that blood and urine fatty acids
id levels.
Key words: Fatty acids; gas chromatography; sweat
RESUMEN
En el metabolismo del ganado, los ácidos grasos son componentes
biológicos básicos que satisfacen las necesidades energéticas
del organismo y se utilizan en importantes procesos metabólicos.
En este estudio, se tomaron muestras de sudor, orina y sangre de
vacas y se determinaron los ácidos grasos de las muestras mediante
cromatografía de gases. Se encontraron menos ácidos grasos en
muestras de sudor que en muestras de sangre y orina (14 en el sudor,
25 en la sangre y 19 en la orina). En el análisis de correlación, hubo una
(P<0,01) entre los ácidos grasos del sudor y los ácidos grasos de la
P
los ácidos grasos en la sangre. Como resultado del análisis, se vio
que el cambio de ácidos grasos en el sudor y la orina podría explicar
el 81% del cambio en la sangre. También se determinó que existía una
correlación moderada en los ácidos grasos de la orina. El cambio en
los ácidos grasos de la orina podría explicar el 79% de los cambios
en los ácidos grasos del sudor. Se determinó que los cambios en los
ácidos grasos de la sangre estaban relacionados con los cambios en
los ácidos grasos del sudor y la orina. Por lo tanto, se concluyó que los
estimar observando los niveles de ácidos grasos en el sudor.
Palabras clave: Ácidos grasos; cromatografía de gases; sudor
Ratio of fatty acids in sweat, blood and urine in cattle
Proporción de ácidos grasos en sudor, sangre y orina en ganado bovino
Özgül Anitaş
1
* , Serap Göncü
1
, Fatma Hepsağ
2
, Yeşim Özoğul
3
1
University of Cukurova, Faculty of Agriculture, Department of Animal Science. Adana, Türkiye.
2
University of Osmaniye Korkut Ata, Faculty of Applied Sciences, Department of Food Technology. Kadirli Campus, Osmaniye, Türkiye.
3
University of Çukurova, Faculty of Fisheries, Department of Seafood Processing Technology. Adana, Türkiye.
*Corresponding author: ozgulanitas01@gmail.com
Fatty acids of sweat and body fluids in cattle / Anitaş et al. _________________________________________________________________________
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INTRODUCTION
Cattle sweat has been the subject of studies for different purposes
in the world in recent years. Researchers have reported that chemical
changes occur in the blood, urine or milk of cattle, depending on
clinical diseases and problems in the body, starting years ago [1].
This awareness, combined with the advances in technology that
enables practical measurements at cow and herd levels through
the tests carried out on the farm, has rapidly gained importance
among dairy farm managers, veterinarians and other herd consultants
[2, 3]. In addition to the diagnostic methods that were constantly
used due to the discomfort caused by taking blood with needles for
diagnosis and treatment in bovine, studies have been focused on
developing alternative methods. One of these alternative methods
was to determine the compounds and their proportions by performing
sweat analysis. It constitutes an important advantage because the
sweat of animals can be easily collected from the body surfaces and
even mixed with molecules in the air [4
the less researched biological sample group [5].
Each animal has its unique body structure, nutrition, circumstances,
breed, age, and illness status, which might change depending on
body metabolism, however the majority of information obtained
about body metabolism functioning was based on results in serum/
blood metabolism and urine and fecal metabolism. However, the
number of detailed studies on the metabolism of sweat in animals
and their functions in the body, and in which situations the sweat–
forming substances change, was very limited [6]. Therefore, in
order to understand the health and disease states of animals, it was
necessary to understand sweat metabolism and its relationship with
sweat formation, sweat components, functional roles and physical
properties will provide useful information for animal health [7]. One of
these parameters is the fatty acid ratios determined in animal sweat.
Fatty acids (FAs) are biological molecules that are primarily used as
metabolic fuels and are involved in important metabolic processes.
