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Topic:Protozoa

Protozoa are single-celled eukaryotic organisms that can interact with horses in various ways, including as parasites. In equine health, protozoal infections can lead to significant diseases, such as equine protozoal myeloencephalitis (EPM), which is caused by the protozoan Sarcocystis neurona. These organisms can affect the central nervous system and other bodily systems, leading to a range of clinical symptoms in horses. Understanding the life cycle, transmission, and impact of protozoa is important for managing and preventing infections. This page compiles peer-reviewed research studies and scholarly articles that explore the biology, pathogenesis, and diagnostic approaches related to protozoal interactions in horses.
A review of Sarcocystis neurona and equine protozoal myeloencephalitis (EPM).
Veterinary parasitology    February 27, 2001   Volume 95, Issue 2-4 89-131 doi: 10.1016/s0304-4017(00)00384-8
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...
Direct agglutination test for the detection of antibodies to Sarcocystis neurona in experimentally infected animals.
Veterinary parasitology    February 27, 2001   Volume 95, Issue 2-4 179-186 doi: 10.1016/s0304-4017(00)00389-7
Lindsay DS, Dubey JP.Equine protozoal myeloencephalitis (EPM) is a serious neurological disease of horses in the Americas. The apicomplexan protozoan most commonly associated with EPM is Sarcocystis neurona. A direct agglutination test (SAT) was developed to detect antibodies to S. neurona in experimentally infected animals. Merozoites of the SN6 strain of S. neurona collected from cell culture were used as antigen and 2-mercaptoethanol was added to the antigen suspension to destroy IgM antibodies when mixed with test sera. Mice fed sporocysts of S. speeri or S. falcatula-like sporocysts from opossums did not sero...
Comparison of Sarcocystis neurona isolates derived from horse neural tissue.
Veterinary parasitology    February 27, 2001   Volume 95, Issue 2-4 167-178 doi: 10.1016/s0304-4017(00)00388-5
Mansfield LS, Schott HC, Murphy AJ, Rossano MG, Tanhauser SM, Patterson JS, Nelson K, Ewart SL, Marteniuk JV, Bowman DD, Kaneene JB.Sarcocystis neurona is a protozoan parasite that can cause neurological deficits in infected horses. The route of transmission is by fecal-oral transfer of sporocysts from opossums. However, the species identity and the lifecycle are not completely known. In this study, Sarcocystis merozoites from eight isolates obtained from Michigan horses were compared to S. neurona from a California horse (UCD1), Sarcocystis from a grackle (Cornell), and five Sarcocystis isolates from feral opossums from Michigan. Comparisons were made using several techniques. SDS-PAGE analysis with silver staining showed...
Initiation of a Sarcocystis neurona expressed sequence tag (EST) sequencing project: a preliminary report.
Veterinary parasitology    February 27, 2001   Volume 95, Issue 2-4 233-239 doi: 10.1016/s0304-4017(00)00418-0
Howe DK.To accelerate genetic and molecular characterization of Sarcocystis neurona, the primary causative agent of equine protozoal myeloencephalitis (EPM), a sequencing project has been initiated that will generate approximately 7000-8000 expressed sequence tags (ESTs) from this apicomplexan parasite. Poly(A)(+) RNA was isolated from culture-derived S. neurona merozoites, and a cDNA library was constructed in a unidirectional lambda phage cloning vector. Sixty phage clones were randomly picked from the library, and the cDNA inserts were amplified from these clones using the T3 and T7 primers that fl...
In vitro quantitative analysis of (3)H-uracil incorporation by Sarcocytis neurona to determine efficacy of anti-protozoal agents.
Veterinary parasitology    February 27, 2001   Volume 95, Issue 2-4 241-249 doi: 10.1016/s0304-4017(00)00403-9
Marsh AE, Mullins AL, Lakritz J.Parasite-specific incorporation of (3)H-uracil was used to assess the replication of Sarcocystis neurona, a protozoal parasite associated with equine protozoal myeloencephalitis (EPM). Anti-protozoal drugs, pyrimethamine (0.01, 0.1 and 1.0microg/ml PYR), sulfadiazine (5microg/ml; SDZ), sulfamethoxazole (5microg/ml; SMZ), diclazuril (100ng/ml; DCZ), atovaquone (0.04ng/ml; ATQ), tetracycline (5microg/ml; TET) and the herbicide glyphosate (1.5 and 4.5mM; GLY) were studied with varying S. neurona parasite densities (2x10(1)-1.2x10(6)merozoites/well). A microtiter plate format was used to test thes...
