FREE SHIPPING over $40
OVER 10000 FIVE-STAR REVIEWS
5% OFF ALL SUBSCRIPTIONS
100% HAPPINESS GUARANTEE
Analyze Diet
0
Journal of veterinary internal medicine2024; 39(1); e17282; doi: 10.1111/jvim.17282

Effect of refrigeration, room temperature, and processing time on serum immunofluorescent antibody titers for Sarcocystis neurona.

Abstract: 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 were retested on Days 5 and 10 after collection. A paired t test was used to compare IFA titers at different time points. Results: There was no significant difference between IFA titers from baseline with those stored at room temperature at Days 5 (P = .741, 95% CI [-56.83, 78.65]), 10 (P = .677, 95% CI [-50.01, 75.46]), and between 5 and 10 days (P = 0.949, 95% CI [-57.50, 61.14]). There was no significant difference from baseline with those stored at 4°C for Days 5 (P = .964, 95% CI [-81.81, 85.45]), 10 (P = 0.573, 95% CI [-109.4, 62.15]), and between 5 and 10 days (P = .5, 95% CI [-102.6, 51.67]). There was no statistical difference between samples stored at room temperature and 4°C (P = .688, CI [-55.51, 37.33]) on Days 5 and 10 (P = .104, CI [-80.8, 8.07]). Conclusions: Immunofluorescent antibody test titers for S. neurona are stable for up to 10 days at room temperature and 4°C.
© 2024 The Author(s). Journal of Veterinary Internal Medicine published by Wiley Periodicals LLC on behalf of American College of Veterinary Internal Medicine.
Get Full Text
Publication Date: 2024-12-23 PubMed ID: 39715359PubMed Central: PMC11665959DOI: 10.1111/jvim.17282Google Scholar: Lookup
The Equine Research Bank provides access to a large database of publicly available scientific literature. Inclusion in the Research Bank does not imply endorsement of study methods or findings by Mad Barn.
LinkedInCopy
  • Journal Article

Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

This research investigates the reliability of immunofluorescent antibody test titers for Sarcocystis neurona, which are used for diagnosing equine protozoal myeloencephalitis, under different storage conditions (room temperature and 4°C) and times up to 10 days. The study finds these test titers remain stable regardless of the storage conditions and times tested.

Study Objective and Methodology

  • The objective of this research was to study the impact of storage at room temperature and refrigeration, along with the time of storage, on the immunofluorescent antibody (IFA) titers for S. neurona. This is an important aspect to evaluate, as ensuring samples’ timely and proper refrigeration may not always be achievable.
  • The study utilized twenty-two serum samples and designed a prospective longitudinal study using these samples. Two serum aliquots, each 1mL, were stored – one at room temperature (20-23.3°C) and the other at 4°C (refrigeration temperature).
  • The IFA titers for the unrefrigerated sample were tested immediately, while both aliquots were retested on 5th and 10th day after collection.
  • A paired t test was deployed to compare the IFA titers at these different time points.

Key Findings

  • Results indicated no significant difference in the IFA titers from baseline compared to those stored at room temperature at 5 days and 10 days. The same was the case for samples stored at 4°C at these two intervals. In other words, the IFA titers showed stability over time, irrespective of the storage conditions.
  • Additionally, there was no significant statistical difference observed in samples stored at room temperature versus those kept at 4°C on Days 5 and 10. This result further points to the temperature-insensitivity of the IFA titers for S. neurona.

Conclusion

  • Overall, the research concludes that the immunofluorescent antibody test titers for S. neurona remain stable for up to 10 days, regardless of whether the samples are stored at room temperature or 4°C. This stability might imply the feasibility of conducting accurate S. neurona tests even in scenarios where immediate refrigeration of serum samples is not possible.

Cite This Article

APA
Valderrama-Martinez C, Packham A, Zheng S, Smith W, Plancarte M, Aleman M. (2024). Effect of refrigeration, room temperature, and processing time on serum immunofluorescent antibody titers for Sarcocystis neurona. J Vet Intern Med, 39(1), e17282. https://doi.org/10.1111/jvim.17282

Publication

Journal of veterinary internal medicine
ISSN: 1939-1676
NlmUniqueID: 8708660
Country: United States
Language: English
Volume: 39
Issue: 1
Pages: e17282
PII: e17282

Researcher Affiliations

Valderrama-Martinez, Claudia
  • Departamento de Salud Animal, Facultad de Medicina Veterinaria y Zootecnia, Universidad Nacional de Colombia, Bogotá, Colombia.
Packham, Andrea
  • One Health Institute, School of Veterinary Medicine, University of California Davis, Davis, California, USA.
Zheng, Shichen
  • Department of Medicine and Epidemiology, School of Veterinary Medicine, University of California Davis, Davis, California, USA.
Smith, Woutrina
  • One Health Institute, School of Veterinary Medicine, University of California Davis, Davis, California, USA.
Plancarte, Magdalena
  • One Health Institute, School of Veterinary Medicine, University of California Davis, Davis, California, USA.
  • Karen C. Drayer Wildlife Health Center, School of Veterinary Medicine, University of California, Davis, California, USA.
Aleman, Monica
  • Department of Medicine and Epidemiology, School of Veterinary Medicine, University of California Davis, Davis, California, USA.

