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Effect of time and autologous serum addition on the analysis of cerebrospinal fluid in horses.
Abstract: Cerebrospinal fluid (CSF) is highly labile and delayed processing might alter results of analysis. Objective: To determine the effects of time and addition of autologous serum on cytological evaluation of CSF. Methods: Ten client-owned adult horses requiring euthanasia. Methods: Prospective study. Serum and CSF were collected from each horse before and within 10 minutes after euthanasia. CSF samples were divided into 15 aliquots (2 mL each); 1 aliquot was submitted for routine CSF analysis within 60 minutes of collection. Four drops of autologous serum were added to 7 of the aliquots, and stored at 4°C (serum group); the remaining 7 samples were stored unaltered at 4°C (control group). Total nucleated cell count (TNCC) and cell morphology score were done at T4, T8, T12, T24, T48, T72, and T96 hours after collection. Protein concentration was measured in the control group at T0 and T96 hours. Results: The cell morphology scores were significantly different in the control group at T48 (median 2, range 0-4), T72 (2, 0-4), and T96 (3, 0-4) in comparison to T0 (1). No change was observed in the serum group. TNCC remained stable over time in both groups. No statistically significant difference in CSF protein concentration was found between T0 and T96. Conclusions: The addition of autologous serum to an aliquot of CSF sample before shipping improves the preservation of cell morphology up to 96 hours after collection.
© 2023 The Authors. Journal of Veterinary Internal Medicine published by Wiley Periodicals LLC on behalf of American College of Veterinary Internal Medicine.
Publication Date: 2023-01-24 PubMed ID: 36692189PubMed Central: PMC10061166DOI: 10.1111/jvim.16637Google 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.
- Journal Article
Summary
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The study aims to assess the impact of time and the addition of autologous serum on the cytological evaluation of cerebrospinal fluid (CSF) from euthanized horses. It revealed that adding autologous serum to a CSF sample before sending it for testing enhances the preservation of cell morphology for up to 96 hours post-collection.
Methods
This research was carried out using a prospective study framework. The samples required for the study were CSF and serum, which were gathered from ten adult horses that were due for euthanasia. Both CSF and serum were collected before and within ten minutes after euthanasia.
The collected CSF samples were divided into 15 aliquots, each of 2 mL. One of these aliquots was sent for routine CSF analysis within an hour after collection. The remaining aliquots were divided into two groups: a ‘serum group’ and a ‘control group’. The serum group consisted of seven aliquots to which four drops of autologous serum were added and subsequently stored at 4°C. The rest, which formed the control group, were left unaltered and stored at the same temperature.
Analysis
The Total Nucleated Cell Count (TNCC) and cell morphology score for both groups were determined at specific time intervals: T4, T8, T12, T24, T48, T72, and T96 hours post-collection. Protein concentration was measured in the control group at T0 and T96 hours.
Results
The findings showed significant differences in the cell morphology scores for the control group at T48, T72, and T96 hours when compared to T0. However, no noticeable changes were observed in the serum group over time. The TNCC remained stable in both groups over a given timeframe. Also, no noteworthy differences were recorded in the CSF protein concentration between T0 and T96.
Conclusion
The research concluded that the addition of autologous serum to a CSF sample before shipment can enhance the preservation of cell morphology up to 96 hours after collection. Therefore, this method appears to be more effective for maintaining the condition of CSF samples set for cytological evaluation.
Cite This Article
APA
Quattrini C, Scalco R, Vernau W, Dini P, Aleman M.
(2023).
Effect of time and autologous serum addition on the analysis of cerebrospinal fluid in horses.
J Vet Intern Med, 37(2), 713-717.
https://doi.org/10.1111/jvim.16637 Publication
Researcher Affiliations
- William R. Pritchard Veterinary Medical Teaching Hospital (Quatrini) and Department of Medicine and Epidemiology (Scalco, Aleman), Pathology, Microbiology and Immunology (Vernau), Population, Health and Reproduction (Dini), School of Veterinary Medicine, University of California Davis, Davis, California, USA.
- William R. Pritchard Veterinary Medical Teaching Hospital (Quatrini) and Department of Medicine and Epidemiology (Scalco, Aleman), Pathology, Microbiology and Immunology (Vernau), Population, Health and Reproduction (Dini), School of Veterinary Medicine, University of California Davis, Davis, California, USA.
- William R. Pritchard Veterinary Medical Teaching Hospital (Quatrini) and Department of Medicine and Epidemiology (Scalco, Aleman), Pathology, Microbiology and Immunology (Vernau), Population, Health and Reproduction (Dini), School of Veterinary Medicine, University of California Davis, Davis, California, USA.
- William R. Pritchard Veterinary Medical Teaching Hospital (Quatrini) and Department of Medicine and Epidemiology (Scalco, Aleman), Pathology, Microbiology and Immunology (Vernau), Population, Health and Reproduction (Dini), School of Veterinary Medicine, University of California Davis, Davis, California, USA.
- William R. Pritchard Veterinary Medical Teaching Hospital (Quatrini) and Department of Medicine and Epidemiology (Scalco, Aleman), Pathology, Microbiology and Immunology (Vernau), Population, Health and Reproduction (Dini), School of Veterinary Medicine, University of California Davis, Davis, California, USA.
MeSH Terms
- Horses
- Animals
- Prospective Studies
- Time Factors
- Cell Count / veterinary
- Cerebrospinal Fluid
Grant Funding
- V435804 / Gifts from anonymous donors towards the Equine and Comparative Neurology Research Group
Conflict of Interest Statement
Authors declare no conflict of interest.
References
This article includes 14 references
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Citations
This article has been cited 1 times.- 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.
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