Multiple DNA markers differentiate Sarcocystis neurona and Sarcocystis falcatula.
Abstract: 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 between S. neurona and S. falcatula. Useful sequence heterogeneity between the 2 organisms was identified, creating potential markers to distinguish these Sarcocystis spp. These markers were used to characterize Sarcocystis isolates from opossum (Didelphis virginiana) feces. Our data suggest that S. neurona and S. falcatula can be differentiated with these markers and that multiple Sarcocystis spp., including S. neurona and S. falcatula, are shed by opossums.
Publication Date: 1999-04-29 PubMed ID: 10219299
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- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
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The research article discusses the use of multiple DNA markers to distinguish between two similar species, Sarcocystis neurona and Sarcocystis falcatula, which both occur in the same definitive host and are associated with equine protozoal myeloencephalitis.
Understanding the Problem
- The paper highlights the hurdle faced by researchers when studying the agent of equine protozoal myeloencephalitis – a considerable resemblance between Sarcocystis neurona and other Sarcocystis spp. found in the same host.
- This similarity complicates the understanding of the life cycle of the disease-causing agent, and the development of effective control and treatment strategies.
Methods Used
- To address this problem, the researchers employed random-amplified polymorphic DNA (RAPD) techniques to amplify DNA from isolates of S. neurona and S. falcatula.
- The amplified DNA was then subjected to sequence analysis of the polymerase chain reaction (PCR) products. This was done to design PCR primers that could amplify specific DNA products of the individual Sarcocystis species.
- Additionally, the internal transcribed spacer (ITS) of ribosomal RNA was amplified and compared between S. neurona and S. falcatula.
Findings
- The DNA sequence analysis and the comparison of ribosomal RNA ITS led to the identification of useful sequence diversity between S. neurona and S. falcatula. This heterogeneity provides potential markers for distinguishing these two Sarcocystis species.
- These markers were then applied to characterize Sarcocystis isolates obtained from feces of opossums (Didelphis virginiana).
- The results suggest that these DNA markers are effective in distinguishing S. neurona and S. falcatula. They also provide evidence that multiple Sarcocystis species, including both S. neurona and S. falcatula, are shed by opossums.
The Implication of the Study
- This study contributes to the scientific understanding of the complexities of the life cycle of the agents causing equine protozoal myeloencephalitis.
- The developed DNA markers could be beneficial in the diagnosis, prevention, and treatment of the disease by allowing precise identification of the causative species.
Cite This Article
APA
Tanhauser SM, Yowell CA, Cutler TJ, Greiner EC, MacKay RJ, Dame JB.
(1999).
Multiple DNA markers differentiate Sarcocystis neurona and Sarcocystis falcatula.
J Parasitol, 85(2), 221-228.
Publication
Researcher Affiliations
- Department of Pathobiology, College of Veterinary Medicine, University of Florida, Gainesville 32611-0880, USA.
MeSH Terms
- Animals
- Bird Diseases / parasitology
- Birds / parasitology
- DNA, Protozoan / analysis
- Encephalitis / parasitology
- Encephalitis / veterinary
- Genetic Markers
- Horse Diseases / parasitology
- Horses
- Opossums / parasitology
- Phylogeny
- Polymerase Chain Reaction / methods
- Random Amplified Polymorphic DNA Technique
- Restriction Mapping
- Sarcocystis / classification
- Sarcocystis / genetics
- Sarcocystis / isolation & purification
- Sarcocystosis / parasitology
- Sarcocystosis / veterinary
Citations
This article has been cited 19 times.- Enriquez CK, Morrow JK, Graves A, Johnson A. Evaluation of real-time polymerase chain reaction for the diagnosis of protozoal myeloencephalitis in horses using cerebrospinal fluid.. J Vet Intern Med 2023 Sep-Oct;37(5):1893-1898.
- Llano HAB, Zavatieri Polato H, Borges Keid L, Ferreira de Souza Oliveira TM, Zwarg T, de Oliveira AS, Sanches TC, Joppert AM, Gondim LFP, Martins Soares R. Molecular screening for Sarcocystidae in muscles of wild birds from Brazil suggests a plethora of intermediate hosts for Sarcocystis falcatula.. Int J Parasitol Parasites Wildl 2022 Apr;17:230-238.
- Borges-Silva W, de Jesus RF, Ferreira R, Gondim LFP. Reactivity of Horse Sera to Antigens Derived From Sarcocystis falcatula-Like and Sarcocystis neurona.. Front Vet Sci 2020;7:573016.
