A005 1

Liver, ovary, serum, and yolk concentrations of total superoxide dismutase (T-SOD, A001-1), glutathione peroxidase (GPX, A005-1), total antioxidant capacity (T-AOC, A015-1), and malondialdehyde (MDA, A003-1) concentrations were obtained according to manufacturer’s instructions and kit (Nanjing Jiancheng Bioengineering Institute, Nanjing, China) using biochemical methods.

The contents of superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), total antioxidant capacity (T-AOC), and malondialdehyde (MDA) in the serum and jejunum were detected using kits (Kits A001-3, A015-2, A005-1, A003-1, respectively; Nanjing Jiancheng Bioengineering Institute, Nanjing, China). The methods entailed the use of the xanthine-oxidase–xanthine reaction, reduced glutathione, ammonium molybdate, and 2-thiobarbituric acid for the determination of SOD, GSH-Px, CAT, and MDA, respectively.

Liver tissue samples weighing approximately 0.5 g from part of the left lobe (n = 6 layers each group) were subjected to homogenization in 4.5 mL of phosphate buffer saline. The samples were then centrifuged at 3,000 r/min for 10 min at 4 °C. The resulting supernatant was collected, yielding a 10% liver homogenate. Liver homogenates were utilized to measure the activity or content of antioxidant substrates, including glutathione peroxidase (GSH-Px), superoxide dismutase (SOD), catalase (CAT), malondialdehyde (MDA) as the operation manual described (A005-1, A001-3, A007-1-1 and A003-1, Nanjing Jiancheng Bioengineering Institute, Nanjing, China). GSH-Px activities were assessed by colorimetric assay, while SOD, CAT, and MDA activities were determined using by microplate assay.
The 10% liver homogenate was centrifuged at 2,000 r/min for 10 min. Subsequently, the supernatant was isolated by further centrifugation at 10,000 r/min for 15 min to obtain precipitate, which is liver mitochondria. ATPase activities, including Ca²⁺Mg²⁺-ATPase, Mg²⁺-ATPase, Ca²⁺-ATPase, and Na⁺K⁺-ATPase, were assayed colorimetrically on the obtained liver mitochondria using manufacturer's instructions (A016-2, Nanjing Jiancheng Bioengineering Institute, Nanjing, China).