https://doi.org/10.52973/rcfcv-e33290
Received: 12/07/2023 Accepted: 25/08/2023 Published: 25/09/2023
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Revista Científica, FCV-LUZ / Vol. XXXIII, rcfcv-e33290
ABSTRACT
This study was carried out to determine the effect of castration
procedure on hemogram and leukogram values and the levels of
oxidative stress factors in serum tissue in cats. Preoperative and
postoperative 10th day hemogram/leukogram values and total oxidant/
antioxidant capacities of 19 castrated cats were evaluated in the
study. When the hemogram and leukogram values were examined,
it was determined that there was a statistically signicant increase
in the amount of eosinophils (P<0.05). Although the increase in white
blood cells (WBC), neutrophil and lymphocyte levels and the decrease
in red blood cells (RBC) and hematocrit (HCT) levels were remarkable,
it was not found to be statistically signicant (P>0.05). When the
levels of oxidative stress factors in the serum tissues of the cats
were examined, it was determined that there was a decrease in the
total antioxidant status (TAS) and an increase in the total oxidant
status (TOS), but the difference in the results was not statistically
signicant (P>0.05). In conclusion, in this study, it was determined that
the castration procedure in cats did not cause a signicant difference
in hemogram and leukogram parameters and the level of oxidative
stress factors in serum tissue, except for the amount of eosinophils.
Key words: Castration; cat; eosinophil; hemogram; oxidative stress
RESUMEN
Este estudio se llevó a cabo para determinar el efecto del
procedimiento de castración sobre los valores de hemograma y
leucograma y los niveles de factores de estrés oxidativo en el tejido
sérico de gatos. En el estudio se evaluaron los valores de hemograma/
leucograma preoperatorios y posoperatorios al décimo día y las
capacidades oxidantes/antioxidantes totales 19 gatos castrados.
Cuando se examinaron los valores de hemograma y leucograma en
el estudio, se determinó que hubo un aumento estadísticamente
signicativo en la cantidad de eosinólos (P<0,05). Aunque el aumento
en los niveles de glóbulos blancos (WBC), neutrólos y linfocitos y
la disminución en los niveles de glóbulos rojos (RBC) y hematocrito
(HCT) fueron notables, no se encontró que fueran estadísticamente
signicativos (P>0,05). Cuando se examinaron los niveles de factores
de estrés oxidativo en los tejidos séricos de los gatos, se determinó
que hubo una disminución en el estado antioxidante total (TAS) y
un aumento en el estado oxidante total (TOS), pero la diferencia
en resultados no fue estadísticamente signicativa (P<0,05). En
conclusión, en este estudio se determinó que el procedimiento de
castración en gatos no provocó una diferencia signicativa en los
parámetros del hemograma, leucograma y el nivel de factores de
estrés oxidativo en el tejido sérico, excepto la cantidad de eosinólos.
Palabras clave: Castración; eosinólos; estrés oxidativo; gato;
hemograma
Investigation of changes in Hematological parameters and levels of
Oxidative stress factors in castrated Cats
Investigación de cambios en parámetros hematológicos y niveles
de factores de estrés oxidativo en gatos castrados
Eren Polat
1
* ,Tutku Can Acisu
2
, Mustafa SÖnmez
2
, Emine Ünsaldi
1
, Önder Otlu
3
, Murat Tanrisever
1
, Aslıhan Çakır
2
, Yasin Baykalir
4
1
Firat University, Faculty of Veterinary Medicine, Department of Veterinary Surgery. Elazig, Turkey.
2
Firat University, Faculty of Veterinary Medicine, Department of Reproduction and Articial Insemination. Elazig, Turkey.
3
Turgut Ozal University, Faculty of Medicine, Department of Medical Biochemistry. Malatya, Turkey.
4
Balikesir University, Faculty of Veterinary Medicine, Department of Veterinary Biostatistics. Balikesir, Turkey.
