https://doi.org/10.52973/rcfcv-e34325
Received: 25/09/2023 Accepted: 04/10/2023 Published: 06/02/2024
1 of 7
Revista Científica, FCV-LUZ / Vol. XXXIV, rcfcv-e34325
ABSTRACT
In order to determine the presence of Helicobacter in shelter Dogs
in Yozgat Province of
Türkiye
by fecal antigen test and ELISA and to
treat the infection during the initial period. As the material of the
study, faecal antigen test, faecal samples for ELISA, blood samples
for haematological and biochemical analyses were collected from 82
dogs in Sorgun animal shelter where food and water bowls were kept
in compartments. Infected and control groups were formed according
to the results of the fecal antigen test. As a result of fecal antigen test,
6 of 82 animals were antigen positive and 76 were antigen negative.
According to ELISA analysis; 53 dogs were antibody positive and
between the groups in terms of age and gender in both tests (P>0.05).
Dogs in the infected group were treated according to a protocol
known as triple therapy, which is used in Helicobacter infections.
WBC, NEU and MON counts, RDW and % NEU values of the infected
group were higher in the hematological examination, while the %
biochemical examination, the concentrations TNF–α (21.17 pg·mL
-1
vs.
48.21 pg·mL
-1
), IL–1β (73.41 pg·mL
-1
vs. 37.60 pg·mL
-1
) (P<0.01) and CRP
(644.0 mg·dL
-1
vs. 234.01 mg·dL
-1
) (P<0.001) were found to be higher
than those of the control group. As a result, it was determined in the
study that the presence of Helicobacter was intense in dogs that were
in one–to–one contact with each other, including the feeding of dogs in
shelter conditions. With the methods used in the study, it is predicted
that the presence of Helicobacter can be detected and treated in the
early stages, thus preventing transmission of Helicobacter in dogs.
Key words: Acute phase response; dog; ELISA; Helicobacter;
faecal antigen test
RESUMEN
Con el objeto de determinar la presencia de Helicobacter en perros de
refugio de la provincia turca de Yozgat mediante la prueba del antígeno
fecal y ELISA y tratar la infección durante el periodo inicial. Como
material del estudio, se recogieron pruebas de antígenos fecales,
muestras fecales para ELISA, muestras de sangre para análisis
hematológicos y bioquímicos de 82 perros en el refugio de animales
de Sorgun, donde los cuencos de comida y agua se guardaban en
compartimentos. Se formaron grupos infectados y de control según
los resultados de la prueba del antígeno fecal. Como resultado de
la prueba del antígeno fecal, 6 de 82 animales fueron positivos al
antígeno y 76 negativos. Según el análisis ELISA; 53 perros fueron
positivos a anticuerpos y 29 perros fueron negativos a anticuerpos. No
y sexo en ambas pruebas (P>0,05). Los perros del grupo infectado
fueron tratados según un protocolo conocido como terapia triple, que
se utiliza en las infecciones por Helicobacter. Los recuentos de WBC,
NEU y MON, los valores de RDW y % NEU del grupo infectado fueron
superiores en el examen hematológico, mientras que los valores de
En el examen bioquímico, se observó que las concentraciones de
TNF–α (21,17 pg·mL
-1
frente a 48,21 pg·mL
-1
), IL–1β (73,41 pg·mL
-1
frente
a 37,60 pg·mL
-1
) (P<0,01) y PCR (644,0 mg·dL
-1
-1
)
(P<0,001) eran superiores a las del grupo de control. Como resultado,
se determinó en el estudio que la presencia de Helicobacter era
intensa en perros que estaban en contacto uno a uno entre sí, incluida
la alimentación de perros en condiciones de refugio. Con los métodos
utilizados en el estudio, se prevé que la presencia de Helicobacter
pueda detectarse y tratarse en las primeras fases, evitando así la
transmisión de Helicobacter en perros.