Sporocyst size of isolates of Sarcocystis shed by the Virginia opossum (Didelphis virginiana).
Veterinary parasitology    February 27, 2001   Volume 95, Issue 2-4 305-311 doi: 10.1016/s0304-4017(00)00396-4
Cheadle MA, Dame JB, Greiner EC.The Virginia opossum (Didelphis virginiana) is a definitive host for multiple Sarcocystis species including Sarcocystis neurona, one of the causative agents of equine protozoal myeloencephalitis (EPM), a severe, neuromuscular disease of horses. Size and morphologic characteristics of isolates of Sarcocystis shed by the opossum were examined to determine if differences were useful in discriminating between the isolates and/or species. Collections of sporocysts from 17 opossums were molecularly characterized and measured using an ocular micrometer. The mean sporocyst size of isolates of S. neuro...
A random amplified polymorphic DNA polymerase chain reaction technique that differentiates between Neospora species.
The Journal of parasitology    February 24, 2001   Volume 86, Issue 6 1366-1368 doi: 10.1645/0022-3395(2000)086[1366:ARAPDP]2.0.CO;2
Spencer JA, Witherow AK, Blagburn BL.Neospora caninum is a recently described coccidial parasite that was first isolated from a dog in 1988 and has subsequently been shown to infect a wide range of mammals. Neospora hughesi, a new species of this genus, has recently been isolated from the spinal cord of horses showing clinical signs of equine protozoal myeloencephalitis. The random amplified polymorphic DNA polymerase chain reaction technique is capable of differentiating between N. caninum and N. hughesi.
Completion of the life cycle of Sarcocystis neurona.
The Journal of parasitology    February 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...
Equine protozoal myeloencephalitis: mystery wrapped in enigma.
Parasitology research    November 30, 2000   Volume 86, Issue 11 940-943 doi: 10.1007/pl00008517
Dame JB, Cutler TJ, Tanhauser S, Ellison S, Greiner EC, MacKay RJ.No abstract available
Evaluation of risk factors associated with clinical improvement and survival of horses with equine protozoal myeloencephalitis.
Journal of the American Veterinary Medical Association    October 24, 2000   Volume 217, Issue 8 1181-1185 doi: 10.2460/javma.2000.217.1181
Saville WJ, Morley PS, Reed SM, Granstrom DE, Kohn CW, Hinchcliff KW, Wittum TE.To investigate risk factors for use in predicting clinical improvement and survival of horses with equine protozoal myeloencephalitis (EPM). Methods: Longitudinal epidemiologic study. Methods: 251 horses with EPM. Methods: Between 1992 and 1995, 251 horses with EPM were admitted to our facility. A diagnosis of EPM was made on the basis of neurologic abnormalities and detection of antibody to Sarcocystis neurona or S neurona DNA in CSF. Data were obtained from hospital records and through telephone follow-up interviews. Factors associated with clinical improvement and survival were analyzed, us...
Determination of the activity of ponazuril against Sarcocystis neurona in cell cultures.
Veterinary parasitology    August 18, 2000   Volume 92, Issue 2 165-169 doi: 10.1016/s0304-4017(00)00280-6
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.
Inoculation of Sarcocystis neurona merozoites into the central nervous system of horses.
Veterinary parasitology    August 18, 2000   Volume 92, Issue 2 157-163 doi: 10.1016/s0304-4017(00)00281-8
Lindsay DS, Dykstra CC, Williams A, Spencer JA, Lenz SD, Palma K, Dubey JP, Blagburn BL.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.
Veterinary parasitology    August 18, 2000   Volume 92, Issue 2 119-128 doi: 10.1016/s0304-4017(00)00279-x
Walsh CP, Duncan RB, Zajac AM, Blagburn BL, Lindsay DS.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...
Cross-sectional study of faecal shedding of Giardia duodenalis and Cryptosporidium parvum among packstock in the Sierra Nevada Range.
