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.
Bethel M.EPM remains particularly rampant in racing populations due to transport stress, young age, and congregate housing conditions. For compounding pharmacists, understanding that we're managing chronic parasite suppression - not achieving cure - fundamentally changes our approach to formulation strategy, stability assessment, and client counseling. The two formulations presented separately represent years of refinement, demonstrating compounding's essential role when commercial products fall short.
Qin S, Kulabieke T, Mizhamuhan D, Zhang M, Jin M, Abula G, Pi M, Wang H, Zhang Y, Guo Q.Equine theileriosis, caused by the protozoan parasite , represents a significant economic threat to the equine industry. In Xinjiang, China, an endemic area for this disease, systematic research on the genetic diversity of has been notably lacking. The aim of this study was to obtain molecular epidemiological data pertaining to the parasite in selected regions of Xinjiang, China, and analyze the genetic characteristics (including rRNA, , and genes) and genotype distribution patterns of isolates from these regions, providing a scientific basis for developing targeted prevention and control s...
Alsultan A, Karim SM, Al-Saadi M, Alsallami D, Ben Said M, Belkahia H.Equine piroplasmosis (EP), caused by the intracellular protozoa Theileria equi, Babesia caballi, and Theileria haneyi, represents a major health and economic threat to the equine industry worldwide. Existing diagnostic methods, including PCR, serology, and microscopy, are constrained by their dependence on specialized equipment, lengthy protocols, and the requirement for skilled personnel. Objective: This study aimed to develop a rapid, accurate, and field-deployable molecular diagnostic assay for T. equi. Methods: A nucleic acid-based diagnostic platform combining recombinase polymerase ampli...
Soliman AM, Elhawary NM, Helmy NM, El-Seify MA, Amer MM, Mohamed S, Memon FU, Rashid MHO, Gadelhaq SM.Equine piroplasmosis represents one of the main and serious health problems affecting the equines industry globally, caused by a tick-borne protozoa called and . This study aimed to identify and genotype within the equine population from Giza Governorate, Egypt, by comparing the obtained results using the available diagnostic methods. Unassigned: We collected 116 apparently healthy horses from the study area during the first half of 2019 to identify using real-time PCR (qPCR), targeting the gene. The results were compared with those from microscopic examination of Giemsa-stained blood sme...
Machado RZ, André MR, Pereira JG, Oliveira MDSC, Ribeiro LSDS, Toledo CZP, Gonçalves LR.Equine Piroplasmosis (EP) is a tick-borne disease caused by the protozoan parasites Babesia caballi, Theileria equi, and Theileria haneyi, characterized by intravascular hemolysis and associated systemic illness. Although T. equi and B. caballi have been widely reported in some regions of Brazil, data from other states are limited. Additionally, despite reports of T. equi genotype C, currently recognized as T. haneyi, has been identified in Brazil, there are no investigations using T. haneyi-specific molecular tools. This study assessed the presence of these three agents in horses from Baixada...
Rocha IA, McPeek JL, Hulverson MA, Choi R, Orr K, Sponseller BT, Kennedy T, Ojo KK, Barrett LK, Ruby R, Janes J, Reed SM, Arnold SLM, Van Voorhis WC....To determine the systemic and CNS distribution of bumped-kinase inhibitors (BKIs) in healthy horses, assess potential side effects, and identify a candidate compound for a clinical trial in equine protozoal myeloencephalitis (EPM) cases. Unassigned: 9 pharmacokinetic (PK) experiments were conducted from March 2021 through November 2024. Bumped-kinase inhibitors 1708, 1748, and 1841 were screened in Sarcocystis neurona growth assays and IV PKs, followed by investigation of a lead BKI compound by single-dose or multiday oral administration. Serial plasma collections were performed to assess syst...
Krücken J, Diekmann I, Andreotti S, Bredtmann CM, Mbedi S, Sparmann S, Schmidt JS, de Almeida Borges F, de Freitas MG, Sallé G, Hofer H....Equines are parasitized by complex communities of Strongylidae (Nematoda) comprising multi-species infections. Currently, Cyathostominae are most prevalent, while Strongylus species are only rarely detected. Since eggs and, in most cases, infective larvae cannot be differentiated to species level, except for Strongylus spp., species-specific knowledge of the pathology, epidemiology and ecology of these parasitic nematodes is limited. Reference sequence data for several cyathostomin species are limited or missing. Deep amplicon sequencing of internal transcribed spacer 2 (ITS-2) regions of nema...
