Fig 1.
No differences in fecal microbiota alpha richness and diversity among groups.
A) Boxplots of observed OTUs for control (non-obese) and obese horses. The horizontal line is the median, and the top and bottom of the box extend to the 75% and 25% percentiles, respectively. The horizontal lines extending outside the box represent multiples of 1.75 times their respective interquartile range There was no statistical difference between groups (P = 0.34). B) Boxplot of Shannon diversity index for control (non-obese) and obese horses. The horizontal line within the box is the median, and the top and bottom of the box extend to the 75% and 25% percentiles, respectively. The horizontal lines extending outside the box represent multiples of 1.75 times their respective interquartile range. There was no statistical difference between groups (P = 0.62).
Fig 2.
Principal coordinate analysis plots.
A) PCoA based on unweighted UniFrac distance metric revealing no visual evidence of clustering by obese (circle) and non-obese(triangle) horses colored by farm B) PCoA based on Bray Curtis distance metric revealing no visual evidence of clustering of obese (circle) and non-obese (triangle) horses colored by farm.
Fig 3.
Unsupervised principal coordinate analysis (PCA) of fecal metabolites.
A) Unsupervised PCA segregated according to body condition score (BCS) from 3 to 8. Fecal metabolites did not cluster by BCS. B) Unsupervised PCA segregated according based on farm identification (Farm 1–7). Fecal metabolites visually clustered by farm.
Fig 4.
Intermediates of the mitochondrial tricarboxylic acid (TCA) cycle were altered in obese horses.
Box plots of the scaled intensity (y-axis) of the TCA cycle intermediates citrate (A), malate (B), isocitrate (C), and aconitate (D) comparing control (blue) and case (orange) (x-axis). The horizontal line of the boxplot is the median, and the top and bottom of the box extend to the 75% and 25% percentiles, respectively. The mean values are indicated by the plus (+) and the filled circles outside the thin horizontal line represent outliers. These TCA cycle intermediates were increased (P≤0.05) in the feces of obese horses. E) A graphical depiction of the TCA cycle is included for reference.
Fig 5.
Alterations in Vitamin E of obese horses.
Box plots of the scaled intensity (y-axis) of the vitamin E analogs α-tocopherol acetate (A), β-tocopherol (B), delta-tocopherol (C), α-tocotrienol (D), and gamma-tocotrienol (E) comparing control (blue) and case (orange) (x-axis). The horizontal line of the boxplot is the median, and the top and bottom of the box extend to the 75% and 25% percentiles, respectively. The mean values are indicated by the plus (+) and the filled circles outside the thin horizontal line represent outliers. These fecal metabolites were decreased (P≤0.05) in the feces of obese horses.
Fig 6.
Unsupervised principal coordinate analysis (PCA) of serum lipids.
A) Unsupervised PCA segregated according to body condition score (BCS) from 4 to 8, demonstrates no significant visual difference by BCS, B) Unsupervised PCA segregated according based on farm identification (Farm 1–7).
Fig 7.
Free fatty acids (FFA) in case and control horses.
Box plots of the scaled intensity (y-axis) of total FFA (A) including myristic acid (B), palmitoleic acid (C), oleic acid (D), linoleic acid (E), and α-linoleic acid (F) comparing control (blue) and case (orange) (x-axis). The horizontal line of the boxplot is the median, and the top and bottom of the box extend to the 75% and 25% percentiles, respectively. The mean values are indicated by the plus (+) and the filled circles outside the thin horizontal line represent outliers. These serum fatty acids were increased (P≤0.05) in the serum of obese horses.