Microsporidia Detection and Genotyping Study of Human Pathogenic E. bieneusi in Animals from Spain

Ana Luz Galván-Díaz,Francisco Ponce Gordo,Soledad Fenoy,Fernando Izquierdo,Carmen Del Águila,Angela Magnet,María Haro,Guadalupe Miró,Nuno Henriques-Gil,Elizabeth Didier

doi:10.1371/journal.pone.0092289

Ana Luz Galván-Díaz, Francisco Ponce Gordo + Show 8 more

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https://doi.org/10.1371/journal.pone.0092289

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Abstract

Microsporidia are ubiquitous parasites infecting all animal phyla and we present evidence that supports their zoonotic potential. Fecal samples taken from domestic (cats and dogs), farm (pigs, rabbits and ostriches) and wild animals (foxes) from different provinces of Spain were evaluated for microsporidia infection by light microscopy and PCR. After Microsporidia species identification, E. bieneusi genotypes were additionally studied by sequence analysis of the ITS region. Eighty-five samples out of 159 exhibited structures that were compatible with microsporidia spores by Webeŕs stain with 37 of them being confirmed by PCR. Microsporidia species identified included E. bieneusi, E. intestinalis and A. algerae. We report the first diagnosis of E. intestinalis and E. bieneusi in ostriches and A. algerae in pigs. We also provide new information on the molecular characterization of E. bieneusi isolates both in rabbits and ostriches. All of the E. bieneusi genotypes identified belonged to the zoonotic group of genotypes (Group I) including genotypes A (dogs), I (pigs), D (rabbits and foxes) and type IV (ostriches). Our results demonstrate that microsporidia are present in domestic, farm and wild animals in Spain, corroborating their potential role as a source of human infection and environmental contamination.

Highlights

Microsporidia are a ubiquitous group of obligate intracellular parasites that infect all major animal groups
At least 14 microsporidian species are considered to be pathogenic for humans, Enterocytozoon bieneusi being the most prevalent species in humans worldwide followed by Encephalitozoon intestinalis [1,5,6,7]
Eighty-five samples of 159 exhibited structures that were compatible with microsporidia spores by Webers stain with 37 of them being confirmed by PCR

Summary

Introduction

Microsporidia are a ubiquitous group of obligate intracellular parasites that infect all major animal groups (both vertebrates and invertebrates). In the case of E. bieneusi, sequence analysis of the internal transcribed spacer (ITS) region has revealed a considerable genetic variation within E. bieneusi isolates of human and animal origins, with more than 100 genotypes described so far [7,9,10,11]. These studies have identified E. bieneusi genotypes host-associated to several animals and some genotypes with no host specificity, which are considered zoonotic [12,13,14]. The occasional sequences included in the divergent groups II, III, and IV have so far been restricted to specific hosts such as raccoons (III) and cats and dogs (IV)

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