Angwin CJ, de Assis Rocha I, Reed SM, Morrow JK, Graves A, Howe DK.Equine protozoal myeloencephalitis (EPM) caused by Sarcocystis neurona is one of the most important neurological diseases of horses in the Americas. While seroprevalence of S. neurona in horses is high, clinical manifestation of EPM occurs in less than 1 % of infected horses. Antemortem diagnosis has proven challenging as serum antibodies against S. neurona are an indicator of infection but not necessarily disease. Factors governing the occurrence of EPM are largely unknown, although horse immunity might contribute to EPM pathogenesis. Immunoglobulin G is the predominant antibody class in eq...
Valderrama-Martinez C, Packham A, Smith W, Mendoza-Flores JE, Zheng S, Chigerwe M, Plancarte M, Aleman M.Long-term freezing storage can alter the stability of proteins, thereby compromising accurate determination of indirect fluorescent antibody test (IFAT) titers that support the diagnosis of equine protozoal myeloencephalitis. Objective: Assess the effect of long-term storage at -80°C on IFAT against S. neurona and N. hughesi in equine serum and cerebrospinal fluid (CSF). Methods: Paired serum and CSF (n = 46), and serum only (n = 25) samples. Methods: Prospective study of samples stored 6-12, 13-18, and 19-24 months. Comparing antibody titers across time points, McNemar and Wilcoxon...
Duaso J, Perez-Ecija A, Navarro A, Martínez E, De Las Heras A, Mendoza FJ.Equine piroplasmosis (EP), caused by and , is a worldwide tick-borne disease with severe economic, commercial, and sanitary implications for equids. Although diagnosis is based on direct (blood smear or PCR) or indirect (serology) methods, these techniques are expensive, laborious, and false-negative and false-positive results can be yielded. Biochemistry blood profiles are routinely performed in horses. Biochemical parameters and ratios could be a reliable complementary diagnostic tool to assist clinicians in EP diagnosis, mainly in endemic areas, or for discarding similar disorders (piro-li...
Nadal C, Chanet C, Delaunay C, Pitel PH, Marsot M, Bonnet SI.Equine piroplasmosis, caused by the protozoan parasites Babesia caballi and Theileria equi, is endemic in Europe's Mediterranean basin, creating significant health and economic challenges for the equine sector. With no available vaccine, an eco-epidemiological approach is essential in order to identify and implement effective preventive measures. With this aim in view, we identified risk factors associated with B. caballi and T. equi infections and tick infestation for draught horses in the Auvergne-Rhône-Alpes region of France, known for its high piroplasmosis seroprevalence. During the spri...
Valderrama-Martinez C, Packham A, Zheng S, Smith W, Plancarte M, Aleman M.Evaluating antibody titers for Sarcocystis neurona for the diagnosis of equine protozoal myeloencephalitis from serum samples is a common practice. However, ensuring timely and proper refrigeration is not always possible. Objective: To evaluate immunofluorescent antibody (IFA) titers for S. neurona from serum samples stored at room temperature and 4°C. Methods: Twenty-two serum samples. Methods: Prospective longitudinal study. Two serum aliquots of 1 mL each were stored at room temperature (20-23.3°C) and 4°C. The unrefrigerated aliquot was immediately tested for IFA titers. Both aliquots...
de Albuquerque CV, da Silva Andrade M, de Freitas MS, Paulino PG, Santos HA, de Tarso Landgraf Botteon P.Equine Piroplasmosis (EP) and Equine Granulocytic Anaplasmosis (EGA) are diseases that affect horses, transmitted by ixodid ticks, causing a nonspecific febrile syndrome. Equine Piroplasmosis is endemic in Brazil, and most horses are in enzootic stability. Serological and molecular studies carried out on horses in Brazil have shown the presence of Anaplasma phagocytophilum, however, the clinical relevance of this infection has not yet been established. The present study aims to evaluate the importance of Babesia caballi, Theileria equi, and A. phagocytophilum as etiological agents in horses wi...
Adam E, Scollay M, Howard B, Taormina J, Hartmann P, Knych H.Levamisole is a regulated substance sometimes administered to racehorses to treat equine protozoal myelitis. Metabolites include compound II, aminorex, and pemoline. Aminorex and pemoline are Horseracing Integrity and Safety Authority-banned substances. Previous studies have examined single doses of the drug. This study examined the disposition of levamisole after 7 days of dosing. Methods: 6 healthy Thoroughbred geldings. Methods: Horses were treated with 500 mg (approx 0.91 to 1 mg/kg) of compounded levamisole hydrochloride paste PO every 12 hours for a total of 13 doses over 7 days. Serum a...
