Scientia Agricultura Sinica ›› 2022, Vol. 55 ›› Issue (4): 796-806.doi: 10.3864/j.issn.0578-1752.2022.04.014


Effects of 2-Hydroxy-4-(Methylthio)-Butanoic Acid on Rumen Fermentation and Microbiota in Holstein Female Calves

KONG FanLin1(),LI Yuan1,FU Tong2,DIAO QiYu1,TU Yan1()   

  1. 1Feed Research Institute, Chinese Academy of Agricultural Sciences/Beijing Key Laboratory for Dairy Cow Nutrition, Beijing 100081
    2College of Animal Science and Veterinary Medicine, Henan Agricultural University, Zhengzhou 450000
  • Received:2020-06-30 Accepted:2021-12-30 Online:2022-02-16 Published:2022-02-23
  • Contact: Yan TU;


【Objective】2-Hydroxy-4-(Methylthio)-Butanoic Acid (HMBi) is widely used to satisfy the absent of methionine (Met) in ruminal diet. Although the characteristic of HMBi is a rumen protected product, there is still an amount of HMBi degraded in rumen, which should be taken seriously. Hence, this study was conducted to evaluate the effects of HMBi on rumen fermentation and microbiota. 【Method】The experiment was conducted for 97 days with 36 Holstein female calves aged about 84 day-old with (101±10) kg body weight, and those calves were allocated to 2 groups, including PC group (0.40% Met) and PCMet group (0.28% Met). The treatment was achieved by deducting HMBi in diet of PCMet group and made Met level 30% lower than that of PC group. The first 7 d were an adaptation to the diets and the next 90 d for sampling. The body weight was measured at 0 d and 90 d, respectivley. The dry matter intake was recorded daily throughout the whole trial period. The serum and rumen fluid samples from five calves in each group were sampled on day 90 to determine rumen fermentation parameters and microbial communities. 【Result】(1) Compared with PC group, the growth performance of PCMet group was not changed (P>0.05). The Met in serum of PCMet group had trend to be significantly decreased when compared with PC group (0.05<P<0.1); (2) The molar proportion of acetate and microprotein concentration in PCMet group was significantly decreased by Met deduction (P<0.05). There were no significant differences on concentrations of total volatile fatty acid and ammonia nitrogen between two groups (P>0.05). (3) The Shannon index of microbiota in PCMet group was lower than that in PC group (P<0.05). The PCoA and PREANOVA analysis showed the significant distinction between microbiota in two groups (P<0.05). Furthermore, the relative abundance of Firmicutes in PCMet group was decreased and the relative abundance of Bacteroidetes was increased when compared with PC group (P<0.05). At genus level, the relative abundance of Olsenella, [Ruminococcus] gauvreauii group, Acetitomaculum, [Eubacterium] nodatum group, and Coprococcus 1 were decreased in PCMet group (P<0.05). The correlation analysis showed that [Eubacterium] nodatum group and Acetitomaculum were significantly correlated with acetate and [Ruminococcus] gauvreauii group was significantly correlated with MCP (P<0.05, r>0.7). 【Conclusion】 The ruminal microbiota was inhibited by HMBi deduction, which led to the decrease of MCP and Shannon index. Among them, the acetogen was sensitive with HMBi. In conclusion, although HMBi was a rumen protected product, the part of HMBi degraded in rumen still had the ability to regulate rumen fermentation.