Akbar et al. [8] stated that fats, fatty acids and metabolic products
formed after the use of fatty acids in the body have important roles
in metabolism. Among these duties of fatty acids were to create
resistance to the stresses and damages that may come from external
factors, to provide the necessary energy for the body, and being
the precursor of hormone–like eicosanoid compounds such as
thromboxane, leukotrienes and prostaglandins [9]. However, since
information regarding events that occur directly within the body.
Therefore, compounds in sweat were generally expected to contain
clinical biomarkers detected in the blood. According to Nunome et
al. [10] the fatty acids in human sweat were converted into sweat by
blood as a result of the breakdown of these fatty acids in adipose
tissue, particularly during stressful conditions. it was determined that
the concentration of fatty acids in sweat is related to the increase in
fatty acids in plasma [10]. Lack of studies on sweat fatty acid content,
fatty acid composition determined in the sweat of farm animals will
provide important information about the metabolism of the body [11].
This study was carried out in order to determine the concentration
fatty acids of sweat, blood and urine samples, which were the body
concentration of sweat fatty acids corresponding to the concentration
of blood and urine fatty acids in terms of cattle breeding.
MATERIALS AND METHODS
The present study was conducted at a research and experimental
farm located at Faculty of Agriculture, Çukurova University, Adana,
Türkiye. This study was approved by Cukurova University Animal
Experiments Local Ethics Board (Approval no: 26.02.2018/2). For
the study, sweat, blood and urine samples were obtained from 6
Holstein cows. Healthy cows with similar characteristics were used
(weight 600–670 kg (Tartimsan, Cattle Livestock Scale, Türkiye),
body condition according to score 2.5–3, age 3–5 years, at least one
delivery, no reproductive issues, 45–60 days postpartum). Based on
anamnesis data and clinical examinations, the enterprise veterinarian
concluded that none of the trial animals had any diseases. Sweat (10
septum) were obtained in accordance with animal welfare regulations
without harming the animals. There were 160 dairy cows on the farm,
50 of which were healthy and had similar characteristics. The number
of animals used has been determined by considering that the smallest
dairy cows to be used in the study was determined to be 10% of the
total number of
The experimental animals were milked twice a day using an
automated milking system (Sezer Milking Machines, Çanakkale,
Türkiye) at the central milking center. Cows were fed with a total
mixture ration (TMR) with a concentrate: roughage ratio of 60:40.
TMR consists of concentrate fed, alfalfa, wheat straw and corn silage
(18% crude protein and 2650 kcal/metabolic energy (ME)/kg) and was
given at 07.30 and 16.00.
Urine, blood and sweat samples were taken from experimental
fatty acids.
Data collection and method
Urine samples were collected into 10 mL tubes by manual
stimulation of the perineal areas of cows. For sweat samples, the
animal’s nose area was washed with water, then dried with a paper
towel and the sweat collected on the nose area was taken into tubes.
Sweat sample was placed in 10 mL tubes (Aromel, Konya, Türkiye)
for analysis [7et al. [12] were taken
as they stated. Blood samples were collected early in the morning.
Samples were prepared by mixing a 10 mL aliquot of blood with 10%
septum (Obstitech ApS, Denmark). Sweat, urine, and blood samples
Chromatography device.
Gas chromatography analysis
The method used by Bligh and Dyer [13] in their studies was applied
for the analysis of lipid extraction from samples taken from animals in
certain amounts. Methyl esters were prepared by Trans Methylation
using 2 M KOH in methanol and n–hexane according to the method
reported by Pollard and Shachar–Hill [14] 10 mg of oil obtained from the
analysis was dissolved in 2 mL of hexane, then 4 mL of 2 M methanol
KOH was added. The tube was then vortexed (Ika vortex 3, IKA Turkey
Laboratory and Process Technologies Inc., Türkiye) for 2 min at room
temperature to mix the liquid in it thoroughly. After centrifuging
FIGURE 1. Comparison of sweat, blood and urine fatty acids of cows
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for fatty acid compositions with an auto sampler (Perkin Elmer, Clarus
to 220°C and 280°C, respectively, the oven temperature was kept at
140°C for 5 min. The oven temperature was raised to 200°C at a rate
of 4°C·min
-1
and then to 220°C at a rate of 1°C·min
-1
. Samples (standard
helium) were sized to 1 μL and carrier gas operated at 16 psi. Partition
each sample while identifying the fatty acids, and the results were
presented as the mean value and standard deviation by calculating the
inferential statistical methods. With correlation analysis and multiple
regression analysis, it was tried to determine whether there was any
relationship between the variables, and if there was, the direction,
the relationship.