*Corresponding author: erenpolat@rat.edu.tr
Oxidative Stress in castrated cats / Polat et al. ______________________________________________________________________________________
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INTRODUCTION
Castration is the process of eliminating the source of hormones
that create sex–related physical and behavioral characteristics in
order to prevent reproduction in male cats (Felis catus) and dogs
(Canis lupus familiaris). Surgical removal of testicles is the most widely
used castration technique in all mammalian species [1, 2]. In many
Countries (such as USA, England), castration processes are used
to prevent the uncontrolled population growth of cats and dogs.
However, in some Countries (such as Germany, Norway, Sweden),
castration is seen as an unethical practice [1, 3].
It has been stated by some international organizations (American
Veterinary Medical Association, American Animal Hospital Association,
British Small Animal Veterinary Association) that castration in
the prepubertal period is acceptable. In some studies [1, 4], it has
been stated that castrations performed at an early age in cats and
dogs have no or very few side effects. However, many veterinarians
oppose castration in the prepubertal period due to the orthopedic and
oncological risks that may occur in later ages [1, 5]. In addition, it also
causes problems such as the narrowing of the urethra, which prevents
the urinary catheter from passing, and the increase in obesity.
Today, there are discussions about the advantages and
disadvantages of the castration process. With the castration process,
the excessive increase in cat and dog populations are prevented, the
risk of formation of some genital system diseases (some prostate
pathologies, some testicular tumors) and the control of unwanted
behaviors (aggression, urinating in inappropriate places) are provided
[1, 2]. Again, some studies report that the life span of castrated cats
and dogs is prolonged [6, 7]. Castration has both advantages and
disadvantages. Some studies have found an increased incidence
of osteosarcoma and prostate tumors in castrated cats [1, 8, 9, 10].
Again, some studies report an increased risk of obesity [11, 12, 13],
some orthopedic problems [14, 15], and diabetes mellitus (especially
Burmese breed) [1, 16] in cats after castration.
It is known that the castration process has an effect on some
hematological parameters and the levels of oxidative stress factors
[2, 17, 18]. Aengwanich et al. [2] reported that there were signicant
changes in the neutrophil/lymphocyte ratio in dogs after castration
(surgical method). Surgical operations are one of the important stress
factors for living beings. The deterioration of the oxidant/antioxidant
balance in the body in favor of oxidants under the inuence of any
stress factor is called oxidative stress. Oxidative stress causes the
body's resistance to degenerative diseases to decrease. In addition
to causing cardiovascular system diseases and neurodegenerative
central nervous system diseases, it also causes oncological diseases by
damaging the Deoxyribonucleic acid (DNA) helix [2, 19, 20, 21, 22, 23, 24].
In this study, changes in some blood parameters and oxidative stress
factors in preoperative and postoperative periods were evaluated in
castrated cats. Thus, by investigating the effect of castration on the
formation of oxidative stress, it was aimed to evaluate the decrease
or increase in body resistance against some degenerative diseases in
the body. At the same time, it was determined whether the castration
process would cause a change in some blood parameters, and its
effect on the health status of cats was investigated.
MATERIALS AND METHODS
Ethical approval and study plan
This study was carried out in accordance with ethical principles
with the approval of Fırat University Animal Experiments Local Ethics
Committee (dated: 22.03.2021, numbered: 28617). The study was
carried out on 19 cats brought to Fırat University Animal Hospital
Surgery Clinic for castration. In the study, the changes in the levels
of some blood parameters and oxidative stress factors in the
preoperative and postoperative period of the cats were evaluated.
Collecting blood samples
For the study, 5 mL blood was collected from cats both in tubes with
10% Ethylenediaminetetraacetic acid (EDTA) and in tubes with vacuum
gel and clot activator on the preoperative and postoperative 10th day. The
cephalic vein in the forearm was used when blood samples were taken.
