Biochemical and Immune Profiles in Serum Samples

The biochemical parameters and hormone levels influencing the levels of calcium (Ca), phosphorus (P), blood urea nitrogen (BUN), cholesterol (CHO), glucose (GLU), triglyceride (TG), albumin (ALB), progesterone (P4), estrogen (E), and cortisol hormone in serum samples were determined using an Olympus AU400 chemistry analyzer (Olympus Optical Co., Ltd., Tokyo, Japan) following the manufacturer’s instructions. Immune activities, including alternative complement hemolytic 50 (ACH50) activities, total immunoglobulin (total Ig), and lysozyme activities (LZY), were determined using a method previously described by Incharoen et al. [13 (link)] with slight modifications. SOD activities were also determined using a modification of the mentioned method [13 (link)], and the enzymatic activity was expressed as the percent inhibition rate. The catalase activity (CAT; EC1.11.1.6) was determined using a catalase assay kit (ab83464; Abcam, Cambridge, UK) according to the manufacturer’s standard procedures. The activity of glutathione peroxidase (GPx; EC.1.11.1.9) was determined using a GPx assay kit (ab102530; Abcam, Cambridge, UK) following the manufacturer’s standard protocols. The activity of glutathione reductase (GR; EC1.8.1.7) was measured using a commercially available assay kit (ab83461; Abcam, Cambridge, UK) according to the manufacturer’s standard processes.