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Infection and Immunity, June 2006, p. 3448-3454, Vol. 74, No. 6
0019-9567/06/$08.00+0     doi:10.1128/IAI.01215-05
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Genetic Variation among Isolates of Sarcocystis neurona, the Agent of Protozoal Myeloencephalitis, as Revealed by Amplified Fragment Length Polymorphism Markers

H. M. Elsheikha,1,{dagger} H. C. Schott II,1 and L. S. Mansfield1,2*

Department of Large Animal Clinical Sciences,1 Department of Microbiology and Molecular Genetics, College of Veterinary Medicine, Michigan State University, East Lansing, Michigan 488242

Received 28 July 2005/ Returned for modification 2 September 2005/ Accepted 4 March 2006

Sarcocystis neurona causes serious neurological disease in horses and other vertebrates in the Americas. Based on epidemiological data, this parasite has recently emerged. Here, the genetic diversity of Sarcocystis neurona was evaluated using the amplified fragment length polymorphism (AFLP) method. Fifteen S. neurona taxa from different regions collected over the last 10 years were used; six isolates were from clinically diseased horses, eight isolates were from wild-caught opossums (Didelphis virginiana), and one isolate was from a cowbird (Molothrus ater). Additionally, four outgroup taxa were also fingerprinted. Nine primer pairs were used to generate AFLP patterns, with a total number of amplified fragments ranging from 30 to 60, depending on the isolate and primers tested. Based on the presence/absence of amplified AFLP fragments and pairwise similarity values, all the S. neurona isolates tested were clustered in one monophyletic group. No significant correlation could be found between genomic similarity and host origin of the S. neurona isolates. AFLP revealed significant intraspecific genetic variations, and S. neurona appeared as a highly variable species. Furthermore, linkage disequilibrium analysis suggested that S. neurona populations within Michigan have an intermediate type of population structure that includes characteristics of both clonal and panamictic population structures. AFLP is a reliable molecular technique that has provided one of the most informative approaches to ascertain phylogenetic relationships in S. neurona and its closest relatives, allowing them to be clustered by relative similarity using band matching and unweighted pair group method with arithmetic mean analysis, which may be applicable to other related protozoal species.

* Corresponding author. Mailing address: 181 National Food Safety and Toxicology Center, Michigan State University, East Lansing, MI 48824. Phone: (517) 432-3100, ext. 119. Fax: (517) 432-2310. E-mail: mansfie4{at}cvm.msu.edu.

Editor: W. A. Petri, Jr.

{dagger} Present address: Department of Parasitology, College of Veterinary Medicine, Mansoura University, Mansoura 35516, Egypt.

Infection and Immunity, June 2006, p. 3448-3454, Vol. 74, No. 6
0019-9567/06/$08.00+0     doi:10.1128/IAI.01215-05
Copyright © 2006, American Society for Microbiology. All Rights Reserved.





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