Immunoreactive insulin stability in horses at risk of insulin dysregulation.
Abstract: Diseases associated with insulin dysregulation (ID), such as equine metabolic syndrome and pituitary pars intermedia dysfunction, are of interest to practitioners because of their association with laminitis. Accurate insulin concentration assessment is critical in diagnosing and managing these diseases. Objective: To determine the effect of time, temperature, and collection tube type on insulin concentrations in horses at risk of ID. Methods: Eight adult horses with body condition score >6/9. Methods: In this prospective study, subjects underwent an infeed oral glucose test 2 hours before blood collection. Blood samples were divided into ethylenediaminetetraacetic acid, heparinized, or serum tubes and stored at 4 or 20°C. Tubes were centrifuged and analyzed for insulin by a chemiluminescent assay over 8 days. Changes in insulin concentrations were compared with a linear mixed effects model. Results: An overall effect of time, tube type and temperature was identified (P = .01, P = 0.001, and P = 0.001, respectively). Serum and heparinized samples had similar concentrations for 3 days at 20°C and 8 days at 4°C; however, after 3 days at 20°C, heparinized samples had significantly higher insulin concentrations (P = .004, P = .03, and P = .03 on consecutive days). Ethylenediaminetetraacetic acid samples had significantly lower insulin concentrations regardless of time and temperature (P = .001 for all comparisons). Conclusions: These results suggest an ideal protocol to determine insulin concentrations involves using serum or heparinized samples with analysis occurring within 3 days at 20°C or 8 days at 4°C.
© 2019 The Authors. Journal of Veterinary Internal Medicine published by Wiley Periodicals, Inc. on behalf of the American College of Veterinary Internal Medicine.
Publication Date: 2019-10-16 PubMed ID: 31617618PubMed Central: PMC6872612DOI: 10.1111/jvim.15629Google Scholar: Lookup
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- Journal Article
- Adult Horses
- Blood Analysis
- Body Condition Score
- Clinical Study
- Diagnosis
- Disease Diagnosis
- Disease Management
- Equine Health
- Equine Metabolic Syndrome
- Glucose Tolerance Test
- Heparin
- Horses
- Insulin
- Laboratory Methods
- Metabolic Health
- Pituitary Pars Intermedia Dysfunction
- Serum
- Temperature
- Veterinary Medicine
- Veterinary Research
Summary
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The research evaluates how time, temperature, and type of collection tube affects insulin concentrations in horses at risk of insulin dysregulation (ID), a condition associated with diseases like equine metabolic syndrome and pituitary pars intermedia dysfunction.
Research Objectives and Methods
- The primary aim of this research is to ascertain the effect of factors such as time, temperature, and the type of collection tube on insulin concentrations in horses at risk of ID.
- The researchers undertook a prospective study involving eight adult horses which had body condition scores greater than 6 out of 9. These horses were subjected to an oral glucose test about 2 hours before blood collection.
- The collected blood samples were then divided into three types of tubes – ethylenediaminetetraacetic acid, heparinized, or serum tubes. These samples were stored at temperatures of either 4 or 20°C.
- Once stored, the tubes were centrifuged and then analyzed for insulin via a chemiluminescent assay for a total time of 8 days. The resulting changes in insulin concentrations were compared using a statistical tool known as a linear mixed-effects model.
Research Findings
- The analysis found a significant effect of time, tube type, and temperature on insulin concentrations (P-values were 0.01, 0.001, and 0.001, respectively).
- More specifically, the serum and heparinized samples displayed similar concentrations for the first 3 days at 20°C and 8 days at 4°C. After 3 days at 20°C though, the concentration of insulin in the heparinized samples was higher.
- The tubes containing ethylenediaminetetraacetic acid consistently displayed lower insulin concentrations, regardless of the time or the temperature condition, when compared to the other types of tubes used.
Conclusion
- Based on the results, it’s suggested that using serum or heparinized samples with an analysis occurring within a three-day period at 20°C or an eight-day period at 4°C is the ideal protocol. This protocol will support accurate determination of insulin concentrations.
- This study contributes valuable insights for managing and diagnosing diseases linked with insulin dysregulation, enhancing our understanding of the effects of time, temperature, and collection tube types on insulin stability.