Equine veterinary journal    June 3, 2000   Volume 32, Issue 3 247-252 doi: 10.2746/042516400776563545
Atwill ER, McDougald NK, Perea L.Faecal specimens from 305 horses and mules used as packstock at one of 17 commercial or governmental (National Park Service, US Forest Service) operations were examined for Giardia duodenalis and Cryptosporidium parvum using immunofluorescent microscopy. Fourteen packstock (4.6%) were shedding G. duodenalis cysts, with herd-level prevalences ranging 0-22%. Number of packstock in the corral, size of corral and density of packstock in the corral were associated with the odds of shedding G. duodenalis cysts. None of the horses had detectable C. parvum oocysts. Assuming a sensitivity of at least 4...
Improvement of western blot test specificity for detecting equine serum antibodies to Sarcocystis neurona. Rossano MG, Mansfield LS, Kaneene JB, Murphy AJ, Brown CM, Schott HC, Fox JC.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.
Journal of veterinary pharmacology and therapeutics    January 29, 2000   Volume 22, Issue 6 374-379 doi: 10.1046/j.1365-2885.1999.00232.x
Dirikolu L, Lehner F, Nattrass C, Bentz BG, Woods WE, Carter WG, Karpiesiuk W, Jacobs J, Boyles J, Harkins JD, Granstrom DE, Tobin T.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 ...
Prevalence of antibodies to Neospora sp. in horses from Alabama and characterisation of an isolate recovered from a naturally infected horse [corrected].
International journal for parasitology    December 23, 1999   Volume 29, Issue 10 1537-1543 doi: 10.1016/s0020-7519(99)00140-x
Cheadle MA, Lindsay DS, Rowe S, Dykstra CC, Williams MA, Spencer JA, Toivio-Kinnucan MA, Lenz SD, Newton JC, Rolsma MD, Blagburn BL.An IFAT was used to determine the prevalence of Neospora-specific IgG antibodies in serum from Alabama horses. Serum samples (n = 536) were from asymptomatic horses routinely submitted for equine infectious anaemia virus infection testing. We also subjected a 13-year-old horse with CNS disease to necropsy examination for isolation and in vitro cultivation of protozoal organisms. In antemortem tests, this horse was positive for antibodies to Neospora sp. in the IFAT and western immunoblot. Results of the prevalence survey indicated that IgG antibodies to Neospora were present in 62 (11.5%) of t...
Simplified technique for isolation, excystation, and culture of Sarcocystis species from opossums.
The Journal of parasitology    November 30, 1999   Volume 85, Issue 5 979-981 
Murphy AJ, Mansfield LS.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...
Hepatic sarcocystosis in a horse.
The Journal of parasitology    November 30, 1999   Volume 85, Issue 5 965-968 
Davis CR, Barr BC, Pascoe JR, Olander HJ, Dubey JP.Hepatic sarcocystosis was diagnosed in a horse in association with refractory bacterial osteomyelitis and plasma cell tumor of the maxilla and hepatic salmonellosis. Gross lesions included pleural, pericardial, and peritoneal effusions, hepatomegaly, gastric ulceration, colonic edema, and proliferative tissues filling 2 maxillary dental alveoli. Histologically, liver was characterized by severe suppurative, necrotizing, periportal hepatitis, and severe periacinar necrosis. Hepatocytes frequently contained protozoal schizonts in various stages of development. In mature schizonts, merozoites wer...
Serologic prevalence of Sarcocystis neurona, Toxoplasma gondii, and Neospora caninum in horses in Brazil.
Journal of the American Veterinary Medical Association    October 8, 1999   Volume 215, Issue 7 970-972 
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 ...
Prevalence of antibodies to Sarcocystis neurona, Toxoplasma gondii and Neospora caninum in horses from Argentina.
Veterinary parasitology    September 17, 1999   Volume 86, Issue 1 59-62 doi: 10.1016/s0304-4017(99)00127-2
Dubey JP, Venturini MC, Venturini L, McKinney J, Pecoraro M.Sera from 76 horses from Argentina were examined for antibodies to Sarcocystis neurona, Toxoplasma gondii and Neospora caninum. Antibodies to S. neurona were found in 27 (35.5%) of 76 horses using immunoblots with culture derived merozoites as antigen. Antibodies to T. gondii were found in 10 (13.1%) of 76 horses by using the modified agglutination test with formalin-fixed tachyzoites and mercaptoethanol; titers were 1:25 (two horses), 1:50 (six horses), 1:100 (two horses), and 1:200 (one horse). Antibodies to N. caninum were not found in any of the 76 horses by the use of N. caninum agglutina...
