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Molecular Identification of Cryptosporidium spp. in Iranian Dogs Using Seminested PCR: A First Report
Reza Ranjbar1, Hossein Mirhendi2, Morteza Izadi3
11 Molecular Biology Research Center, Baqiyatallah University of Medical Sciences , Tehran, Iran .
Abstract:
Cryptosporidium is a parasitic protozoon with a wide range of vertebrate hosts. Cryptosporidiosis has been reported from numerous countries, including Iran. Molecular identification can be applied to characterize Cryptosporidium, of which there are over 30 species and 50 genotypes. Herein, we report the genetic diversity of Cryptosporidium spp. in Iranian dogs for the first time based on 18S ribosomal RNA gene sequencing. One hundred forty fecal samples of herd dogs were collected from Isfahan, central Iran. The samples were concentrated using sucrose flotation and subjected to Kinyoun staining. DNA extraction of positive samples was performed, and molecular diagnosis was carried out using highly specific seminested PCR for the characterization of Cryptosporidium species. Finally, sequencing and DNA analysis were performed to identify Cryptosporidium species. A total of 2.14% of herd dogs were positive for cryptosporidiosis in both microscopy and molecular methods. In all cases, the causative agent was identified as Cryptosporidium parvum. Dogs associated with positive samples had been in close relationship with livestock. Cryptosporidiosis in the herd dogs in Isfahan could be due to their close contact with animals, particularly cattle and sheep. Given that dogs with cryptosporidiosis lack clinical symptoms, they are a potential source of zoonotic transmission of this disease as they are companion animals for humans. Dogs with cryptosporidiosis are a potential source of the zoonotic transmission of this disease.
Insights
Cryptosporidiosis was identified in Iranian herd dogs for the first time. Cryptosporidium parvum was found, indicating dogs can be a source of zoonotic transmission.
Area of Science:
- Veterinary Parasitology
- Molecular Biology
- Zoonotic Diseases
Background:
- Cryptosporidium is a protozoan parasite infecting diverse hosts globally.
- Cryptosporidiosis is a known issue in Iran, with various species and genotypes identified.
- Dogs can act as reservoirs for Cryptosporidium, posing a zoonotic risk.
Purpose of the Study:
- To investigate the genetic diversity of Cryptosporidium species in Iranian dogs.
- To identify Cryptosporidium species in herd dogs from Isfahan, Iran, using molecular methods.
- To assess the potential of dogs as a source of zoonotic Cryptosporidium transmission.
Main Methods:
- Collection and processing of 140 fecal samples from herd dogs in Isfahan.
- Microscopic examination (Kinyoun staining) and molecular diagnosis (seminested PCR) for Cryptosporidium.
- 18S ribosomal RNA gene sequencing for species identification and genetic analysis.
Main Results:
- 2.14% of herd dogs tested positive for cryptosporidiosis via microscopy and molecular methods.
- Cryptosporidium parvum was identified as the sole causative agent in all positive cases.
- Positive dogs were in close contact with livestock, suggesting a transmission link.
Conclusions:
- Herd dogs in Isfahan are infected with Cryptosporidium parvum, likely through contact with livestock.
- Asymptomatic infected dogs represent a potential zoonotic transmission source for humans.
- This study highlights the importance of monitoring Cryptosporidium in dogs for public health.
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