Traditional Chinese herbal formulas modulate inflammatory mediators, antioxidant enzyme levels, and ruminal microbiota composition in postpartum female Yaks

Abstract

Traditional Chinese Medicine (TCM) is an emerging area due to increased antimicrobial resistance (AMR). The objective of this research was to explore the antioxidant, and anti-inflammatory potential of three traditional Chinese herbal formulas (TCHFs), along with variations in rumen bacteria. In this study, forty postpartum (80 ± 15) female yaks after the calves had weaned (PWFs) were divided into three experimental groups, which were offered basal feed with 5% (95% basal diet) TCHF1 (DE group), 5% TCHF2 (DF group) and 5% TCHF3 (DG group), and fourth, control group (DH group), fed only a basal diet for 30 days. Following blood and rumen fluid samples on the 15th and 30th day, ELISA testing was performed to check antioxidant enzyme levels and inflammatory mediators. The results indicated that TCHF2 significantly upregulated the interleukin-10 (IL-10) (p < 0.05). Additionally, 16 S rRNA sequencing results showed that TCHF2 significantly enhanced Firmicutes to Bacteroidetes ratio (F/B) at the phylum level. On day 15th, phylum Actinobacteria, SR1, Cyanobacteria, and Armatimonadetes were found to be significantly (p < 0.05) different, while, at the genus level, Butyrivibrio, CF231, YRC22, Moraxella, Clostridium, etc. were significantly different (p < 0.05). On day 30, phylum SR1, Armatimonadetes, Chlorobi, and genus Coprococcus, Oscillospira, Selenomonas, L7A_E11, Clostridium, etc. were found to be significantly different (p < 0.05). This study concluded that TCHF2 is the most effective one among all.

Keywords: Bos grunniens; 16S rRNA sequencing; Antioxidant; Herbal; Microbiome; Rumen.

Fig. 1

Effects of TCHFs on oxidative stress and inflammation in PWFs. (A–D) indicate the serum antioxidant-related indicators, including SOD, T-AOC, GSH-Px, and MDA. (E–H) represent the serum inflammatory mediators including IL-6, TNF-α, IL-1β, and IL-10. Data are represented as means ± SD. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.

Fig. 2

Alpha diversity analysis of rumen bacteria after provision with TCHFs in different groups. (A,D) alpha-diversity index of rumen bacteria from the group provided with TCHFs and sampled on days 15 and 30 of the study respectively. )B,E) Species accumulation curves of all samples collected on the days 15 and 30 of the study respectively. (C,F) Rank abundance curve of different samples collected on the days 15 and 30 of the study, respectively.

Fig. 3

Venn diagram of bacterial OTUs distribution of rumen fluid, collected from different groups fed with TCHFs on the 15th (A) and 30th day (B) respectively. Each ellipse stands for a sample group (DEZ1, DFZ1, DGZ1 and DHZ1 sampled on the 15th day; DEZ2, DFZ2, DGZ2, and DHZ2 sampled on the 30th day). Numbers indicate the count and percentage of unique and shared elements within each dataset.

Fig. 4

Beta diversity analysis of rumen bacteria from different groups treated with TCHFs. (A, C) PCoA analysis of bacterial OTU distribution of rumen fluid collected from different TCHFs fed groups on the 15th and 30th days respectively. (B, D) NMDS analysis of bacterial OTU distribution of rumen fluid, collected from different groups fed with TCHFs on the 15th and 30th day respectively.

Fig. 5

The relative abundances and distribution of significant rumen bacteria in different TCHFs treated groups, collected on the 15th (A–E) and 30th days (F–J) respectively. Rumen bacterial composition at the phylum (A, F), class (B, G), order (C, H), family (D, I), and genus (E, J) levels.

Fig. 6

Metastatic analysis of the rumen bacteria collected on the 15th day in the different groups treated with TCHFs (DEZ1, DFZ1, and DGZ1) compared with the control group (DHZ1) at the phylum and genus levels, showing the significantly different phylum and genera among four groups at day 15. Data are represented as means ± SD. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.

Fig. 7

Metastatic analysis of the rumen bacteria collected on the 30th day in the different groups (DEZ2, DFZ2, and DGZ2) compared with the control group (DHZ2) at the phylum and genus levels, showing the significantly different phylum and genera among four groups at day 30. Data are represented as means ± SD. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.

Fig. 8

LefSe analysis of rumen fluid bacteria collected from different groups provided with TCHFs on the 15th (A, C) and 30th (B, D) days respectively. (A, B) Cladograms showing taxa distribution across groups. (C, D) LDA scores highlight differentially abundant taxa within each group. LDA scores > 2 were considered significantly different.

Fig. 9

This diagram illustrates the key stages in the reproductive cycle of yaks over a year, offering a comprehensive overview of yak breeding and management. The cycle includes periods of estrus (heat), mating, pregnancy, calving, and weaning, along with the peak calving period and most productive time of the year. Mating occurs following the estrus period in late autumn. Pregnancy in yaks lasts approximately 250–260 days. The calving peak occurs in spring, followed by the weaning of calves.The period between summer and early autumn represents the most productive time for yaks.

Fig. 10

The inset map demonstrates the location of Lhasa within Tibet, underscoring the regional context. The main panel presents a detailed topographic map of Lhundup County, delineated by a black border, highlighting the mountainous terrain and geographical positioning of the study area.