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Topic:Equine Protozoal Myeloencephalitis
Equine Protozoal Myeloencephalitis (EPM) is a neurological disease affecting horses, caused by the protozoan parasites Sarcocystis neurona or, less commonly, Neospora hughesi. The disease occurs when these parasites infect the central nervous system, leading to a range of neurological symptoms. Horses with EPM may exhibit signs such as ataxia, muscle weakness, and incoordination. Diagnosis can be challenging and typically involves a combination of clinical assessment, serological testing, and sometimes cerebrospinal fluid analysis. This page compiles peer-reviewed research studies and scholarly articles that explore the pathogenesis, diagnostic approaches, and management strategies for Equine Protozoal Myeloencephalitis in horses.
Inoculation of Sarcocystis neurona merozoites into the central nervous system of horses. Equine protozoal myeloencephalitis (EPM) is a neurologic syndrome in horses from the Americas and is usually caused by infection with the apicomplexan parasite, Sarcocystis neurona. A horse model of EPM is needed to test the efficacy of chemotherapeutic agents and potential vaccines. Five horses that were negative for antibodies to S. neurona in their serum and cerebrospinal fluid (CSF) were injected in the subarachnoid space with living merozoites of the SN2 isolate of S. neurona. None of the horses developed clinical disease or died over a 132-day observation period. All five horses develope...
Neospora hughesi: experimental infections in mice, gerbils, and dogs. Neospora hughesi is a recently described cause of equine protozoal myeloencephalitis (EPM). A rodent model for pathogenicity would facilitate development of therapies to be used in horses. In the present study, we examined the susceptibility of BALB/c gamma-interferon gene knockout (gamma-INFKO), BALB/c, CD-1, and C57BL/6 strains of mice and gerbils to infection with tachyzoites of the Nh-A1 strain of N. hughesi isolated from a horse from AL, USA. Only the gamma-IFNKO mice developed severe clinical disease following infection with N. hughesi and died 19-25 days after infection and exhibited se...
Cerebrospinal fluid and blood concentrations of toltrazuril 5% suspension in the horse after oral dosing. Toltrazuril 5% suspension (Baycox, Bayer Canada, Ontario, Canada) was administered to six adult horses followed by blood collection and assay to determine the concentration of toltrazuril and its principal metabolites, toltrazuril sulfone and toltrazuril sulfoxide. From this data, the maximum concentration (C(max)), elimination half-life (T 1/2), and mean residence times of the plasma were determined from standard pharmacokinetic formulas. After a single oral dose of 10 mg/kg body weight a rapid absorption was found, with a mean peak serum concentration of 11.17 mg/L at 18 hours. Elimination w...
Determination of the activity of diclazuril against Sarcocystis neurona and Sarcocystis falcatula in cell cultures. Diclazuril is a benzeneacetonitril anticoccidial that has excellent activity against the extraintestinal stages of Toxoplasma gondii and Neospora caninum. It also is highly active against intestinal coccidia of poultry. The present study examined the efficacy of diclazuril in inhibiting merozoite production of Sarcocystis neurona and Sarcocystis falcatula in bovine turbinate cell cultures. Diclazuril inhibited merozoite production by more than 80% in cultures of S. neurona or S. falcatula treated with 0.1 ng/ml diclazuril and greater than 95% inhibition of merozoite production was observed whe...
Improvement of western blot test specificity for detecting equine serum antibodies to Sarcocystis neurona. Equine protozoal myeloencephalitis (EPM) is a neurological disease of horses and ponies caused by the apicomplexan protozoan parasite Sarcocystis neurona. The purposes of this study were to develop the most stringent criteria possible for a positive test result, to estimate the sensitivity and specificity of the EPM Western blot antibody test, and to assess the ability of bovine antibodies to Sarcocystis cruzi to act as a blocking agent to minimize false-positive results in the western blot test for S. neurona. Sarcocystis neurona merozoites harvested from equine dermal cell culture were heat ...
Diclazuril in the horse: its identification and detection and preliminary pharmacokinetics. Diclazuril (4-chlorophenyl [2,6-dichloro-4-(4,5-dihydro-3H-3,5-dioxo-1,2,4-triazin-2-yl)pheny l] acetonitrile), is a benzeneacetonitrile antiprotozoal agent (Janssen Research Compound R 64433) marketed as Clinacox . Diclazuril may have clinical application in the treatment of Equine Protozoal Myeloencephalitis (EPM). To evaluate its bioavailability and preliminary pharmacokinetics in the horse we developed a sensitive quantitative high-pressure liquid chromatography (HPLC) method for diclazuril in equine biological fluids. MS/MS analysis of diclazuril in our HPLC solvent yielded mass spectral ...
