7600 automatic biological analyzer

Duplicate ingredient and diet samples were analyzed according to the AOAC (2007) procedure [15 ], including dry matter, crude protein, total phosphorus, calcium, and ether extract [16 (link)]. Neutral detergent fiber and acid detergent fiber were determined using filter bags and fiber analyzer equipment (Fibre Analyzer; Ankom Technology, Macedon, NY, USA), following a modified procedure [17 (link)]. Analysis of amino acid (AA) content included in the ingredients and diets was conducted according to a reported method [18 (link)].
After the frozen serum samples were thawed at 4 °C, serum levels of glucose, total protein, albumin, globin, triglyceride, total cholesterol, and urea nitrogen were quantified using a biological analyzer (7600 Automatic Biological Analyzer; Hitachi, Tokyo, Japan) at the Kangjia Hongyuan Biotech Company (Beijing, China). Serum heat stress protein 70 (HSP 70) concentration was measured using a microplate reader (Multiskan MK3 Microplate Reader, Waltham, MA, USA) at the Kangjia Hongyuan Biotech Company (Beijing, China).

Chemical analyses of all samples were conducted in duplicate. The diet was analyzed including dry matter (DM), crude protein (CP), calcium, total phosphorus (16 ) and ether extract (EE) (17 (link)). Neutral detergent fiber (NDF) and acid detergent fiber (ADF) were measured with filter bags using a Fiber Analyzer (Ankom Technology, Macedon, NY, USA). Gross energy (GE) was determined by an automatic adiabatic oxygen bomb calorimeter (Parr 1281, Automatic Energy Analyzer; Moline, IL, USA) according to previous study (18 (link)).
Blood samples were centrifuged (Biofuge22R; Heraeus, Hanau, Germany) at 3,000 × g for 10 min at 4°C, then the supernatant was transferred to storage tubes and stored at−80°C. After the frozen samples were thawed at 4°C, plasma levels of triglyceride, total cholesterol, high-density lipoprotein, low-density lipoprotein, total protein, albumin, and urea nitrogen were quantified using a biological analyzer (7600 Automatic Biological Analyzer; Hitachi, Tokyo, Japan) at the Kangjia Hongyuan Biotech Company (Beijing, China). Sample preparation for metabolomics, UPLC-MS analysis and data mining and processing according to previous study (1 (link)).