Comparison of Sarcocystis neurona isolates derived from horse neural tissue.
Abstract: Sarcocystis neurona is a protozoan parasite that can cause neurological deficits in infected horses. The route of transmission is by fecal-oral transfer of sporocysts from opossums. However, the species identity and the lifecycle are not completely known. In this study, Sarcocystis merozoites from eight isolates obtained from Michigan horses were compared to S. neurona from a California horse (UCD1), Sarcocystis from a grackle (Cornell), and five Sarcocystis isolates from feral opossums from Michigan. Comparisons were made using several techniques. SDS-PAGE analysis with silver staining showed that Sarcocystis spp. from the eight horses appeared the same, but different from the grackle isolate. One Michigan horse isolate (MIH6) had two bands at 72 and 25kDa that were more prominent than the UCD1 isolate and other Michigan horse isolates. Western blot analysis showed that merozoites of eight of eight equine-derived isolates, and the UCD1 S. neurona isolate had similar bands when developed with serum or CSF of an infected horse. Major bands were seen at 60, 44, 30, and 16kDa. In the grackle (Cornell) isolate, bands were seen at 60, 44, 29, and 16kDa. DNA from merozoites of each of the eight equine-derived isolates and the grackle-derived isolate produced a 334bp PCR product (Tanhauser et al., 1999). Restriction fragment length polymorphism (RFLP) analysis of these horse isolates showed banding patterns characteristic for S. neurona. The grackle (Cornell) isolate had an RFLP banding pattern characteristic of other S. falcatula species. Finally, electron microscopy examining multiple merozoites of each of these eight horse isolates showed similar morphology, which differed from the grackle (Cornell) isolate. We conclude that the eight Michigan horse isolates are S. neurona species and the grackle isolate is an S. falcatula species.
Publication Date: 2001-02-27 PubMed ID: 11223197DOI: 10.1016/s0304-4017(00)00388-5Google Scholar: Lookup
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- Comparative Study
- Journal Article
- Research Support
- Non-U.S. Gov't
- Research Support
- U.S. Gov't
- Non-P.H.S.
Summary
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The research paper discusses a study comparing the isolates of a protozoan parasite, Sarcocystis neurona, derived from neural tissues of horses. The aim was to understand the differences and similarities between these isolates and possibly identify the species and lifecycle of this parasite that primarily affects horses.
Comparison Methods
- The researchers employed several comparison techniques on isolates derived from eight Michigan horses, a California horse, a grackle and five isolates from feral opossums from Michigan.
- SDS-PAGE analysis with silver staining was used to visually see proteins, providing insights into the protein makeup difference between species.
- Western blot analysis was done to verify the proteins identified by SDS-PAGE.
- PCR of DNA from merozoites (life stage parasites) to produce a specific 334bp PCR product following a previous study.
- Restriction Fragment Length Polymorphism (RFLP) was conducted to analyze the genetic variations in different isolates.
- Electron microscopy was also used to examine the morphology of different merozoites.
Key Findings
- Results demonstrated similar protein band patterns on SDS-PAGE and Western blot, among all eight equine-derived isolates, and the S. neurona isolate derived from the California horse.
- One band pattern from the Michigan horse isolates showed more prominent bands than the UCD1 isolate and other Michigan horse isolates, hinting at a potential differentiation.
- The DNA PCR product of 334bp was consistently found in all horse-derived isolates, including the grackle-derived isolate.
- RFLP analysis showed a band pattern characteristic for S. neurona in the horse isolates, while the grackle had patterns characteristic of another species, S. falcatula.
- The morphology of the merozoites from the horse isolates, observed through electron microscopy, was uniform and distinct from that of the grackle isolate.
Conclusion
Judging from the protein bands, DNA PCR product, RFLP band patterns and morphological characteristics, the scientists concluded that the eight Michigan-derived horse isolates were of S. neurona species, and the grackle isolate was of S. falcatula species.
Cite This Article
APA
Mansfield LS, Schott HC, Murphy AJ, Rossano MG, Tanhauser SM, Patterson JS, Nelson K, Ewart SL, Marteniuk JV, Bowman DD, Kaneene JB.
(2001).
Comparison of Sarcocystis neurona isolates derived from horse neural tissue.
Vet Parasitol, 95(2-4), 167-178.
https://doi.org/10.1016/s0304-4017(00)00388-5 Publication
Researcher Affiliations
- Department of Large Animal Clinical Sciences, College of Veterinary Medicine, Michigan State University, East Lansing 48824, USA. mansfie4@cvm.msu.edu
MeSH Terms
- Animals
- Blotting, Western / veterinary
- Electrophoresis, Polyacrylamide Gel / veterinary
- Encephalomyelitis / parasitology
- Encephalomyelitis / veterinary
- Horse Diseases / parasitology
- Horses
- Molecular Weight
- Nervous System / parasitology
- Opossums / parasitology
- Sarcocystis / classification
- Sarcocystis / isolation & purification
- Sarcocystosis / parasitology
- Sarcocystosis / veterinary
- Songbirds / parasitology
Citations
This article has been cited 7 times.- Dubey JP, Howe DK, Furr M, Saville WJ, Marsh AE, Reed SM, Grigg ME. An update on Sarcocystis neurona infections in animals and equine protozoal myeloencephalitis (EPM).. Vet Parasitol 2015 Apr 15;209(1-2):1-42.
- Kersh GJ, Lambourn DM, Self JS, Akmajian AM, Stanton JB, Baszler TV, Raverty SA, Massung RF. Coxiella burnetii infection of a Steller sea lion (Eumetopias jubatus) found in Washington State.. J Clin Microbiol 2010 Sep;48(9):3428-31.
- Elsheikha HM, Schott HC 2nd, Mansfield LS. Genetic variation among isolates of Sarcocystis neurona, the agent of protozoal myeloencephalitis, as revealed by amplified fragment length polymorphism markers.. Infect Immun 2006 Jun;74(6):3448-54.
- Elsheikha HM, Lacher DW, Mansfield LS. Phylogenetic relationships of Sarcocystis neurona of horses and opossums to other cyst-forming coccidia deduced from SSU rRNA gene sequences.. Parasitol Res 2005 Nov;97(5):345-57.
- Elsheikha HM, Murphy AJ, Mansfield LS. Phylogenetic congruence of Sarcocystis neurona Dubey et al., 1991 (Apicomplexa: Sarcocystidae) in the United States based on sequence analysis and restriction fragment length polymorphism (RFLP).. Syst Parasitol 2005 Jul;61(3):191-202.
- Elsheikha HM, Mansfield LS. Sarcocystis neurona major surface antigen gene 1 (SAG1) shows evidence of having evolved under positive selection pressure.. Parasitol Res 2004 Dec;94(6):452-9.
- Elsheikha HM, Murphy AJ, Fitzgerald SD, Mansfield LS, Massey JP, Saeed MA. Purification of Sarcocystis neurona sporocysts from opossum (Didelphis virginiana) using potassium bromide discontinuous density gradient centrifugation.. Parasitol Res 2003 Jun;90(2):104-9.
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