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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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    Öğe
    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.
  • [ X ]
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    Preliminary investigation of the prevalence and genotype distribution of Cryptosporidium spp., and Giardia duodenalis in cats in Siirt, Turkey
    (Sciendo, 2023) Aslan Celik, Burcak; Celik, Ozgur Yasar; Ayan, Adnan; Akyildiz, Gurkan; Orunc Kilinc, Ozlem; Oktay Ayan, Ozge; Ercan, Kerem
    Cryptosporidium spp. and Giardia duodenalis are protozoan parasites found in humans and many animal species worldwide. The aim of this study was to determine the prevalence and genotypes of Cryptosporidium spp. and Giardia duodenalis in cats and to evaluate the zoonotic potential of these agents. The animal material of the study consisted of a total of 40 cats brought to the Veterinary Faculty. Fresh fecal samples taken from the cats were placed in individual sample containers. All samples were examined under the microscope by Kinyoun Acid Fast staining for Cryptosporidium spp. and by the native-Lugol method for Giardia duodenalis. Nested PCR and sequence analyses were then performed. As a result of microscopic and nested PCR analyses for Cryptosporidium spp., no positivity was found in any sample. The prevalence of Giardia duodenalis was 2.5% in both microscopic examination and nested PCR analyses. When the DNA sequences of the beta-Giardin gene obtained in the study were compared with the database in NCBI Basic Local Alignment Search Tool, it was determined that one sample overlapped with Assemblage B samples. As a result of this study, the prevalence of Cryptosporidium spp. and Giardia duodenalis in cats was determined and the presence of Assemblage B was revealed. It is recommended that repetitive studies should be carried out as much as possible to determine the possible role of these parasites in the transmission of these parasites to humans.
  • [ X ]
    Öğe
    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.
  • [ X ]
    Öğe
    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.

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