Pandit PS, Smith WA, Finno CJ, Aleman M, Conrad PA, Packham A, Plancarte M, Woolard K, Marsh A, Pusterla N.Equine protozoal myeloencephalitis (EPM) is a challenging disease to diagnose in horses with neurological signs. To optimize contemporary diagnostic testing, including the use of serum:CSF antibody ratios, the SarcoFluor antibody test for Sarcocystis neurona requires revalidation. The SarcoFluor, a previously validated immunofluorescent antibody test (IFAT) for the detection of antibodies specific to S. neurona in serum and cerebrospinal fluid (CSF) of naturally infected horses was analyzed using recent data and considering a serum:CSF antibody ratio threshold. Utilization of serum and CSF pho...
Palmisano M, Kulp J, Bender S, Stefanovski D, Robinson M, Johnson A.Eight-hydroxy-2'-deoxyguanosine (8-OHdG), a biomarker of oxidative damage evaluated in human neurodegenerative disease, has potential to correlate with postmortem diagnosis of neuroaxonal dystrophy/degenerative myeloencephalopathy (NAD/DM) in horses. Objective: We hypothesized that 8-OHdG will be higher in CSF and serum from NAD/DM horses compared with horses with other neurologic diseases (CVSM, EPM) and a control group of neurologically normal horses. We also hypothesized that 8-OHdG will be higher in CSF compared with serum from NAD/DM horses. Methods: Fifty client-owned horses with postmor...
Enriquez CK, Morrow JK, Graves A, Johnson A.Equine protozoal myeloencephalitis (EPM) caused by Sarcocystis neurona remains an antemortem diagnostic challenge in some horses. Recent work suggested the use of real-time PCR (rtPCR) on cerebrospinal fluid (CSF) as a promising diagnostic tool. Objective: To evaluate the sensitivity and specificity of S. neurona rtPCR on CSF for EPM diagnosis using horses with EPM and S. neurona-seropositive horses with other neurologic conditions. Methods: Ninety-nine horses with neurologic disease that underwent complete neurologic examination, CSF collection, and, if euthanized, necropsy including the cent...
Equine protozoal myeloencephalitis (EPM) is a neurological disease caused by Sarcocystis neurona. Immunofluorescence antibody tests (IFATs) have been widely used to identify exposure of horses to S. neurona in Brazil. Here we used IFAT to search for IgG antibodies against Sarcocystis falcatula-like (Dal-CG23) and S. neurona (SN138) in sera from 342 horses sampled in Campo Grande, Mato Grosso do Sul state (Midwestern), and São Paulo, São Paulo state (Southeastern), Brazil. The 1:25 cutoff value was chosen to maximize sensitivity of the test. IgG antibodies against S. neurona were detected in ...
Bernardino PN, Pusterla N, Conrad PA, Packham AE, Tamez-Trevino E, Aleman M, James K, Smith WA.Among the recognized neurologic diseases in horses, equine protozoal myeloencephalitis (EPM) has been reported around the world and still presents challenges in diagnosis and treatment. Horses can present with clinical neurologic signs consistent with EPM while testing negative for the two main causative agents, Sarcocystis neurona or Neospora hughesi, and may still be clinically responsive to anti-parasitic drug therapy. This context led to our hypothesis that another protozoal parasite, Toxoplasma gondii, which is known to cause toxoplasmosis in other mammalian species, is a potential pathog...
Pusterla N, Vaala W, Bain FT, Chappell DE, Craig B, Schneider C, Barnett DC, Gaughan E, Papich MG.Equine protozoal myeloencephalitis (EPM) has remained a devastating neurological disease of the Americas, especially in young performance horses. Prophylactic treatment strategies with diclazuril have shown to reduce seroprevalence and titer levels to Sarcocystis neurona in healthy horses continuously exposed to the apicomplexan parasite. The goal of this study was to determine if the FDA-labeled dose of 1 mg/kg of 1.56% diclazuril (Protazil) given once weekly to healthy adult horses would achieve steady-state concentrations in plasma known to be inhibitory to S. neurona in cell culture. Five ...
Journée HL, Journée SL.Depending on the localization of the lesion, spinal cord ataxia is the most common type of ataxia in horses. Most prevalent diagnoses include cervical vertebral stenotic myelopathy (CVSM), equine protozoal myeloencephalitis (EPM), trauma and equine degenerative myeloencephalopathy (EDM). Other causes of ataxia and weakness are associated with infectious causes, trauma and neoplasia. A neurologic examination is indispensable to identify the type of ataxia. In addition, clinical neurophysiology offers tools to locate functional abnormalities in the central and peripheral nervous system. Clinical...
MacKay RJ, Howe DK.Advances in the understanding of equine protozoal myeloencephalitis (EPM) are reviewed. It is now apparent that EPM can be caused by either of 2 related protozoan parasites, Sarcocystis neurona and Neospora hughesi, although S neurona is the most common etiologic pathogen. Horses are commonly infected, but clinical disease occurs only infrequently; the factors influencing disease occurrence are not well understood. Epidemiologic studies have identified risk factors for the development of EPM, including the presence of opossums and prior stressful health-related events. Attempts to reproduce EP...
