## 2 Materials and Methods
### Experimental Design and Dietary Treatment
\begin{table}
\begin{tabular}{l c c c} \hline
**Ingredients (\%, Otherwise as Indicated)** & **Control Diet** & **AM Diet** & **PM Diet** \\ \hline Soybean meal & 12.71 & 15.60 & 11.86 \\ Barley & 10.00 & 10.00 & 10.00 \\ Wheat & 51.86 & 50.20 & 46.71 \\ Canada meal & 10.00 & 10.00 & 10.00 \\ Canada oil & 3.71 & 3.60 & 4.00 \\ Limestone & 10.72 & 7.60 & 11.71 \\ Salt & 0.16 & 0.33 & 0.19 \\ Monocalcium phosphate & 0.39 & 0.18 & 0.98 \\ Sodium bicarbonate & 0.24 & 0.00 & 0.20 \\ L-lysine HCl & 0.060 & 0.062 & 0.007 \\ D,L-methionine & 0.137 & 0.173 & 0.092 \\ L-threonine & 0.010 & 0.019 & 0.000 \\ Choline chloride 60\% & 0.027 & 0.000 & 0.000 \\ Layer vitamin–mineral premix 1 & 0.100 & 0.100 & 0.100 \\ Pigment red & 0.004 & 0.004 & 0.004 \\ Pigment yellow & 0.003 & 0.003 & 0.003 \\ Xylanase (Axtra XB) 2 & 0.010 & 0.010 & 0.010 \\ Phytase (Axtra Phy) 3 & 0.010 & 0.010 & 0.010 \\ Bentonite & 0.000 & 2.200 & 4.100 \\ \hline \multicolumn{4}{l}{Calculated nutrient composition} \\ \hline AMEn, kcal/kg & 2780 & 2980 & 2580 \\ CP, \% & 18.8 & 20.1 & 17.5 \\ Crude fat, \% & 5.3 & 6.7 & 3.6 \\ Crude fiber, \% & 2.9 & 3.0 & 2.8 \\ Dig. Arg, \% & 1.013 & 1.097 & 0.926 \\ Dig. Lys, \% & 0.810 & 0.900 & 0.760 \\ Dig. Met, \% & 0.440 & 0.511 & 0.410 \\ Dig. Cys, \% & 0.288 & 0.303 & 0.274 \\ Dig. Met + Cys, \% & 0.735 & 0.820 & 0.691 \\ Dig. Trp, \% & 0.214 & 0.229 & 0.198 \\ Dig. Ile, \% & 0.670 & 0.720 & 0.619 \\ Dig. Thr, \% & 0.570 & 0.630 & 0.527 \\ Dig. Val, \% & 0.774 & 0.826 & 0.720 \\ Calcium, \% & 4.10 & 2.50 & 5.60 \\ Available phosphorus, \% & 0.45 & 0.45 & 0.45 \\ Sodium, \% & 0.17 & 0.17 & 0.17 \\ Chloride, \% & 0.23 & 0.23 & 0.23 \\ Choline, mg/kg & 1400 & 1400 & 1400 \\ Linoleic acid, \% & 1.67 & 2.05 & 1.25 \\ \hline \end{tabular} \({}^{\dagger}\) Vitamin–mineral premix included the following per kilogram of diet: 10,000 IU of vitamin A, 3000 IU of vitamin D, 20 mg of vitamin E, 3 mg of vitamin K, 35 mg of nicotinic acid (niacin), 12 mg of folic acid, 6 mg of riboflavin (B2), 0.02 mg of cyanocobalamin (B12), 0.1 mg of biotin, 5 mg of pyridoxine (Rb6), 2 mg of thiamine (B1), 8 mg of copper as copper sulphate pentohydrate, 0.2 mg of cobalt as cobalt sulphate 21%, 0.5 mg of molybdenum as sodium molybdate, 1 mg of iodine as potassium iodide 68%, 0.3 mg of selenium as selenium 2%, 60 mg of iron as iron sulphate 30%, 60 mg of zinc as zinc sulphate 35%, 90 mg of manganese as manganous oxide 60%, and 20 mg of antioxidant. 2 Xylanase: Aktra XB TPT 201, Danisco Animal Nutrition (IFF); 3 \({}^{\dagger}\)[PERSON]: Aktra PHY Gold, Danisco Animal Nutrition (IFF).  
\end{table} 
Table 1: Ingredients and nutrient composition of the control diet, AM diet, and PM diet. such as dry matter (DM), apparent metabolisable energy (AMEn), CP, crude fat, crude fibre, and total and digestible amino acids, as well as mineral and ash contents, were analysed using near-infrared reflectance spectroscopy (Foss NIR 6500, Hillerod, Denmark), standardised with Evonik AMINONIR Advanced calibration. These results were then utilised in diet formulation. Additionally, the nutrient composition of the final diets, including DM, gross energy (GE), CP, mineral, and ash contents, was analysed using standard methods [27] to ensure the nutrient content was as expected. These results are presented in Table 2.