中国畜牧兽医2025，Vol.52Issue(6)：2569-2581,13.DOI:10.16431/j.cnki.1671-7236.2025.06.011
基于体外产气法探究不同假单胞菌添加量对人工瘤胃内环境的影响
Exploring the Effect of Different Pseudomonas Additions on the Artificial Rumen Internal Environment Based on in vitro Gas Production Method
摘要
Abstract
[Objective]This study was aimed to investigate the effect of different Pseudomonas additions on the artificial rumen internal environment,and screen the appropriate Pseudomonas additions.[Method]Four high-yielding dairy cows were selected as the donors of rumen fluid,and the total mixed ration(TMR)of donor cows was used as the fermentation substrate.A one-way four-level experimental design was used,the fermentation substrate in control group(CON)was supplemented with 0.5 g TMR,and fermentation substrate in experimental groups(A-D)were supplemented with 1×109,2×109,4 × 109 and 8 × 109 CFU/mL Pseudomonas,respectively,three replicates were set up for each level.pH and gas production were recorded at 2,4,6,8,10,12 and 24 h of fermentation,and the contents of ammoniacal nitrogen(NH3-N),microbial protein(MCP)and volatile fatty acid(VFA)were measured at the end of fermentation.16S rRNA and internal transcribed spacer(ITS)were used for macro-genome sequencing to analyze the changes in the microbiota of rumen fluid.[Result]①At 10 h of fermentation,the pH of rumen fermentation broth in groups C and D was significantly lower than that in CON group(P＜0.05),and there were no significant changes in gas production,NH3-N and MCP contents among all treatment groups at each time point(P＞0.05).② At 10 h of fermentation,the contents of butyric acid and total VFA in rumen fermentation broth of group B were significantly or extremely significantly higher than those of CON group(P＜0.05 or P＜0.01).At 12 h of fermentation,the isobutyric acid content of rumen fermentation broth in groups A and B were significantly higher than that in group D(P＜0.05).At 24 h of fermentation,the contents of propionic acid,butyric acid,isovaleric acid,acetic acid/propionic acid and total VFA in rumen fermentation broth of each experimental group were significantly or extremely significantly higher than those of CON group(P＜0.05 or P＜0.01).③The Chao1 index and Ace index of 16S rRNA and ITS in rumen fermentation broth of groups C and D were significantly or extremely significantly lower than those of CON group(P＜0.05 or P＜0.01).④At the level of phylum and genus of 16S rRNA,compared with CON group,the relative abundance of Proteobacteria and Klebsiella in rumen fermentation broth of group D was significantly increased(P＜0.05),and the relative abundance of Succinivibrionaceae_UCG-001 and Rikenellaceae_RC9_gut_group in groups C and D were significantly or extremely significantly decreased(P＜0.05 or P＜0.01).The relative abundance of Succiniclasticum in rumen fermentation broth in group B was significantly higher than that in groups C and D(P＜0.05),the relative abundance of norank_F082 was significantly higher than that in group D(P＜0.05).⑤At the level of phylum and genus of ITS,compared with CON group,the relative abundance of Kodamaea in rumen fermentation broth of groups B,C and D was significantly increased(P＜0.05),and the relative abundance of Anaeromyces in group B was significantly increased(P＜0.05).[Conclusion]Under the condition of this experiment,2 × 109 CFU/mL Pseudomonas had no side effects on rumen fermentation parameters in dairy cows,which increased the production of VFA,and also promoted the proliferation of fiber-degrading bacteria.Therefore,the appropriate amount of Pseudomonas to be added was 2×109 CFU/mL.关键词
体外产气法/假单胞菌/瘤胃发酵液/微生物区系Key words
in vitro gas production method/Pseudomonas/rumen fermentation broth/microbiota分类
畜牧业引用本文复制引用
韩一铭,皇甫和平,王金明,张照杰,高宇凤,杨文华,任晓丽,宋超,石冬梅,章越,马梦辉..基于体外产气法探究不同假单胞菌添加量对人工瘤胃内环境的影响[J].中国畜牧兽医,2025,52(6):2569-2581,13.基金项目
河南省科技攻关项目(212102110377) （212102110377）