MacKay RJ, Howe DK.Advances in the understanding of equine protozoal myeloencephalitis (EPM) are reviewed. It is now apparent that EPM can be caused by either of 2 related protozoan parasites, Sarcocystis neurona and Neospora hughesi, although S neurona is the most common etiologic pathogen. Horses are commonly infected, but clinical disease occurs only infrequently; the factors influencing disease occurrence are not well understood. Epidemiologic studies have identified risk factors for the development of EPM, including the presence of opossums and prior stressful health-related events. Attempts to reproduce EP...
Abisheva A, Abishov A, Khairullaeva K, Shynybayev K, Kalissynov B, Maikhin K, Kydyrmanov A, Karamendin K, Valdovska A, Syrym N.Equine rhinopneumonitis is an acute, highly contagious disease found virtually worldwide. The purpose of the studies presented in this paper is to develop a technology for the manufacture of a cell-derived equine rhinopneumonitis vaccine, as well as to assess the safety and immunogenicity of the newly developed vaccine in laboratory animals model. The object of the studies was the AK-2011 strain isolated from the horses suffering from rhinopneumonitis during an outbreak of abortions. The viability of the AK-2011 strain was assessed using a continuous line of calf trachea cells, a continuous li...
Hay AN, Wagner B, Leeth CM, LeRoith T, Cecere TE, Lahmers KK, Andrews FM, Werre SR, Johnson AL, Clark CK, Pusterla N, Reed SM, Lindsay DS, Taylor S....Equine protozoal myeloencephalitis (EPM) is a debilitating neurologic disease affecting horses across the Americas. Gaps in understanding the inflammatory immune response in EPM-affected horses create difficulties with diagnosis and treatment, subsequently negatively impacting the prognosis of affected horses. The purpose of the current study was to evaluate circulating levels of the inflammatory immune marker soluble CD14 (sCD14), in horses with EPM (n = 7) and determine if they differed from healthy neurologically normal horses (n = 6). Paired sera and cerebrospinal fluid (CSF) samples were ...
Hay AN, Potter A, Lindsay D, LeRoith T, Zhu J, Cashwell S, Witonsky S, Leeth C.Sarcocystis neurona is the predominant etiological agent of the infectious equine neurologic disease, equine protozoal myeloencephalitis (EPM), which is prevalent in the United States. A wealth of knowledge about S. neurona biology and its life cycle has accumulated over the last several decades. However, much remains unknown about the aberrant equine host's immune response to S. neurona and the relatively high prevalence of exposure to the protozoa but relatively infrequent occurrence of clinical neurologic disease. Mouse models simulating EPM are commonly used to study the disease due to num...
Ruppert S, Lee JK, Marsh AE.A 6-year-old female captive zebra (Equus zebra) had a three-year history of slow progressive neurologic signs that recently worsened with hind limb ataxia, head tilt, and circling. Gross examination including the brain and spinal cord were unremarkable. On histopathology, the brain and brainstem had multiple random areas of severe lymphoplasmacytic meningoencephalitis associated with numerous 15-25 μm in diameter protozoal cysts with a discernible outer wall containing numerous 2 × 4 μm oval to crescent-shaped organisms. Immunohistochemistry and PCR identified the presence of Neospora or...
Llano HAB, Soares RM, Acevedo-Gutierrez LY, Rodas JD, Polo G, Borges-Silva W, Jesus RF, Gondim LFP.Sarcocystis neurona and Neospora spp. are related protozoa that can cause equine protozoal myeloencephalitis (EPM). The present study aimed to determine the frequency of antibodies to these parasites in 649 equids (351 horses, 267 donkeys, and 31 mules) from six departments in the North and Northwest of Colombia. For this purpose, the indirect fluorescent antibody test (IFAT) was used for detecting antibodies against S. neurona and Neospora spp. with a cut-off point of 1:20 and 1:50, respectively. A binomial logistic regression model was selected to predict variables associated with exposure. ...
Bernardino PN, Smith WA, Conrad PA, Packham AE, Tamez-Trevino E, Barnum S, Pusterla N.An ante-mortem diagnosis of equine protozoal myeloencephalitis (EPM) is presently based on clinical presentation, immunodiagnostics performed on serum and cerebrospinal fluid (CSF), and ruling out other neurological disorders. Molecular techniques introduce a novel and promising approach for the detection of protozoal agents in CSF. Hypothesizing that real-time PCR (rtPCR) can be a useful complement to EPM diagnostics, 210 CSF samples from horses suspected of neurological disease with EPM included as a differential diagnosis were tested using rtPCR to detect Sarcocystis neurona DNA and immunod...