Anesthesia and castration procedure
Before the castration procedure, 2 mg·kg
-1
Xylazine hydrochloride
was administered intramuscularly to the cats. After 10 min, anesthesia
was achieved by administering Ketamine hydrochloride at a dose of
10 mg·kg
-1
intramuscularly [25].
After shaving and disinfection of the scrotum of the cats, the
operation area was limited to the operation cover. The testicles were
limited and the scrotum skin was tightened (FIG. 1A). An incision
long enough for the testicles to protrude was made into the skin of
the scrotum, parallel to the raphe scroti (FIG. 1B,1C). After the soft
tissues and tunica vaginalis were cut, the testis was taken out and
separated from all its connections (FIG. 1D). After the hemostatic
forceps were placed on the part of the testicular cord towards the
body, a ligature was applied by transxation method using absorbable
suture material (FIG. 1E,1F). The same procedure was performed
for the second testis. In the postoperative period, Povidin–iodine
solution was applied to the operation wound for 3 days (d) and
parenteral antibiotics (Sülcid, 0.25mg) were administered for 3 d.
For postoperative pain management, 0.2 mg·kg
-1
meloxicam was
administered subcutaneously to all cats.
The cats that made up the material of the study were from different
breeds, aged 12–36 months. The surgeons performing and assisting
the operations were the same in all operations. Each of the operations
lasted an average of 3–4 min. All cats included in this study were
castrated during the summer months.
Measurement of hematological parameters
Hemogram and leukogram values of the cats were done with whole
blood analyzer (Hasvet Mindray BC–5000 Vet branded, Turkey). In order
to determine the oxidative stress factors, blood samples in tubes with
vacuum gel and clot activator were centrifuged at 3000 G for 10 min.
Determination of total oxidant status (TOS) activity
TOS activity of serum tissue was studied using Total Oxidant Status
kit (Rel Assay Diagnostics). 250 µL of buffer solution was added to all
wells. 37 µL of standard sample was added to the standard well and
37 µL of sample was added to the sample well. The rst reading was
made at 530 nm. 12 µL of substrate solution was added to all wells and
incubated at room temperature (20 – 25°C) for 10 min with shaking.
AA BB CC
DD EE FF
FIGURE 1. Stages of the surgical castration procedure (A–F)
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At the end of the incubation, a second reading was made at 530 nm
and the results were calculated [26].
Determination of total antioxidant status (TAS) activity
Serum tissue TAS activity was studied using the Total Antioxidant
Status kit (Rel Assay Diagnostics). 250 µL of buffer solution was
added to all wells. 15 µL of standard sample was added to the
standard well and 15 µL of sample was added to the sample well.
The rst reading was taken at 660 nm. 37 µL of 2,2'–azino–bis(3–
ethylbenzothiazoline–6–sulfonic acid (ABTS) radical solution was
added to all wells and incubated at room temperature (20 – 25°C) for
10 min with shaking. At the end of the incubation, a second reading
was made at 660 nm and the results were calculated [26].
Determination of oxidative stress index
Oxidative Stress Index (OSI), which is an indicator of oxidative
stress load, was obtained by dividing the TOS value by the TAS value.
Arbitrary Unit (AU) was used as the unit of OSI.
()
(, )
(, )
·
·
OSIArbitraryUnit
TASmolTrolox eq L
TOSmolHO eq L
1
1
22
n
n
=
-
-
Statistical analysis
All data on the preoperative and postoperative 10th d were evaluated
in the IBM SPSS 22 package program. The Paired–Samples t–test was
used because the data were continuous and obtained as a result
of pre– and post–operative observations obtained from the same
individuals. Data are presented as mean ± standard error. Statistical
signicance was accepted when P≤0.05 [27].
RESULTS AND DISCUSSIONS
Hemogram and leucogram results
In the study, the results of hemogram and leukograms of 19 male
cats before the castration operation and on the 10th day after the
castration operation were evaluated. In the examinations, it was
determined that there was only a statistically signicant increase
in the amount of eosinophils. The changes in the hemogram and
leukogram results are presented in TABLE I and FIGS. 2, 3 and 4.
