基于奶牛瘤胃菌群分析四君子散健脾效应的作用机制

doi:10.11843/j.issn.0366-6964.2025.12.049

摘要/Abstract

摘要： 本试验旨在研究四君子散对产后奶牛瘤胃菌群及其发酵能力的调节作用，以揭示其健脾效应的作用机制。试验将16头体重(600 kg±30 kg)、体况(3.0~3.5)、胎次相近的待产荷斯坦奶牛随机均分为试验组和对照组。试验组奶牛待分娩后于每天晨饲前灌服300 g四君子散，对照组奶牛灌服等体积干净饮用水，连续灌服7 d。在第8天晨饲前采集两组奶牛瘤胃液，采用ELISA法检测瘤胃发酵参数，气相色谱法测定瘤胃挥发性脂肪酸含量，16S rRNA高通量测序技术分析瘤胃菌群数量和结构变化。结果表明：灌服四君子散后，瘤胃pH无显著性变化(P＞0.05)，氨态氮(NH3-N)、微生物蛋白(MCP)、纤维素酶和α-淀粉酶含量增加(P＜0.01)，脲酶活性升高(P<0.05)；瘤胃丙酸和异丁酸的含量降低(P<0.05)，正丁酸的含量增高(P＜0.01)；瘤胃优势菌群在门水平无显著变化(P＞0.05)，仍为拟杆菌门(Bacteroidota)和厚壁菌门(Firmicutes)，但在属水平上，普雷沃氏菌属、理研菌科RC9菌属增多，Muribaculaceae菌属减少，另外10种非优势菌群在丰度上也发生了显著改变，6种有益菌属显著上调(P＜0.05)，包括[Eubacterium]_ruminantium_group、Rothia、Bacteroidales_RF16_group、Limnohabitans、hgcI_clade和Escherichia-Shigella，4种有害菌属显著下调(P＜0.05)，包括UCG-001、Clostridium_sensu_stricto_1、Dubosiella和Parvimonas。相关性分析显示，Rothia与α-淀粉酶、丙酸和异丁酸呈正相关(P＜0.05)；Limnohabitans与脲酶、纤维素酶、α-淀粉酶、NH3-N、MCP、丙酸、异丁酸含量呈正相关(P＜0.05)，与pH呈负相关(P＜0.001)；HgcI_clade与NH3-N呈正相关(P＜0.05)；Clostridium_sensu_stricto_1与pH呈负相关(P＜0.001)；Dubosiella与MCP、NH3-N和丙酸呈负相关(P＜0.05)；Parvimonas与MCP和NH3-N呈负相关(P＜0.05)。结果提示，四君子散健脾效应可能与调节瘤胃菌群丰度和增强瘤胃内消化酶活性密切相关。

关键词: 四君子散, 瘤胃发酵, 挥发性脂肪酸, 瘤胃微生物, 16S rRNA高通量测序

Abstract: The objective of this study was to explore how Sijunzi San influences the rumen microbial community and fermentation processes in postpartum dairy cows, to investigate the potential mechanisms of its spleen-invigorating effects. Sixteen multiparous Holstein cows with similar body weight (600 kg±30 kg) and body condition score (3.0-3.5) were randomly assigned to either an experimental or a control group. The experimental group received an oral administration of 300 grams of Sijunzi San following calving, whereas the control group was given an equal volume of clean drinking water, both for a duration of 7 consecutive days. On the eighth day, rumen fluid was collected from both groups prior to morning feeding. Rumen fermentation parameters were measured using ELISA, the content of volatile fatty acids was determined by gas chromatography, and changes in the quantity and structure of the rumen microbiota were analyzed using 16S rRNA amplicon high-throughput sequencing. The results indicated that following the administration of Sijunzi San, the rumen pH remained relatively stable and did not exhibit a significant change (P>0.05). However, the levels of ammonia nitrogen (NH₃-N), microbial protein (MCP), cellulase, and α-amylase experienced a notable increase (P<0.01), and urease activity also rose (P<0.05). Conversely, the concentrations of propionic acid and isobutyric acid in the rumen saw a decrease (P<0.05), whereas the content of n-butyric acid surged (P<0.01). At the phylum level, Bacteroidota and Firmicutes maintained their predominance as the two dominant bacterial phyla, showing no statistically significant alterations in their relative abundances (P>0.05). However, genus-level analysis revealed distinct shifts in rumen microbiota composition: the relative abundances of Prevotella and Rikenellaceae_RC9_gut_group exhibited a significant increase, while Muribaculaceae showed a marked reduction in abundance. Additionally, the abundance of 10 non-dominant microbiota changed significantly, with 6 beneficial genera significantly upregulated (P<0.05), including the Eubacterium ruminantium group, Rothia, Bacteroidales RF16 group, Limnohabitans, hgcI clade, and Escherichia Shigella. Four harmful genera were significantly downregulated (P<0.05), including UCG-001, Clostridium_sensu_stricto_1, Dubosiella, and Parvimonas. Rothia exhibited a positive correlation with α-amylase, propionic acid, and isobutyric acid (P<0.05); Limnohabitans was positively associated with urease, cellulase, α-amylase, NH₃-N, MCP, propionic acid, and isobutyric acid (P<0.05), and negatively correlated with pH (P<0.001); HgcI_clade had a positive correlation with NH₃-N (P<0.05); Clostridium_sensu_stricto_1 was negatively correlated with pH (P<0.001); Dubosiella was negatively associated with MCP, NH₃-N, and propionic acid (P<0.05); and Parvimonas was negatively correlated with MCP and NH₃-N (P<0.05). The results suggest that the spleen-invigorating effect of Sijunzi San may be closely related to the regulation of rumen microbiota abundance and the enhancement of digestive enzyme activity in the rumen.

Key words: Sijunzi San, rumen fermentation, volatile fatty acids, rumen microbiota, 16S rRNA high-throughput sequencing

中图分类号:

S853.74

杨文慧, 王菲菲, 李晨雷, 孙宇, 秦俊杰, 朱浩, 郭延生. 基于奶牛瘤胃菌群分析四君子散健脾效应的作用机制[J]. 畜牧兽医学报, 2025, 56(12): 6487-6501.

YANG Wenhui, WANG Feifei, LI Chenlei, SUN Yu, QIN Junjie, ZHU Hao, GUO Yansheng. Study on the Mechanism of Sijunzi San to Strengthen Spleen based on Rumen Flora of Dairy Cows[J]. Acta Veterinaria et Zootechnica Sinica, 2025, 56(12): 6487-6501.

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