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    A Molecular survey of Hepatozoon canis in dogs in the Siirt province of Turkey
    (Veterinarni A Farmaceuticka Univerzita Brno, 2022) Celik, Burcak Aslan; Celik, Ozgur Yasar; Ayan, Adnan; Yilmaz, Ali Bilgin; Kilinc, Ozlem Orunc; Ayan, Ozge Oktay
    This study aimed to determine Hepatozoon canis prevalence in dogs in the Siirt province of Turkey by the molecular method. The animal material of the study consisted of a total of 75 dogs that appeared clinically healthy. Two ml of blood sample were taken from the vena cephalica antebrachii. Then, DNA extraction was performed. Polymerase chain reaction (PCR) was performed to amplify the 666 bp 18S rRNA gene region of Hepatozoon canis. Two positive PCR products were purified and sequenced. As a result of Nested-PCR, H. canis specific bands in 666 bp size were obtained in 7 (9.33%) out of 75 dogs. The result of sequence analysis, the nucleotide sequence was registered in the NCBI GenBank database with accession numbers OL467380.1-OL467538.1. Hepatozoon canis registered in GenBank of sequence OL467380.1 was found to be similar with other H. canis strains of registration numbers MW684292.1 with 99.69% and MH615006.1-MK091085.1-MF797806.1 with 99.53% rates; and the sequence with registration number OL467538.1 was found to be similar to the series MW684291.1 with 99.09% and MH615006.1-MK091085.1-KX 818220.1 with 99.08% rates by BLAST analysis. Hepatozoon canis prevalence of dogs in the Siirt province was determined as a result of this study. It is of great importance to take preventive measures, especially to fight ticks with appropriate acaricides, since there is no vaccine to prevent the disease.
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    Cryptosporidium spp. in Dogs - Prevalence and Genotype Distribution
    (Univ Fed Rio Grande Do Sul, 2023) Celik, Ozgur Yasar; Kochan, Akin; Celik, Burcak Aslan; Ayan, Adnan; Akyildiz, Gurkan; Kilinc, Ozlem Orunc; Ercan, Kerem
    Background: Cryptosporidium spp. is a zoonotic protozoan parasite that affects the gastrointestinal tract of humans and animals. The disease can cause acute and chronic diarrhoea and even death in both humans and animals. In this study, it was aimed to determine the prevalence and genotype distribution of Cryptosporidiosis in shelter dogs in Diyarbakir province located in the Southeastern Anatolia Region of Turkey. Materials, Methods & Results: The animal material of the study consisted of 100 dogs of different breeds and sexes. Faecal samples were collected from the rectum with disposable latex gloves and placed in individual sample containers. All of the samples were examined for Cryptosporidium spp. by Kinyoun Acid Fast and Nested PCR methods. In the Kinyoun Acid Fast staining method, firstly, smear preparations were prepared from fresh faecal samples, fixed in pure methanol for 1 min and allowed to dry. The slides were kept in Kinyoun Carbol-Fuxin for 5 min, dipped in 50% ethyl alcohol, shaken, washed in tap water, kept in 1% sulphuric acid for 2 min and washed in tap water. The slides were kept in methylene blue for 1 min, washed in tap water and allowed to dry. After drying, immersion oil was dripped and examined under a microscope at 100 magnification. DNA extraction was performed from all samples using GeneMATRIX Stool DNA Purification Kit according to the manufacturer's protocol. After Nested PCR analysis was performed. In the PCR step, primers 5'-TTCTAGAGCTAATACATGCG-3' and 5'- CCCATTTCCTTCCTTCGAAACAGGA-3' were used to amplify the 1325 bp gene region. In the nested PCR step, primers 5'- GGAAGGGTTGTATTTATTTATTAGATAAAG-3' and 5'-AAGGAG-TAAGGAACAACCTCCA-3' were used to amplify the 826-864 bp gene region. As a result of both methods, a prevalence of 3% was determined. The infection rate was higher in males (3.57%) than females (2.27%) and in younger than 1 year (5.56%) than in older than 1 year (1.56%). The DNA sequences obtained from the sequence analysis of 3 positive PCR samples were analysed in BioEdit software. A phylogenetic tree was constructed with the data set created by using the 18s rRNA gene sequences obtained from the NCBI genbank database and the DNA sequences obtained as a result of the study, and it was shown which Cryptosporidium species the study samples were related to. Today, many Cryptosporidium species have been identified and most of these species have host adaptation. Although C. canis is the most common species in dogs, C. muris, C. meleagridis, and C. parvum have also been detected. Among these species, C. parvum is recognized as a zoonotic species infecting a wide range of mammals. In this study, DNA sequencing of nested PCR positive samples revealed that 3 samples were zoonotic C. parvum. Discussion: This suggests that dogs may be a reservoir for zoonotic transmission of Cryptosporidium. Consequently, it is recommended that people should be informed about the potential for transmission of this protozoan to humans and animals and that control programmes should be implemented, including the prevention of free entry of stray dogs into public places and homes.
