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Investigation of Cryptosporidium Oocyst Contamination in Water from Zabol and Zahedan: A Molecular and Microscopic Analysis
Reza Shahraki1 , Mohammadreza Beheshtizadeh2 , Mahdi khoshsima Shahraki3 , Mansour Dabirzadeh4
1- Zabol University of Medical Sciences, Medical School, Department of Parasitology,
2- Group of molecular biology, Azma Elixir Pajooh, Tehran, Iran.
3- Department of Pathology faculty of veterinary medicine, Shahid Bahonar University of Kerman, Kerman, Iran
4- Department of Pathology faculty of veterinary medicine, Shahid Bahonar University of Kerman, Kerman, Iran , mdabirzadeh20002000@gmail.com
Abstract:   (1859 Views)
Abstract
Background: The presence of Cryptosporidium oocysts in water represents a significant public health concern due to its potential impact on human health. This study aims to detect and characterize Cryptosporidium in water samples from Zabol and Zahedan cities using microscopic examination and molecular methods.
Material and methods: 180 water samples were collected and analyzed at the Parasitology Laboratory of Zabol Medical School. Ziehl-Neelsen and Trichrome staining followed initial direct microscopic examinations, identifying contamination through the presence of Cryptosporidium oocysts at 100x magnification.
Results: Among these samples, 35 tested positive, 142 negative, and 3 were deemed suspicious. In Zahedan, the distribution was 9 positive, 2 suspicious, and 89 negative samples, while in Zabol, 26 were positive, 1 suspicious, and 53 negative.
PCR amplification targeted the ITS1 gene, and electrophoresis on a 1% agarose gel showed band patterns ranging from 871 to 961 base pairs. For genotype differentiation, restriction enzymes RsaI and AluI were used, with AluI generating distinct patterns featuring bands at 410, 457, 480, 760, and 850 bp. BLAST analysis revealed a 98.84% sequence identity with C. parvum. Phylogenetic analysis further indicated a genetic relationship with known C. parvum isolates.
Conclusion: This study provides comprehensive detection and molecular characterization of Cryptosporidium in the examined water samples. The results highlight the need for continuous monitoring and improvements in water treatment processes to mitigate associated public health risks.
 
Keywords: Cryptosporidium, water contamination, molecular methods, PCR, DNA sequencing, Zabol, Zahedan
Full-Text [PDF 587 kb]   (374 Downloads)    
Research Article: Original Paper | Subject: Parasitology
Received: 2024/12/14 | Accepted: 2025/01/1
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