Comparison of Sarcocystis neurona isolates derived from horse neural tissue

L. S. Mansfield; H. C. Schott; A. J. Murphy; M. G. Rossano; S. M. Tanhauser; J. S. Patterson; K. Nelson; S. L. Ewart; J. V. Marteniuk; D. D. Bowman; J. B. Kaneene

doi:10.1016/S0304-4017(00)00388-5

Comparison of Sarcocystis neurona isolates derived from horse neural tissue

L. S. Mansfield, H. C. Schott, A. J. Murphy, M. G. Rossano, S. M. Tanhauser, J. S. Patterson, K. Nelson, S. L. Ewart, J. V. Marteniuk, D. D. Bowman, J. B. Kaneene

Animal and Food Sciences

Research output: Contribution to journal › Article › peer-review

23 Scopus citations

Abstract

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 that Sarcocystis spp. from the eight horses appeared the same, but different from the grackle isolate. One Michigan horse isolate (MIH6) had two bands at 72 and 25 kDa that were more prominent than the UCD1 isolate and other Michigan horse isolates. Western blot analysis showed that merozoites of eight of eight equine-derived isolates, and the UCD1 S. neurona isolate had similar bands when developed with serum or CSF of an infected horse. Major bands were seen at 60, 44, 30, and 16 kDa. In the grackle (Cornell) isolate, bands were seen at 60, 44, 29, and 16 kDa. DNA from merozoites of each of the eight equine-derived isolates and the grackle-derived isolate produced a 334 bp PCR product (Tanhauser et al., 1999). Restriction fragment length polymorphism (RFLP) analysis of these horse isolates showed banding patterns characteristic for S. neurona. The grackle (Cornell) isolate had an RFLP banding pattern characteristic of other S. falcatula species. Finally, electron microscopy examining multiple merozoites of each of these eight horse isolates showed similar morphology, which differed from the grackle (Cornell) isolate. We conclude that the eight Michigan horse isolates are S. neurona species and the grackle isolate is an S. falcatula species.

Original language	English
Pages (from-to)	167-178
Number of pages	12
Journal	Veterinary Parasitology
Volume	95
Issue number	2-4
DOIs	https://doi.org/10.1016/S0304-4017(00)00388-5
State	Published - Feb 26 2001

Bibliographical note

Funding Information:
Thanks are given to Drs. Patricia Conrad and Antoinette Marsh for providing the UCD-1 strain of S. neurona for Western blot analysis. We thank Dr. J. Carl Fox of Oklahoma State University for the kind gift of a sample of bovine Sarcocystis. We would like to thank Sally Burns for her expertise in electron microscopy and Ms. Shannon Lynch for her expert technical work. The authors would like to thank Kate and Charles Baird for donating opossums with S. neurona and for their untiring enthusiasm and efforts to help provide wildlife for the project. Finally, we give sincere thanks and sympathy to the people who donated the bodies of their horses so that this work could be accomplished. This work was supported by an Animal Health Initiative Grant from USDA distributed through the State of Michigan (71-9419), by a Michigan State University Intellectual Property Office Grant (71-4668), by a donation from the Michigan Quarter Horse Association (31-1258), and by donations from Great Lakes Standardbred Sales Company.

Keywords

Apicomplexan protozoan parasite
Equine protozoal myeloencephalitis
Neurological disease
Opossums
Sarcocystis neurona

ASJC Scopus subject areas

Parasitology
General Veterinary

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10.1016/S0304-4017(00)00388-5

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title = "Comparison of Sarcocystis neurona isolates derived from horse neural tissue",

abstract = "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 that Sarcocystis spp. from the eight horses appeared the same, but different from the grackle isolate. One Michigan horse isolate (MIH6) had two bands at 72 and 25 kDa that were more prominent than the UCD1 isolate and other Michigan horse isolates. Western blot analysis showed that merozoites of eight of eight equine-derived isolates, and the UCD1 S. neurona isolate had similar bands when developed with serum or CSF of an infected horse. Major bands were seen at 60, 44, 30, and 16 kDa. In the grackle (Cornell) isolate, bands were seen at 60, 44, 29, and 16 kDa. DNA from merozoites of each of the eight equine-derived isolates and the grackle-derived isolate produced a 334 bp PCR product (Tanhauser et al., 1999). Restriction fragment length polymorphism (RFLP) analysis of these horse isolates showed banding patterns characteristic for S. neurona. The grackle (Cornell) isolate had an RFLP banding pattern characteristic of other S. falcatula species. Finally, electron microscopy examining multiple merozoites of each of these eight horse isolates showed similar morphology, which differed from the grackle (Cornell) isolate. We conclude that the eight Michigan horse isolates are S. neurona species and the grackle isolate is an S. falcatula species.",

keywords = "Apicomplexan protozoan parasite, Equine protozoal myeloencephalitis, Neurological disease, Opossums, Sarcocystis neurona",

author = "Mansfield, {L. S.} and Schott, {H. C.} and Murphy, {A. J.} and Rossano, {M. G.} and Tanhauser, {S. M.} and Patterson, {J. S.} and K. Nelson and Ewart, {S. L.} and Marteniuk, {J. V.} and Bowman, {D. D.} and Kaneene, {J. B.}",