Dubey JP, Lindsay DS, Saville WJ, Reed SM, Granstrom DE, Speer CA.Equine protozoal myeloencephalitis (EPM) is a serious neurological disease of horses in the Americas. The protozoan most commonly associated with EPM is Sarcocystis neurona. The complete life cycle of S. neurona is unknown, including its natural intermediate host that harbors its sarcocyst. Opossums (Didelphis virginiana, Didelphis albiventris) are its definitive hosts. Horses are considered its aberrant hosts because only schizonts and merozoites (no sarcocysts) are found in horses. EPM-like disease occurs in a variety of mammals including cats, mink, raccoons, skunks, Pacific harbor seals, p...
Blazejewski T, Nursimulu N, Pszenny V, Dangoudoubiyam S, Namasivayam S, Chiasson MA, Chessman K, Tonkin M, Swapna LS, Hung SS, Bridgers J....Sarcocystis neurona is a member of the coccidia, a clade of single-celled parasites of medical and veterinary importance including Eimeria, Sarcocystis, Neospora, and Toxoplasma. Unlike Eimeria, a single-host enteric pathogen, Sarcocystis, Neospora, and Toxoplasma are two-host parasites that infect and produce infectious tissue cysts in a wide range of intermediate hosts. As a genus, Sarcocystis is one of the most successful protozoan parasites; all vertebrates, including birds, reptiles, fish, and mammals are hosts to at least one Sarcocystis species. Here we sequenced Sarcocystis neurona, th...
Reed SM, Furr M, Howe DK, Johnson AL, MacKay RJ, Morrow JK, Pusterla N, Witonsky S.Equine protozoal myeloencephalitis (EPM) remains an important neurologic disease of horses. There are no pathognomonic clinical signs for the disease. Affected horses can have focal or multifocal central nervous system (CNS) disease. EPM can be difficult to diagnose antemortem. It is caused by either of 2 parasites, Sarcocystis neurona and Neospora hughesi, with much less known about N. hughesi. Although risk factors such as transport stress and breed and age correlations have been identified, biologic factors such as genetic predispositions of individual animals, and parasite-specific factors...
Fenger CK, Granstrom DE, Langemeier JL, Stamper S, Donahue JM, Patterson JS, Gajadhar AA, Marteniuk JV, Xiaomin Z, Dubey JP.Sarcocystis neurona is an apicomplexan that causes equine protozoal myeloencephalitis (EPM) in North and South America. Horses appear to be an aberrant host, because the merozoites continually divide in the central nervous system, without encysting. The natural host species has not previously been identified. The small subunit ribosomal RNA (SSURNA) gene of S. neurona was compared to those of Sarcocystis muris, Sarcocystis cruzi, Toxoplasma gondii, and Cryptosporidium parvum to identify a unique region suitable for a species-specific amplification primer. The S. neurona SSURNA primer was used ...
Dubey JP, Howe DK, Furr M, Saville WJ, Marsh AE, Reed SM, Grigg ME.Equine protozoal myeloencephalitis (EPM) is a serious disease of horses, and its management continues to be a challenge for veterinarians. The protozoan Sarcocystis neurona is most commonly associated with EPM. S. neurona has emerged as a common cause of mortality in marine mammals, especially sea otters (Enhydra lutris). EPM-like illness has also been recorded in several other mammals, including domestic dogs and cats. This paper updates S. neurona and EPM information from the last 15 years on the advances regarding life cycle, molecular biology, epidemiology, clinical signs, diagnosis, treat...
Stock ML, Elazab ST, Hsu WH.Triazines are relatively new antiprotozoal drugs that have successfully controlled coccidiosis and equine protozoal myeloencephalitis. These drugs have favorably treated other protozoal diseases such as neosporosis and toxoplasmosis. In this article, we discuss the pharmacological characteristics of five triazines, toltrazuril, ponazuril, clazuril, diclazuril, and nitromezuril which are used in veterinary medicine to control protozoal diseases which include coccidiosis, equine protozoal myeloencephalitis, neosporosis, and toxoplasmosis.
Dubey JP, Davis SW, Speer CA, Bowman DD, de Lahunta A, Granstrom DE, Topper MJ, Hamir AN, Cummings JF, Suter MM.Sarcocystis neuronan n. sp. is proposed for the apicomplexan taxon associated with myeloencephalitis in horses. Only asexual stages of this parasite presently are known, and they are found within neuronal cells and leukocytes of the brain and spinal cord. The parasite is located in the host cell cytoplasm, does not have a parasitophorous vacuole, and divides by endopolygeny. Schizonts are 5-35 microns x 5-20 microns and contain 4-40 merozoites arranged in a rosette around a prominent residual body. Merozoites are approximately 4 x 1 micron, have a central nucleus, and lack rhoptries. Schizonts...
