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Evaluation of the effects of time and ambient temperature on the concentration stability of β-hydroxybutyrate and total protein in serum and plasma samples of sheep: a preliminary study about technical method

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Abstract

The extrinsic factors affecting the concentration of biochemical parameters such as transportation and storage at different temperatures are still wellspring of interest. This study is aimed at investigating the effects of time and environmental temperature on the concentration of β-hydroxybutyrate and total protein in ewe serum and plasma samples. The serum and plasma were separated from jugular vein blood samples of 80 ewes (2–3 years old). After being stored at temperatures of 4, 22–24, and 37 °C for 24 h, the serum and plasma samples were evaluated to measure their β-hydroxybutyrate and total protein concentration at zero time (T0) and 24 h (T24) after storage. The results showed that the concentration of β-hydroxybutyrate and total protein in these samples significantly increased after 24 h of storage at 37 °C compared to T0 (P < 0.05). However, the concentration of β-hydroxybutyrate and total protein in the samples at 4 °C and 22–24 °C decreased significantly as compared to 37 °C (P < 0.05). Moreover, the total protein concentration in serum and plasma decreased significantly after 24-h storage at 4 °C and 22–24 °C, while a significant increase occurred at 37 °C compared to T0 (P < 0.05). The findings also showed a significant reduction in the total protein concentration in serum and plasma at 4 °C and 22–24 °C compared to 37 °C (P < 0.05). However, there was no significant reduction in the plasma and total protein concentration at 4 °C compared to 22–24 °C (P > 0.05). According to the results, the concentration of β-hydroxybutyrate and total protein in the samples was not stable 24 h after sampling. Besides, the total protein concentration in the serum samples at 4 °C and 22–24 °C was not acceptable.

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The datasets generated during and/or analysed during the current study are available from the corresponding author on reasonable request.

References

  • Abraham RA (2019) Effect of temperature and time delay in centrifugation on stability of select biomarkers of nutrition and non-communicable diseases in blood samples. Biochem Med (zagreb) 29(2):020708

    Article  PubMed  Google Scholar 

  • Alberghina D, Casella S, Giannetto C, Marafioti S, Piccione G (2013) Effect of storage time and temperature on the total protein concentration and electrophoretic fractions in equine serum. Can J Vet Res 77:293–296

    CAS  PubMed  PubMed Central  Google Scholar 

  • Bobby L, Boyanton JR, Kenneth E (2002) Stability studies of twenty-four analytes in human plasma and serum. Clin Chem 48:2242–2247

    Article  Google Scholar 

  • Constable P, Hinchcliffe KW, Done S, Gruenberg W (2016) Veterinary medicine: a textbook of the disease of cattle, horses, sheep, pigs and goats. 17: Metabolic and Endocrine Diseases. Philadelphia, PA, USA. (11th ed.). Saunders Elsevier

  • Cuhadar S, Atay A, Koseoglu M, Dirican A, Hur A (2012) Stability studies of common biochemical analytes in serum separator tubes with or without gel barrier subjected to various storage conditions. Biochem Med 22:202–214

    Article  CAS  Google Scholar 

  • Custer EM, Myers JL, Poffenbarger PL, Schoen I (1983) The storage stability of 3-hydroxybutyrate in serum, plasma, and whole blood. Ame J Clin Patho 80(3):375–380

    Article  CAS  Google Scholar 

  • Dirar A, Dalael AA, Abdelsalam AK (2008) Effect of storage time and temperature on some serum analytes. Int J Patho 8:68–71

    Google Scholar 

  • Ehsani A, Afshari A, Bahadori H, Mohri M, Seifi HA (2008) Serum constituents analyses in dairy cows: effects of duration and temperature of the storage of clotted blood. Res Vet Sci 85:473–475

    Article  CAS  PubMed  Google Scholar 

  • Kaplan LA (1999) Determination and application of desirable analytical performance goals. Clin Lab Invest 59:479–482

    Article  CAS  Google Scholar 

  • Laffel L (1999) Ketone bodies: a review of physiology, pathophysiology and application of monitoring to diabetes. Diabetes Metab Res Rev 15:412–426

    Article  CAS  PubMed  Google Scholar 

  • LeBlanc S (2010) Monitoring metabolic health of dairy cattle in the transition period. J Rep Dev 56:29–35

    Article  Google Scholar 

  • Lippi G, Chance JJ, Church S, Dazzi P, Fontana R, Giavarina D (2011) Preanalytical quality improvement: from dream to reality. Clin Chem Lab Med 49:1113–1126

    Article  CAS  PubMed  Google Scholar 

  • Lotfollahzadeh S, Zakian A, Tehrani-Sharif M, Watson DG (2016) Assessment the alterations of some biochemical parameters in Afshari sheep with possible metabolic disorders. Small Rum Res 145:58–64

    Article  Google Scholar 

  • Mahrt A, Burfeind O, Heuwieser W (2014) Effects of time and sampling location on concentrations of b-hydroxybutyric acid in dairy cows. J Dairy Sci 97:291–298

    Article  CAS  PubMed  Google Scholar 

  • Marianne OM, Wenche O, Sigve B, Tore F (2018) Effects of storage duration and temperature conditions on biochemical analysts in porcine clotted, uncentrifuged blood samples. J Dairy Vet Anim Res 7(1):00181

    Google Scholar 

  • McNeil CA, Pramfalk C, Humphreys SM, Hodson L (2014) The storage stability and concentration of acetoacetate differs between blood fractions. Clin Chim Acta 433:278–283

