Effects of Formic Acid and Wood Vinegar on Bacterial Community Composition of Alfalfa Silage

doi:10.11924/j.issn.1000-6850.casb2021-0082

Abstract

Abstract:

To investigate the effects of inhibitory silage additives on bacterial community composition of alfalfa silage, the experiment was conducted with formic acid and wood vinegar as the additives, alfalfa silage was made by mixing alfalfa with dry matter content of (23.02±0.70) % with the additive of 6 mL/kg, the treatment with equivalent distilled water replacing additive was used as the control, and the bacterial community was analyzed through High-throughput sequencing method after 45 days. The results showed that in the control group, the Enterobacteriaceae unknown genus of Proteobacteria was dominant, followed by Enterococcus of Firmicutes; in the wood vinegar treatment, the Enterococcus of Firmicutes was dominant, followed by Enterobacteriaceae unknown genus of Proteobacteria; while in the formic acid treatment, the dominant bacteria community was unknown family and genus of Streptophyta in Cyanobacteria. Compared with the control, the relative abundance of Enterococcus was increased significantly in the wood vinegar treatment, but decreased significantly in the formic acid treatment; both of the two additives significantly reduced the relative abundance of Enterobacteriaceae, and the decrease of formic acid treatment was significantly higher than that of wood vinegar treatment; the formic acid treatment significantly increased the abundance indices (Chao1 and ACE) of bacterial community but decreased the dominance index (Simpson). According to the principal component analysis, the indices of bacteria community and main nutrition and fermentation quality could explain 88.786% information of alfalfa silage. From the distribution of samples, the distribution of wood vinegar treatment was between the formic acid treatment and the control but closer to the control. In conclusion, according to the bacterial community analysis, both the formic acid and wood vinegar could improve the bacterial community composition of alfalfa silage, in which formic acid could mainly inhibite bacterial fermentation but wood vinegar is helpful to promote lactic acid bacteria fermentation.

Key words: formic acid, wood vinegar, alfalfa, bacteria, community composition

CLC Number:

WANG Lixue, HAN Jing, CHEN Longbin, YU Xinyue, LIU Jingxi, MA Yi, HUO Wenjuan. Effects of Formic Acid and Wood Vinegar on Bacterial Community Composition of Alfalfa Silage[J]. Chinese Agricultural Science Bulletin, 2022, 38(2): 92-101.