The Journal of parasitologyFebruary 24, 2001
Volume 86, Issue 6 1276-1280 doi: 10.1645/0022-3395(2000)086[1276:COTLCO]2.0.CO;2
Dubey JP, Saville WJ, Lindsay DS, Stich RW, Stanek JF, Speert CA, Rosenthal BM, Njoku CJ, Kwok OC, Shen SK, Reed SM.Sarcocystis neurona is the most important cause of a neurologic disease in horses, equine protozoal myeloencephalitis (EPM). The complete life cycle of S. neurona, including the description of sarcocysts and intermediate hosts, has not been completed until now. Opossums (Didelphis spp.) are definitive hosts, and horses and other mammals are aberrant hosts. In the present study, laboratory-raised domestic cats (Felis domesticus) were fed sporocysts from the intestine of a naturally infected opossum (Didelphis virginiana). Microscopic sarcocysts, with a maximum size of 700 x 50 microm, developed...
Tanhauser SM, Yowell CA, Cutler TJ, Greiner EC, MacKay RJ, Dame JB.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...
Cheadle MA, Tanhauser SM, Dame JB, Sellon DC, Hines M, Ginn PE, MacKay RJ, Greiner EC.The nine-banded armadillo (Dasypus novemcinctus) is an intermediate host of at least three species of Sarcocystis, Sarcocystis dasypi, Sarcocystis diminuta, and an unidentified species; however, life cycles of these species have not been determined. Following feeding of armadillo muscles containing sarcocysts to the Virginia opossum (Didelphis virginiana), the opossums shed sporulated Sarcocystis sporocysts in their faeces. Mean dimensions for sporocysts were 11.0x7.5 microm and each contained four sporozoites and a residual body. Sporocysts were identified as Sarcocystis neurona using PCR and...
Ojo KK, Dangoudoubiyam S, Verma SK, Scheele S, DeRocher AE, Yeargan M, Choi R, Smith TR, Rivas KL, Hulverson MA, Barrett LK, Fan E, Maly DJ....Sarcocystis neurona is the most frequent cause of equine protozoal myeloencephalitis, a debilitating neurological disease of horses that can be difficult to treat. We identified SnCDPK1, the S. neurona homologue of calcium-dependent protein kinase 1 (CDPK1), a validated drug target in Toxoplasma gondii. SnCDPK1 shares the glycine "gatekeeper" residue of the well-characterized T. gondii enzyme, which allows the latter to be targeted by bumped kinase inhibitors. This study presents detailed molecular and phenotypic evidence that SnCDPK1 can be targeted for rational drug development. Recombinant ...
Reed SM, Howe DK, Morrow JK, Graves A, Yeargan MR, Johnson AL, MacKay RJ, Furr M, Saville WJ, Williams NM.Recent work demonstrated the value of antigen-specific antibody indices (AI and C-value) to detect intrathecal antibody production against Sarcocystis neurona for antemortem diagnosis of equine protozoal myeloencephalitis (EPM). Objective: The study was conducted to assess whether the antigen-specific antibody indices can be reduced to a simple serum : cerebrospinal fluid (CSF) titer ratio to achieve accurate EPM diagnosis. Methods: Paired serum and CSF samples from 128 horses diagnosed by postmortem examination. The sample set included 44 EPM cases, 35 cervical-vertebral malformation (CVM) ca...
Hawkins E, Kock R, McKeever D, Gakuya F, Musyoki C, Chege SM, Mutinda M, Kariuki E, Davidson Z, Low B, Skilton RA, Njahira MN, Wamalwa M, Maina E.The role of equine piroplasmosis as a factor in the population decline of the Grevy's zebra is not known. We determined the prevalence of Babesia caballi and Theileria equi in cograzing Grevy's zebras (Equus grevyi) and donkeys (Equus africanus asinus) in northern Kenya and identified the associated tick vectors. Blood samples were taken from 71 donkeys and 16 Grevy's zebras from March to May 2011. A nested PCR reaction using 18s ribosomal (r)RNA primers on 87 blood spots showed 72% (51/71; 95% confidence interval [CI] 60.4-81.0%) of donkeys and 100% (16/16; 95% CI, 77.3-100%) of Grevy's zebra...
Fenger CK, Granstrom DE, Gajadhar AA, Williams NM, McCrillis SA, Stamper S, Langemeier JL, Dubey JP.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...