Cite This Article
APA
Leschke DH, Muir GS, Hodgson JK, Coyle M, Horn R, Bertin FR.
(2019).
Immunoreactive insulin stability in horses at risk of insulin dysregulation.
J Vet Intern Med, 33(6), 2746-2751.
https://doi.org/10.1111/jvim.15629 Publication
Researcher Affiliations
- School of Veterinary Science, The University of Queensland, Gatton, Queensland, Australia.
- School of Veterinary Science, The University of Queensland, Gatton, Queensland, Australia.
- School of Veterinary Science, The University of Queensland, Gatton, Queensland, Australia.
- School of Agriculture and Food Sciences, The University of Queensland, Gatton, Queensland, Australia.
- School of Veterinary Science, The University of Queensland, Gatton, Queensland, Australia.
- School of Veterinary Science, The University of Queensland, Gatton, Queensland, Australia.
MeSH Terms
- Animals
- Blood Specimen Collection / veterinary
- Female
- Horse Diseases / blood
- Horses
- Insulin / blood
- Insulin / metabolism
- Male
- Metabolic Syndrome / veterinary
- Pituitary Diseases / blood
- Pituitary Diseases / veterinary
- Pituitary Gland, Intermediate
Grant Funding
- John and Mary Kibble Trust
- The University of Queensland, Early Career Researcher Award
Conflict of Interest Statement
François‐René Bertin has consulted for and received funding from Boehringer Ingelheim Pty Ltd for his research.
References
This article includes 28 references
- Ireland JL, McGowan CM. Epidemiology of pituitary pars intermedia dysfunction: A systematic literature review of clinical presentation, disease prevalence and risk factors.. Vet J 2018 May;235:22-33.
- Hoffman RM, Boston RC, Stefanovski D, Kronfeld DS, Harris PA. Obesity and diet affect glucose dynamics and insulin sensitivity in Thoroughbred geldings.. J Anim Sci 2003 Sep;81(9):2333-42.
- Welsh CE, Duz M, Parkin TDH, Marshall JF. Prevalence, survival analysis and multimorbidity of chronic diseases in the general veterinarian-attended horse population of the UK.. Prev Vet Med 2016 Sep 1;131:137-145.
- Frank N, Geor RJ, Bailey SR, Durham AE, Johnson PJ. Equine metabolic syndrome.. J Vet Intern Med 2010 May-Jun;24(3):467-75.
- Horn R, Bamford NJ, Afonso T, Sutherland M, Buckerfield J, Tan RHH, Secombe CJ, Stewart AJ, Bertin FR. Factors associated with survival, laminitis and insulin dysregulation in horses diagnosed with equine pituitary pars intermedia dysfunction.. Equine Vet J 2019 Jul;51(4):440-445.
- Bertin FR, de Laat MA. The diagnosis of equine insulin dysregulation.. Equine Vet J 2017 Sep;49(5):570-576.
- Tadros EM, Fowlie JG, Refsal KR, Marteniuk J, Schott HC 2nd. Association between hyperinsulinaemia and laminitis severity at the time of pituitary pars intermedia dysfunction diagnosis.. Equine Vet J 2019 Jan;51(1):52-56.
- de Laat MA, McGowan CM, Sillence MN, Pollitt CC. Equine laminitis: induced by 48 h hyperinsulinaemia in Standardbred horses.. Equine Vet J 2010 Mar;42(2):129-35.
- Bertin FR, Sojka-Kritchevsky JE. Comparison of a 2-step insulin-response test to conventional insulin-sensitivity testing in horses.. Domest Anim Endocrinol 2013 Jan;44(1):19-25.
- Smith S, Harris PA, Menzies-Gow NJ. Comparison of the in-feed glucose test and the oral sugar test.. Equine Vet J 2016 Mar;48(2):224-7.
- Rendle DI, Litchfield E, Gough S, Cowling A, Hughes KJ. The effects of sample handling and N-phenylmaleimide on concentration of adrenocorticotrophic hormone in equine plasma.. Equine Vet J 2015 Sep;47(5):587-91.
- Prutton JS, Kass PH, Watson JL, Pusterla N. Pre-analytical stability of adrenocorticotrophic hormone from healthy horses in whole blood, plasma and frozen plasma samples.. Vet J 2015 Apr;204(1):123-4.