Determination of the activity of pyrimethamine, trimethoprim, sulfonamides, and combinations of pyrimethamine and sulfonamides against Sarcocystis neurona in cell cultures.
Veterinary parasitology    May 29, 1999   Volume 82, Issue 3 205-210 doi: 10.1016/s0304-4017(99)00020-5
Lindsay DS, Dubey JP.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...
[Equine protozoal myeloencephalitis (EPM) in the Netherlands?].
Tijdschrift voor diergeneeskunde    May 27, 1999   Volume 124, Issue 9 288-289 
Boersema JH.No abstract available
Are Sarcocystis neurona and Sarcocystis falcatula synonymous? A horse infection challenge.
The Journal of parasitology    April 29, 1999   Volume 85, Issue 2 301-305 
Cutler TJ, MacKay RJ, Ginn PE, Greiner EC, Porter R, Yowell CA, Dame JB.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.
The Journal of parasitology    April 29, 1999   Volume 85, Issue 2 221-228 
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...
Neospora caninum-associated equine protozoal myeloencephalitis.
Veterinary parasitology    December 1, 1998   Volume 79, Issue 4 269-274 doi: 10.1016/s0304-4017(98)00178-2
Hamir AN, Tornquist SJ, Gerros TC, Topper MJ, Dubey JP.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...
Description of a new Neospora species (Protozoa: Apicomplexa: Sarcocystidae).
The Journal of parasitology    October 30, 1998   Volume 84, Issue 5 983-991 
Marsh AE, Barr BC, Packham AE, Conrad PA.Neospora hughesi n. sp. was isolated from the central nervous system tissue of an adult equine (Equus caballus) from California. The tachyzoites are crescent-shaped, approximately 2 x 5 microm (1.8-3.0 x 4.0-7.0 microm), with characteristic apical complex structures consisting of an anterior polar ring, conoid, numerous rhoptries filled with a uniform electron-dense material, and 22 microtubules extending posteriorly from the polar ring. Comparison of N. hughesi to canine and bovine Neospora caninum isolates showed phenotypic differences in immunoreactive proteins. Molecular analysis of the sm...
Amebic meningoencephalitis caused by Balamuthia mandrillaris (leptomyxid ameba) in a horse. Kinde H, Visvesvara GS, Barr BC, Nordhausen RW, Chiu PH.No abstract available
Effect of sodium methyl arsinate and imidocarb dipropionate antiprotozoal drugs on the pharmacokinetic of gentamicin in equines.
DTW. Deutsche tierarztliche Wochenschrift    August 11, 1998   Volume 105, Issue 7 274-276 
Soliman GA.The pharmacokinetic behaviour of gentamicin sulphate (3.4 mg/kg bwt) was studied following its intramuscular injection to a group of horses and to another group of horses premedicated with sodium methyl arsinate (2 mg/kg bwt) or imidocarb dipropionate (4.8 mg/kg bwt). Considerable differences were observed in the pharmacokinetics of gentamicin in pre-medicated horses and in horses which had received the antibiotic alone. Peak serum concentration of gentamicin (9.85 +/- 0.05 and 11.15 +/- 0.15 micrograms/ml) were attained within 1.45 +/- 0.05 and 0.92 +/- 0.04 h in arsinate and imidocarb-medica...
Giardia and Cryptosporidium in Canadian farm animals.
Veterinary parasitology    March 1, 1997   Volume 68, Issue 4 375-381 doi: 10.1016/s0304-4017(96)01072-2
Olson ME, Thorlakson CL, Deselliers L, Morck DW, McAllister TA.Giardia intestinalis and Cryptosporidium spp. are commonly identified intestinal pathogens in humans and animals. In light of the clinical disease, production losses and zoonotic potential of both Giardia and Cryptosporidium infections, a study was undertaken to investigate the prevalence of these parasites in cattle, sheep, pigs and horses in Canadian farms at different geographical locations. A total of 104 cattle, 89 sheep, 236 pigs and 35 horses were sampled from 15 different Canadian geographical locations. Fecal samples were examined after concentration and immunofluorescent staining. Gi...