Supplementary Materials - Serum 1H NMR-based metabolomics of sole lesion development in Holstein cows

Supplementary Methods. Details regarding NMR spectra acquisition and processing, and approach to statistical analysis.

Supplementary Table 1. Details of the spectral bins selected as representative of each annotated metabolite and used in statistical analysis, including the Bovine Metabolome database identification number (BMDB ID).

Supplementary Table 2. Details of the unlabeled spectral bins (not annotated to a metabolite) used in the statistical analysis.

Supplementary Table 3. Results of univariable analysis with Wilcoxon signed-rank tests. Data were analyzed in 17 prespecified subsets split by time point, parity, and outcome definition; the control group in all cases were animals without sole lesions, outcomes were either cases of sole new hemorrhage (New SH), cases of new sole ulcers (New SU), those two groups combined (New SH/SU) or all cases of sole ulcers (All SU).

Supplementary Table 4. Variable selection stability results including the mean log (base 2) fold-change (Log2FC) and the baseline stability thresholds (T99 and T100, equivalent to an expected 1% and 0% false positive rate, respectively). Data were analyzed in 17 prespecified subsets split by time point, parity, and outcome definition; the control group in all cases were animals without sole lesions, outcomes were either cases of sole new hemorrhage (New SH), cases of new sole ulcers (New SU), those two groups combined (New SH/SU) or all cases of sole ulcers (All SU).

补充方法。涉及核磁共振波谱的采集与处理细节，以及统计分析方法的阐述。

补充表 1. 详述了作为每个注释代谢物代表并用于统计分析的波谱区间，包括牛代谢组数据库识别号（BMDB ID）。

补充表 2. 详述了在统计分析中使用的未标记波谱区间（未标注为代谢物）的细节。

补充表 3. 单变量分析结果，采用 Wilcoxon 符号秩检验。数据在 17 个预设子集中进行分析，这些子集根据时间点、产次和结果定义进行划分；在所有情况下，对照组均为无单一损伤的动物，结果包括单一新出血（New SH）、新单一溃疡（New SU）、这两组合并（New SH/SU）或所有单一溃疡病例（All SU）。

补充表 4. 变量选择稳定性结果，包括平均对数（以 2 为底）变化倍数（Log2FC）和基线稳定性阈值（T99 和 T100，分别相当于预期的 1% 和 0% 假阳性率）。数据在 17 个预设子集中进行分析，这些子集根据时间点、产次和结果定义进行划分；在所有情况下，对照组均为无单一损伤的动物，结果包括单一新出血（New SH）、新单一溃疡（New SU）、这两组合并（New SH/SU）或所有单一溃疡病例（All SU）。