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Journal of veterinary internal medicine2023; 37(2); 713-717; doi: 10.1111/jvim.16637

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.
Publication Date: 2023-01-24 PubMed ID: 36692189PubMed Central: PMC10061166DOI: 10.1111/jvim.16637Google Scholar: Lookup
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  • 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

ISSN: 1939-1676
NlmUniqueID: 8708660
Country: United States
Language: English
Volume: 37
Issue: 2
Pages: 713-717

Researcher Affiliations

Quattrini, Camilla
  • 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.
Scalco, Rebeca
  • 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.
Vernau, William
  • 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.
Dini, Pouya
  • 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.
Aleman, Monica
  • 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
  1. Mayhew IG, Whitlock RH, Tasker JB. Equine cerebrospinal fluid: reference values of normal horses.. Am J Vet Res 1977 Aug;38(8):1271-4.
    pubmed: 911095
  2. Vernau W, Vernau KA, Bailey CS. Cerebrospinal fluid. In: Kaneko JJ, Harvey JW, Bruss ML, eds. Clinical Biochemistry of Domestic Animals. 6th ed. San Diego, CA: Elsevier; 2008:769‐819.
  3. Mayhew IG. Collection of cerebrospinal fluid from the horse.. Cornell Vet 1975 Oct;65(4):500-11.
    pubmed: 1192748
  4. Aleman M, Borchers A, Kass PH, Puchalski SM. Ultrasound-assisted collection of cerebrospinal fluid from the lumbosacral space in equids.. J Am Vet Med Assoc 2007 Feb 1;230(3):378-84.
    pubmed: 17269870doi: 10.2460/javma.230.3.378google scholar: lookup
  5. Pease A, Behan A, Bohart G. Ultrasound-guided cervical centesis to obtain cerebrospinal fluid in the standing horse.. Vet Radiol Ultrasound 2012 Jan-Feb;53(1):92-5.
  6. D'Angelo A, Miniscalco B, Bellino C, Bormida S, Borrelli A, Maurella C, Riondato F, Biolatti C, Cagnasso A. Analysis of cerebrospinal fluid from 20 calves after storage for 24 hours.. Vet Rec 2009 Apr 18;164(16):491-3.
    pubmed: 19377088doi: 10.1136/vr.164.16.491google scholar: lookup
  7. Fry MM, Vernau W, Kass PH, Vernau KM. Effects of time, initial composition, and stabilizing agents on the results of canine cerebrospinal fluid analysis.. Vet Clin Pathol 2006 Mar;35(1):72-7.
  8. Bienzle D, McDonnell JJ, Stanton JB. Analysis of cerebrospinal fluid from dogs and cats after 24 and 48 hours of storage.. J Am Vet Med Assoc 2000 Jun 1;216(11):1761-4.
    pubmed: 10844967doi: 10.2460/javma.2000.216.1761google scholar: lookup
  9. Bellino C, Miniscalco B, Bertone I, Cagnasso A, Occhiena E, Gianella P, D'Angelo A. Analysis of cerebrospinal fluid from cattle with central nervous system disorders after storage for 24 hours with autologous serum.. BMC Vet Res 2015 Aug 13;11:201.
    pmc: PMC4542111pubmed: 26268228doi: 10.1186/s12917-015-0502-xgoogle scholar: lookup
  10. Leary S, Underwood W, Anthony R. AVMA guidelines for the euthanasia of animals. Am Vet Med Assoc 2020;2020:1‐121.
  11. Peterson LN, Christian JA, Bentley RT, Moore GE, Thomovsky SA. Evaluation of the hydroxyethyl starch stabilizing agent, Vetstarch, in the preservation of canine cerebrospinal fluid samples.. Vet Clin Pathol 2020 Mar;49(1):95-99.
    pubmed: 32190921doi: 10.1111/vcp.12834google scholar: lookup
  12. Reed SM, Furr M, Howe DK, Johnson AL, MacKay RJ, Morrow JK, Pusterla N, Witonsky S. Equine Protozoal Myeloencephalitis: An Updated Consensus Statement with a Focus on Parasite Biology, Diagnosis, Treatment, and Prevention.. J Vet Intern Med 2016 Mar-Apr;30(2):491-502.
    pmc: PMC4913613pubmed: 26857902doi: 10.1111/jvim.13834google scholar: lookup
  13. Aleman M. Cerebrospinal fluid. In: Pusterla N, Higgins J, eds. Interpretation of Equine Laboratory Diagnostics. 1st ed. Singapore: Wiley Blackwell; 2018:393‐400.
  14. Young KAS, Hepworth-Warren KL, Dembek KA. Comparison of Fluid Analysis and Cytologic Findings of Cerebrospinal Fluid Between Three Collection Sites in Adult Equids With Neurological Disease.. Front Vet Sci 2022;9:821815.
    pmc: PMC9087559pubmed: 35558896doi: 10.3389/fvets.2022.821815google scholar: lookup

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