MeSH Terms

  • Sarcocystis / immunology
  • Animals
  • Refrigeration / veterinary
  • Horses
  • Temperature
  • Sarcocystosis / veterinary
  • Sarcocystosis / blood
  • Sarcocystosis / diagnosis
  • Sarcocystosis / parasitology
  • Antibodies, Protozoan / blood
  • Prospective Studies
  • Horse Diseases / blood
  • Horse Diseases / parasitology
  • Horse Diseases / immunology
  • Horse Diseases / diagnosis
  • Specimen Handling / veterinary
  • Longitudinal Studies
  • Fluorescent Antibody Technique / veterinary
  • Time Factors
  • Encephalomyelitis / veterinary
  • Encephalomyelitis / parasitology
  • Encephalomyelitis / blood
  • Encephalomyelitis / immunology

Grant Funding

  • PD1001 / The Seed Grants for International Activities Program (Aleman), University of California Davis
  • V435AM2 / Equine and Comparative Neurology Research Group (Aleman), University of California Davis

Conflict of Interest Statement

Authors declare no conflict of interest.

References

This article includes 29 references
  1. Dubey JP. Invited review: Sarcocystis neurona, Neospora spp. and Toxoplasma gondii infections in horses and equine protozoal myeloencephalitis (EPM): five decades of personal experience, perspectives, and update.. Parasitology 2022;149:1‐44.
    pubmed: 35260209
  2. Dubey JP, Howe DK, Furr M. An update on Sarcocystis neurona infections in animals and equine protozoal myeloencephalitis (EPM).. Vet Parasitol 2015;209:1‐42.
    pmc: PMC4461864pubmed: 25737052
  3. Fayer R, Mayhew IG, Baird JD. Epidemiology of equine protozoal myeloencephalitis in North America based on histologically confirmed cases. A report.. J Vet Intern Med 1990;4:54‐57.
    pubmed: 2342022
  4. More G, Vissani A, Pardini L. Seroprevalence of Sarcocystis neurona and its association with neurologic disorders in Argentinean horses.. J Equine Vet Sci 2014;34:1051‐1054.
  5. Llano HAB, Soares RM, Acevedo‐Gutierrez LY. Seroepidemiology of Sarcocystis neurona and Neospora spp. in horses, donkeys, and mules from Colombia.. Acta Trop 2021;220:105970.
    pubmed: 34033805
  6. Wobeser BK, Godson DL, Rejmanek D, Dowling P. Equine protozoal myeloencephalitis caused by Neospora hughesi in an adult horse in Saskatchewan.. Can Vet J 2009;50:851‐853.
    pmc: PMC2711471pubmed: 19881924
  7. Alvarado‐Esquivel C, Howe DK, Yeargan MR, Alvarado‐Esquivel D, Alfredo Zamarripa‐Barboza J, Dubey JP. Seroepidemiology of Sarcocystis neurona and Neospora hughesi infections in domestic donkeys (Equus asinus) in Durango, Mexico.. Parasite 2017;24:27.
    pmc: PMC5520387pubmed: 28730993
  8. Yeargan MR, Alvarado‐Esquivel C, Dubey JP, Howe DK. Prevalence of antibodies to Sarcocystis neurona and Neospora hughesi in horses from Mexico.. Parasite 2013;20:29.
    pmc: PMC3767921pubmed: 24016396
  9. Brown CM, Morrow JK, Carleton CL. Persistence of serum antibodies to Sarcocystis neurona in horses moved from North America to India.. J Vet Intern Med 2006;20:994‐997.
    pubmed: 16955828
  10. Reed SM, Furr M, Howe DK. Equine protozoal myeloencephalitis: an updated consensus statement with a focus on parasite biology, diagnosis, treatment, and prevention.. J Vet Intern Med 2016;30:491‐502.
    pmc: PMC4913613pubmed: 26857902
  11. Packham AE, Conrad PA, Wilson WD. Qualitative evaluation of selective tests for detection of Neospora hughesi antibodies in serum and cerebrospinal fluid of experimentally infected horses.. J Parasitol 2002;88:1239‐1246.
    pubmed: 12537119
  12. Renier AC, Morrow JK, Graves AJ. Diagnosis of equine protozoal myeloencephalitis using indirect fluorescent antibody testing and enzyme‐linked immunosorbent assay titer ratios for Sarcocystis neurona and Neospora hughesi .. J Equine Vet Sci 2016;36:49‐51.
  13. Bernardino PN, Pusterla N, Conrad PA. Evidence of intrathecally‐derived antibodies against Toxoplasma gondii in horses suspected of neurological disease consistent with equine protozoal myeloencephalitis.. Vet Parasitol 2023;318(109919):109911‐109915.
    pubmed: 37043865
  14. James KE, Smith WA, Packham AE, Conrad PA, Pusterla N. Toxoplasma gondii seroprevalence and association with equine protozoal myeloencephalitis: a case‐control study of Californian horses.. Vet J 2017;224:38‐43.