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    First Report of Zoonotic Cryptosporidium parvum Subtype IIaA15G2R1 in Dogs in Türkiye
    (Pakistan Veterinary Journal, 2024) Ayan, Adnan; Celik, Burcak Aslan; Celik, Ozgur Yasar; Akyildiz, Gurkan; Kilinc, Ozlem Orunc; Ayan, Ozge Oktay; Uslu, Ugur
    Cryptosporidium (C.) is an opportunistic protozoan causing gastrointestinal illness in both humans and animals, leading to acute or chronic diarrhea and even death. The study aimed to investigate the prevalence and subtyping of Cryptosporidium spp. in shelter dogs in Van province, Türkiye. For microscopic identification of this parasite, a total of 300 fecal samples were collected and stained with Kinyoun's acid-fast method. For molecular analysis, the positive samples were subjected to DNA extraction and SSU rRNA gene of Cryptosporidium spp. was amplified using nested PCR. The microscopic examination revealed a 4.67% prevalence of Cryptosporidium spp. Sequence analysis indicated all samples were positive to C. parvum. In addition, GP60 gene was also amplified and C. parvum subtypes IIaA15G2R1 was confirmed by analyzing the obtained sequences. All the sequences of SSU rRNA and GP60 were deposited in GenBank. To our knowledge, Cryptosporidium parvum subtypes IIaA15G2R1 have been reported first time in dogs in Türkiye. It is recommended to implement control strategies by awareness campaign, preventing stray dogs from freely entering public areas, and proper disposal of dog feces.
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    Investigation of Toxoplasma gondii and Neospora caninum in different tissues of aborted foetuses of sheep in Van Province, Turkiye: Analysis by nested PCR, histopathological and immunohistochemical methods
    (Veterinarni A Farmaceuticka Univerzita Brno, 2023) Kilinc, Ozlem Orunc; Ayan, Adnan; Yumusak, Nihat; Kömüroglu, Ahmet Ufuk; Aslan, Burcak; Celik, Ozgur Yasar; Göz, Yasar
    Toxoplasma gondii and Neospora caninum are protozoon parasites from the intracellular apicomplexan family. Toxoplasma gondii is the cause of health and economic problems in the sheep industry worldwide. Neospora caninum is usually reported in cows and leads to infections causing abortions; however, its prevalence in sheep is not clear. The present study aimed to investigate the prevalence and pathology of T. gondii and N. caninum by PCR, histopathological and immunehistochemical methods in aborted sheep foetuses collected at different sheep flocks in the Van Province, Turkiye, in 2021. Firstly, the DNA of T. gondii and N. caninum were investigated by PCR in the brain, heart, and peritoneal fluid samples from 42 sheep foetuses. Toxoplasma gondii DNA was proved in 35.7% (15/42) of foetuses whereas N. caninum DNA was not determined in any of the samples. Histopathologically, all T. gondii positive brain tissue samples showed lymphohistiocytic multifocal encephalomyelitis and additional findings included necrotizing myocarditis in the positive heart samples. Toxoplasma gondii tachyzoites were identified in the lesions (diffuse or focal mononuclear cell infiltration in the meninges, and microglia proliferation, myocarditis with oedema) by anti-T. gondii antibodies by the immunohistochemical method. Based on our results, we can conclude that T. gondii is an important agent in sheep abortions and the PCR method is a suitable method for diagnosis which can also be used in heart tissue in pathological studies.
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    MOLECULAR DETECTION OF MYCOPLASMA HAEMOCANIS AND CANDIDATUS MYCOPLASMA HAEMATOPARVUM IN RHIPICEPHALUS SANGUINE US TICK SPECIES COLLECTED FROM DOGS IN ADANA PROVINCE OF TURKEY
    (Parlar Scientific Publications (P S P), 2021) Sababoglu, Ezgi; Ayanz, Adnan; Kilinc, Ozlem Orunc; Yilmaz, Ali Bilgin; Tekindal, Mustafa Agah; Akkaya, Hayrettin; Celik, Burcak Aslan
    Mlycoplasma haemocanis (Mhc) and Candidatus Mycoplasma haematoparvum (CMhp) arc hemo-tropic mycoplasma species known to be specific to dogs. Rhipicephalus sanguineus (sensu lam) ticks have been suggested to bc the main vector in transmission; but a clear relationship between tick infestation and the disease has yet to be established under field conditions. This study aimed to investigate the presence of Mhc and CMhp in R. sanguineus (s.1.) s.ticks that arc considered to be a possible vector for hemoplasma infections, Using the species -specific polymerase chain reaction (PCR) test, the presence of Mhc. and CMhp was investigated in total of 312 archived adult tick samples collected from 63 pet dogs kept in the municipal dog shelter of Adana province located in the Mediterranean region of Turkey. Although R. sanguineus (s.1.) ticks tested negative for Mhc, 2.56% (n = 8) were positive for CMhp. There was a statistically significant difference in the Mhc and CMhp detection rates in R. sanguineus (s.1.) ticks (2.56%; 95% CI: 0.81 - 4.31; P < 0.05). To our knowledge, this is the first study to reveal the presence of CAlhp in R. sanguineus (s.l.) ticks collected from dogs in Turkey.