Palabras clave: Respuesta de fase aguda; Prueba de antígeno en
heces; Perro; ELISA; Helicobacter
Treatment and Determination of the Presence of Helicobacter in Shelter
dogs by Faecal Antigen Testing and Enzyme–Linked Immunosorbent Assay
Tratamiento y determinación de la presencia de Helicobacter en perros de refugio
mediante pruebas de antígenos en heces y ensayo inmunoenzimático
Güvenç Gökalp
1
* , Akın Kırbaş
2
, Emre Sayar
1
, Emre Tüfekçi
3
, Nevzat Emre Aslan
4
1
Yozgat Bozok University, Faculty of Veterinary Medicine, Department of Internal Medicine. Yozgat, Türkiye.
2
Atatürk University, Faculty of Veterinary Medicine, Department of Internal Medicine, Erzurum, Türkiye.
3
Erciyes University, Faculty of Veterinary Medicine, Department of Internal Medicine. Kayseri, Türkiye.
4
Yozgat Bozok University, Faculty of Veterinary Medicine, Department of Surgery. Yozgat, Türkiye.
*Corresponding Author: guvenc.gokalp@yobu.edu.tr
Detection of Helicobacter in Shelter dogs / Gökalp et al. ____________________________________________________________________________
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INTRODUCTION
Helicobacter is a genus of gram–negative, microaerophilic, spiral
bacteria containing more than 40 species. Non–Helicobacter pylori
helicobacteria (NHPHs) such as Helicobacter heilmannii sensu stricto
(s.s.), Helicobacter bizzozeronii and Helicobacter felis have been
frequently detected in the stomach of dogs (Canis lupus familiaris)
and cats (Felis catus) [1]. The main types of NHPH found in dogs are H.
heilmannii s.s., H. bizzozeronii, H. salomonis, H. felis and H. canis [2].
Most of the NHPHS may also cause illness in humans. Attention should
be paid to the zoonotic importance of NHPHS due to the increased
risk of occurrence in people who are in close and intense contact with
animals [3]. Since Helicobacter pathogens can be excreted through
defecation, expectorating, and vomiting, they present faecal–oral,
oral–oral and gastro–oral transmission routes [4]. Dogs are the natural
hosts of NHPH and harbour these bacteria in their gastric mucosa,
are possible sources of transmission of these bacteria to humans [2].
Invasive and non–invasive methods are used in the diagnosis of
Helicobacter infection. Among invasive methods with endoscopic
biopsy, histological examination, culture, rapid urease test (RUT),
and polymerase chain reaction (PCR) are shown; while among non–
invasive methods, faecal antigen test, urea breath test (UBT), and
serological tests are named [5]. Although gastroscopy, which is within
than other methods, it should be noted that many veterinarians do not
have the opportunity and ability to perform gastroscopy, in addition to
the risks of gastroscopy and associated anaesthesia. For this reason,
non–invasive methods such as serology, culture or PCR, which can
be made from blood, saliva and faecal samples, are preferred [6]. In
addition, it was reported in a study that PCR analyses alone would not
state in the diagnosis of Helicobacter, and this should be supported
by histopathological analyses [7].
The faecal antigen test is highly accurate and a rapid test that
and after treatment. The simplicity of the present method does not
require prior preparation of the patient, but it is recommended not
to apply a proton pump inhibitor (PPI), a 4–week antibiotic treatment,
and bismuth compounds two weeks before the test is performed [8].
Erythrocyte synthesis is regulated by many factors such as
erythropoietin, B
12
, folic acid, and vitamin C. Helicobacter infections
have been associated with vitamin B
12
causing the persistence of vitamin B
12
9]. In addition, it is
known that high levels of TNF–alpha and interleukin levels are present
in Helicobacter
cause structural changes in the gastric epithelial cells [10].
