Unicel dxc 600

All participants had a blood sample collection on second day after admission to the rehabilitation clinic. Venous blood samples were drawn after a minimum 12 h overnight fast for the evaluation of thyroid axis hormones, NT-pro-BNP, hs-CRP and IL-6 concentrations. Blood was centrifuged and serum was frozen at –40 °C. Serum concentrations of T3, fT3, T4, free T4 (fT4), TSH were analysed using automated enzyme immunoassay analyser AIA 600/21/1800 (Tosoh corporation, US) and radioimmunoassay kit RIA (R-EW-125, Belgium) for reverse T3 (rT3). Normal range for T3, from 70 to 170 ng/dL; fT3, from 2.0 to 4.0 pg/mL; rT3, from 9 to 35 nd/dl; T4, from 4,5 to 12.0 μg/dL; fT4, from 7.0 to 17.0 pg/mL; TSH, from 0.25 to 4.5 μIU/mL. Serum concentration of NT-pro-BNP and IL-6 were assessed using radio-immunoassay method (Roche Cobas analyser, Roche Diagnostics, UK). Normal serum concentration of NT-pro-BNP and IL-6 are <157 ng/L and from 0 to 7 pg/mL, respectively. Serum hs-CRP concentrations were assessed using the chemiluminescent immunoassay method (Beckman Coulter Unicel DXC 600) with normal values of ≤0.3 mg/dL.

The plasma lipase activity was evaluated using specific commercial kit (SYNCHRON LX^® Systems Enzyme Validator Set) on an automatic biochemistry analyzer (UniCel^®DxC 600, Beckman Coulter) in the Habib Bourguiba hospital of Sfax City, Tunisia. The plasma lipase activity in each group was expressed in U/L.

Without any further processing, the urine samples were stored at -20°C until analysis. The concentrations of sodium (cNa⁺ (u)), potassium (cK⁺ (u)), calcium (cCa²⁺ (u)) and chloride (cCl^- (u)) were obtained by indirect ion-selective potentiometry. Magnesium (cMg (u)) was measured with an automatic analyzer (UniCel DxC 600, Beckman Coulter; timed endpoint method). The pH of urine (pH (u)) was assessed by pH-meter at room temperature (22±2 °C, air-conditioning). Additionally, the net acid-base excretion (cNABE (u)) with base excess (cBE (u)), acid excess (cAE (u)), the concentration of ammonium (cAmm (u)) and the base-acid ratio (BAR (u)) were determined by the titrimetric method as described by Lachmann and Schäfer [2 ]. Shortly, 10 ml of urine were titrated first with hydrochloric acid to reduce pH to 4.0 whereas the required amount of acid corresponded to cBE. Second, the sample was titrated with sodium hydroxide to raise pH again to 7.0, and the used amount of lye resulted in cAE. Last, formyl aldehyde was used to precipitate ammonium ions, pH was titrated with lye to 7.0 again in order to assess the cAmm in urine. After this, cNABE and BAR were calculated by the following Eqs 1 and 2:
$\mathrm{c}\mathrm{N}\mathrm{A}\mathrm{B}\mathrm{E}=\mathrm{c}\mathrm{B}\mathrm{E}–(\mathrm{c}\mathrm{A}\mathrm{E}+\mathrm{c}\mathrm{A}\mathrm{m}\mathrm{m})$
$\mathrm{B}\mathrm{A}\mathrm{R}=\mathrm{c}\mathrm{B}\mathrm{E}/(\mathrm{c}\mathrm{A}\mathrm{E}+\mathrm{c}\mathrm{A}\mathrm{m}\mathrm{m})$

Blood analysis was performed twice: once before (0 weeks) and once after the end of the study (8 weeks). After an overnight fast of at least 8 h, blood samples of 5 mL were collected and coagulated for at least 30 min at room temperature before being centrifuged at 3500 rpm for 7 min. For each sample, 1 mL of separated serum was placed in a microtube and frozen. The frozen serum was analyzed by the Buraydah Central Hospital Laboratories using a Chemistry Analyzer (Beckman Coulter UniCel DxC 600; Beckman Coulter, Brea, CA, USA). In accordance with Choi et al. [25 (link)], the analysis included the total cholesterol, triglyceride, LDL-C, and HDL-C levels.

Information about signalment, clinical signs, serum biochemistry profile (SBP), and CBC within the first 24 hours after admission, treatments during hospitalization, and bacteriological and necropsy results were collected retrospectively when available.
Point‐of‐care blood testing upon admission (microhematocrit; manual refractometry; blood gas analysis by using the ABL80 Flex Co‐ox, Radiometer America, Brea, California; blood l‐lactate concentration using the Lactate Pro, Arkray Factory Inc., Shiga, Japan; and blood glucose concentration using the glucometer, Breeze 2, Ascensia Diabetes Care, Basel, Switzerland ) was chosen when available for statistical analysis instead of delayed results from our medical laboratory (hematology: Advia 120, Siemens Healthineers, Erlangen, Germany; serum biochemistry: Unicel DxC 600, Beckman Coulter, Brea, California).
Follow‐up examinations by phone were carried out in February 2018 by a single investigator. The owners of the calves were asked about long‐term survival of the calves and, if they died, the time between discharge and death. Three attempts were made to reach the owner; otherwise, the data were declared missing.