Neutrophils Lymphocyte Monocyte Eosinophil Basophil
0
10
20
30
40
50
44.51
46.31
0.85
8.24
0.15
46.67
45.49
1.00
6.81
0.06
(%)
Preoperative Postoperative (10th day)
Neutrophils Lymphocyte Monocyte Eosinophil Basophil
0
2,000
4,000
6,000
8,000
5,83×10
3
4,85×10
3
1,19×10
-1
8,39×10
-1
1,60×10
-2
4,31×10
3
4,29×10
3
8,50×10
-2
6,57×10
-1
5,00×10
-3
Preoperative
Postoperative (10th day)
P<0.05
WBCRBC HCTHGB MCVPLT WBCRBC HCTHGB MCVPLT
0
50,000
100,000
150,000
200,000
250,000
1,165×10
4
1,084×10
4
4,015×10
1
1,460×10
1
3,732×10
1
2,367×10
5
9,288×10
3
1,108×10
4
4,055×10
1
1,471×10
4
3,725×10
1
2,366×10
5
Preoperative
Postoperative (10th day)
FIGURE 2. Graph of percentage of white blood cells before castration and on
the 10th postoperative day
FIGURE 4. Graph of the amounts of some hemogram and leukogram parameters
before castration and on the tenth postoperative day
FIGURE 3. Graph of the amount of white blood cells before castration and on
the 10th postoperative day
Oxidative Stress in castrated cats / Polat et al. ______________________________________________________________________________________
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TABLE I
Some blood parameters before and after the operation in castrated male cats
Parameter Preoperative
Postoperative
(10th day)
P–value
Neutrophils (cells·μL
-1
) 4,311 ± 392
a
5,832 ± 1,590
a
0.334
Neutrophil (%) 46.671 ± 4.081
a
44.512 ± 4.139
a
0.509
Lymphocyte (cells·μL
-1
) 4,285 ± 522
a
4,851 ± 580
a
0.185
Lymphocyte (%) 45.488 ± 4.457
a
46.306 ± 4.009
a
0.798
Monocyte (cells·μL
-1
) 0.085 ± 0.015
a
0.119 ± 0.044
a
0.400
Monocyte (%) 1.000 ± 0.203
a
0.853 ± 0.124
a
0.430
Eosinophil (cells·μL
-1
) 0.657 ± 0.101
a
0.839 ± 0.120
b
0.015
Eosinophil (%) 6.806 ± 0.763
a
8.241 ± 1.108
a
0.078
Basophil (cells·μL
-1
) 0.005 ± 0.002
a
0.016 ± 0.004
a
0.066
Basophil (%) 0.060 ± 0.022
a
0.150 ± 0.058
a
0.204
WBC (cells·μL
-1
) 9,288 ± 594
a
11,651 ± 1,549
a
0.154
RBC (cells×10
3
·μL
-1
) 11,075 ± 589
a
10,838 ± 427
a
0.615
HCT (%) 40.553 ± 1.485
a
40.153 ± 1.430
a
0.809
HGB (g·dL
-1
) 14.706 ± 0.542
a
14.600 ± 0.419
a
0.850
MCV (fL) 37.253 ± 1.143
a
37.324 ± 0.961
a
0.921
PLT (platelet·μL
-1
) 236,590 ± 24,747
a
236,706 ± 28,732
a
0.996
WBC: White Blood Cell, RBC: Red Blood Cell, HCT: Hematocrit Value, HGB: Hemoglobin
Value, MCV: Mean Corpuscular Volume, PLT: Platelet Value.
a,b
: Different letters on
the same line mean statistical signicance
Total antioxidant–oxidant (TAS–TOS) results
In the study, TAS and TOS activity and OSI results of 19 male cats
were evaluated before the castration operation and on the 10th day
after the castration operation. According to the obtained results,
it was determined that there was a decrease in TAS activity and
an increase in TOS activity, but these results were not statistically
signicant. The changes in TAS and TOS activities and OSI ratio are
presented in TABLE II and FIG. 5.