It has been reported that all Helicobacter species are highly sensitive to
and neomycin, and over 90% of dogs with Helicobacter–induced gastritis
are relieved by using a combination of metronidazole, amoxicillin and
famotidine. This treatment procedure, known as triple therapy, lasts at least
21 days, increasing the chance of eradication [11]. The aim of the study was
to determine the presence of Helicobacter in dogs by fecal antigen test and
ELISA methods and to provide early treatment, to have information about
α, IL–1β and C–reactive
protein concentrations and to measure the effects of the presence of the
agent on folic acid (FA) and cobalamin (COB) concentrations.
MATERIALS AND METHODS
Study population and experimental design
The population consisted of 82 dogs in the Temporary Animal
Shelter of the Municipality of Sorgun. Priority was given to those
sampled dogs which showed signs of vomiting and gastritis. Sampling
was done from 82 dogs in the form of scanning. The samples were
collected from different groups of breed, gender and age. Sampling
was done by screening from 82 dogs. Samples were collected from
different breeds, sex and age groups. Samples were composed of 47
dogs from the 0 > 2 age group, 35 dogs from the 2 > 4 age group, 45
dogs from the female animal group and 37 dogs from the male animal
group. According to faecal antigen test results, 2 (4,25%) dogs from
0 > 2 age group and 4 (11,4%) dogs from 2 > 4 age group were positive
for Helicobacter antigen. These dogs were formed the infected group
(n=6) and the antigen negative dogs (n=6) were formed the control
group. The study counted with the approval of the Erciyes University
Animal Experiments Local Ethics Committee (10.09.2020, 20/128).
Clinical examination
The clinical examination of the 82 dogs was performed. The animals
were examined for signs such as loss of appetite, vomiting, abdominal
pain, and increased urge to drink water with dehydration.
Blood and faecal collection
Clinical symptoms frequently encountered in infections originating
from Helicobacter were considered while sampling. 82 dogs with
of sampling were included in the study. For faecal antigen testing, faecal
samples were collected from the rectal region. Blood samples were
taken via vena jugularis into EDTA tubes (Vacusel, Konya, Türkiye) for
haematological analysis; into vacuum tubes (BD Vacutainer, Plymouth,
United Kingdom) for biochemical and serological analysis. The blood
samples obtained were centrifuged in a cooled centrifuge (Hettich
Universal 320R, Germany) at 4.000 g·5 min
-1
. After centrifugation, the
serum was stored at -80°C (ESCO, ESC–UUS–480A, Singapore) until
biochemical and serological analysis.
Treatment protocol
It was paid attention that no antimicrobial treatment was
administered to the dogs for the previous week and that the dogs
sampled were at least one year old. In the dogs to be sampled, those
showing symptoms of vomiting and gastritis were prioritized, and
scanning was performed on 82 dogs that formed the material of the
study. Faecal antigen tests were performed on the faecal samples.
According to the results of faecal antigen tests, positivity (n=6) and
negativity (n=6) were determined. Those found positive were accepted
as infected group and treatment was applied. The infected group
was treated according to the protocol known as triple therapy used
in Helicobacter infections. Amoxicillin–Clavulanate (Croxilex–BID 625
mg tablet, Turkey) 20 mg·kg
-1
PO once a day, Metronidazole (Flagyl
500 mg tablet, France) 15 mg·kg
-1
PO once a day, Ranitidine (Ranitab
150 mg tablet, Turkey) 4 mg·kg
-1
PO once a day. After treatment, stool
samples were collected with sterile test apparatus for 3 weeks and
stool antigen test was repeated.
_____________________________________________________________________________Revista Cientifica, FCV-LUZ / Vol. XXXIV, rcfcv-e34325
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Serological analyses
• Faecal antigen test
Faecal samples were taken with rectal swabs from 82 dogs. The
Helicobacter Antigen Test (Dia Pro Diagnostic Bioprobes, San Giovanni,
Italy), which enables the qualitative detection of Helicobacter antigen
in faecal on an immunochromatographic basis, was applied. The
plastic bottle was opened from the screwed part, and an amount of
faeces sample of approximately the size of lentils was introduced.