Clinical pharmacology of nervous system diseases. The well-developed defense barriers of the CNS and the expense of drug therapy limit the pharmacologic options for the treatment of neurologic diseases in horses. New approaches to controlling inflammation in the CNS are improving the outcomes of bacterial meningitis. The appropriate treatment of EPM remains controversial. More research is needed to evaluate the pharmacokinetics and pharmacodynamics of drugs in the CNS of the horse. Behavioral pharmacology has become fashionable in human and small animal medicine, but it needs to be evaluated for the potential of unethical use in performance h...
Simplified technique for isolation, excystation, and culture of Sarcocystis species from opossums. Sarcocystis neurona is a protozoan parasite that causes a neurological disease in horses called equine protozoal myeloencephalitis. The route of transmission is speculated to be by fecal-oral transfer of sporocysts shed from opossums. Controversy exists regarding both the natural life cycle for this parasite as well as the species identity of opossum Sarcocystis. To provide stage-specific material for species comparison, 27 opossums from southern Michigan were screened for Sarcocystis spp. sporocysts. Seven opossums were positive for Sarcocystis sporocysts by fecal flotation. A simplified, eff...
Characterization of a Sarcocystis neurona isolate (SN6) from a naturally infected horse from Oregon. An isolate of Sarcocystis neurona (SN6) was obtained from the spinal cord of a horse from Oregon with neurologic signs. The parasite was isolated in cultures of bovine monocytes and equine spleen cells. The parasite divided by endopolygeny and completed at least one asexual cycle in cell cultures in three days. Two gamma interferon knockout mice inoculated with cell culture-derived merozoites became ill 35 d later and S. neurona schizonts and merozoites were found in encephalitic lesions. The parasite in tissue sections of mice reacted with S. neurona-specific antibodies and S. neurona was rei...
Serologic prevalence of Sarcocystis neurona, Toxoplasma gondii, and Neospora caninum in horses in Brazil. To determine serologic prevalence of Sarcocystis neurona, Toxoplasma gondii, and Neospora caninum in horses in Brazil. Methods: Prevalence survey. Methods: 101 Thoroughbreds in Brazil. Methods: Blood samples were obtained from horses and tested for serum antibodies against S neurona by use of an immunoblot procedure with culture-derived S neurona merozoites as antigen, and for serum antibodies against T gondii and N caninum by use of a modified agglutination test with formalin-preserved tachyzoites and mercaptoethanol. Results: Antibodies against S neurona and T gondii were detected in 36 and ...
Sudden death of two horses associated with pulmonary aspergillosis. The sudden death of two horses was attributed to the rapid and acute development of pulmonary aspergillosis. One horse was making excellent postoperative progress after a jejunal resection and anastomosis for intestinal adhesions. The other horse was being treated routinely for equine protozoal myeloencephalitis (EPM). Signs of fever and an increased respiratory rate were detected shortly before death in the first horse, but no premonitory clinical signs characteristic of pulmonary infection were detected in the horse being treated for EPM. Both horses developed rapidly debilitating, acute pul...
Determination of the activity of pyrimethamine, trimethoprim, sulfonamides, and combinations of pyrimethamine and sulfonamides against Sarcocystis neurona in cell cultures. Equine protozoal myeloencephalitis (EPM) is a neurologic syndrome in horses from the Americas and is usually caused by infection with the apicomplexan parasite, Sarcocystis neurona. The activities of pyrimethamine, trimethoprim, sulfachloropyridazine, sulfadiazine, sulfadimethoxine, sulfamethoxazole, sulfamethazine, and sulfathiazole were examined against developing S. neurona merozoites in bovine turbinate cell cultures. A microtiter plate host cell lesion based assay was used to determine the effects of agents on developing merozoites. A cell culture flask assay was used to determine if sele...