Key words: HMBi, female calves, microbiota, rumen fermentation parameters

Table 1

Composition and nutrient levels of basal diet (dry matter basis, %)"

日粮组成 Ingredient 含量Content (%) 营养成分Nutrient 2) 营养水平Nutrient level
玉米 Corn 45.67 干物质 DM
(风干物质基础 Air-dry basis)
苜蓿 Alfalfa 25.00
小麦麸 Wheat bran 15.00 总能 GE(Mcal·kg-1 16.76
大豆粕 Soybean meal 11.97 代谢能 ME(Mcal·kg-1 12.71
细石粉 CaCO3 1.06 有机物 OM 93.32
预混料1% Premix1) 1.00 粗蛋白质 CP 19.22
食盐 NaCl 0.30 粗脂肪 EE 4.56
合计 Total 100.00 中性洗涤纤维 NDF 20.08
酸性洗涤纤维 ADF 11.43
钙 Ca 1.07
磷 P 0.49
赖氨酸 Lys 0.73
蛋氨酸 Met 0.11
苏氨酸 Thr 0.41

Table 2

Dietary rumen protected amino acid and amino acid levels (dry matter basis, %)"

组别 Group
基础日粮蛋氨酸 Met in basal diet 0.11 0.11
基础日粮赖氨酸 Lys in basal diet 0.73 0.73
过瘤胃蛋氨酸添加量 Addition of HMBi1) 1.32 0.77
过瘤胃赖氨酸添加量 Addition of rumen protected Lys2) 1.68 1.68
日粮蛋氨酸水平 Met level in test diet 0.40 0.28
日粮赖氨酸水平 Lys level in test diet 1.21 1.21

Table 3

Effects of partial deduction of methionine on growth performance and serum methionine level of female calves"

组别 Group SEM P
P value
初始体重 Initial BW (kg) 101.75 101.25 1.790 0.89
结束体重 Final BW (kg) 180.62 176.92 2.207 0.41
日增重 ADG (g·d-1) 876.31 840.78 18.109 0.34
干物质采食量 DMI (kg·d-1) 3.20 3.25 0.029 0.42
饲料转化比 Feed effeiciency, gain/feed 0.28 0.26 0.007 0.21
血清蛋氨酸 Serum Met (μg·mL-1) 4.19 3.44 0.212 0.07

Fig. 1

Effects of partial deduction of methionine on rumen fermentation parameters of female calves"

Fig. 2

Comparison of alpha diversity indices between groups"

Fig. 3

Cluster analysis by principal coordinate analysis"

Fig. 4

Composition and comparison of abundances at the phylum and genus level in the PC and PCMet group Significant differences in relative abundance between different treatments were marked with star. * indicates P<0.05, ** indicates P<0.01"

Table 4

Effects of partial deduction of methionine on KEGG pathway of bacterial gene function in rumen"

KEGG Pathway
组别 Group SEM P
P value
前5通路 Top 5 pathway
转运蛋白 Transporters 7.01 5.96 0.277 0.06
一般功能基因 General function prediction only 3.63 3.60 0.035 0.76
ABC转运蛋白 ABC transporters 3.42 2.94 0.136 0.08
DNA修复和重组蛋白DNA repair and recombination proteins 2.89 2.99 0.043 0.28
核糖体 Ribosome 2.50 2.56 0.040 0.46
蛋氨酸代谢相关通路 Methionine metabolism pathway
氨酰-tRNA生物合成 Aminoacyl-tRNA biosynthesis 1.27 1.26 0.013 0.63
半胱氨酸和蛋氨酸代谢 Cysteine and methionine metabolism 0.97 0.99 0.011 0.37
甘氨酸、丝氨酸和苏氨酸代谢Glycine, serine and threonine metabolism 0.82 0.87 0.015 0.08
氮代谢 Nitrogen metabolism 0.66 0.69 0.008 0.13
谷胱甘肽代谢 Glutathione metabolism 0.18 0.20 0.008 0.28
牛磺酸和亚牛磺酸代谢 Taurine and hypotaurine metabolism 0.10 0.11 0.004 0.78

Fig. 5

Spearman’s correlation between the rumen bacterial communities and phenotypic variables * indicates that correlations were statistically significant with strong correlations (P<0.05, |r|>0.7). The strength of the correlation between variables is indicated by the color intensity of the squares. The colors denote whether the correlation is positive (Blue squares) or negative (Red squares)"

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