Mahami Oskouei Mahmoud, Fallah Esmaeil, Ahmadi Mahmoud, Safaiyan Abdolrasoul, Bakhtiyari Salar, Naserifar Razi, Dousti Majid
Department of Parasitology, Faculty of Medicine, Tabriz University of Medical Sciences, Tabriz, Iran.
International Branch of Tabriz University of Medical Sciences (Aras), Tabriz, Iran.
Iran J Parasitol. 2014 Sep;9(3):435-40.
Cryptosporidiosis is one of the most important parasitic infections in human and animals. This study was designed for survey on the prevalence of Cryptosporidium infection in farms of Ilam, west of Iran, using parasitology method and genotyping by Nested PCR-RFLP.
Fecal samples of 217 cattle were collected fresh and directly from the rectum of cattle. All of the samples were examined by microscopic observation after staining with modified Ziehl-Neelsen (MZN). Genomic DNA extracted by using EURx DNA kit. A Nested PCR-RFLP protocol amplifying 825 bp fragment of 18s rRNA gene conducted to differentiate species and genotyping of the isolates using SspI and VspI as restriction enzymes.
The prevalence of Cryptosporidium infection in cattle using both methods is 3.68%. Most of the positive cattle were calves under six months. Species diagnosis carried out by digesting the secondary PCR product with SspI that C. parvum generated 3 visible bands of 448, 247 and 106 bp and digested by VspI restriction enzyme generated 2 visible bands of 628 and 104bp. In this investigation all of the positive samples were Cryptosporidium parvum.
C. parvum (bovine genotype) detected in all positive cattle samples in Ilam, west of Iran. The results of the present study can help for public health care systems to prevention and management of cryptosporidiosis in cattle and the assessment of cattle cryptosporidiosis as a reservoir for the human infection.
隐孢子虫病是人和动物最重要的寄生虫感染之一。本研究旨在采用寄生虫学方法和巢式PCR-RFLP基因分型技术，对伊朗西部伊拉姆省养殖场隐孢子虫感染率进行调查。
从217头牛的直肠直接采集新鲜粪便样本。所有样本经改良齐-尼氏（MZN）染色后进行显微镜观察。使用EURx DNA试剂盒提取基因组DNA。采用巢式PCR-RFLP方案扩增18s rRNA基因的825bp片段，以SspI和VspI作为限制性内切酶对分离株进行种属鉴别和基因分型。
两种方法检测牛隐孢子虫感染率均为3.68%。大多数阳性牛为6个月以下的犊牛。通过用SspI消化二次PCR产物进行种属诊断，微小隐孢子虫产生448、247和106bp的3条可见带，用VspI限制性内切酶消化产生628和104bp的2条可见带。在本次调查中，所有阳性样本均为微小隐孢子虫。
在伊朗西部伊拉姆省所有阳性牛样本中均检测到微小隐孢子虫（牛基因型）。本研究结果有助于公共卫生保健系统预防和管理牛隐孢子虫病，并评估牛隐孢子虫病作为人类感染源的情况。