- Gondim LFP, Soares RM, Tavares AS, Borges-Silva W, de Jesus RF, Llano HAB, Gondim LQ. Sarcocystis falcatula-like derived from opossum in Northeastern Brazil: In vitro propagation in avian cells, molecular characterization and bioassay in birds.. Int J Parasitol Parasites Wildl 2019 Dec;10:132-137.
- Nazir MM, Ayaz MM, Ahmed AN, Maqbool A, Ashraf K, Oneeb M, Yasin G, Subhani A, Ali MA, Nazir N, Sajid MA. Prevalence of Toxoplasma gondii, Neospora caninum, and Sarcocystis Species DNA in the Heart and Breast Muscles of Rock Pigeons (Columbia livia).. J Parasitol Res 2018;2018:6264042.
- Gallo SSM, Lindsay DS, Ederli NB, Matteoli FP, Venancio TM, de Oliveira FCR. Identification of opossums Didelphis aurita (Wied-Neuweid, 1826) as a definitive host of Sarcocystis falcatula-like sporocysts.. Parasitol Res 2018 Jan;117(1):213-223.
- Zitzer NC, Marsh AE, Burkhard MJ, Radin MJ, Wellman ML, Jugan M, Parker V. Parasitemia due to Sarcocystis neurona-like infection in a clinically ill domestic cat.. Vet Clin Pathol 2017 Sep;46(3):526-532.
- Dubey JP, Howe DK, Furr M, Saville WJ, Marsh AE, Reed SM, Grigg ME. An update on Sarcocystis neurona infections in animals and equine protozoal myeloencephalitis (EPM).. Vet Parasitol 2015 Apr 15;209(1-2):1-42.
- Rejmanek D, Miller MA, Grigg ME, Crosbie PR, Conrad PA. Molecular characterization of Sarcocystis neurona strains from opossums (Didelphis virginiana) and intermediate hosts from Central California.. Vet Parasitol 2010 May 28;170(1-2):20-9.
- Bisby TM, Holman PJ, Pitoc GA, Packer RA, Thompson CA, Raskin RE. Sarcocystis sp. encephalomyelitis in a cat.. Vet Clin Pathol 2010 Mar;39(1):105-12.
- Elsheikha HM, Schott HC 2nd, Mansfield LS. Genetic variation among isolates of Sarcocystis neurona, the agent of protozoal myeloencephalitis, as revealed by amplified fragment length polymorphism markers.. Infect Immun 2006 Jun;74(6):3448-54.
- Elsheikha HM, Lacher DW, Mansfield LS. Phylogenetic relationships of Sarcocystis neurona of horses and opossums to other cyst-forming coccidia deduced from SSU rRNA gene sequences.. Parasitol Res 2005 Nov;97(5):345-57.
- Elsheikha HM, Murphy AJ, Mansfield LS. Phylogenetic congruence of Sarcocystis neurona Dubey et al., 1991 (Apicomplexa: Sarcocystidae) in the United States based on sequence analysis and restriction fragment length polymorphism (RFLP).. Syst Parasitol 2005 Jul;61(3):191-202.
- Elsheikha HM, Rosenthal BM, Mansfield LS. Dexamethasone treatment induces susceptibility of outbred Webster mice to experimental infection with Besnoitia darlingi isolated from opossums (Didelphis virginiana).. Parasitol Res 2005 Apr;95(6):413-9.
- Sellon DC, Knowles DP, Greiner EC, Long MT, Hines MT, Hochstatter T, Tibary A, Dame JB. Infection of immunodeficient horses with Sarcocystis neurona does not result in neurologic disease.. Clin Diagn Lab Immunol 2004 Nov;11(6):1134-9.
- Elsheikha HM, Murphy AJ, Mansfield LS. Prevalence of Sarcocystis species sporocysts in Northern Virginia opossums (Didelphis virginiana).. Parasitol Res 2004 Aug;93(5):427-31.
- Elsheikha HM, Fitzgerald SD, Mansfield LS, Saeed AM. Sarcocystis inghami n. sp. (Sporozoa: Sarcocystidae) from the skeletal muscles of the Virginia opossum Didelphis virginiana in Michigan.. Syst Parasitol 2003 Sep;56(1):77-84.
- Elsheikha HM, Saeed MA, Fitzgerald SD, Murphy AJ, Mansfield LS. Effects of temperature and host cell type on the in vitro growth and development of Sarcocystis falcatula.. Parasitol Res 2003 Sep;91(1):22-6.
- Elsheikha HM, Murphy AJ, Fitzgerald SD, Mansfield LS, Massey JP, Saeed MA. Purification of Sarcocystis neurona sporocysts from opossum (Didelphis virginiana) using potassium bromide discontinuous density gradient centrifugation.. Parasitol Res 2003 Jun;90(2):104-9.
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