The bottle was then shaken to mix the extraction buffer with the
sample and waited for about 30 s. 4 drops of the mixture containing
faeces and buffer solution were dripped onto the test card. Results
were read within 5 min. In the presence of Helicobacter antigen in
the test card, double lines were observed in the positive samples,
while a single line was in the negative samples.
• Enzyme–linked immunosorbent assay (ELISA)
In the sera from dogs (n=82), animals positive for Helicobacter
antibody were detected using the Canine Helicobacter IgG Elisa kit
(Sunlong Biotech, Zhejiang, China) according to the instructions of
the manufacturer. Biotek ELX800 (Wermont, USA) device was used
for ELISA measurement. ELISA plates were read at 450 nm.
• Haematological parameters
Hematological parameters (Red blood cell, White blood cell, Platelet
indices) were determined with a blood count device (Mindray BC–2800,
China) in the blood samples of the infected and healthy groups.
Biochemical analyses
• Inammatory parameters
TNF–α, IL–1β and CRP concentrations in the serums were determined
according to the manufacturer's instructions, commercial ELISA test
kit (Sunlong Biotech, Zhejiang, China).
• Vitamins
Cobalamin (B
12
) and Folic acid (B
9
) concentrations were determined
in serums of the infected and control groups according to the
manufacturer's instructions using a commercial ELISA assay kit
(Sunlong Biotech, Zhejiang, China).
Statistical analysis
IBM SPSS Statistics 25,0 (IBM Corp. Armonk, New York, USA)
package program was used for the statistical analysis. The
suitability of the data to the normal distribution was evaluated by the
histogram, Q–Q plots, and the Shapiro–Wilk tests and was shown as
mean±standard deviation or median (min–max). The data presenting a
normal distribution were evaluated with the independent two–sample
T–test, and the data lacking a normal distribution were evaluated with
the Mann–Whitney U test. The relationship between the parameters of
the groups was investigated with the help of the Spearman Correlation
P<0.05.
RESULTS AND DISCUSSION
Serological ndings
• Faecal antigen test ndings
antigen tests were repeated for the presence of Helicobacter 3
weeks after the treatment was applied to the dogs in the infected
group. Negativity was detected in 5 of the infected dogs. According
the treatment, complete recovery was observed in 5 (85%) of the
infected dogs. Antigen positivity was detected in 5 of 45 female
dogs and in one of 37 male dogs. Antigen positivity was detected in
2 of 47 dogs in the 0 > 2 age group and in 4 of 35 dogs in the 2 > 4 age
group. When evaluated in terms of age and gender, it was determined
that antigen positivity rates in faecal antigen tests were higher in
the 2 > 4 age group compared to the 0 > 2 group; and in female dogs
compared to male dogs. However, these results were not statistically
P
TABLE I
Distribution of antigen positivity by age and gender
according to the Feacal antigen test (n=82)
Parameters Antigen positive (n=6) Antigen negative (n=76)
Age
0 > 2 (n) 2 (4.25%) 45 (95.75%)
2 > 4 (n) 4 (11.4%) 31 (88.6%)
Gender
Female (n=45) 5 (11.0%) 40 (89.0%)
Male (n=37) 1 (2,7%) 36 (97.3%)
• ELISA test ndings
According to the Helicobacter IgG results in the serum of the
sampled dogs, significant differences were determined in the
concentrations of Helicobacter IgG in control (n=6) and infected dogs
(n=6). According to the normal range of Helicobacter IgG (6.93–8.80),
results with a value below 6.93 mg·dL
-1
were considered seronegative,
and results with a value of 8.80 mg·dL
-1
and above were considered
seropositive. The mean IgG concentration was 5.93 mg·dL
-1
in the
control group and 10.83 mg·dL
-1
in the infected group. Seropositivity
was detected in 28 of 45 female dogs (62.5%) and 25 of 37 male dogs
(67.5%). 53 dogs were positive by ELISA. Antibody positivity was
detected in 30 (63.5%) of 47 dogs aged 0 > 2 years and 23 (65.5%)
of 35 dogs aged 2 > 4 years. When evaluated in terms of age and
gender, it was determined that the seropositivity rates in the ELISA
test were higher in the 0 > 2 age group compared to the 2 > 4 age
group and in male dogs compared to female dogs. However, these
P>0.05). The distribution of
seropositivity according to Helicobacter IgG ELISA test according to
age and gender is shown in TABLE II.
