Molecular diagnosis of microsporidia strains in slaughtered cows of southwest of Iran

Abstract

Microsporidia is often considered as an opportunistic infection in patients with impaired immune systems such as patients with acquired immune deficiency syndrome, transplant recipients, children and old people. Due to the ability of the parasite to transmit from animals to human as well as the increasing prevalence of parasitic infections and immune deficiency diseases; therefore, the aim of this study was to evaluate molecular diagnosis of microsporidia strains in slaughtered cows of southwest of Iran. Initially, 256 stool samples of cows were collected from 5 regions of Khuzestan province, southwest of Iran and stained by modified trichrome (weber). Then, the parasite spores were examined by optical microscope. Total DNA was extracted from samples using DNA extraction kit for stool (Bioneer) and evaluated by multiplex/nested PCR method. The products of nested-PCR were explored by RFLP method using restriction enzyme MnlI. For genotyping, positive samples of RFLP were sequenced. Of 256, 21 and 48 samples were found positive by the staining and nested PCR tests, respectively. Of 48, 36 samples were Enterocytozoon bieneusi with genotype frequency D (22), J (9) and M (5). Also 9 were detected as Encephalitozoon species among which 2 were E. cuniculi, 6 were E. intestinalis and 1 was E. hellem Eventually, 3 samples were found positive for both Enterocytozoon and Encephalitozoon. The results showed that cows can be a source of microsporidia infections. Due to the zoonotic importance of this parasite and its ability to transmit from animals to humans; the detection and species determination of the parasite seems essential. The highest risk of infection is for individuals with impaired immune systems.