- McDonald TJ, Perry MH, Peake RW, Pullan NJ, O'Connor J, Shields BM, Knight BA, Hattersley AT. EDTA improves stability of whole blood C-peptide and insulin to over 24 hours at room temperature.. PLoS One 2012;7(7):e42084.
- Carslake H, Karikoski N, Pinchbeck G, McGowan C. Serum insulin concentration in horses: Effect of storage and handling.. Vet J 2016 May;211:94-6.
- Carslake HB, Pinchbeck GL, McGowan CM. Evaluation of a Chemiluminescent Immunoassay for Measurement of Equine Insulin.. J Vet Intern Med 2017 Mar;31(2):568-574.
- Warnken T, Huber K, Feige K. Comparison of three different methods for the quantification of equine insulin.. BMC Vet Res 2016 Sep 9;12(1):196.
- Kraft JR. IMx (Abbott) Immunoassay of Insulin: A Practical Alternative to RIA Hyperinsulinemia Identification in Idiopathic Neurootology and Other Hyperinsulin Metabolic Disorders.. Int Tinnitus J 1997;3(2):113-116.
- Henneke DR, Potter GD, Kreider JL, Yeates BF. Relationship between condition score, physical measurements and body fat percentage in mares.. Equine Vet J 1983 Oct;15(4):371-2.
- Bertin FR, Taylor SD, Bianco AW, Sojka-Kritchevsky JE. The Effect of Fasting Duration on Baseline Blood Glucose Concentration, Blood Insulin Concentration, Glucose/Insulin Ratio, Oral Sugar Test, and Insulin Response Test Results in Horses.. J Vet Intern Med 2016 Sep;30(5):1726-1731.
- Kutzner-Mulligan J, Eisemann J, Siciliano P, Smith J, Hewitt K, Sharlette J, Pratt-Phillips S. The effect of different feed delivery methods on time to consume feed and the resulting changes in postprandial metabolite concentrations in horses.. J Anim Sci 2013 Aug;91(8):3772-9.
- Chevenne D, Letailleur A, Trivin F, Porquet D. Effect of hemolysis on the concentration of insulin in serum determined by RIA and IRMA.. Clin Chem 1998 Feb;44(2):354-6.
- Ho EN, Wan TS, Wong AS, Lam KK, Stewart BD. Doping control analysis of insulin and its analogues in equine urine by liquid chromatography-tandem mass spectrometry.. J Chromatogr A 2011 Feb 25;1218(8):1139-46.
- Cross L. Serum insulin or plasma-insulin?. Lancet 1973 Mar 3;1(7801):491-2.
- Thorell JI, Lanner A. Influence of heparin-plasma, EDTA-plasma, and serum on the determination of insulin with three different radioimmunoassays.. Scand J Clin Lab Invest 1973 Mar;31(2):187-90.
- Conlon JM. Evolution of the insulin molecule: insights into structure-activity and phylogenetic relationships.. Peptides 2001 Jul;22(7):1183-93.
- Oddoze C, Lombard E, Portugal H. Stability study of 81 analytes in human whole blood, in serum and in plasma.. Clin Biochem 2012 Apr;45(6):464-9.
- Chameroy KA, Frank N, Elliott SB, Boston RC. Effects of a supplement containing chromium and magnesium on morphometric measurements, resting glucose, insulin concentrations and insulin sensitivity in laminitic obese horses.. Equine Vet J 2011 Jul;43(4):494-9.
- Reimers TJ, McCann JP, Cowan RG, Concannon PW. Effects of storage, hemolysis, and freezing and thawing on concentrations of thyroxine, cortisol, and insulin in blood samples.. Proc Soc Exp Biol Med 1982 Sep;170(4):509-16.
Citations
This article has been cited 2 times.- de Tonnerre DJ, Medina Torres CE, Stefanovski D, Robinson MA, Kemp KL, Bertin FR, van Eps AW. Effect of sirolimus on insulin dynamics in horses.. J Vet Intern Med 2023 Mar;37(2):703-712.
- Hicks GR, Fraser NS, Bertin FR. Changes Associated with the Peri-Ovulatory Period, Age and Pregnancy in ACTH, Cortisol, Glucose and Insulin Concentrations in Mares.. Animals (Basel) 2021 Mar 20;11(3).
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