Detection of Helicobacter in Shelter dogs / Gökalp et al. ____________________________________________________________________________
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Haemotological parameters
P<0.001), MON and RDW–SD
(P
P
%, RBC, HGB, HCT, MCV, MCH, MCHC, RDW–CV, PLT, MPV, PDW and
PCT values (P>0.05).
Biochemical ndings
• Inammatory markers (TNF–α, IL–1β, and CRP)
Helicobacter IgG ELISA, vitamin B
12
and folic acid concentrations
of the control and infected groups are presented in TABLE II. The
concentrations of TNF–α (21.17 pg·mL
-1
vs. 48.21 pg·mL
-1
), IL–1β (73.41
pg·mL
-1
vs. 37.60 pg·mL
-1
) (P<0.01) and CRP (644.0 mg·dL
-1
vs. 234.01
mg·dL
-1
) (P<0.001) were found to be higher than in the control group.
• Vitamins B
12
and B
9
(folic acid)
Serum cobalamin (9.54 pg·dL
-1
, vs. 17.18 pg·dL
-1
) (P<0.05) and folic
acid (2004.40 ng·mL
-1
, vs. 870.85 ng·mL
-1
) (P<0.01) concentrations of
TABLE II
Distribution of seropositivity by age and gender
according to Helicobacter IgG ELISA test (n=82)
Parameters n=82 Seronegative Seropositive
Average value
(6,93–8,80) mg·dL
-1
Age
0 > 2 (n=47) 17 (36,5%) 30 (63,5%) 7,75
2 > 4 (n=35) 12 (34,5%) 23 (65,5%) 8,01
Gender
Female (n=45) 17 (37,5%) 28 (62,5%) 8,11
Male (n=37) 12 (32,5%) 25 (67,5%) 7,75
TABLE III
The levels of hematological parameters in infected and control dogs
Parameters Control (n=6) Infected (n=6) P
WBC (10
3
·µL
-1
)
12.08 ± 4.10 20.93 ± 2.36 0.001
NEU (10
3
·µL
-1
)
8.00 ± 2.93 16.92 ± 2.30 <0.001
LYM (10
3
·µL
-1
)
2.53 ± 0.91 1.60 ± 1.01 0.130
MON (10
3
·µL
-1
)
1.08 ± 0.47 1.76 ± 0.53 <0.05
EOS (10
3
·L
-1
)
0.48 ± 0.24 0.64 ± 0.63 0.570
NEU %
66.20 ± 6.03 80.93 ± 7.23 <0.01
LYM %
21.00 ± 4.31 7.63 ± 4.74 <0.001
MON %
8.73 ± 1.24 8.38 ± 2.15 0.737
EOS %
4.07 ± 2.31 3.63 ± 2.77 0.775
RBC (10
6
·µL
-1
)
6.06 ± 0.54 5.66 ± 0.82 0.350
HGB (g·dL
-1
)
14.23 ± 0.83 13.75 ± 1.85 0.577
HCT %
41.88 ± 3.74 39.13 ± 6.83 0.407
MCV (fL)
70.00 ± 1.99 69.07 ± 6.59 0.747
MCH (pg)
23.83 ± 1.36 24.38 ± 1.75 0.557
MCHC (g·L
-1
)
340.67 ± 17.58 356.67 ± 48.81 0.477
RDW–CV %
14.08 ± 0.74 16.03 ± 2.22 0.069
RDW–SD (fL)
37.67 ± 2.29 42.63 ± 3.36 <0.05
PLT (10
3
·µL
-1
)
260.83 ± 55.48 343.83 ± 102.14 0.111
MPV (fL)
10.03 ± 1.08 9.38 ± 1.44 0.397
PDW (fL)
15.47 ± 0.37 15.46 ± 0.53 1.00
PCT (mL·L
-1
)
2.60 ± 0.53 3.22 ± 0.96 0.193
Data were expressed as mean ± standard deviation
TABLE IV