Are Sarcocystis neurona and Sarcocystis falcatula synonymous? A horse infection challenge. Equine protozoal myeloencephalitis (EPM) is a debilitating neurologic disease of the horse. The causative agent. Sarcocystis neurona, has been suggested to be synonymous with Sarcocystis falcatula, implying a role for birds as intermediate hosts. To test this hypothesis, opossums (Didelphis virginiana) were fed muscles containing S. falcatula sarcocysts from naturally infected brown-headed cowbirds (Molothrus ater). Ten horses were tested extensively to ensure no previous exposure to S. neurona and were quarantined for 14 days, and then 5 of the horses were each administered 10(6) S. falcatula...
Multiple DNA markers differentiate Sarcocystis neurona and Sarcocystis falcatula. Studies designed to investigate the causative agent of equine protozoal myeloencephalitis and its life cycle have been hampered by the marked similarity of Sarcocystis neurona to other Sarcocystis spp. present in the same definitive host. Random-amplified polymorphic DNA techniques were used to amplify DNA from isolates of S. neurona and Sarcocystis falcatula. DNA sequence analysis of polymerase chain reaction (PCR) products was then used to design PCR primers to amplify specific Sarcocystis spp. DNA products. The ribosomal RNA internal transcribed spacer was also amplified and compared betwee...
Spinal muscle pathology. Clinical signs that are easily referred to spinal muscle pathology include atrophy of epaxial muscles, focal swelling and palpable tenderness, as well as enlarged muscles with increased tone. Less easily recognized signs include rigidity of the spine, shortened stride, hindlimb lameness, and indicators of poor performance. Muscle biopsy is one option in evaluating sore backs and is best used when physical examination and imaging procedures do not reveal a likely diagnosis or when conventional treatments are unsuccessful. Rhabdomyolysis of spinal muscles may be due to nutritional myodegeneratio...
Neospora caninum-associated equine protozoal myeloencephalitis. Equine protozoal myeloencephalitis (EPM) was clinically diagnosed in a 20-year-old horse with severe ataxia. The cerebrospinal fluid was positive for Sarcocystis neurona antibodies by western blot. The horse was administered corticosteroids to facilitate in vitro culture of S. neurona from its spinal cord following necropsy. Microscopic lesions of EPM were present in the brain and in the spinal cord, including multifocal inflammatory cellular infiltrates and several large groups of protozoa. Immunohistochemical, and light and electron microscopic examinations revealed that the protozoa were Ne...
Evidence that surface proteins Sn14 and Sn16 of Sarcocystis neurona merozoites are involved in infection and immunity. Sarcocystis neurona is the etiologic agent of equine protozoal myeloencephalitis (EPM). Based on an analysis of 25,000 equine serum and cerebrospinal fluid (CSF) samples, including samples from horses with neurologic signs typical of EPM or with histologically or parasitologically confirmed EPM, four major immunoblot band patterns have been identified. Twenty-three serum and CSF samples representing each of the four immunoblot patterns were selected from 220 samples from horses with neurologic signs resembling EPM and examined for inhibitory effects on the infectivity of S. neurona by an in vi...
Congenital defects in newborn foals of mares treated for equine protozoal myeloencephalitis during pregnancy. Three weak, recumbent neonatal foals with skin lesions, including a thin wooly coat, were born to mares being treated for equine protozoal myeloencephalitis. Mares received sulfadiazine or sulfamethoxazole-trimethoprim, pyrimethamine, folic acid, and vitamin E orally. Foals were anemic, leukopenic, azotemic, hyponatremic, and hyperkalemic. Serum folate concentrations in the 3 foals and 2 mares were lower than those reported in the literature for clinically normal brood mares. Treatment was unsuccessful. For each foal, necropsy revealed lobulated kidneys with thin cortices and a pale medulla, a...
Management of headshaking in three horses by treatment for protozoal myeloencephalitis. Unlike the incidence of equine protozoal myeloencephalitis (EPM), which appears to be increasing, headshaking is an uncommon problem for horses in Missouri and the adjacent states. Equine protozoal myeloencephalitis was incriminated in three horses examined for the treatment of headshaking on the basis of a neurological examination, an analysis of cerebrospinal fluid and their response to treatment. The headshaking and stereotypical behaviour associated with EPM was successfully treated with potentiated sulphonamides and pyrimethamine.