Statistical analysis
The statistical analyzes were made in the SPSS 16.0 package
program by determining fatty acids from sweat, urine and blood
samples taken from animals. Following the application of analysis of
variance (ANOVA) to the collected data, multiple comparison analysis
was done using the P
model was under the effect of one dependent variable and more than
one independent variable. In statistical theory, this multiple regression
relationship was generally expressed as follows [15]:
α + β
1
X
1
+ β
2
X
2
+ . . . + β
K
X
K
+ ε
In the above equation, the dependent variable was determined by
a linear combination of Y: X1, X2,…,Xk independent variables [16]. In
the equation, k: the number of independent variables, α: the constant
term, and βwere used.
In our study, the variables of fatty acids found in sweat, urine and
blood were used, and each variable was considered separately as a
dependent variable and its relationship was determined by performing
multiple regression analysis.
RESULTS AND DISCUSSION
TABLE I.
The fatty acids detected according to the analysis results of
samples were shown in detail in TABLE I (14 in sweat, 25 in blood and
19 in urine). Although margaric acid, alpha linolenic acid, eicosadienoic
acid, dihomo–γ–linolenic acid, behenic acid and erucic acid were
detected in blood and urine, they were not detected in sweat. Despite
the fact that methyl pentadecanoate, heptadecenoic acid, vaccenic
acid, linolenic acid, and lignoceric acid were found in blood, they were
not found in sweat or urine. In addition, as a result of the analysis,
the highest fatty acid content in animals was 24% palmitic acid and
17% myristic acid in sweat; 19% palmitic acid, 20% linoleic acid and
21% oleic acid in blood; In the urine, it was seen that 19% oleic acid
examined, 14 fatty acids were detected in sweat, 25 in blood and 19
in urine, and it was seen that less fatty acids were detected in sweat
compared to blood and urine (TABLE I).
TABLE I
Saturated fatty acids, monounsaturated fatty acids and polyunsaturated fatty acids in sweat, blood and urine
Fatty acids (%) Carbon Number Molecular formula
Molecular weight
(g·mol
–1
)
Sweat (%)* Blood (%)* Urine (%)*
Butyric acid C4:0 C
4
H
8
O
2
88.11 1.48 ± 0.42 ND** 2.00 ± 0.50
Caproic acid C6:0 C
6
H
12
O
2
116.15 9.34 ± 2.41 0.33 ± 0.15 0.77 ± 0.30
Caprylic acid C8:0 C
8
H
16
O
2
144.21 3.05 ± 1.33 0.15 ± 0.05 0.48 ± 0.08
Capric acid C10:0 C
10
H
20
O
2
172.26 3.48 ± 1.17 0.51 ± 0.11 ND
Laurik acid C12:0 C
12
H
24
O
2
200.31 2.45 ± 0.65 0.49 ± 0.10 ND
Myristic acid C14:0 C
14
H
28
O
2
228.37 17.82 ± 2.97 4.33 ± 1.65 2.68 ± 0.83
Pentadecanoic acid C15:0 C
15
H
30
O
2
242.40 1.20 ± 0.13 0.48 ± 0.11 0.96 ± 0.14
Palmitic acid C16:0 C
16
H
32
O
2
256.42 24.57 ± 3.07 19.80 ± 4.28 12.29 ± 2.68
Margaric acid C17:0 C
17
H
34
O
2
270.45 ND 0.78 ± 0.05 0.24 ± 0.12
Stearic acid C18:0 C
18
H
36
O
2
280.44 9.97 ± 1.72 14.78 ± 1.49 9.88 ± 2.10
Arachidic acid C20:0 C
20
H
40
O
2
312.53 1.23 ± 0.05 0.40 ± 0.09 1.23 ± 0.38
Behenic acid C22:0 C
22
H
44
O
2
340.58 ND 0.10 ± 0.05 0.20 ± 0.04
Lignoceric acid C24:0 C
24
H
48
O
2
368.64 ND 0.37 ± 0.09 ND