The concentrations of inammatory markers and
vitamins in infected and control dogs
Parameters Control (n=6) Infected (n=6) P
TNF–α (pg·mL
-1
) 21.17 48.21 <0.01
IL–1β (pg·mL
-1
) 37.60 ± 8.40 73.41 ± 20.29 <0.01
CRP (mg·dL
-1
) 234.01 ± 74.99 644.04 ± 203.94 <0.001
Helicobacter IgG (mg·dL
-1
) 5.93 10.83 <0.01
B
12
(pg·dL
-1
) 17.18 ± 5.14 9.54 ± 2.21 <0.05
Folic acid (ng·mL
-1
) 2004.40 ± 355.92 870.85 ± 324.47 <0.001
Data were expressed as mean ± standard deviation
Correlation ndings
Spearman correlation analysis was applied to TNF–α, IL–1β,
Helicobacter IgG, CRP, B
12
and folic acid parameters in control and
were determined between Helicobacter IgG and TNF–α, IL–1β and CRP.
found between Helicobacter IgG and B
12
and folic acid. Statistically
TNF–α and B
12
positive and very high correlation between TNF–α and CRP.
observed between CRP and B
12
positive and high correlations were found, as well, between CRP and
IL–1β and between B
12
and folic acid. Correlation results between
parameters in control and infected dogs are given in TABLE V.
Helicobacter agents in shelter dogs by faecal antigen test and the
ELISA method, along with the application of the treatment in the
early period of the infection, were accomplished. There are many
methods for the diagnosis of the presence of Helicobacter of gastric
origin. However, each of these methods has both advantages and
disadvantages. The use of one or more tests, the accessibility of
the tests, the equipment needed in the laboratories, and the clinical
conditions of the sick animals to be sampled are determinative for the
methods chosen in the diagnosis [8]. Most immunoassay methods rely
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on the detection of Helicobacter antibodies in the serum. In the case
of active infection, IgM levels can be detected, followed by an increase
in IgG and IgA levels, which remain consistently high until the infection
clears [5]. In the presented study, faecal antigen test and Helicobacter
IgG ELISA test in serum, which are among the non–invasive methods,
were used to diagnose the presence of Helicobacter.
Hong et al. [12] applied the Helicobacter faecal antigen test [HpSA]
and faecal PCR tests to identify Helicobacter spp. from faecal samples
taken from eight dogs showing signs of gastritis and detected
positivity in only two dogs with the faecal antigen test. Faten et al.
