Ultrastructural and molecular confirmation of the development of Sarcocystis neurona tissue cysts in the central nervous system of southern sea otters (Enhydra lutris nereis)

M. A. Miller, B. C. Barr, R. Nordhausen, E. R. James, S. L. Magargal, M. Murray, Patricia A Conrad, S. Toy-Choutka, D. A. Jessup, M. E. Grigg

Research output: Contribution to journalArticle

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Abstract

In 2004, three wild sea otters were diagnosed with putative Sarcocystis neurona-associated meningoencephalitis by histopathology and immunohistochemistry. Schizonts, free merozoites and tissue cysts were observed in the brains of all three infected animals. Tissue cysts walls from sea otter 1 (SO1) stained positively using anti-S. neurona polyclonal antiserum. However, positive staining does not preclude infection by closely related or cross-reactive tissue cyst-forming coccidian parasites. Two immature tissue cysts in the brain of SO1 were examined using transmission electron microscopy. Ultrastructural features included cyst walls with thin villous projections up to 1 μm long with tapered ends and a distinctive, electron-dense outer lining layer composed of linearly-arranged, semi-circular structures with a "hobnailed" surface contour. Small numbers of microtubules extended down through the villi into the underlying granular layer. Metrocytes were short and plump with an anterior apical complex, 22 sub-pellicular microtubules, numerous free ribosomes and no rhoptries. Some metrocytes appeared to be dividing, with two adjacent nuclear profiles. Collectively these ultrastructural features were compatible with developing protozoal cysts and were similar to prior descriptions of S. neurona tissue cysts. Panspecific 18S rDNA primers were utilized to identify protozoa infecting the brains of these otters and DNA amplification and additional sequencing at the ITS1 locus confirmed that all three otters were infected with S. neurona. No other Sarcocystis spp. were detected in the brains or skeletal muscles of these animals by immunohistochemistry or PCR. We believe this is the first ultrastructural and molecular confirmation of the development of S. neurona tissue cysts in the CNS of any animal.

Original languageEnglish (US)
Pages (from-to)1363-1372
Number of pages10
JournalInternational Journal for Parasitology
Volume39
Issue number12
DOIs
StatePublished - Oct 2009

Fingerprint

Otters
Central Nervous System Cysts
Sarcocystis
Cysts
Brain
Microtubules
Immunohistochemistry
Schizonts
Merozoites
Meningoencephalitis
Ribosomal DNA
Transmission Electron Microscopy
Ribosomes
Immune Sera
Parasites
Skeletal Muscle
Electrons
Staining and Labeling
Polymerase Chain Reaction

Keywords

  • 18S rDNA
  • Brain
  • Central nervous system
  • ITS1
  • Sarcocystis neurona
  • Sea otter
  • Tissue cyst
  • Ultrastructure

ASJC Scopus subject areas

  • Parasitology
  • Infectious Diseases

Cite this

Ultrastructural and molecular confirmation of the development of Sarcocystis neurona tissue cysts in the central nervous system of southern sea otters (Enhydra lutris nereis). / Miller, M. A.; Barr, B. C.; Nordhausen, R.; James, E. R.; Magargal, S. L.; Murray, M.; Conrad, Patricia A; Toy-Choutka, S.; Jessup, D. A.; Grigg, M. E.

In: International Journal for Parasitology, Vol. 39, No. 12, 10.2009, p. 1363-1372.

Research output: Contribution to journalArticle

Miller, M. A. ; Barr, B. C. ; Nordhausen, R. ; James, E. R. ; Magargal, S. L. ; Murray, M. ; Conrad, Patricia A ; Toy-Choutka, S. ; Jessup, D. A. ; Grigg, M. E. / Ultrastructural and molecular confirmation of the development of Sarcocystis neurona tissue cysts in the central nervous system of southern sea otters (Enhydra lutris nereis). In: International Journal for Parasitology. 2009 ; Vol. 39, No. 12. pp. 1363-1372.
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abstract = "In 2004, three wild sea otters were diagnosed with putative Sarcocystis neurona-associated meningoencephalitis by histopathology and immunohistochemistry. Schizonts, free merozoites and tissue cysts were observed in the brains of all three infected animals. Tissue cysts walls from sea otter 1 (SO1) stained positively using anti-S. neurona polyclonal antiserum. However, positive staining does not preclude infection by closely related or cross-reactive tissue cyst-forming coccidian parasites. Two immature tissue cysts in the brain of SO1 were examined using transmission electron microscopy. Ultrastructural features included cyst walls with thin villous projections up to 1 μm long with tapered ends and a distinctive, electron-dense outer lining layer composed of linearly-arranged, semi-circular structures with a {"}hobnailed{"} surface contour. Small numbers of microtubules extended down through the villi into the underlying granular layer. Metrocytes were short and plump with an anterior apical complex, 22 sub-pellicular microtubules, numerous free ribosomes and no rhoptries. Some metrocytes appeared to be dividing, with two adjacent nuclear profiles. Collectively these ultrastructural features were compatible with developing protozoal cysts and were similar to prior descriptions of S. neurona tissue cysts. Panspecific 18S rDNA primers were utilized to identify protozoa infecting the brains of these otters and DNA amplification and additional sequencing at the ITS1 locus confirmed that all three otters were infected with S. neurona. No other Sarcocystis spp. were detected in the brains or skeletal muscles of these animals by immunohistochemistry or PCR. We believe this is the first ultrastructural and molecular confirmation of the development of S. neurona tissue cysts in the CNS of any animal.",
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AU - Murray, M.

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