ucks were fed a corn oybean basal diet program formulated in accordance with the National Research Council (1994) (Table S2) and 500 mg kg-1 curcumin was added inside the basal diets for ducks inside the T500 + AFB1 group. On the 70th days, ducks have been fasted for 12 h and 15 have been selected from each group, oral administration of phosphate-buffered saline (PBS) (T0 ), and of 60 of AFB1 kg-1 physique weight (AFB1 was dissolved in PBS, for each T0 + AFB1 group and T500 + AFB1 group). All animal care and therapy regimens had been performed in strict accordance together with the regulation of the National Study Council Guide (1996) and Ethical and Animal Welfare Committee of Heilongjiang province, China (revised in 2016). The protocols employed within this study were authorized by the Institutional Animal Care and Use Committee of Northeast Agricultural University (protocol number: Northeast Agricultural University (NEAU)-[2011]-9). two.3. Sample Collection Complete blood samples have been obtained from duck wing veins 12 h immediately after AFB1 administration and were then centrifuged (1000g for 15 min at four C) and stored at -80 C. The liver was washed 3 instances in ice-cold phosphate-buffered saline (PBS, Beyotime Biotechnology BD2 Source Shanghai, China; pH = 7.2.4), then instantly and individually stored at -80 C for antioxidant enzymes activity and Genuine time quantitative PCR (qRT-PCR) analyses.Foods 2021, ten,3 of2.4. Histopathological Observation About 0.125 cm3 of liver was rapidly harvested and fixated with 4 paraformaldehyde for pathological research. Immediately after paraffin embedding, the samples were cut and stained with hematoxylin and eosin (H E) and observed using a light microscope (Nikon Eclipse Ci-L, Tokyo, Japan). The liver samples, at the degree of 1 mm3 , was fixed with two.5 glutaraldehyde and 1 osmic acid, dehydrated and embedded in resin. A final examination using the transmission electron microscopy (TEM, H-7650, Hitachi, Tokyo, Japan) was performed just after staining with uranyl acetate and lead citrate. two.five. Assay of CYP450 Content, AFB1-DNA Adducts Level and Antioxidant Capacity in Liver Liver samples have been homogenized in a pre-cooled 0.9 stroke-physiological saline answer (four C, 0.9 NaCl, pH = 7.two.4) and centrifuged at four C (5000g, ten min) to receive the supernatant. The contents of CYP450 and AFB1-DNA adducts in the liver have been determined by a competitive enzyme linked immune sorbent assay (ELISA) approach, in accordance with the manufacturer’s instructions (Nanjing Jiancheng CD40 web Bioengineering Institute, Nanjing, China). The activity or content material of total antioxidant capacity (T-AOC, U/mg protein), catalase (CAT, U/mg protein), total superoxide dismutase (T-SOD, U/mg protein), reductive glutathione glutathione S-transferase (GSH, ol/mg protein), Glutathione S-transferase (GST, U/mg protein), hydrogen peroxide (H2 O2 , mmol/mg protein), and hydrogen peroxide (MDA, nmol/mg protein) of liver homogenates was measured utilizing commercial kits (Nanjing Jiancheng Bioengineering Institute, Nanjing, China) as outlined by the manufacturer’s instructions. 2.6. Plasma Biochemical Assay Hematological and biochemical parameters were determined working with an automatic biochemical analyzer. The content material or activity of total protein (TP, g/L), albumin (ALB, g/L), globulin (GLB, g/L), ALB/GLB (A/G), total bilirubin (TBIL, ol/L), alkaline phosphatase (ALP, U/L), ALT (alanine aminotransferase, U/L), AST (alanine aminotransferase, U/L), and AST/ALT in the plasma was assessed with commercial kits (Nanjing Jiancheng Bioengineering Institute,