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    MOLECULAR IDENTIFICATION OF EHRLICHIA CANIS IN RHIPICEPHALUS SANGUINEUS TICKS FROM SIIRT PROVINCE
    (Sciendo, 2021) celik, Burcak Aslan; Ayan, Adnan; Yilmaz, Ali Bilgin; celik, Ozgur Yasar; Kilinc, Ozlem Orunc; Ayan, Ozge Oktay
    This study was performed on Ehrlichia canis positive ticks collected from dogs to perform sequencing of their 16S rRNA genetic section using the PCR method. The collection of ticks was performed from a total of 60 dogs in the Siirt province, Turkey. A total of 250 ticks were collected and morphologically investigated. All ticks were identified as Rhipicephalus sanguineus sensu lato (s.l). Ehrlichial DNA was detected by the PCR method performed on 38 (15.2 %) of the ticks. The E. canis strains obtained as a result of the sequence analysis were found to be 100% identical to the American Texas (MH620194), Indian (KX766395), and Egyptian (MG564254) strains. This study thereby has identified a zoonotic agent from the R. sanguineus ticks collected from the dogs in the Siirt province.
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    Molecular Identification of Hepatozoon canis in Ticks from Dogs in Siirt, Turkey
    (Natl Information Documentation Centre, 2022) Celik, Burcak Aslan; Ayan, Adnan; Yilmaz, Ali Bilgin; Celik, Ozgur Yasar; Kilinc, Ozlem Orunc; Ayan, Ozge Oktay
    HEPATAZOON species are tick borne protozoan parasites classified in the Heptazoidae family, and they are closely related to hemosporinids and piroplasms. In this work, the 18S rRNA genetic section of Hepatozoon canispositive ticks removed from dogs was sequenced using the PCR technique. Ticks were collected from a total of 80 dogs in Siirt, Turkey.A total of 300 collected ticks were morphologically identified to the species level and all ticks identified as Rhipicephalussanguineus (s.l.). H. canis DNA was detected in 12 (%4) out of 300 in R. sanguineusticks by PCR. The phylogenetic tree created via comparison of amplified 18S rRNA region sequences of H. caniswith MT107097.1, MH595911.1, KT215377.1, KT 215376.1, KC 584780.1, KC 584777.1, KC 584775.1, and KC 584774.1. The results obtained will provide important reference material for both veterinary cliniciansand dog owners in terms of managing canine hepatozoonosis.
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    Prevalence and genotypes of Giardia duodenalis in shelter dogs of southeastern Türkiye
    (Urmia Univ, 2023) Celik, Burcak Aslan; Celik, Ozgur Yasar; Kochan, Akin; Ayan, Adnan; Kilinc, Ozlem Orunc; Akyildiz, Guerkan; Irak, Kivanc
    Giardia duodenalis is a protozoan parasite found in humans and several mammals. This parasite spreads worldwide and is generally recognized as a zoonotic agent being reported to be one of the most common causes of diarrhea in humans and animals. In this study, it was aimed to determine the prevalence and genotypes of G. duodenalis in shelter dogs in Diyarbakir province being located in the southeastern Anatolia region of Turkiye. Native-Lugol method and nested polymerase chain reaction analyses of 100 fecal samples showed a prevalence of 3.00 and 4.00%, respectively. The prevalence was higher in females and in those younger than 1 year. Sequence analysis revealed the presence of zoonotic assemblage B, assemblage D and assemblage E. The detection of zoonotic assemblage B in this study suggests that dogs may be a reservoir for human giardiasis. Further molecular research is needed to determine the genotype diversity of Giardia as well as its possible role in the transmission of this parasite to humans.(c) 2023 Urmia University. All rights reserved.
