Evidence for geographic clusters: Molecular genetic differences among strains of Pythium insidiosum from Asia, Australia and the Americas are explored.

Twenty-eight isolates of Pythium insidiosum and P. destruens from Asia, Australia and the Americas were compared on the basis of restriction fragment-length polymorphisms of the amplified ribosomal intergenic spacer. Comparison of band profiles yielded three distinct clusters and an isolate that did not fall into any of the clusters. Cluster I consisted of 16 isolates, all from the Americas (Costa Rica, Brazil, Haiti, United States). Cluster II consisted of seven isolates from Asia (India, Thailand, Japan, Papua New Guinea) and Australia, including the two isolates of P. destruens. This cluster also included a United States isolate from a human who might have contracted an infection of P. insidiosum by contact with food from the Middle East. Cluster III was most distantly related to the other two clusters and consisted of two isolates from Thailand and one from the United States. The isolate excluded from all three clusters was from a spectacled bear in a zoo in the United States. These results indicate that all the isolates are more closely related to each other than to any other Pythium species and thus indeed might be one species, but they also point to geographical variants. Cluster III and Isolate M18 are so distant from the others that they might prove to be separate species. Knowledge of intraspecific variability in P. insidiosum might be important for the management of pythiosis in mammals.