Molecular Diagnosis of Enterocytozoon bieneusi and Encephalitozoon spp. in Wild Rats of Southwest of Iran

Abstract

Background: Microsporidiosis is considered as an opportunistic infection in immunodeficient patients. Objectives: Due to the increasing prevalence of parasitic infections and immunodeficiency diseases as well as the transmission risk of microsporidia from animals to human, the aim of this study was to evaluate molecular diagnosis of Enterocytozoon bieneusi and Encephalitozoon spp. by the multiplex/nested polymerase chain reaction (PCR) and staining methods in wild rats of Ahvaz city, southwest of Iran. Methods: Initially, 160 stool samples were collected from wild rats in different parts of Ahvaz city. The samples were stained by the modified trichrome staining and explored microscopically. The DNA was extracted using the DNA stool kit and examined by multiplex/nested PCR. For differentiating the species of Encephalitozoon, the multiplex/nested PCR products were explored by the restriction fragment length polymorphism (RFLP) method using the restriction enzyme of Mnl1. Results: Of 160, 26 cases were suspected positive for microsporidia spore by the staining. Also, of 160, 18 cases were positive by the multiplex/nested PCR method, that 14 and 4 samples were detected as E. bieneusi and E. intestinalis, respectively. Of 14 E. bieneusi samples, 12 and 2 cases were detected as genotype D and M, respectively. Conclusions: The findings revealed a relatively high prevalence of microsporidia infection in wild rats of the city and these animals can be a source of microsporidiosis. Due to the zoonotic potential of the microorganisms, high -risk individuals should be receiving the information about the risk of direct and indirect contact with the infected animals.

First detection and genotyping of Enterocytozoon bieneusi in pet fancy rats (Rattus norvegicus) and guinea pigs (Cavia porcellus) in China

“…To our knowledge, this is the first report of E. bieneusi infection in pet rats and pet guinea pigs worldwide. To date, there have been three studies focusing on E. bieneusi infection in wild rats in Iran and China [24,30,34] (Table 3). In this study, the prevalence of E. bieneusi in pet fancy rats was slightly higher than that (4.0%-8.9%) in wild rats in the above-mentioned reports.…”

“…For pet guinea pigs, two genotypes were identified, including the predominant genotype S7 (n = 30, 85.7%) and a novel genotype (named PGP, n = 5) ( Table 2). Until now, molecular studies of E. bieneusi in rats have been limited to three reports in wild rats (R. norvegicus and R. rattus) in Iran and China, and a total of six genotypes were identified, including genotypes D, CD6 (synonyms: CHG14), Peru6, M, BEB6, and CHG2 [24,30,34] (Table 3). For guinea pigs, only one survey conducted in Peru identified genotype peru16 from household guinea pigs (Table 3) [3], and this genotype was not detected in the present study.…”

“…Genotyping based on the internal transcribed spacer (ITS) region of the rRNA gene has identified 11 major phylogenetic groups and more than 470 genotypes of E. bieneusi from various hosts [17]. To date, more than 60 E. bieneusi genotypes have been identified in rodents worldwide [6,8,12,17,24,26,30,34]. For rats (Rattus spp.…”

“…For rats (Rattus spp. ), only three surveys have focused on the molecular characterization of E. bieneusi in wild rats in Iran and China, and six genotypes (D, M, Peru6, CD6, BEB6, and CHG2) have been identified [24,30,34]. Only one published article has reported genotype peru16 from household guinea pigs in Peru [3].…”

The First Report and Molecular Analysis of Enterocytozoon bieneusi from Raccoon (Procyon lotor) in North of Iran

“…), and free range rats (Tavalla et al. 2018a), there are no data about the distribution of microsporidia in wildlife animals in Iran.…”