∑SFA 74.59 42.52 30.73
Myristoleic acid C14:1 C
14
H
26
O
2
226.36 ND 0.25 ± 0.04 ND
Methyl pentadecanoate C15:1 C
16
H
30
O
2
254.41 ND 0.24 ± 0.03 ND
Palmitoleic acid C16:1 C
16
H
30
O
2
254.41 2.00 ± 0.51 1.67 ± 0.46 1.57 ± 0.43
Heptadecenoic acid C17:1 C
17
H
32
O
2
268.44 ND 0.38 ± 0.05 ND
Vaccenic acid C18:1n7c C
18
H
34
O
2
282.46 ND 1.02 ± 0.02 ND
Oleic acid C18:1n9c C
18
H
34
O
2
282.46 7.38 ± 1.54 21.57 ± 4.15 19.47 ± 6.34
Eicosanoic acid C20:1 C
20
H
40
O
2
312.5304 ND ND 2.18 ± 0.78
Erucic acid C22:1
C
22
H
42
O
2
338.60 ND 2.77 ± 0.08 0.38 ± 0.16
∑MUFA 9.38 27.90 23.60
Linoleic acid C18:2n6 C
18
H
32
O
2
280.44 2.06 ± 0.40 20.09 ± 7.36 14.57 ± 3.35
Linolenic acid C18:3n6 C
18
H
30
O
2
278.43 ND 0.27 ± 0.11
Alpha Linolenic acid C18:3n3 C
18
H
30
O
2
278.43 ND 0.84 ± 0.48 2.63 ± 0.33
Eicosadienoic acid C20:2n6 C
20
H
36
O
2
308.50 ND 0.10 ± 0.06 9.67 ± 1.83
Dihomo–γ–linolenic acid C20:3n6 C
20
H
34
O
2
306.48 ND 0.30 ± 0.16 0.43 ± 0.02
Docosahexaenoc acid C22:6n3 C
22
H
32
O
2
328.48 1.02 ± 0.59 0.31 ± 0.13 2.99 ± 0.44
∑PUFA 3.08 21.91 30.29
MUFA·SFA–1 0.13 0.66 0.77
PUFA·SFA–1 0.04 0.52 0.99
PUFA/MUFA 0.33 0.79 1.28
∑n6 2.06 20.76 24.67
∑n3 1.02 1.15 5.62
n6/n3 2.02 18.05 4.39
*The ratios are shown as the mean ± standard deviation (SD). **ND: Not detected. Total SFA: all saturated fatty acids (without any double bond. 4:0 to 24:0). Total MUFA:
all monounsaturated fatty acids with a single double bond (14:1 to 22:1). Total PUFA: all polyunsaturated fatty acids. Total n–6 polyunsaturated fatty acids (PUFA): 18:2n6;
18:3n6; 20:2n6; 20:3n6. Total n–3 polyunsaturated fatty acids (PUFA): 18:3n3; 22:6n3.
Fatty acids of sweat and body fluids in cattle / Anitaş et al. _________________________________________________________________________
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FIGURE 2. Comparison of sweat, blood and urine fatty acids SFA, MUFA and
PUFA (%) of cow
TABLE II
Correlation between sweat, blood and urine fatty acids in cows
n: Compound number r P–value
Sweat–blood 28 0.384** 0.008
Sweat–urine 28 0.236 0.133
Blood–urine 28 0.855** 0.000
**: Correlation (r) is signicant at the 0.01 level (2–tailed)
TABLE III
Multiple regression analysis of sweat, blood and urine fatty acids in cows
Dependent
Variable
Independent
Variable
R R² Adjusted R² P-value
Sweat
Blood
0.379 0.143 0.094 0.067
Urine
Blood
Sweat
0.902 0.813 0.802 0.000
Urine
Urine
Sweat
0.894 0.799 0.788 0.000
Blood
TABLE IV
Coecients and signicance value of the regression model
of sweat, blood and urine fatty acids in cows
Dependent
Variable
– Sweat
B P
Dependent
Variable
– Blood
B P
Dependent
Variable
– Urine
B P
Constant
term
6.620 0.001
Constant
term
-550 0.630
Constant
term
1.496 0.84
Blood 0.585 0.045 Urine 1.150 0.000 Sweat -0.093 0.209
Urine -0.483 0.209 Sweat 0.188 0.045 Blood 0.688 0.000
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cows; It was seen that saturated fatty acids values were high in sweat,
low in urine, monounsaturated fatty acids values were high in blood,
low in sweat, and polyunsaturated fatty acids values were high in
urine and low in sweat.
