Erba chem 7

Rams were given saline water daily for nine months. All sheep had their blood extracted through the jugular vein, and 5 ml of the blood was collected in a dry, clean vacuum tube (Biomedica Alex.Co., Alexandria, Egypt). Clear, non-hemolyzed serum samples were held at -20 °C until further examination after blood was clotted at room temperature for 20 min and centrifuged at 1008 g for 10 min. The serum samples were used to measure nitric oxide (NO), malondialdehyde (MDA), glutathione (GSH), and superoxide dismutase (SOD) using commercial kits provided by Spectrum Diagnostics (Obour City, Cairo, Egypt). The Clinical Chemistry Analyzer ERBA CHEM 7, ERBA (Mannheim, Germany) was used. The FSH levels were measured using an enzyme-linked immunosorbent ELISA kit (MyBioSource, Southern California, San Diego, USA) [24 (link)]. LH concentration was assessed using an ELISA kit (MyBiosourse, Southern California, San Diego, USA), while testosterone concentration was evaluated using an ELISA kit (MyBiosourse, Southern California, San Diego, USA) [25 (link)]. The cortisol concentration was assessed using the RIA method [26 (link)].

Blood samples (10 mL) were withdrawn from jugular vein of lambs at 3 h after morning feeding. Blood was clotted at room temperature for 20 min and centrifuged at 3000 rpm for 10 min to obtain the serum. Serum samples were immediately separated, transported, and stored at − 20 ⁰C until the subsequent analysis. Blood serum samples were analyzed for total protein, albumin, alanine aminotransferase (ALT), aspartate aminotransferase (AST), urea, and creatinine using commercial test kits provided by (Spectrum Diagnostics, Egypt) and a spectrophotometer (Clinical Chemistry Analyzer ERBA CHEM 7, ERBA, Mannheim, Germany). The serum globulin concentration was calculated by subtracting the obtained albumin value from the total protein value.