Response of serum biochemical profile, antioxidant enzymes, and gut microbiota to dietary Hong-bailanshen supplementation in horses

Abstract

Background: Traditional Chinese medicine (TCM) is widely used in humans and animals, which is very important for health. TCM affects the body 's immunity and changes in intestinal flora. This study was conducted to investigate the effects of dietary Hong-bailanshen (HBLS) supplementation in horses on serum biochemical profile, antioxidant enzymes and gut microbiota.

Methods: In this study, five horses were selected. On day 0, 14, 28, blood samples and feces were collected on days 0, 14, and 28 to analyse gut microbiota, serum biochemical and redox indexes.

Results: The results showed that the addition of HBLS to horse diets significantly decreased the level of alanine aminotransferase, alkaline phosphatase, creatine kinase and malondialdehyde (p < 0.05, p < 0.01) and significantly increased the activity of total antioxidant capacity, superoxide dismutase and catalase (p < 0.05, p < 0.01). Compared with day 14, the levels of alanine aminotransferase, alkaline phosphatase and creatine kinase were significantly decreased; however, the level of catalase was significantly increased in the horses continuously fed with HBLS for 28 days (p < 0.05, p < 0.01). Alpha diversity analysis was performed that chao1 (p < 0.05), observed_specicies, faith'pd and goods_coverage upregulated in the horses fed HBLS. A total of 24 differential genera were detected adding HBLS to diet increased the abundance of Bacillus, Lactobacillaceae, Leuconostocaceae, Christensenellaceae, Peptostreptococcaceae, Faecalibacterium, Erysipelotrichaceae, Pyramidobacter, Sphaerochaeta, WCHB1-25, Bacteria, Oscillospira, and Acetobacteraceae, while reduced Aerococcus, EtOH8, Syntrophomonas, Caulobacter, Bradyrhizobiaceae, W22, Succinivibrionaceae, and Desulfovibrio (p < 0.05, p < 0.01).

Conclusion: Adding HBLS to the diet could be a potentially effective strategy to improve horses' health.

Keywords: HBLS; antioxidant enzymes; biochemical profile; gut microbiota; horse.

Figure 1

Study design for the experiment.

Figure 2

Effect of HBLS on the serum biochemical profile.

Figure 3

Effect of HBLS on antioxidant enzymes.

Figure 4

HBLS changed the structure and diversity of the gut microbiota of horses. (A) Sequencing data statistical analysis, (B) length distribution of sequencing data, (C) rank abundance curve (D) alpha diversity index analysis, (E) sample rarefaction curves, (F) beta diversity analysis, and (G) group difference analysis. Significance is presented as ^*p < 0.05, ^**p < 0.01. Data are presented as the mean ± SEM (n = 5).

Figure 5

Effect of dietary HBLS on the relative abundance of gut microbiota in different taxa levels.

Figure 6

Species composition analysis of gut microbiota of horses. (A) Classification levels tree diagram, (B) GraPhlAn evolutionary tree diagram, and (C) Krona species composition diagram.

Figure 7

Different species and marker species analysis in horses. (A) Venn diagram, (B) PCA, (C) OPLS-DA, (D) bar chart of ASV/OTU numbers in different regions of the Venn diagram, (E) genera composition heatmap, and (F) LEfSe analysis.

Figure 8

Random forests. (A) Model accuracy, (B) family, and (C) genus.

Figure 9

Comparing genera differences in horses’ microbiota. Significance is presented as ^∗p < 0.05, ^∗∗p < 0.01. All data were presented as mean ± SD (n = 5).

Figure 10

Potential function prediction analysis of horses’ microbiota. (A) KEGG, (B) MetaCyc, and (C) species composition of metabolic pathways.