en انگل شناسی Parasitology تحقيقي Original Paper <span style="font-size:11pt"><span style="line-height:150%"><span calibri="" style="font-family:"><b><span lang="EN" style="font-size:12.0pt"><span style="line-height:150%"><span new="" roman="" style="font-family:" times="">Abstract</span></span></span></b></span></span></span><br> <span style="font-size:11pt"><span style="line-height:150%"><span calibri="" style="font-family:"><b>Background</b>: The presence of <i>Cryptosporidium</i> oocysts in water represents a significant public health concern due to its potential impact on human health. This study aims to detect and characterize <i>Cryptosporidium</i> in water samples from Zabol and Zahedan cities using microscopic examination and molecular methods.<br> <b>Material and methods:</b> 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 <i>Cryptosporidium</i> oocysts at 100x magnification.<br> <b>Results: </b>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.<br> 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 <i>RsaI </i>and <i>AluI </i>were used, with <i>AluI</i> generating distinct patterns featuring bands at 410, 457, 480, 760, and 850 bp. BLAST analysis revealed a 98.84% sequence identity with <i>C. parvum</i>. Phylogenetic analysis further indicated a genetic relationship with known <i>C. parvum</i> isolates.<br> <b>Conclusion:</b> This study provides comprehensive detection and molecular characterization of <i>Cryptosporidium</i> 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.</span></span></span><br> &nbsp; Cryptosporidium, water contamination, molecular methods, PCR, DNA sequencing, Zabol, Zahedan 0 0 http://mlj.goums.ac.ir/browse.php?a_code=A-10-1853-1&slc_lang=en&sid=1 Reza Shahraki predator4tear@gmail.com 100319475328460032531 100319475328460032531 No Zabol University of Medical Sciences, Medical School, Department of Parasitology, Mohammadreza Beheshtizadeh dmrbz@hotmail.com 100319475328460032532 100319475328460032532 No Group of molecular biology, Azma Elixir Pajooh, Tehran, Iran. Mahdi khoshsima Shahraki mail.khoshsima6144@vet.uk.ac.ir 100319475328460032533 100319475328460032533 No Department of Pathology faculty of veterinary medicine, Shahid Bahonar University of Kerman, Kerman, Iran Mansour Dabirzadeh mdabirzadeh20002000@gmail.com 100319475328460032534 100319475328460032534 Yes Department of Pathology faculty of veterinary medicine, Shahid Bahonar University of Kerman, Kerman, Iran