Identification and characterization of threeEncephalitozoon cuniculistrains

Abstract

SUMMARYMicrosporidia are increasingly recognized as causing opportunistic infections in immunocompromised individuals.Encephalitozoon cuniculiis probably the most studied mammalian microsporidian that infects insects and mammals, including man. In this study, 8E. cuniculiisolates were compared and were found to fall into 3 strains. Strain type I includes the rabbit type isolate, as well as isolates from an additional rabbit, a dwarf rabbit, and a mouse. Strain type II includes 2 murine isolates and strain type III includes 2 isolates obtained from domestic dogs. By SDS-PAGE, the 3 strains differ primarily in the molecular weight range of 54–59 kDa where strain type I displays an apparent broad singlet at 57 kDa, strain type II displays an apparent doublet at 54 and 58 kDa, and strain type III displays an apparent broad band at 59 kDa. Antigenic differences were detected in the molecular weight regions of 54–58 kDa as well as 28–40 kDa by Western blot immunodetection using murine antisera raised againstE. cuniculi, Encephalitozoon hellem,and theEncephalitozoon-like Septata intestinalis.Polymerase chain reaction (PCR) products containing only small subunit rDNA sequences from the differentE. cuniculiisolates formed homoduplexes whereas PCR products containing intergenic rRNA gene sequences formed heteroduplexes in mobility shift analyses.FokI digestion of the PCR products containing the intergenic rRNA gene region resulted in unique restriction fragment length polymorphism patterns, and DNA sequencing demonstrated that in the intergenic spacer region, the sequence 5'-GTTT-3' was repeated 3 times in strain type I, twice in strain type II, and 4 times in strain type III. This study indicates that there exist at least 3E. cuniculistrains which may become important in the epidemiology of humanE. cuniculiinfections. Furthermore, as additionalE. cuniculiisolates are characterized, these strains will be named or reclassified once the criteria for taxonomy and phylogenetic tree construction for microsporidia become better defined.