Furr M, Kennedy T, MacKay R, Reed S, Andrews F, Bernard B, Bain F, Byars D.Equine protozoal myeloencephalitis (EPM) is a neurologic disease of horses most commonly caused by the protozoan parasite Sarcocystis neurona. Until recently the only treatment option was the combination of a sulfonamide with pyrimethamine. The present study was performed to assess the efficacy of ponazuril, an anticoccidial triazine-based compound, as a treatment for naturally occurring EPM. One hundred one horses with EPM were randomly allocated to treatment with ponazuril 15% oral paste at either 5 or 10 mg/kg body weight for 28 consecutive days. Horses were evaluated clinically and by anal...
Hoane JS, Yeargan MR, Stamper S, Saville WJ, Morrow JK, Lindsay DS, Howe DK.Neospora hughesi is a recently identified cause of equine protozoal myeloencephalitis. However, the significance of this parasite is poorly understood. An enzyme-linked immunosorbent assay (ELISA) with a recombinant form of the N. hughesi 29-kDa surface antigen (rNhSAG1) was developed for serodiagnosis of equine N. hughesi infections. Parallel ELISA analysis showed that animals immunized or infected with N. hughesi exhibited greater antibody reactivity with rNhSAG1 than with the Neospora caninum homolog, rNcSAG1. The rNhSAG1 ELISA showed 94.4% sensitivity and 95.0% specificity when compared wi...
Dubey JP, Kerber CE, Granstrom DE.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 ...
Lindsay DS, Dubey JP, Kennedy TJ.The present study examined the efficacy of ponazuril in inhibiting merozoite production of Sarcocystis neurona in cell cultures. Ponazuril inhibited merozoite production by more that 90% in cultures of S. neurona treated with 1.0 microg/ml ponazuril and greater than 95% inhibition of merozoite production was observed when infected cultures were treated with 5.0 microg/ml ponazuril. Ponazuril may have promise as a therapeutic agent in the treatment of S. neurona induced equine protozoal myeloencephalitis (EPM) in horses.
Speer CA, Dubey JP.The ultrastructure of Sarcocystis neurona schizonts and merozoites was studied in specimens derived from cell culture and from the brains of infected mice. Schizonts and merozoites were located in the host cell cytoplasm without a parasitophorous vacuole at any stage of development. Merozoites divided by endopolygeny. Fully formed merozoites had a pellicle, numerous polysomes and ribosomes, smooth and rough endoplasmic reticulum, 22 subpellicular microtubules, 9-16 dense granules, 25-75 micronemes, a plastid, a Golgi complex, 1-3 mitochondria, a conoid, 2 apical rings, 2 polar rings, 0-6 lipid...
Saville WJ, Reed SM, Granstrom DE, Hinchcliff KW, Kohn CW, Wittum TE, Stamper S.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...
Gondim LSQ, Jesus RF, Ribeiro-Andrade M, Silva JCR, Siqueira DB, Marvulo MFV, Aléssio FM, Mauffrey JF, Julião FS, Savani ESMM, Soares RM, Gondim LFP.Sarcocystis neurona and Neospora spp. are protozoan parasites that induce neurological diseases in horses and other animal species. Opossums (Didelphis albiventris and Didelphis virginiana) are definitive hosts of S. neurona, which is the major cause of equine protozoal myeloencephalitis (EPM). Neospora caninum causes abortion in cattle and infects a wide range of animal species, while N. hughesi is known to induce neurologic disease in equids. The aims of this study were to investigate S. neurona and N. caninum in tissues from opossums in the northeastern Brazil, and to isolate Brazilian stra...
Saville WJ, Stich RW, Reed SM, Njoku CJ, Oglesbee MJ, Wunschmann A, Grover DL, Larew-Naugle AL, Stanek JF, Granstrom DE, Dubey JP.Neurologic disease in horses caused by Sarcocystis neurona is difficult to diagnose, treat, or prevent, due to the lack of knowledge about the pathogenesis of the disease. This in turn is confounded by the lack of a reliable equine model of equine protozoal myeloencephalitis (EPM). Epidemiologic studies have implicated stress as a risk factor for this disease, thus, the role of transport stress was evaluated for incorporation into an equine model for EPM. Sporocysts from feral opossums were bioassayed in interferon-gamma gene knockout (KO) mice to determine minimum number of viable S. neurona ...
The Journal of parasitologyJuly 26, 2003
Volume 89, Issue 3 604-606 doi: 10.1645/0022-3395(2003)089[0604:CKACAP]2.0.CO;2
Bork S, Yokoyama N, Matsuo T, Claveria FG, Fujisaki K, Igarashi I.The antifungal agents clotrimazole (CLT) and ketoconazole (KC) and the herbicide clodinafop-propargyl (CP) inhibit growth of Plasmodium sp., Toxoplasma sp., and Trypanosoma sp. In the present study, we evaluated these drugs against the in vitro growth of the equine protozoan parasites Babesia equi and B. caballi. Clotrimazole (IC50: 2 and 17 microM), KC (IC50: 6 and 22 microM), and CP (IC50: 450 and 354 microM) were effective growth inhibitors. Interestingly, intraerythrocytic KC-treated Babesia sp. were observed to be in immediate contact with the plasma fraction of the blood in electron micr...