Figures/Tables 5

References 28

[1]	王媛, 王雁萍, 郝湘妹, 等. 苜蓿青贮乳酸菌的筛选及其对苜蓿青贮发酵的影响[J]. 草业科学, 2020, 37(2):377-382.
[2]	SARICICEK B Z, KILIC U. Effect of different additives on the nutrient composition, in vitro gas production and silage quality of alfalfa silage[J]. Asian journal of animal and veterinary advances, 2011(6):618-626.
[3]	马召稳, 李元晓, 梁含, 等. 苜蓿青贮中微生物种群分析及主要菌种的筛选鉴定[J]. 草业科学, 2019, 36(11):2980-2988.
[4]	王旭哲, 张凡凡, 马春晖, 等. 同/异型乳酸菌对青贮玉米开窖后品质及微生物的影响[J]. 农业工程学报, 2018, 34(10):296-304.
[5]	RAJABI R, TAHMASBI R, DAYANI O, et al. Chemical composition of alfalfa silage with waste date and its feeding effect on ruminal fermentation characteristics and microbial protein synjournal in sheep[J]. Journal of Animal Physiology and Animal Nutrition, 2017, 101(3):466-474. doi: 10.1111/jpn.2017.101.issue-3 URL
[6]	CHEN Lei, LI J F, DONG Z H, et al. Effects of lactic acid bacteria inoculants and fibrolytic enzymes on the fermentation quality, in vitro degradability, ruminal variables and microbial communities of high-moisture alfalfa silage[J]. Grassland science, 2019, 65:216-225. doi: 10.1111/grs.v65.4 URL
[7]	AGARUSSI M C N, PEREIRA O G, SILVA V P, et al. Fermentative profile and lactic acid bacterial dynamics in non-wilted and wilted alfalfa silage in tropical conditions[J]. Molecular biology reports, 2019, 46:451-460. doi: 10.1007/s11033-018-4494-z URL
[8]	YUAN X J, LI J F, DONG Z H, et al. The reconstitution mechanism of napier grass microiota during the ensiling of alfalfa and their contributions to fermentation quality of silage[J]. Bioresource technology, 2019, 297:122391. doi: 10.1016/j.biortech.2019.122391 URL
[9]	张玲, 席琳乔, 张凡凡, 等. 残次枣粉添加青贮苜蓿中微生物、发酵和营养品质动态规律[J]. 新疆农业科学, 2019, 56(10):1929-1938. doi: 10.6048/j.issn.1001-4330.2019.10.019
[10]	WANG C, HE L W, XING Y Q, et al. Fermentation quality and microbial community of alfalfa and stylo silage mixed with Moringa oleifera leaves[J]. Bioresource technology, 2019, 284:240-247. doi: 10.1016/j.biortech.2019.03.129 URL
[11]	SILVA V P, PEREIRA O G, LEANDRO E S, et al. Effects of lactic acid bacteria with bacteriocinogenic potential on the fermentation profile and chemical composition of alfalfa silage in tropical conditions[J]. Journal of dairy sence, 2016, 99(3):1895-1902.
[12]	ZHENG M L, NIU D Z, JIANG D, et al. Dynamics of microbial community during ensiling direct-cut alfalfa with and without LAB inoculant and sugar[J]. Journal of applied microbiology, 2017, 122:1456-1470. doi: 10.1111/jam.2017.122.issue-6 URL
[13]	WU B Y L, ZHANG Q, LIU Z K, et al. Bacterial communities in alfalfa and corn silages produced in large-scale stack and bunker silos in China[J]. Grassland science, 2014, 60(4):247-251. doi: 10.1111/grs.2014.60.issue-4 URL
[14]	卢强, 撒多文, 都帅, 等. 盐碱地苜蓿青贮品质及微生物群落的研究[J]. 黑龙江畜牧兽医, 2020(2):96-100,150.
[15]	Li D X, Ni K K, Zhang Y C, et al. Influence of lactic acid bacteria, cellulase, cellulase-producing Bacillus pumilus and their combinations on alfalfa silage quality[J]. Journal of integrative agriculture, 2018, 17(12):172-186.
[16]	OGUNADE I M, JIANG Y, CERVANTES A A P, et al. Bacterial diversity and composition of alfalfa silage as analyzed by Illumina MiSeq sequencing: Effects of Escherichia coli O157:H7 and silage additives[J]. Journal of dairy ence, 2017, 101(3):2048-2059.
[17]	闫晶, 陆冰圆, 席华悦, 等. 外源添加剂对黄贮小麦秸秆产甲烷潜力及微生物群落的影响[J]. 农业工程学报, 2020, 36(15):252-260.
[18]	包万华, 卜登攀, 周凌云, 等. 青贮饲料添加剂应用的研究进展[J]. 中国畜牧兽医, 2012, 39(8):124-128.
[19]	马春晖, 夏艳军, 韩军, 等. 不同青贮添加剂对紫花苜蓿青贮品质的影响[J]. 草业学报, 2010, 19(1):128-133.
[20]	MUNGKUNKAMCHAO T, KESMALA T, PIMRATCH S, et al. Wood vinegar and fermented bioextracts: Natural products to enhance growth and yield of tomato (Solanum lycopersicum L.)[J]. Scientia horticulturae, 2013, 154:66-72. doi: 10.1016/j.scienta.2013.02.020 URL
[21]	王丽学, 何峰, 韩静, 等. 不同肥料和青贮添加剂对苜蓿青贮的影响[J]. 农学学报, 2018(5):48-54.
[22]	YUAN X J, Wen A Y, DESTA S T, et al. Effects of four short-chain fatty acids or salts on dynamics of fermentation and microbial characteristics of alfalfa silage[J]. Journal of the science of food and agriculture, 2017, 97(9):2759-2766. doi: 10.1002/jsfa.2017.97.issue-9 URL
[23]	李宝明, 杨织瑞. 乳酸菌添加剂对青贮饲料中微生物菌群的影响[J]. 畜牧兽医杂志, 2016, 35(1):13-17.
[24]	杨艳, 智健飞, 严学兵, 等. 添加乳酸菌剂后苜蓿青贮料发酵的动态变化[J]. 中国畜牧杂志, 2014, 50(11):41-45.
[25]	NI K K, MINH T T, TU T T M, et al. Comparative microbiota assessment of wilted Italian ryegrass, whole crop corn, and wilted alfalfa silage using denaturing gradient gel electrophoresis and next-generation sequencing[J]. Applied microbiology & biotechnology, 2017, 101:1385-1394.
[26]	王凤林, 张莉娟, 孟媛, 等. 不同比例微生物组合对玉米秸秆青贮品质的影响[J]. 基因组学与应用生物学, 2017, 36(11):4701-4706.
[27]	乔江涛, 郭荣波, 袁宪正, 等. 玉米秸秆厌氧降解复合菌系的微生物群落结构[J]. 环境科学, 2013, 34(4):1531-1539.
[28]	王雅雅. 几种典型农业废弃物高含固率厌氧共发酵产气性能与协同机理研究[D]. 北京:中国农业大学, 2018.