TABLE IV displays the data values derived from the fatty acid
regression analysis. When the dependent variable sweat was chosen,
P value
was stated as 0.001, and the constant term was found to be important
in estimating the amount of sweat fatty acid content (P<0.05).
The TABLE II shows the results of the correlation test between
sweat fatty acids and blood and urine fatty acids of animals. The
correlation between sweat, blood, and urine fatty acids was found
to be statistically P<0.01).
When the values in TABLE III are examined, the R² value in the
dependent variable, sweat, was calculated as 0.143 and the adjusted
variables, urine and blood, explain 0.94% of the variation in sweat,
which was the dependent variable. When blood was chosen as the
dependent variable, the adjusted R
2
said that there was a high level of positive correlation and that the
independent variables, sweat and urine, could explain 81% of the
variance in the dependent variable, blood. Furthermore, the adjusted
correlated and could explain 79% of the variability in blood and sweat,
the independent variables.
The lack of studies on fatty acid analysis in cow sweat does not
allow comparison of these data. However, Klous et al. [17] stated in
their study with human sweat that eccrine and apocrine sweat glands
have different functions. They stated that sweat was produced in
the secretory cells in these glands, that the necessary components
of the sweat are reabsorbed as the produced sweat passes through
the excretory channels, and the remaining liquid was secreted to the
skin surface as sweat. When the values in TABLE I were examined, it
was understood that the fatty acid content in cattle sweat was lower
et al. [17].
The fatty acids with the greatest rate in the blood, according to
TABLE I, were oleic acid (21.57%), linoleic acid (20.09%), palmitic
acid (19.80%), and stearic acid (14.78%). Selionova et al. [18] stated
that the compounds with the highest fatty acid ratio in the blood
of cows were oleic acid 29.63%, palmitic acid 22.68%, stearic acid
20.33% and linoleic acid 18.34%. TABLE I shows that the proportions
of Selionova et al. [18].
alpha linolenic acid (C18: 3 n3) were two essential fatty acids required
for growth, structural health of the skin and reproduction in animals.
The researchers also stated that fatty acids are one of the most
important structural components that make up cell membranes, and
when fatty acids in cell membranes are included in phospholipids, they
Fatty acids of sweat and body fluids in cattle / Anitaş et al. _________________________________________________________________________
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activity of membrane–bound enzymes and signals occurring in the
cell [19, 20]. Studies have shown that these fatty acids are found in
the functioning of red blood cells, immune cells [21], atherosclerotic
plaques [22], heart tissue [23] and other cells in the body. When Table
1 is examined, linoleic acid, one of the important fatty acids, was
found to be 2.06% in sweat, 20.09% in blood and 14.57% in urine. It
was seen that linoleic acid is present in the blood at a rate of 0.27%,
and this fatty acid was not detected in sweat and urine. The absence
of linolenic acid in sweat and urine can be interpreted as an indicator
body metabolism.