MacKay RJ, Granstrom DE, Saville WJ, Reed SM.Recent advances in the understanding of the parasite life cycle, epidemiology, clinical signs, diagnosis, treatment, and prevention of EPM are reviewed. The NAHMS Equine '98 study and a controlled retrospective study from The Ohio State University College of Veterinary Medicine identified a number of risk factors associated with development of the disease. The national annual incidence of EPM was 1% or less depending on the primary use of the animals. Increased disease risk was associated with age (1-5 and > 13 years of age), season (lowest in winter months and increasing with ambient tempe...
Rejmanek D, Miller MA, Grigg ME, Crosbie PR, Conrad PA.Sarcocystis neurona is a significant cause of neurological disease in horses and other animals, including the threatened Southern sea otter (Enhydra lutris nereis). Opossums (Didelphis virginiana), the only known definitive hosts for S. neurona in North America, are an introduced species in California. S. neurona DNA isolated from sporocysts and/or infected tissues of 10 opossums, 6 horses, 1 cat, 23 Southern sea otters, and 1 harbor porpoise (Phocoena phocoena) with natural infections was analyzed based on 15 genetic markers, including the first internal transcribed spacer (ITS-1) region; the...
Lindsay DS, Mitchell SM, Vianna MC, Dubey JP.Equine protozoal myeloencephalitis is a major cause of neurological disease in horses from the Americas. Horses are considered accidental intermediate hosts. The structure of sporocysts of the causative agent, Sarcocystis neurona, has never been described. Sporocysts of S. neurona were obtained from the intestines of a laboratory-raised opossum fed skeletal muscles from a raccoon that had been fed sporocysts. Sporocysts were 11.3 by 8.2 microm and contained 4 sporozoites. The appearance of the sporocyst residuum was variable. The residuum of some sporocysts was composed of many dispersed granu...
Mullaney T, Murphy AJ, Kiupel M, Bell JA, Rossano MG, Mansfield LS.Opossums (Didelphis spp.) are the definitive host for the protozoan parasite Sarcocystis neurona, the causative agent of equine protozoal myeloencephalitis (EPM). Opossums shed sporocysts in feces that can be ingested by true intermediate hosts (cats, raccoons, skunks, armadillos and sea otters). Horses acquire the parasite by ingestion of feed or water contaminated by opossum feces. However, horses have been classified as aberrant intermediate hosts because the terminal asexual sarcocyst stage that is required for transmission to the definitive host has not been found in their tissues despite...
Ellison SP, Omara-Opyene AL, Yowell CA, Marsh AE, Dame JB.A gene encoding a major 29 kDa surface antigen from Sarcocystis neurona, the primary causative agent of equine protozoal myeloencephalitis (EPM), was cloned, sequenced, and expressed as a recombinant protein. A cDNA library was prepared in the expression vector lambda ZAP from polyA+mRNA isolated from S. neurona merozoites cultivated in vitro. Random sequencing of 96 clones identified a clone of an abundant transcript having a translated amino acid sequence with 30% identity to the 31-kDa surface antigen of Sarcocystis muris cyst merozoites. Southern blot analysis indicated that the correspond...
Furr M, Howe D, Reed S, Yeargan M.Diagnosis of equine protozoal myeloencephalitis (EPM) remains a challenge for equine practitioners. Current utilized methods have inadequate sensitivity and specificity, because of a high number of false positive results. Objective: Evaluation of antibody indices to Sarcocystis neurona should provide high sensitivity and specificity for diagnosis of EPM. Methods: Archived samples from 29 clinical patients. Methods: Archived serum and cerebrospinal fluid (CSF) samples from clinical patients with either EPM (14) or cervical vertebral compressive myelopathy (CVM) (15) were examined and tested for...
Johnson AL, Morrow JK, Sweeney RW.Recent research suggests that serum : CSF titer ratios could provide the most accurate antemortem diagnosis of equine protozoal myeloencephalitis (EPM) caused by Sarcocystis neurona. Objective: The purpose of this study was to assess the utility of two commercially available tests, the indirect fluorescent antibody test (IFAT) and the surface antigen 2, 4/3 ELISA (SAG2, 4/3 ELISA), using archived paired serum and CSF samples. Methods: Samples were obtained from 4 types of clinical patients. Confirmed positive cases (n = 9 horses; 11 sample sets) had neurologic deficits and postmortem lesions c...