[13] applied the faecal antigen test in 25 dogs without clinical signs
of gastritis; likewise, Haggag et al. [14] in 50 dogs, and no positivity
was detected in both studies. In the present study, the faecal antigen
test was applied to a total of 82 dogs showing clinical symptoms, and
seropositivity was found in 6 of them. Samples were collected from
shelter dogs showing similar clinical symptoms at the same time
period and under the same conditions. Fecal antigen test and ELISA
were performed on a total of 82 dogs. As a result of faecal antigen
test, positivity was found in 6 of them (7.5%). Seropositivity was
found in a large number of samples by ELISA test. Seropositivity was
found in 30 out of 47 (63.5%) in the 0 > 2 age group and in 23 out of 35
(65.5%) in the 2 > 4 age group by ELISA method. It was determined
that ELISA test was able to detect positivity at higher levels compared
to faecal antigen test. However, these results were not statistically
P>0.05). Waheeb et al. [5] found 20,5% seropositivity
in 18 of the samples taken from 88 dogs by Helicobacter IgG ELISA
method. Moussa et al. [15] applied Helicobacter IgG ELISA to the blood
serum of 30 dogs owned by people with gastritis symptoms and found
seropositivity in all of the samples. In this study, a seropositivity
rate of 64.6% was detected in 53 of the samples taken from 82 dogs
living in shelter conditions.The fact that the rate of Helicobacter
IgG seropositivity in dogs owned by people with gastritis symptoms
is higher than that in dogs living in shelter conditions reveals the
zoonotic feature of the Helicobacter agent. It was determined that
faecal antigen positivity was more common in the 0 > 2 age group
compared to the 2 > 4 age group and in male dogs compared to female
(P>0.05).
In studies related to Helicobacter spp. in dogs, some haematological
changes have been reported [16,17,18,19,20,21,22]. It has been
reported that lymphocytosis due to lymphofollicular hyperpyloplasia
occurs in dogs with Helicobacter infection [16]. Since Helicobacter
agents can cause lymphoplastic, eosinophilic or chronic gastritis,
neutrophilic leukocytosis is encountered in the blood in such cases
[17]. Red blood cell (RBC) synthesis is regulated by many factors, such
as erythropoietin, iron, vitamin B
12
, folic acid, and vitamin C. Decreases
in erythropoietin, iron, vitamin B
12
, folic acid levels and, therefore,
a decrease in RBC, can be observed in Helicobacter infections [9].
Meral et al. [18] reported a decrease in erythrocyte and hemoglobin
counts, an increase in platelet counts and a decrease in hematocrit
ratios in infected dogs. Testault et al. [19] reported an increase in
leukocyte, neutrophil and granulocyte counts and a decrease in
erythrocyte and hemoblobin counts in infected dogs. Baan et al.
[20] determined a decrease in hematocrit rates in infected dogs
compared to healthy dogs in their study. Meral et al. [17] determined
an increase in leukocyte and lymphocyte counts and a decrease in
hemoglobin count in infected dogs.
Patel et al. [21
neutrophil count in infected dogs. Papagiannakis et al. [22] reported
a decrease in platelet concentrations in Helicobacter–induced
gastrointestinal diseases. There are similarities between the results
of these studies and the results obtained in the present study.
Acute and chronic inflammation caused by Helicobacter can
necrosis/apoptosis. IL–1 and TNF–α
cytokines in this process [23]. Helicobacter colonization causes
by neutrophils and mononuclear cells and is characterized by
increases in various cytokines such as TNF–α and IL–1β [24]. TNF–α
widely and constitutively expressed by activated immune cells
25]. The increase in TNF–α concentration
in Helicobacter–infected chronic atrophic gastritis (CAG) is associated
(TNFRs) have been shown to be actively produced in Helicobacter–
infected gastric mucosa. High levels of IL–1β secretion by infected
cells suggesting cellular damage, whatever the cause, may act
synergistically with Helicobacter infection to exacerbate the damage
to the gastric epithelium [22]. Park et al. [26] stated that the TNF–α
level in Helicobacter infection increased 3 times in the infected group
compared to the control group, and the IL–1β level increased 3.5 times
in the infected group compared to the control group. Seim–Wikse
et al. [27] reported that CRP concentrations in infected dogs were
higher than those in healthy dogs.
Lin et al. [28α and IL–1β in
infected dogs and reported that these excessive increases in TNF–α
and IL–1β concentrations may be effective in the inflammatory
response to Helicobacter infections. Thus, they demonstrated that
the pathogenesis of Helicobacter infection is associated with high
levels of cytokinins such as TNF–α and IL–1β. In the present study,
serum TNF–α, IL–1β
in the infected group (P<0.01).