Studies have reported that polyunsaturated fatty acids (PUFA)
positively affect the functioning of the reproductive system in
animals [24]. In addition, some researchers have stated that PUFA
supplementation to feeds increases the number of precursors for
the synthesis of steroid hormones (estradiol, progesterone) and
α) and decreases mortality [25, 26]. Therefore,
interpreting whether they were low for the body was important for
that the PUFA ratio was very high in urine and very low in sweat, that
the body may form a defense mechanism to prevent
Didara et al. [24] found SFA, MUFA, PUFA and n–6/n–3 values in
the plasma fatty acid analysis of animals as 32.82, 13.11, 51.73 and
16.55%, respectively. In the current study, SFA and MUFA rates were
determined to be higher than the rates determined by Klous et al. [17],
and PUFA and n–6/n–3 rates were found to be lower. Selionova et al.
[18] found the SFA rate in cow blood to be 46.07%, the MUFA rate to
be 31.56%, and the PUFA rate to be 25.44%. The SFA, MUFA and PUFA
ratios of the current study were compared with those of Selionova
et al. [18of [18].
Looking at the results in TABLE II, it was clear that there was a
weakly positive, statistically (P
0.384, P
Furthermore, a significantly positive statistically significant
P
urine. The relation between sweat and urine was determined to be
PP
et al. [27] found in their study on
human sweat fatty acids and blood fatty acids that there was a good
association between sweat and blood fatty acids.
28P<0.01)
connection between fatty acids as a consequence of the correlation
study they ran to establish the link between the fatty acid levels
of milk, blood, urine, and feces. They also noted that there was a
strong positive association between urine and blood, which was
P<0.01). TABLE II revealed a high level of
positive correlation between blood and urine, comparable to the
0.855, P
28] determined the dependent variable, the
blood–corrected R
2
rate, to be 0.835 and found a considerable level
that the independent variables milk, feces, and urine fatty acids
could satisfy 83% of the change in the dependent variable blood fatty
acids. They also observed that the dependent variable, the adjusted
R
2
in urine, was 0.613, indicating a moderate association and that
the independent variables milk, blood, and feces could account for
61% of the changes in urine fatty acids, which was the dependent
variable. When TABLE III was examined, it was seen that in the multiple
regression analysis, changes in blood were highly dependent on
changes in sweat and urine, and changes in urine, changes in sweat
and blood. Therefore, it could be said that blood can be predicted
be made.
calculated as 0.585 and the P
P
positive, the rate of change in fatty acids in the blood has a directly
proportional relationship with sweat. Fatty acid ratios detected in
the blood have an important place in estimating the fatty acid ratios
in sweat, which was a dependent variable.
The urine regression analysis was determined as -0.483 and
P
proportional relationship with sweat, and the urine was statistically
P<0.05) in the prediction of sweat and did not help a
lot with estimation.
PP
the rate of fatty acids in the blood was directly proportional to urine
P
P<0.05). In other words, it was seen that the fatty acids found in urine
blood fatty acids ratio, which was the dependent variable (TABLE IV).
If urine was used as the dependent variable, the association
between sweat and blood appeared to be negligible (P<0.05). The
P
indicating that the ratio of fatty acids in the blood was directly related
to the ratio of fatty acids in the urine. The association between urine
P<0.05) as
a consequence of the investigation. In other words, fatty acids in
urine considerably help the model predict the blood fatty acids ratio,
which was the dependent variable. The analytical results reported
and 28].
CONCLUSION
As a result of this study conducted to examine the relationships
between bovine sweat fatty acids and blood and urine, a smaller
number and proportion of fatty acids (14 in sweat, 25 in blood and 19 in
urine) were detected in sweat compared to blood and urine. As a result
found between sweat fatty acids and blood. It was also determined
that there was a statistically high level of relationship between
blood and urine. According to the results of the regression analysis,
this relationship, it was determined that the independent variables
sweat and urine could explain 81% of the variance in the dependent
variable blood. In addition, it was concluded that there is a moderate
correlation in the dependent variable, urine, and it can explain 79%
of the variations in the independent variable, blood and sweat. The
content of fatty acids in the blood can be estimated using correlation
and regression analysis, but it is advisable to conduct a statistical
study on a larger number of samples, and for this it is necessary to
establish a formula with a high degree of precision.
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Ethical statement
This study was approved by the Cukurova University Animal
Experiments Local Ethics Board.
Conict of interest
of interest.
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