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    Prevalence and molecular characterization of Cryptosporidium spp. in calves in the Siirt Province, Turkiye
    (Veterinarni A Farmaceuticka Univerzita Brno, 2023) Celik, Ozgur Yasar; Sahin, Tekin; Celik, Burcak Aslan; Kilinc, Ozlem Orunc; Ayan, Adnan; Akyildiz, Gurkan; Ayan, Ozge Oktay
    Cryptosporidiosis, one of the main protozoan infections of the last century, is especially dangerous for calves and causes significant economic losses. This research was carried out to determine the prevalence of Cryptosporidium spp. by microscopic and molecular methods and to determine subtypes in 100 calves up to 6 months old and with diarrhoea in the Siirt Province, Turkiye. As a result of the microscopic examination (Kinyoun's acid-fast), Cryptosporidium spp. oocysts were found in 8 (8%) of 100 samples. As a result of nested PCR, 826-864 bp specific bands for Cryptosporidium spp. were obtained in 13 (13%) of 100 samples. When the DNA sequences of the SSU rRNA gene were compared with the NCBI Basic Local Alignment Search Tool database, it was determined that eight samples sequence analyses showed 100% similarity with the C. parvum, C. ryanae, and C. bovis samples. The detection of C. parvum, which has zoonotic importance in this study, suggests that calves with diarrhoea may be a source of contamination for other animals and humans. Therefore, animal owners and people in close contact with animals should be informed about the public health of cryptosporidiosis.
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    Prevalence and Subtypes Distribution (ST10, ST14, ST25, ST26) of Blastocystis spp. in Anatolian Water Buffalo (Bubalus bubalis) in Van, Turkiye
    (Wiley, 2024) Ayan, Adnan; Celik, Burcak Aslan; Celik, Ozgur Yasar; Yilmaz, Ali Bilgin; Kilinc, Ozlem Orunc; Ayan, Ozge Oktay
    Background: Blastocystis spp. is one of the most common protozoa worldwide, living in the gastrointestinal tract of humans and many other animals. On the basis of the genetic heterogeneity of small subunit ribosomal RNA, at least 28 subtypes (ST1-ST17, ST21 and ST23-ST32) are reported to exist in mammals and birds. ObjectivesThis study was carried out to determine the prevalence and subtypes of Blastocystis spp. in Anatolian buffaloes (Bubalus bubalis) in Van province in the Eastern Anatolia Region of Turkey. Methods: DNA was extracted using commercial GeneMATRIX Stool DNA Purification Kit and then stored at -20 degrees C until PCR amplification. After PCR amplification of the SSU rRNA gene region positive Blastocystis spp., amplicons from buffalo faeces were sequenced and then deposited in GenBank (OR576949.1, OR576950.1, OR576970.1, OR576971.1, OR577019.1, PP837943.1, PP837940.1, PP837939.1, PP837604.1, PP837937.1, PP837934.1, PP837601.1, PP837936.1 and PP837603.1). Results: PCR analysis of 120 faecal samples showed a total prevalence of 11.67% (14/120). The prevalence was higher in females and young animals (p > 0.05). Sequence analysis revealed Blastocystis spp., ST10, ST14, ST25 and ST26 subtypes. To our knowledge, Blastocystis subtypes ST25 and ST26 in buffaloes were reported for the first time in this study. Conclusion: It is thought that more large-scale studies should be carried out to determine the zoonotic subtype potential of this protozoan in the region.
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    The Investigation of Giardiasis (Foodborne and Waterborne Diseases) in Buffaloes in Van Region, Turkiye: First Molecular Report of Giardia duodenalis Assemblage B from Buffaloes
    (Mdpi, 2023) Kilinc, Ozlem Orunc; Ayan, Adnan; Celik, Burcak Aslan; Celik, Ozgur Yasar; Yuksek, Nazmi; Akyildiz, Gurkan; Oguz, Fatma Ertas
    Giardia duodenalis (G. duodenalis) is an important zoonotic protozoan agent that causes foodborne and waterborne diarrhea in humans and other mammals. Molecular-based tests are critical in diagnosing giardiasis in humans and animals, identifying species, understanding the zoonotic potential and transmission routes, and evaluating taxonomy. Therefore, this study aimed to investigate the molecular characterization of G. duodenalis in buffaloes in the Van region in Turkiye. Buffaloes are a species that has been poorly studied in this regard. For this purpose, 100 fecal samples were collected from buffaloes in the Van region. The DNA extraction was performed using the GeneMATRIX STOOL DNA Purification Kit from stool samples. The nested PCR test was performed with the appropriate primers from the obtained DNA samples. The obtained bands suitable for sequencing were sent for sequence analysis, and the sequence results were aligned bidirectionally and compared with the database of GenBank by BLAST. As a result of the study, an 11% positivity rate for G. duodenalis was found in buffaloes, and assemblage E and assemblage B were isolated. To our knowledge, assemblage B in buffaloes was reported for the first time in this study. As a result, it was concluded that buffaloes are an important reservoir for waterborne and foodborne giardiasis.