Ab105135

Plasma alanine aminotransferase (ALT; ab105134) and aspartate aminotransferase (AST; ab105135) were measured with a colorimetric assay, using reagents from Abcam Inc. (Cambridge, UK). Lactate dehydrogenase (LDH; K726) activity and blood urea nitrogen (BUN; ABIN577679) were analyzed using detection kit according to the manufacturer's instructions, which kits were obtained from BioVision, Inc. (Milpitas, CA, USA) and antibodies‐online GmbH (Aachen, Germany), respectively.

ALT and AST activity was determined using kits (ab105134, ab105135) from Abcam.

Serum glucose was measured using a FreeStyle Optium Neo Blood Glucose Meter (Abbott Laboratories). Commercially available kits were used to determine serum insulin (ALPCO, Cat No. 80-INSMSU-E01), alanine aminotransferase (ALT) and aspartate aminotransferase (AST) (Abcam, ab105134 and ab105135). The serum was diluted 1:25 for insulin analysis and 1:10 for ALT and AST. Experiments were conducted according to the manufacturer’s instructions.

According to the manufacturer’s instructions, plasma immunoglobulin (Ig) A, G, and M levels were measured using commercial ELISA kits (Shanghai Jianglai Biotechnology Co., Ltd., Shanghai, China). The assays limits of detection, limits of quantification, intra-assay CV%, and interassay CV% are presented in Table 3 according to the manufacturer-provided data.
Plasma corticosterone level was quantified using chicken-specific quantitative competitive ELISA kit (MBS701668; MyBioSource, San Diego, CA, USA). According to the manufacturer, the CV% for the intra- and interassay were <8% and <10%, respectively, with a detection range of 0.5–20 ng/mL. Meanwhile, plasma total protein (TP), triglyceride (TG), and total cholesterol levels were quantified according to the kits’ protocols using quantitative colorimetric assays (ab102535, ab65336, and ab65390, respectively; Abcam, Waltham, MA, USA). As for liver enzyme activity, alanine amino transferase (ALT) and aspartate amino transferase (AST) activities were determined using commercial kits applying enzyme activity colorimetric assays (ab241035 and ab105135, respectively; Abcam, Waltham, MA, USA).

The fasting blood glucose level was measured by glucometer weekly through the tail prick method up to six weeks of the experiment. The serum glucose and insulin concentrations were estimated by a rat glucose assay kit (81693, crystal chem, Elk Grove Village, IL, USA) and an insulin ELIZA kit (ELR-insulin, RayBio^®, Peachtree Corners, GA, USA). Glycosylated hemoglobin was estimated using a rat hemoglobin Hb1Ac assay kit (MBS2033689, My BioSure, Grass Valley, CA, USA). The serum triglyceride, high-density lipid (HDL), low-density lipid (LDLs), and cholesterol levels were calculated using commercially available kits (ab65359, ab65336, and ab65390, respectively; Abcam, Cambridge, UK). The serum level of liver function enzymes, i.e., alanine aminotransaminase (ALT) and aspartate aminotransferase (AST), were determined through the kit assay method (ab105134/k752-100 and ab105135, respectively; Abcam, Cambridge, UK).