宏基因组分析方法探究高精料日粮对奶牛瘤胃产甲烷菌的影响

doi:10.19788/j.issn.2096-6369.190105

摘要/Abstract

摘要：

目的本研究旨在探究高精料日粮对瘤胃产甲烷菌区系的影响。方法利用宏基因组检测手段结合生物信息学分析方法,测定高精料日粮和对照日粮处理下瘤胃产甲烷菌区系的变化。结果高精料日粮显著降低奶牛日采食量和产奶量,瘤胃pH以及瘤胃乙酸的含量(P < 0.05);显著提高了瘤胃中丙酸以及氨态氮的含量(P < 0.05)。通过分类学分析,总共鉴定出4纲36属108种产甲烷菌约占瘤胃微生物总量的0.37%~0.47%。其中Methanobacteria为产甲烷菌的主要构成部分,约占产甲烷菌总量的50%~55%,Methanobrevibacter为Methanobacteria的主要组成部分。Thermoplasmata 在所有产甲烷菌中占据第二多的位置,约占产甲烷菌总量的31%。差异分析结果表明：高精料日粮显著降低了产甲烷菌总量、Methanobacteria和Methanopyri的相对含量（P < 0.05）,而对Methanococci 和Methanomicrobia 作用不显著。相关分析结果表明：瘤胃产甲烷菌相对丰度与瘤胃中丙酸含量、氨态氮含量呈负相关关系,与瘤胃乙酸含量呈正相关关系。结论高精料日粮降低瘤胃产甲烷菌的相对丰度,但过多饲喂高精料日粮会降低瘤胃pH,造成亚急性瘤胃酸中毒（SARA）。利用宏基因组方法更加准确全面的鉴定出瘤胃产甲烷菌的种类和数量,使得对于产甲烷菌有了更加准确的认识,同时对于降低甲烷排放,促进奶牛生产具有重要的意义。

关键词: 泌乳奶牛, 高精料日粮, 产甲烷菌, 宏基因组, 生物信息学

Abstract:

[Objective] This study was conducted to investigate the effects of high-concentrate diets on ruminal methanogens. [Method] Metagenomics and bioinformatic methods were used to investigate the impacts of high-concentrate diets on ruminal methanogens in dairy cows. [Results] Daily dry matter intake and milk production of cows offered high-concentrate diets were significantly lower than those of cows offered control diets (P < 0.05). Acetate concentration and ruminal pH decreased significantly while propionate and ammonia nitrogen concentrations were significantly increased in the rumens of cows fed high-concentrate diets, when compared with those in the control group (P < 0.05). Four classes, 36 genera and 108 species of methanogens were identified in this study and the relative abundance of methanogens was about 0.37%-0.47% of the entire rumen microbiome. Of these, Methanobacteria was the dominant genus, accounting for about 50%-55% of all methanogens, while Thermoplasmata was the second most abundant genus, accounting for about 31%. Feeding high-concentrate diets significantly decreased the relative abundance of all methanogens, Methanobacteria and Methanopyri (P<0.05). However, Methanococci and Methanomicrobia were not affected. Moreover, methanogens were positively correlated with acetate concentration and negatively correlated with propionate and ammonia nitrogen concentrations. [Conclusion] Feeding high-concentrate diets could reduce the relative abundance of methanogens; however, overfeeding of such diets may lead to low ruminal pH and subacute ruminal acidosis (SARA). Metagenomic methods used in this study accurately quantified the species and relative abundance of methanogens. The results presented herein will contribute to methane emission mitigation and improved dairy cattle production efficiency.

Key words: Dairy cows, High-concentrate diets, Methanogens, Metagenome, Bioinformatics

中图分类号:

S8-05

薛夫光, 施辉毕, 孙福昱, 罗清尧, 杨亮, 楚康康, 蒋林树, 熊本海. 宏基因组分析方法探究高精料日粮对奶牛瘤胃产甲烷菌的影响[J]. 农业大数据学报, 2019, 1(1): 45-55.

Xue Fuguang, Shi Huibi, Sun Fuyu, Luo Qingyao, Yang Liang, Chu Kangkang, Jiang Linshu, Xiong Benhai. Metagenomic Insights into Effects of High-Concentrate Diets on Ruminal Methanogens in Dairy Cows[J]. Journal of Agricultural Big Data, 2019, 1(1): 45-55.

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项目Items	对照组CON	高精料组HC
原料ingredients( % of DM)
羊草Chinese wildrye	11.0	5.0
玉米青贮Corn silage	34.0	20.0
苜蓿Alfalfa hay	15.0	15.0
玉米Ground corn	10.0	30.0
大豆粕Soybean meal, 43% CP	14.0	14.0
棉籽粕Cottonseed meal, 42%CP	5.0	5.0
干酒糟DDGS,27.5%CP	5.0	5.0
全棉籽Whole cottonseed	3.0	3.0
石粉Limestone meal	1.0	1.0
碳酸氢钙Calcium hydrogen phosphate	0.7	0.7
氯化钠Sodium chloride	0.5	0.5
预混料Premix¹	0.8	0.8
养分含量Nutrient composition (% of DM)
泌乳净能NE_L², MJ/kg	6.61	7.03
粗蛋白CP	18.16	18.10
淀粉Starch	19.95	30.82
中性洗涤纤维NDF	36.18	27.61
酸性洗涤纤维ADF	23.43	17.72
非纤维性碳水化合物NFC³	32.67	45.74
粗脂肪Ether extract	4.61	4.20
灰分Ash	5.04	4.35
钙Calcium	0.88	0.84
磷Phosphorus	0.55	0.55

条目Item	处理组Treatments		SEM	P-value
条目Item	对照组 CON	高精料组 HC	SEM	P-value
平均日采食量Average Daily DMI,kg/d	21.68^a	19.07^b	0.278	0.014
平均日产奶量Average Daily Milk Production,kg/d	27.28^a	22.12^b	1.894	0.001
瘤胃液pH Rumen fluid	6.45^a	5.58^b	0.194	0.016
乙酸Acetate,mmol/L	43.237^a	42.619^b	1.273	0.038
丙酸Propionate,mmol/L	12.899^b	13.849^a	0.632	0.027
丁酸Butyrate,mmol/L	10.77	10.354	0.137	0.356
总挥发酸TVFA,mmol/L	70.323	70.983	1.836	0.156
氨态氮Ammonia-N,mg/100mL	10.495^b	13.863^a	1.711	0.006

条目Item	试验处理组Experimental Treatments		SEM	P-value
条目Item	对照组 CON	高精料组 HC	SEM	P-value
Methanobacteria	0.148^a	0.049^b	0.016	0.011
Methanococci	0.005	0.005	0.002	0.561
Methanomicrobia	0.048	0.051	0.002	0.338
Methanopyri	1.67E^{-05 b}	1.35 E^{-04 a}	<0.001	0.001
合计total	0.200^a	0.105^b	0.037	0.004