Micropreparative high resolution purification of proteins by a combination of sodium dodecyl sulfate-polyacrylamide gel electrophoresis, isoelectric focusing, and membrane blotting. We report a simple, economical, and efficient protocol for protein purification from cells. First, proteins of cell lysates were separated by standard sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and electroblotted to protein-blotting membrane. The blots were stained with Coomassie blue or developed by immunoblotting to visualize specific proteins. The bands corresponding to those visible by immunoblotting were excised from the dye-stained blots and subjected to isoelectric focusing. The focused gel was stained with Coomassie blue. Finally, the stained bands were excise...
Severe polysaccharide storage myopathy in Belgian and Percheron draught horses. A severe myopathy leading to death or euthanasia was identified in 4 Belgian and 4 Percheron draught horses age 2-21 years. Clinical signs ranged from overt weakness and muscle atrophy in 2 horses age 2 and 3 years, to recumbency with inability to rise in 6 horses age 4-21 years. In 5 horses there was mild to severe increases in muscle enzyme levels. Clinical diagnoses included equine motor neuron disease (2 horses), post anaesthetic myopathy (2 horses), exertional myopathy (2 horses), myopathy due to unknown (one horse), and equine protozoal myelitis (one horse). Characteristic histopathology...
Epizootic of equine protozoal myeloencephalitis on a farm. To determine the clinical findings, course of treatment, and long-term outcome of horses on a farm in central Kentucky during an epizootic of equine protozoal myeloencephalitis (EPM). Methods: Cohort study. Methods: 21 horses on a farm in central Kentucky, 12 of which developed clinical signs of EPM. Methods: Horses on the farm were serially examined for signs of neurologic disease and serum and CSF antibodies to Sarcocystis neurona. Horses were considered to have EPM if they had neurologic signs and positive test results for antibodies to S neurona in CSF. Blood values were monitored for evid...
Seroprevalence of antibodies to Sarcocystis neurona in horses residing in Oregon. To determine seroprevalence of antibodies to Sarcocystis neurona in neurologically normal horses residing in 4 regions of Oregon and to describe the effects of age, gender, breed, and housing on seroprevalence within each region. Methods: Prevalence survey. Methods: Serum samples from 334 horses systematically selected by practicing veterinarians. Methods: Antibodies to S neurona were measured in sera, using a western blot. Information including age, gender, breed, housing, geographic location, and duration of residence was obtained for each horse. Data were analyzed, using descriptive statist...
Seroprevalence of antibodies to Sarcocystis neurona in horses residing in Ohio. To determine the seroprevalence of serum antibodies to Sarcocystis neurona in horses residing in Ohio. Methods: Prevalence survey. Methods: Serum from samples from 1,056 horses. Serum was collected on every 36th sample submitted to the Ohio State Diagnostic Laboratory for testing for equine infectious anemia. Methods: Serum was frozen at -80 C and analyzed for antibodies to S neurona, using a western blot. Information regarding blood sample collection, age, breed, sex, and geographic location was recorded for each horse. Data were analyzed, using multivariable logistic regression. Results: Hor...
Seroprevalence of antibodies to Sarcocystis neurona in horses residing in a county of southeastern Pennsylvania. To determine seroprevalence of Sarcocystis neurona-specific antibodies in a population of horses residing in Chester County, Pa. Methods: Prevalence survey. Methods: 117 serum samples from selected members of a population of 580 Thoroughbred horses. Methods: Serum was analyzed for antibodies to Sarcocystic neurona, using a western blot. Information regarding age, sex, and housing of horse was obtained by questionnaire. Data were analyzed, using multivariable logistic regression. Results: Seroprevalence was 45.3% (95% CI, 36.3 to 54.3%). A relationship was not found between seroprevalence and s...
Experimental induction of equine protozoal myeloencephalitis in horses using Sarcocystis sp. sporocysts from the opossum (Didelphis virginiana). 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 labora...
Neosporosis as a cause of equine protozoal myeloencephalitis. Neosporosis was diagnosed in an 11-year-old Quarter Horse gelding with clinical signs and diagnostic test results compatible with equine protozoal myeloencephalitis (EPM). Presumptive postmortem diagnosis of EPM attributable to Sarcocystis neurona infection is generally made on the basis of detecting an antibody titer to S neurona in the CSF or characteristic histologic lesions, even when parasites have not been specifically identified. Neosporosis was confirmed in the horse described here by use of immunohistochemical examination, in vitro culturing, and ultrastructural and molecular characte...