门	A组	B组	C组
厚壁菌门	40.345±4.670b	55.687±9.941c	12.996±6.315a
变形菌门	57.726±5.904c	37.050±9.919b	21.771±4.252a
蓝细菌门	1.710±1.220a	6.047±3.643a	61.775±4.649b
放线菌门	0.170±0.042a	0.982±1.136ab	2.651±1.412b
酸杆菌门	0.021±0.012a	0.114±0.086a	0.239±0.030b
拟杆菌门	0.024±0.013a	0.089±0.045a	0.512±0.140b
绿弯菌门	0.001±0.001a	0.014±0.012ab	0.039±0.022b
梭杆菌门	0.002±0.002a	0.003±0.003a	0.004±0.000a
浮霉菌门	0.000±0.000a	0.001±0.001a	0.002±0.002a

门	A组	B组	C组
厚壁菌门	40.345±4.670b	55.687±9.941c	12.996±6.315a
变形菌门	57.726±5.904c	37.050±9.919b	21.771±4.252a
蓝细菌门	1.710±1.220a	6.047±3.643a	61.775±4.649b
放线菌门	0.170±0.042a	0.982±1.136ab	2.651±1.412b
酸杆菌门	0.021±0.012a	0.114±0.086a	0.239±0.030b
拟杆菌门	0.024±0.013a	0.089±0.045a	0.512±0.140b
绿弯菌门	0.001±0.001a	0.014±0.012ab	0.039±0.022b
梭杆菌门	0.002±0.002a	0.003±0.003a	0.004±0.000a
浮霉菌门	0.000±0.000a	0.001±0.001a	0.002±0.002a

门	属	A组	B组	C组
厚壁菌门	肠球菌属	35.041±1.985b	45.026±7.950c	5.089±4.652a
	芽孢杆菌属	0.074±0.030a	0.289±0.172a	4.325±2.860b
	乳杆菌目未知科属	0.872±0.689a	0.844±0.327a	0.112±0.103a
	乳杆菌科未知属	0.083±0.079a	2.074±1.859a	0.147±0.093a
	乳杆菌属	2.479±2.439a	0.140±0.119a	0.026±0.012a
	片球菌属	0.177±0.106a	1.277±1.065a	0.131±0.090a
	明串珠菌科未知属	0.017±0.014a	3.533±6.052a	0.041±0.047a
	明串珠菌属	0.880±0.824a	0.314±0.268a	0.044±0.012a
	乳球菌属	0.124±0.014a	1.028±0.707b	1.358±0.231b
变形菌门	肠杆菌科未知属	48.302±1.627c	30.798±9.769b	8.444±3.864a
	甲基杆菌属	0.199±0.166a	0.475±0.407a	1.413±1.014a
	农杆菌属	0.012±0.003a	0.075±0.060a	0.648±0.634a
	羽扇豆属	0.000±0.000a	0.002±0.001a	1.172±0.584b
	鞘脂单胞菌属	0.017±0.008a	0.103±0.085a	1.298±0.465b
	肠杆菌属	1.119±1.103a	0.090±0.005a	0.014±0.003a
	特拉布斯氏菌属	7.078±3.044b	2.812±1.765ab	0.139±0.131a
	黄单胞菌科未知属	0.002±0.002a	0.054±0.050a	2.055±2.052a
蓝细菌门	链型植物目未知科属	1.709±1.221a	6.041±3.036a	61.747±4.682b
放线菌门	微杆菌科未知属	0.001±0.000a	0.006±0.001a	0.341±0.006b

门	属	A组	B组	C组
厚壁菌门	肠球菌属	35.041±1.985b	45.026±7.950c	5.089±4.652a
	芽孢杆菌属	0.074±0.030a	0.289±0.172a	4.325±2.860b
	乳杆菌目未知科属	0.872±0.689a	0.844±0.327a	0.112±0.103a
	乳杆菌科未知属	0.083±0.079a	2.074±1.859a	0.147±0.093a
	乳杆菌属	2.479±2.439a	0.140±0.119a	0.026±0.012a
	片球菌属	0.177±0.106a	1.277±1.065a	0.131±0.090a
	明串珠菌科未知属	0.017±0.014a	3.533±6.052a	0.041±0.047a
	明串珠菌属	0.880±0.824a	0.314±0.268a	0.044±0.012a
	乳球菌属	0.124±0.014a	1.028±0.707b	1.358±0.231b
变形菌门	肠杆菌科未知属	48.302±1.627c	30.798±9.769b	8.444±3.864a
	甲基杆菌属	0.199±0.166a	0.475±0.407a	1.413±1.014a
	农杆菌属	0.012±0.003a	0.075±0.060a	0.648±0.634a
	羽扇豆属	0.000±0.000a	0.002±0.001a	1.172±0.584b
	鞘脂单胞菌属	0.017±0.008a	0.103±0.085a	1.298±0.465b
	肠杆菌属	1.119±1.103a	0.090±0.005a	0.014±0.003a
	特拉布斯氏菌属	7.078±3.044b	2.812±1.765ab	0.139±0.131a
	黄单胞菌科未知属	0.002±0.002a	0.054±0.050a	2.055±2.052a
蓝细菌门	链型植物目未知科属	1.709±1.221a	6.041±3.036a	61.747±4.682b
放线菌门	微杆菌科未知属	0.001±0.000a	0.006±0.001a	0.341±0.006b