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Purification of Sarcocystis neurona sporocysts from opossum (Didelphis virginiana) using potassium bromide discontinuous density gradient centrifugation

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Abstract

This report describes a new, inexpensive procedure for the rapid and efficient purification of Sarcocystis neurona sporocysts from opossum small intestine. S. neurona sporocysts were purified using a discontinuous potassium bromide density gradient. The procedure provides a source of sporocyst wall and sporozoites required for reliable biochemical characterization and for immunological studies directed at characterizing antigens responsible for immunological responses by the host. The examined isolates were identified as S. neurona using random amplified polymorphic DNA primers and restriction endonuclease digestion assays. This method allows the collection of large numbers of highly purified S. neurona sporocysts without loss of sporocyst viability as indicated by propidium iodide permeability and cell culture infectivity assays. In addition, this technique might also be used for sporocyst purification of other Sarcocystis spp.

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Fig. 1A, B
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Acknowledgements

We are grateful to Dr. Thomas S. Whittam, Microbial Evolution Laboratory, Dr. Harold C. Schott, Department of Large Animal Clinical Sciences, Michigan State University for their valuable suggestion and critical review of the manuscript. We thank Dr. Shirley Owens, Center for Advanced Microscopy at Michigan State University for her excellent assistance with photomicrography. This study was performed according to All University Committee on Animal Use and Care (AUCAUC), Michigan State University, and United States Department of Agriculture's approved guidelines for Animal Care.

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Authors and Affiliations

  1. Molecular Epidemiology Laboratory, Food Safety and Toxicology Center, Michigan State University, East Lansing, MI 48824, USA

    Hany M. Elsheikha & Mahdi A. Saeed

  2. Animal Health Diagnostic Laboratory, Michigan State University, East Lansing, MI 48824, USA

    Alice J. Murphy & Scott D. Fitzgerald

  3. Department of Microbiology and Molecular Genetics, Michigan State University, East Lansing, MI 48824, USA

    Linda S. Mansfield

  4. Molecular Biology Section, Michigan Department of Community Health, Lansing, MI 48909, USA

    Jeffrey P. Massey

Authors
  1. Hany M. Elsheikha
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  2. Alice J. Murphy
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  3. Scott D. Fitzgerald
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  4. Linda S. Mansfield
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  5. Jeffrey P. Massey
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  6. Mahdi A. Saeed
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Correspondence to Mahdi A. Saeed.

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Elsheikha, H.M., Murphy, A.J., Fitzgerald, S.D. et al. Purification of Sarcocystis neurona sporocysts from opossum (Didelphis virginiana) using potassium bromide discontinuous density gradient centrifugation. Parasitol Res 90, 104–109 (2003). https://doi.org/10.1007/s00436-002-0789-y

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  • DOI: https://doi.org/10.1007/s00436-002-0789-y

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