    pubmed: 28697873
  15. Johnson AL, Morrow JK, Sweeney RW. Indirect fluorescent antibody test and surface antigen ELISAs for antemortem diagnosis of equine protozoal myeloencephalitis.. J Vet Intern Med 2013;27:596‐599.
    pubmed: 23517480
  16. Pandit PS, Smith WA, Finno CJ. A fresh look at the SarcoFluor antibody test for the detection of specific antibodies to Sarcocystis neurona for the diagnosis of equine protozoal myeloencephalitis.. Vet Parasitol 2024;330:110219.
    pubmed: 38897057
  17. Pusterla N, Packham A, Wilson WD. Short communication: evaluation of the kinetics of antibodies against Sarcocystis neurona in serum from seropositive healthy horses without neurological deficits treated with ponazuril paste.. Vet Rec 2013;173:249.
    pubmed: 23893215
  18. Pusterla N, Tamez‐Trevino E, White A. Comparison of prevalence factors in horses with and without seropositivity to Neospora hughesi and/or Sarcocystis neurona .. Vet J 2014;200:332‐334.
    pubmed: 24703324
  19. Gomes FA, Jansen AM, Machado RZ. Serological evidence of arboviruses and coccidia infecting horses in the Amazonian region of Brazil.. PLoS One 2019;14:e0225895.
    pmc: PMC6907776pubmed: 31830142
  20. James KE, Smith WA, Conrad PA. Seroprevalences of anti‐Sarcocystis neurona and anti‐Neospora hughesi antibodies among healthy equids in the United States.. J Am Vet Med Assoc 2017;250:1291‐1301.
    pubmed: 28509641
  21. Hodgkinson VS, Egger S, Betsou F. Preanalytical stability of antibodies to pathogenic antigens.. Cancer Epidemiol Biomarkers Prev 2017;26:1337‐1344.
    pubmed: 28483968
  22. Miller MA, Gardner IA, Packham A. Evaluation of an indirect fluorescent antibody test (IFAT) for demonstration of antibodies to Toxoplasma gondii in the sea otter (Enhydra lutris).. J Parasitol 2002;88:594‐599.
    pubmed: 12099433
  23. Miller MA, Crosbie PR, Sverlow KW. Isolation and characterization of Sarcocystis from brain tissue of a free living southern sea otter (Enhydra lutris nereis) with fatal meningoencephalitis.. Parasitol Res 2001;87:252‐257.
    pubmed: 11293576
  24. Duarte PC, Daft BM, Conrad PA. Evaluation and comparison of an indirect fluorescent antibody test for detection of antibodies to Sarcocystis neurona, using serum and cerebrospinal fluid of naturally and experimentally infected, and vaccinated horses.. J Parasitol 2004;90:379‐386.
    pubmed: 15165063
  25. Braun JP, Bourgès‐Abella N, Geffré A, Concordet D, Trumel C. The preanalytic phase in veterinary clinical pathology.. Vet Clin Pathol 2015;44:8‐25.
    pubmed: 25438659
  26. Rossi G, Richardson A, Jamaludin H, Secombe C. Preanalytical variables affecting the measurement of serum paraoxonase‐1 activity in horses.. J Vet Diagn Invest 2021;33:59‐66.
    pmc: PMC7758704pubmed: 33225843
  27. Gunn‐Christie RG, Flatland B, Friedrichs KR. ASVCP quality assurance guidelines: control of preanalytical, analytical, and postanalytical factors for urinalysis, cytology, and clinical chemistry in veterinary laboratories.. Vet Clin Pathol 2012;41:18‐26.
    pubmed: 22390424
  28. Frohlich M, Knights K, Springer NL. Determination of optimal storage time and temperature for the detection of red blood cell and platelet surface‐associated immunoglobulin by flow cytometry in healthy horses.. Vet Immunol Immunopathol 2023;263:110643.
    pubmed: 37595493
  29. Østergaard M, Sandfeld‐Paulsen B. Preanalytical temperature and storage stability of specific IgE antibodies in serum.. Scand J Clin Lab Invest 2023;83:160‐165.
    pubmed: 36988143

Citations

This article has been cited 1 times.
  1. Valderrama-Martinez C, Packham A, Smith W, Mendoza-Flores JE, Zheng S, Chigerwe M, Plancarte M, Aleman M. Effect of Long-Term Freezing on Indirect Fluorescent Antibody Titers for the Diagnosis of Equine Protozoal Myeloencephalitis. J Vet Intern Med 2025 Sep-Oct;39(5):e70225.
    doi: 10.1111/jvim.70225pubmed: 40873183google scholar: lookup
Subscribe to our newsletter
Join 99,780 horse owners like you who receive our email updates on horse health and nutrition.