Cryptosporidium genotypes and subtypes distribution in river water in Iran M. R. Mahmoudi, E. Nazemalhosseini-Mojarad, B. Kazemi, A. Haghighi, A. Mirzaei, A. Mohammadiha, S. Jahantab, L. Xiao and P. Karanis ABSTRACT Little is known about the diversity and public health significance of Cryptosporidium species in river waters in Iran. In the present study, we determined the genotype and subtype distribution of Cryptosporidium spp. in river water samples in Iran. A total of 49 surface water samples were collected from rivers and surface water in Guilan and Tehran provinces during 2009–2010. Water samples were filtrated through a 1.2-μm pore size membrane filter or by Filta-Max filter followed by immunomagnetic separation or sucrose purification methods. Genotype and subtype of Cryptosporidium were identified by sequence analysis of the 18S rRNA and 60 kDa glycoprotein (gp60) genes, respectively. A total of 24 (48.97%) water samples were positive for Cryptosporidium species by the 18sRNA-based PCR-sequencing technique. DNA sequencing revealed the presence of five species of Cryptosporidium (C. parvum, C. hominis, C. muris, C. andersoni, and C. canis) in the water samples of the study area and, to our knowledge, the first report of C. muris in Iran. The results of GP60 gene analysis showed that all C. parvum and C. hominis isolates belonged to the IId and Id subtype families, respectively. The investigated river water supplies were heavily contaminated by pathogenic species of Cryptosporidium from humans and livestock. There is potential risk of waterborne cryptosporidiosis in humans and animals. M. R. Mahmoudi (corresponding author) Department of Microbiology, Faculty of Medicine, Guilan University of Medical Sciences, Rasht, Iran E-mail: mrmahmoodi2002@yahoo.com E. Nazemalhosseini-Mojarad Gastroenterology and Liver Diseases Research Center, Shahid Beheshti University of Medical Science, Tehran, Iran M. R. Mahmoudi B. Kazemi Research Center of Cellular and Molecular Biology, Shahid Beheshti University of Medical Sciences, Tehran, Iran M. R. Mahmoudi A. Haghighi A. Mohammadiha Department of Medical Parasitology & Mycology, School of Medicine, Shahid Beheshti University of Medical Sciences, Chamran Expressway, Tehran, Iran A. Mirzaei Department of Parasitology, School of Medicine, Iilam University of Medical Sciences, Iilam, Iran S. Jahantab Tehran Province Water & Wastewater Co. (TPWW), Tehran, Iran L. Xiao Division of Foodborne, Waterborne, and Environmental Diseases, National Center for Emerging and Zoonotic Infectious Diseases, Centers for Disease Control and Prevention, Public Health Services, Atlanta, GA, USA P. Karanis Center for Biomedicine and Infectious Diseases, University of Quinghai, Xining, Quinghai, R.P. China Key words | Cryptosporidium spp., environmental samples, Iran, subtype 1 © IWA Publishing 2014 Journal of Water and Health | in press | 2014 doi: 10.2166/wh.2014.234 Uncorrected Proof