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Journal of Clinical Microbiology, April 2005, p. 1865-1868, Vol. 43, No. 4
0095-1137/05/$08.00+0     doi:10.1128/JCM.43.4.1865-1868.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Molecular Evidence for Multiple Host-Specific Strains in the Genus Rhinosporidium

Victor Silva,1,{dagger} Cristiane N. Pereira,1 Libero Ajello,2 and Leonel Mendoza1*

Medical Technology Program, Microbiology and Molecular Genetics, Michigan State University, East Lansing, Michigan 48824-1031,1 Mycotic Diseases Branch, NCID, Centers for Disease Control and Prevention, Atlanta, Georgia 303332

Received 30 August 2004/ Returned for modification 19 September 2004/ Accepted 8 December 2004

The taxonomic relationship of Rhinosporidium seeberi with other organisms remained controversial for over a century. Recently, molecular studies have shown R. seeberi to be a protistal microbe in the newly described class Mesomycetozoea at the animal-fungal boundary. Phylogenetic analyses of R. seeberi using 18S small-subunit (SSU) rRNA genes from several hosts suggested Rhinosporidium as a monotypic genus. To test this hypothesis, the internal transcribed spacer 1 (ITS1), 5.8S, and ITS2 from eight humans, two swans, and a dog with rhinosporidiosis were sequenced. The ITS regions were amplified by PCR using a primer designed from a unique region of R. seeberi's 18S SSU rRNA genes in combination with the ITS4 universal primer. In addition, the universal ITS4 and ITS5 primers were also used. R. seeberi's ITS sequences showed differences in the numbers of nucleotides among strains. For instance, the eight human ITS sequences were uniformly similar with only a few mismatches and ~1,060 bp long. In contrast, sequences from one of the swans and the dog were 1,356 bp and ~1,147 bp long, respectively. Clustal analysis of all of the ITS sequences showed multiple 50- to 60-bp gaps and several mismatches among them. Parsimony analysis placed the Rhinosporidium ITS sequences in three well-supported sister groups according to the hosts' identities. This analysis strongly suggests that the genus Rhinosporidium may possess multiple host-specific strains. No correlation was found between this finding and the phenotypic features of R. seeberi in the studied samples.

* Corresponding author. Mailing address: Medical Technology Program, Michigan State University, 322 North Kedzie Hall, East Lansing, MI 48824-1031. Phone: (517) 353-7800. Fax: (517) 432-2006. E-mail: mendoza9{at}msu.edu.

{dagger} Present address: Microbiology and Mycology Program, Biomedical Science Institute, School of Medicine, University of Chile, Independencia 1027, Santiago, Chile.

Journal of Clinical Microbiology, April 2005, p. 1865-1868, Vol. 43, No. 4
0095-1137/05/$08.00+0     doi:10.1128/JCM.43.4.1865-1868.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



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