    Article  CAS  PubMed  Google Scholar 

  • Megahed AA, Hiew MH, Townsend JR, Constable PD (2017) Characterization of the analytical performance of an electrochemical point-of-care meter for measuring β-hydroxybutyrate concentration in blood and plasma from periparturient dairy cattle. Vet Clin Pathol 46:314–325

    Article  PubMed  Google Scholar 

  • Melendez P, Risco CA (2005) Management of transition cows to optimize reproductive efficiency in dairy herds. Vet Clin Food Anim 21:485–501

    Article  Google Scholar 

  • Oddoze Ch, Lombard E, Portugal H (2012) Stability study of 81 analytes in human whole blood, in serum and in plasma. Clin Biochem 45:464–469

    Article  CAS  PubMed  Google Scholar 

  • Oetzel GR (2004) Monitoring and testing dairy herds for metabolic disease. Vet Clin North Am Food Anim 20(3):651–674 

    Article  PubMed  Google Scholar 

  • Ono T, Kitaguchi K, Takehara M, Shiiba M, Hayami K (1981) Serum-constituents analysis: effect of duration and temperature of storage of clotted blood. Clin Chem 27:35–38

    Article  CAS  PubMed  Google Scholar 

  • Russel A (1991) Body condition scoring of sheep. In: Boden, E. (Ed.), Sheep and goat practice. Bailliere Tindall, Philadelphia, Pp: 3–4

  • Saeed A, Afzal M, Akhtar S (1995) Effect of storage on some constituents of camel serum. Aust Vet J 72(6):2125

    Article  Google Scholar 

  • Simundic AM, Lippi G (2012) Preanalytical phase – a continuous challenge for laboratory professionals. Biochem Med 22:145–219

    Article  Google Scholar 

  • Stokol T, Nydam DV (2005) Effect of anticoagulant and storage conditions on bovine nonstrified fatty acid and B-hydroxybutyrate concentrations in blood. J Dairy Sci 88:3139–3144

    Article  CAS  PubMed  Google Scholar 

  • Young DS, Bermes EW (1999) Specimen collection and processing: sources of biological variation. In: Burtis CA, Ash wood ER. (Ed.), Tietz textbook of clinical chemistry. Philadelphia: WB Saunders company. Pp: 42–72

  • Zakian A, Haji-Hajikolaei MR, Tehrani-Sharif M, Faramarzian K, Safaei P (2019) Evaluation of negative energy balance and some metabolic disorders on peri-partum period in Arabian ewes Khuzestan. Iran Vet J 14(4):31–41

    Google Scholar 

  • Zakian A, Lotfollahzadeh S, Tehrani-Sharif M, Faramarzian K, Gholami N, NematiDelfani S (2015) Correlation among serum β-hydroxybutyric acid, total antioxidant capacity and protein profile pre and post-partum in Shall ewes. J Vet Med Res 2(5):1038

    Google Scholar 

  • Zakian A, Tehrani-Sharif M, Mokhber-Dezfouli MR, Nouri M, Constable PD (2017) Evaluation of a point-of-care electrochemical meter to detect subclinical ketosis and hypoglycaemia in lactating dairy cows. Aust Vet J 95(4):123–128 

    Article  CAS  PubMed  Google Scholar 

  • Zhang DJ, Elswick RK, Miller WG, Bailey JL (1998) Effect of serum-clot contact time on clinical chemistry laboratory results. Clin Chem 44:1325–1333

    Article  CAS  PubMed  Google Scholar 

  • Zivkovic AM, Wiest MM, Nguyen UT, Davis R, Watkins SM, German JB (2009) Effects of sample handling and storage on quantitative lipid analysis in human serum. Metabolomics 5:507–516

    Article  CAS  PubMed  PubMed Central  Google Scholar 

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Acknowledgements

This paper has been extracted from a DVM thesis of Nasim Erfani and the authors would like to express their special thanks and gratitude to the esteemed principal of the Faculty of Veterinary Medicine at Lorestan University and the Vice Dean of Research who gave us this opportunity to conduct this research in the laboratories of the Department of Veterinary Medicine.

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Authors and Affiliations

  1. Faculty of Veterinary Medicine, Lorestan University, Khorramabad, Iran

    Nasim Erfani

  2. Department of Clinical Sciences, Faculty of Veterinary Medicine, Lorestan University, Khorramabad, Iran

    Shahram Maleki, Amir Zakian & Moein Yazdkhasti

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  1. Nasim Erfani
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  2. Shahram Maleki
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  3. Amir Zakian
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  4. Moein Yazdkhasti
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Contributions

Nasim Erfani: conceptualization, methodology, investigation, and writing original draft. Shahram Maleki: project administration, writing, reviewing, and editing. Amir Zakian: writing original draft, writing, reviewing, editing, data curation, and formal analysis. Moein Yazdkhasti: methodology and validation.

Corresponding author

Correspondence to Shahram Maleki.

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Funding

This study was funded in part by Lorestan University (grant No.: LU-9311511035–2020).

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The authors declare no conflict of interest.

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During all stages of our research, all applicable international, national, and/or institutional guidelines for the care and use of animals were followed. In addition, this article does not contain any studies with animals performed by any of the authors and does not contain any studies with human participants or animals performed by any of the authors.

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Erfani, N., Maleki, S., Zakian, A. et al. Evaluation of the effects of time and ambient temperature on the concentration stability of β-hydroxybutyrate and total protein in serum and plasma samples of sheep: a preliminary study about technical method. Comp Clin Pathol 32, 211–216 (2023). https://doi.org/10.1007/s00580-022-03427-0

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