The advantages and disadvantages of castration, which is mostly
done to prevent excessive increase in cat and dog populations and
some gender–related behaviors, are frequently discussed today [1].
There are studies reporting that it has advantages such as a decrease
in the incidence of some genital diseases and prolongation of life span
[1, 6, 7], as well as disadvantages such as obesity and an increase in
the incidence of some orthopedic diseases [11, 12, 13, 14, 15]. There
are studies reporting an increased incidence of osteosarcoma and
prostate tumors in castrated cats [1, 9]. It is known that oxidative
stress is one of the most important causes of cardiovascular system
diseases, neurodegenerative central nervous system diseases and
some oncological diseases [24]. The effect of castration on oxidative
stress is therefore very important. In this study, it was aimed to
establish the basis for new studies on the possibility of castration
to have an effect on the etiology of diseases that may occur in the
future by determining the preoperative and postoperative TAS, TOS
and OSI values of castrated cats.
Surgical operations are one of the most important causes of
stress for living things. Oxidative stress is the deterioration of the
oxidant/antioxidant balance in the body in favor of oxidants, due to
the effect of any stress factor. Oxidative stress causes the body to
TASTOS OSI
0
50
100
150
200
250
300
1,718×10
0
4,314×10
0
2,511×10
2
1,738×10
0
4,294×10
0
2,514×10
2
Preoperative
Postoperative (10th day)
FIGURE 5. Graph of TAS, TOS levels and OSI ratio before castration and on the
tenth postoperative day
______________________________________________________________________Revista Cientifica, FCV-LUZ / Vol. XXXIII, rcfcv-e33290
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decrease its resistance against degenerative diseases [22, 24]. The
hypothesis that the castration process is a surgical operation and
that the metabolic changes that occur after castration may have an
effect on oxidative stress formed the basis of this study.
Aengwanich et al. [2] reported that the total antioxidant capacity
of the dogs on the postoperative third day of the surgical castration
decreased signicantly compared to the postoperative fourteenthd
(P<0.05). At the end of the recovery period, they reported that the
total antioxidant capacity of the dogs was close to the preoperative
period. In the same study, they reported that there was no statistically
signicant change in plasma malondialdehyde (MDA) level before,
during and after castration (P>0.05). Mogheiseh et al. [28] investigated
the effect of Melatonin on oxidative stress level in dogs after
castration. In this study, they reported that the MDA level in dogs
increased statistically signicantly after castration.
In the study, it was reported that the MDA level in dogs treated
with Melatonin (Castration+Melatonin group) decreased signicantly
compared to the dogs in the control and castration groups. They
also reported that antioxidant levels such as catalase, glutathione
peroxidase and superoxide dismutase were statistically signicantly
increased in dogs in the Melatonin–treated group. In the same study,
it was reported that the MDA levels of the dogs in the castration group
increased and their antioxidant levels decreased when compared
to the control group. Mahallingam et al. [17] reported that although
SOD and CAT levels increased postoperatively in dogs neutered by
laparoscopic vasectomy and prescrotal surgical castration, they
decreased to the preoperative level on the fth postoperative day.
In the same study, it was reported that no signicant difference was
found between the CAT and SOD levels of dogs in the laparoscopic
vasectomy group and prescrotal surgical castration groups.
