Fig. 1

Effects of Bohai black cattle control group and body brush intervention group on α-diversity of intestinal microorganismsNote: C2 and N2 were the control group and the body brushing group, respectively. * means significant difference between groups (P < 0.05). The same as below."

Fig. 2

Differences in gut microbial community structure between control and brush-intervention groups of Bohai black cattleNote: Ellipses in the NMDS ordination plot represent confidence intervals. Blue dots (C2) and orange dots (N2) represent samples from the two groups, respectively."

Fig. 3

Stacked bar charts of gut microbiota community composition at the genus and phylum levels of different group"

Fig. 4

Heatmap of genus-level abundanceNote: Z-score scaled heatmap and hierarchical clustering analysis of genus-level microbial abundance. Red indicates higher relative abundance, while blue indicates lower relative abundance. Samples are divided into group C2 (green) and group N2 (purple)."

Fig. 5

Random forest analysis of core gut genera in Bohai black cattleNote: The left bar plot displays the top 15 important genera identified by random forest analysis, while the right bar plot indicates their variable importance in the projection (VIP)."

Fig. 6

Co-occurrence network of gut microbial genera in Bohai black cattleNote: Nodes denote microbial genera."

Fig. 7

Pearson correlation analysis of quality control （QC） samples for serum metabolomics data"

Fig. 8

Orthogonal partial least squares discriminant analysis （OPLS-DA） model"

Fig. 9

Volcano plot analysis of differential metabolitesNote: With red (180) and blue (43) dots representing up- and down-regulated metabolites, respectively; dot size indicates the variable importance in projection (VIP) value."

Fig. 10

Scatter plot of mean metabolite intensitiesNote: Dot color indicates -lg(P-value), dot size represents the VIP value, and key differential metabolites are labeled."

Fig. 11

Pie chart of differential metabolite class distribution"

Fig. 12

Heatmap and clustering analysis of differential metabolite abundancesNote: Z-score scaled heatmap and hierarchical clustering of differential metabolite abundances, showing expression differences between C2 and N2 groups. The right-side columns annotate metabolite ontology, Log₂(FC), -lg(p), and VIP values."

Fig. 13

Pathway enrichment analysis of differential metabolitesNote: The size of each bubble indicates the number of differential metabolites in the pathway, and the color intensity represents enrichment significance (DAscore)."

Fig. 14

Pathway impact bubble plotNote: The ordinate represents pathway enrichment significance (-log₁₀(P)), and the abscissa represents pathway topological importance (Pathway Impact). The size of the dots indicates the pathway impact value, and the color indicates enrichment significance (the redder the color, the smaller the P value and the higher the enrichment level)."

Fig. 15

Box plots of relative abundances for key differential metabolites"

Fig. 16

Bar charts of antioxidant enzyme and oxidative stress marker levels"

Fig. 17

Bar chart of metabolite levels in energy metabolic pathways"

Fig. 18

Correlation network analysis between core gut genera and key differential metabolitesNote: Line color and thickness represent Mantel's P-value and correlation coefficient r, respectively, with a supplementary heatmap of Pearson's r on the right."

Fig. 19

Correlation heatmap between core gut genera and key differential metabolitesNote: Asterisks (*) indicate significant correlations (|r|≥0.5, P≤0.05), with red for positive and blue for negative correlations."