title: Experimental induction of equine protozoal myeloencephalitis in horses using Sarcocystis sp. sporocysts from the opossum (Didelphis virginiana).
abstract: Sarcocystis sp. sporocysts isolated from eight feral opossums (Didelphis virginiana) were pooled and fed to 18 commercially reared budgerigars (Melopsittacus undulatus), 14 wild-caught sparrows (Passer domesticus), one wild-caught slate-colored Junco (Junco hyemalis) and five weanling horses (Equus caballus). All budgerigars died within 5 weeks post inoculation (wpi). Histologic examination revealed meronts within the pulmonary epithelia and typical Sarcocystis falcatula sarcocysts developing in the leg muscles. Sparrows were euthanized 13 and 17 wpi and their carcasses were fed to four laboratory raised opossums. Sporocysts were detected in the feces of two opossums on 15 days post inoculation (dpi) and in a third opossum on 40 dpi. Fecal samples from the fourth opossum remained negative; however, sporocysts were found in intestinal digests from all four opossums. Sporocysts were not found in feces or intestinal digest of an additional opossum that was fed three uninoculated sparrows. Five foals were fed sporocysts (Foals 2, 3, 4, 5, and 7) and two foals were maintained as uninoculated controls (Foals 1 and 6). Sporocysts from two additional feral opossums also were fed to foals. Foal 5 was given 0.05 mg kg-1 dexamethasone sodium phosphate daily beginning 2 days before inoculation for a total of 2 weeks. Horse sera were tested three times per week, and cerebrospinal fluid (CSF) samples were tested biweekly for anti-Sarcocystis neurona antibodies by Western blot analysis. No foals had any S. neurona-specific antibodies by Western blot analysis prior to sporocysts ingestion. Seroconversion occurred in Foals 3, 5, and 7 by 24 dpi, followed by positive CSF tests on 28 dpi. Foals 2 and 4 seroconverted by 40 dpi. Cerebrospinal fluid from Foal 2 tested positive by 42 dpi, but Foal 4 remained seronegative throughout the study. Sera and CSF from control Foals 1 and 6 remained seronegative. All foals with positive CSF developed neurologic clinical signs. Neurologic disease was evident in Foals 2 and 3 by 42 dpi and in Foal 7 by 28 dpi. The severity of clinical signs progressed to marked spasticity, hypermetria and ataxia in Foal 7 by the end of the trial. Necropsy examination of inoculated foals did not reveal gross lesions; however, microscopic lesions consistent with equine protozoal myeloencephalitis (EPM) were found in Foals 2, 3, and 7. Protozoa were not observed in the tissue sections. Microscopic lesions consistent with EPM were not found in Foals 4 and 5 or in uninoculated control Foals 1 and 6. Foal 5 had unilateral non-inflammatory lesions in the cervical and thoracic spinal cord consistent with cord compression. These data indicate that the opossum is a definitive host of S. neurona.
authors.affiliation: Department of Veterinary Science, College of Agriculture, University of Kentucky, Lexington 40546, USA.
authors.forename: C K
authors.initials: CK
authors.lastname: Fenger
authors.forename: D E
authors.initials: DE
authors.lastname: Granstrom
authors.forename: A A
authors.initials: AA
authors.lastname: Gajadhar
authors.forename: N M
authors.initials: NM
authors.lastname: Williams
authors.forename: S A
authors.initials: SA
authors.lastname: McCrillis
authors.forename: S
authors.initials: S
authors.lastname: Stamper
authors.forename: J L
authors.initials: JL
authors.lastname: Langemeier
authors.forename: J P
authors.initials: JP
authors.lastname: Dubey
mesh_terms: D000818:Animals; D000835:Animals, Wild; D001717:Birds; D001933:Brain Stem / Q000469:parasitology / Q000473:pathology; D004679:Encephalomyelitis / Q000469:parasitology / Q000473:pathology / Q000503:physiopathology*; D006736:Horses; D007413:Intestinal Mucosa / Q000469:parasitology*; D008168:Lung / Q000469:parasitology; D018482:Muscle, Skeletal / Q000469:parasitology; D009893:Opossums / Q000469:parasitology*; D010311:Parrots; D012522:Sarcocystis / Q000302:isolation & purification*; D012523:Sarcocystosis / Q000473:pathology / Q000503:physiopathology* / Q000635:transmission