Jana and Samanta [18], evaluated oxidative stress parameters
in male cats neutered by intratesticular administration of different
concentrations of Calcium chloride solutions (5, 10 and 20%). In this
study, it was reported that the MDA level in the testicular content
increased and the antioxidant level decreased in proportion to the
increase in calcium chloride concentration. In the same study, when
the 10% Calcium chloride solution group was compared with the 20%
Calcium chloride solution group, no signicant difference was found
between MDA levels and antioxidant levels (P>0.05). In this study, no
signicant difference was found between pre– and post–operative
TAS, TOS activities and OSI rates of castrated cats. However, it was
determined that there was a decrease in TAS activity and an increase
in TOS activity after castration. In this study, although there was no
statistically signicant difference, the decrease in TAS activity and the
increase in TOS activity in the postoperative period in castrated cats
were associated with the stress occurring during and after the operation.
There are studies reporting signicant changes in some hemogram
and leukogram values after castration. Fazio et al. [29] reported that
they found a decrease in parameters such as RBC, HGB, MCV, and
an increase in WBC and PLT levels in cats and dogs that they had
ovariohysterectomy. Jana and Samanta [18] reported that they did
not detect a signicant change in PCV level in the cats they castrated
by applying different concentrations of calcium chloride solutions
intratesticularly.
Aengwanich et al. [2] reported that the percentage of neutrophils
and lymphocytes increased signicantly on the tenth postoperative
day in castrated dogs. It has been reported that the neutrophil
percentage of the dogs increased signicantly after the recovery
process was completed, and the lymphocyte percentage decreased
signicantly. Türkoglu [30] reported in his study that castration did
not make a signicant difference in the total white blood cell count
of lambs. However, it has been reported that the total white blood
cell count is higher than the control group. In the same study, when
the castrated lambs were compared with the control group, it was
reported that the percentage of lymphocytes decreased and the
percentage of neutrophils increased.
Türkoğlu [30] reported that the most important effect of castration
in lambs (Ovis aries) was the increase in the percentage of eosinophils.
In this study, although the increase in WBC level of castrated cats was
not statistically signicant, the increase in the amount of eosinophils
was statistically signicant (P<0.05). However, although it is not
statistically signicant, the decrease in the percentage of neutrophils
(although there is an increase in the amount of neutrophils) and the
increase in the percentage of lymphocytes after the castration
procedure is remarkable. Although this study is similar to some
studies [30] due to the increase in the amount of eosinophils, it differs
from some studies [2, 30] due to the variability in the percentage of
neutrophils and lymphocytes. There are studies reporting a decrease
in adrenal cortex functions in castrated animals and a corresponding
decrease in the amount of glucocorticoids. In this study, the increase
in the amount of eosinophils in castrated cats may be associated
with the decrease in adrenal cortex activity.
TABLE II
Plasma total antioxidant and oxidant activity and OSI ratio in
castrated male cats at the pre– and postoperative 10th day
Parameter Preoperative
Postoperative
(10th day)
P–value
TAS 1.738 ± 0.013
a
1.718 ± 0.089
a
0.656
TOS 4.294 ± 0.029
a
4.314 ± 0.052
a
0.710
OSI 251.390 ± 0.560
a
251.061 ± 0.112
a
0.583
TAS: Total Antioxidant Status, TOS: Total Oxidant Status, OSI: Oxidative Stress Index.
a
: Different letters on the same line mean statistical signicance.
Oxidative Stress in castrated cats / Polat et al. ______________________________________________________________________________________
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CONCLUSION
Although there are many studies reporting that the castration
process has positive and negative results, when the hemogram/
leukogram results and the levels of oxidative stress factors were
evaluated, no signicant difference could be detected except for
the amount of eosinophils. The signicant increase in the amount of
eosinophils after castration was associated with a possible decrease
in adrenal cortex activity. It is known to cause a decrease in adrenal
cortex functions in neutered animals. With this study, the importance
of conducting more specic studies and detailing similar studies
in order to evaluate the effect of castration on metabolism and the
etiology of diseases has been revealed.
Financial support
This research received no grant from any funding agency/sector.
Ethical statement
This study was approved by the Firat University Animal Experiments
Local Ethics Committee (dated: 22.03.2021, numbered: 28617).
Conict of interest
The authors declared that there is no conict of interest.
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