In the stomach, free cobalamin binds to haptocorrin, which in dogs
is mostly produced by parietal cells in the gastric mucosa. Haptocorrin
is digested by pancreatic proteases in the duodenum, and then free
cobalamin is bound to the intrinsic factor. Although intrinsic factor is
mainly produced by the exocrine pancreas in dogs, it is also produced
in the parietal cells of the stomach. Parietal cells that produce both
haptocorrin and intrinsic factor are typically lost in gastropathies. This
condition leads to hypocobalaminemia in dogs [29]. In the changes
in the microbiota of the gastrointestinal tract, there is a decrease
in the absorption of folic acid from the proximal small intestine and
of cobalamin by the ileum, depending on the density of the bacterial
TABLE V
Spearman correlation results between parameters
in control and infected dogs
Parameters IgG TNF–α IL–1β B
12
CRP Folic acid
IgG
1.000 0.664
*
0.727
**
-0.594
*
0.678
*
-0.832
**
TNF–α
1.000 0.490 -0.741
**
0.811
**
-0.790
**
IL–1β
1.000 -0.350 0.643
*
-0.503
B
12
1.000 -0.685
*
0.650
*
CRP
1.000 -0.741
**
Folic acid
1.000
Detection of Helicobacter in Shelter dogs / Gökalp et al. ____________________________________________________________________________
6 of 7
presence. However, a diet high in folate may result in increased serum
folate concentrations regardless of disease. Unrelated to the source
of infection, pathological conditions in the ileum can also damage
cobalamin receptors and thus lead to cobalamin malabsorption [30].
It was stated by Andres et al. [31] that this condition is caused by
food cobalamin malabsorption syndrome, and stomach diseases
associated with Helicobacter infection are among the most important
causes of this syndrome. Yanik et al. [32] state that Helicobacter
infection, which can become chronic, causes vitamin B
12
and
folate absorption disorder, leading to an increase in circulating
homocysteine levels. Accordingly, increased homocysteine levels
due to Helicobacter infection are thought to play a role in endothelial
et al. [27
Helicobacter
et al.
[33] reported that Helicobacter–derived infections could predispose
patients to pernicious anaemia by inducing autoantibodies against
antigens in the gastric mucosa. It is predicted that this effect may be
due to Helicobacter–induced gastric atrophy and may be due to the
inhibition of B
12
, an important vitamin that needs to be absorbed by the
body, due to disruptions in the intestinal transit of the foods ingested.
Pernicious anaemia, a stage of chronic vitamin B
12
been associated with Helicobacter infection. Helicobacter infection
can cause malabsorption of different micronutrients, including
vitamin B
12
. In patients with vitamin B
12
Helicobacter
eradication is followed by increased serum vitamin B
12
levels and
decreased serum homocysteine levels [34]. Mwafy et al. [35]
cobalamin (B
12
) was found to be 378.2 pg·mL
-1
in the control group
and 262.5 pg·mL
-1
study, serum cobalamin (B
12
) levels were found to be lower in the
infected group than in the control group (P<0.05) (TABLE III). Augustin
et al. [36] think that folate concentrations above the reference range
may be caused by serious bacterial growth in the small intestine
levels, and folate concentrations lower than the reference range
may be caused by infections originating from Helicobacter. In the
present study, it was determined that folic acid concentrations of
group (P<0.001) (TABLE III).
CONCLUSIONS
In conclusion, this study demonstrates that the parameters used
in this study are effective markers in the prognosis and follow–up of
the treatment process of Helicobacter–borne infection in dogs, as
well as the usefulness of fecal antigen testing and ELISA methods in
the early diagnosis of the presence of the causative agent.
ACKNOWLEDGEMENTS
T/20–432].
Conict of interest
interest.
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