note = "Funding Information: Thanks are given to Drs. Patricia Conrad and Antoinette Marsh for providing the UCD-1 strain of S. neurona for Western blot analysis. We thank Dr. J. Carl Fox of Oklahoma State University for the kind gift of a sample of bovine Sarcocystis. We would like to thank Sally Burns for her expertise in electron microscopy and Ms. Shannon Lynch for her expert technical work. The authors would like to thank Kate and Charles Baird for donating opossums with S. neurona and for their untiring enthusiasm and efforts to help provide wildlife for the project. Finally, we give sincere thanks and sympathy to the people who donated the bodies of their horses so that this work could be accomplished. This work was supported by an Animal Health Initiative Grant from USDA distributed through the State of Michigan (71-9419), by a Michigan State University Intellectual Property Office Grant (71-4668), by a donation from the Michigan Quarter Horse Association (31-1258), and by donations from Great Lakes Standardbred Sales Company.",

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doi = "10.1016/S0304-4017(00)00388-5",

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AU - Mansfield, L. S.

AU - Schott, H. C.

AU - Murphy, A. J.

AU - Rossano, M. G.

AU - Tanhauser, S. M.

AU - Patterson, J. S.

AU - Nelson, K.

AU - Ewart, S. L.

AU - Marteniuk, J. V.

AU - Bowman, D. D.

AU - Kaneene, J. B.

N1 - Funding Information: Thanks are given to Drs. Patricia Conrad and Antoinette Marsh for providing the UCD-1 strain of S. neurona for Western blot analysis. We thank Dr. J. Carl Fox of Oklahoma State University for the kind gift of a sample of bovine Sarcocystis. We would like to thank Sally Burns for her expertise in electron microscopy and Ms. Shannon Lynch for her expert technical work. The authors would like to thank Kate and Charles Baird for donating opossums with S. neurona and for their untiring enthusiasm and efforts to help provide wildlife for the project. Finally, we give sincere thanks and sympathy to the people who donated the bodies of their horses so that this work could be accomplished. This work was supported by an Animal Health Initiative Grant from USDA distributed through the State of Michigan (71-9419), by a Michigan State University Intellectual Property Office Grant (71-4668), by a donation from the Michigan Quarter Horse Association (31-1258), and by donations from Great Lakes Standardbred Sales Company.

PY - 2001/2/26

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N2 - 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 that Sarcocystis spp. from the eight horses appeared the same, but different from the grackle isolate. One Michigan horse isolate (MIH6) had two bands at 72 and 25 kDa that were more prominent than the UCD1 isolate and other Michigan horse isolates. Western blot analysis showed that merozoites of eight of eight equine-derived isolates, and the UCD1 S. neurona isolate had similar bands when developed with serum or CSF of an infected horse. Major bands were seen at 60, 44, 30, and 16 kDa. In the grackle (Cornell) isolate, bands were seen at 60, 44, 29, and 16 kDa. DNA from merozoites of each of the eight equine-derived isolates and the grackle-derived isolate produced a 334 bp PCR product (Tanhauser et al., 1999). Restriction fragment length polymorphism (RFLP) analysis of these horse isolates showed banding patterns characteristic for S. neurona. The grackle (Cornell) isolate had an RFLP banding pattern characteristic of other S. falcatula species. Finally, electron microscopy examining multiple merozoites of each of these eight horse isolates showed similar morphology, which differed from the grackle (Cornell) isolate. We conclude that the eight Michigan horse isolates are S. neurona species and the grackle isolate is an S. falcatula species.

AB - 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 that Sarcocystis spp. from the eight horses appeared the same, but different from the grackle isolate. One Michigan horse isolate (MIH6) had two bands at 72 and 25 kDa that were more prominent than the UCD1 isolate and other Michigan horse isolates. Western blot analysis showed that merozoites of eight of eight equine-derived isolates, and the UCD1 S. neurona isolate had similar bands when developed with serum or CSF of an infected horse. Major bands were seen at 60, 44, 30, and 16 kDa. In the grackle (Cornell) isolate, bands were seen at 60, 44, 29, and 16 kDa. DNA from merozoites of each of the eight equine-derived isolates and the grackle-derived isolate produced a 334 bp PCR product (Tanhauser et al., 1999). Restriction fragment length polymorphism (RFLP) analysis of these horse isolates showed banding patterns characteristic for S. neurona. The grackle (Cornell) isolate had an RFLP banding pattern characteristic of other S. falcatula species. Finally, electron microscopy examining multiple merozoites of each of these eight horse isolates showed similar morphology, which differed from the grackle (Cornell) isolate. We conclude that the eight Michigan horse isolates are S. neurona species and the grackle isolate is an S. falcatula species.

KW - Apicomplexan protozoan parasite

KW - Equine protozoal myeloencephalitis

KW - Neurological disease

KW - Opossums

KW - Sarcocystis neurona

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Comparison of Sarcocystis neurona isolates derived from horse neural tissue

Abstract

Bibliographical note

Keywords

ASJC Scopus subject areas

Access to Document

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