title: Ultrastructural and molecular confirmation of the development of Sarcocystis neurona tissue cysts in the central nervous system of southern sea otters (Enhydra lutris nereis).
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 microm 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.
authors.affiliation: Marine Wildlife Veterinary Care and Research Center, California Department of Fish and Game, 1451 Shaffer Road, Santa Cruz, CA 95060, USA. MMiller@ospr.dfg.ca.gov
authors.forename: M A
authors.initials: MA
authors.lastname: Miller
authors.forename: B C
authors.initials: BC
authors.lastname: Barr
authors.forename: R
authors.initials: R
authors.lastname: Nordhausen
authors.forename: E R
authors.initials: ER
authors.lastname: James
authors.forename: S L
authors.initials: SL
authors.lastname: Magargal
authors.forename: M
authors.initials: M
authors.lastname: Murray
authors.forename: P A
authors.initials: PA
authors.lastname: Conrad
authors.forename: S
authors.initials: S
authors.lastname: Toy-Choutka
authors.forename: D A
authors.initials: DA
authors.lastname: Jessup
authors.forename: M E
authors.initials: ME
authors.lastname: Grigg
mesh_terms: D000818:Animals; D001921:Brain / Q000469:parasitology*; D002490:Central Nervous System / Q000469:parasitology*; D003560:Cysts / Q000469:parasitology* / Q000648:ultrastructure; D008297:Male; D046529:Microscopy, Electron, Transmission; D010041:Otters / Q000469:parasitology*; D016133:Polymerase Chain Reaction / Q000662:veterinary; D012337:RNA, Ribosomal, 18S / Q000235:genetics; D012522:Sarcocystis / Q000235:genetics / Q000302:isolation & purification* / Q000648:ultrastructure; D012523:Sarcocystosis / Q000235:genetics / Q000635:transmission* / Q000662:veterinary; D012623:Seawater
chemical_list: D012337:RNA, Ribosomal, 18S