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Dynamics of fermentation profile, bacterial communities and their functional characteristics in red clover

Published online by Cambridge University Press:  17 August 2022

Siran Wang
Affiliation:
Institute of Ensiling and Processing of Grass, College of Agro-Grassland Science, Nanjing Agricultural University, Nanjing 210095, China
Tao Shao
Affiliation:
Institute of Ensiling and Processing of Grass, College of Agro-Grassland Science, Nanjing Agricultural University, Nanjing 210095, China
Junfeng Li
Affiliation:
Institute of Ensiling and Processing of Grass, College of Agro-Grassland Science, Nanjing Agricultural University, Nanjing 210095, China
Jie Zhao
Affiliation:
Institute of Ensiling and Processing of Grass, College of Agro-Grassland Science, Nanjing Agricultural University, Nanjing 210095, China
Zhihao Dong*
Affiliation:
Institute of Ensiling and Processing of Grass, College of Agro-Grassland Science, Nanjing Agricultural University, Nanjing 210095, China
*
Author for correspondence: Zhihao Dong, E-mail: T2020159@njau.edu.cn

Abstract

The bacterial community is important for shaping the fermentation characteristics of silage. This study aimed to investigate the fermentation characteristics, bacterial community and predicted functional characteristics of red clover (Trifolium pratense L.) silage. First-cutting red clover was collected at the early bloom stage, wilted for 5 h and then ensiled in 10 litre-capacity silos. Triplicate silos were sampled after 1, 3, 7, 15, 30 and 60 days of ensiling, respectively. The bacterial communities on days 3 and 60 were assessed through high-throughput sequencing technology, and 16S rRNA-gene predicted functional characteristics were analysed based on the KEGG using Tax4Fun. After 60 days of ensiling, red clover silage was fermented well, as indicated by high lactic acid (~77.3 g/kg DM), and low concentrations of butyric acid (~3.73 g/kg DM) and ammonia nitrogen (~55.0 g/kg TN). During the initial stage of ensiling, fructose and glucose were more preferred than sucrose for microbes. The predominant genus Lactococcus (0.542) on day 3 was replaced by Lactobacillus (0.553) on day 60. The metabolism of amino acids, energy, cofactors and vitamins was inhibited, while the metabolism of nucleotides and carbohydrates was enhanced after ensiling. High-throughput sequencing technology combined with 16S rRNA gene-predicted functional analyses revealed the differences during the early and late stages of red clover silage not only for distinct bacterial compositions but also for specific functional traits. Our results could provide a comprehensive insight into bacterial community and their functional profiles to further improve the silage quality.

Type
Animal Research Paper
Copyright
Copyright © The Author(s), 2022. Published by Cambridge University Press

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References

Aßhauer, KP, Bernd, W, Rolf, D and Peter, M (2015) Tax4fun: predicting functional profiles from metagenomic 16S rRNA data. Bioinformatics 31, 28822884.CrossRefGoogle ScholarPubMed
Bai, J, Ding, Z, Ke, W, Xu, D, Wang, M, Huang, W, Zhang, Y, Liu, F and Guo, X (2021) Different lactic acid bacteria and their combinations regulated the fermentation process of ensiled alfalfa: ensiling characteristics, dynamics of bacterial community and their functional shifts. Microbial Biotechnology 14, 11711182.CrossRefGoogle ScholarPubMed
Bertrand, A, Bipfubusa, M, Castonguay, Y, Rocher, S, Szopinska-Morawska, A and Papadopoulos, Y (2016) A proteome analysis of freezing tolerance in red clover (Trifolium pratense L.). BMC Plant Biology 16, 65.CrossRefGoogle ScholarPubMed
Broderick, GA and Kang, JH (1980) Automated simultaneous determination of ammonia and total amino acids in ruminal fluid and in vitro media. Journal of Dairy Science 63, 6475.CrossRefGoogle ScholarPubMed
Cao, Y, Cai, Y, Takahashi, T, Yoshida, N, Tohno, M, Uegaki, R, Nonaka, K and Terada, F (2011) Effect of lactic acid bacteria inoculant and beet pulp addition on fermentation characteristics and in vitro ruminal digestion of vegetable residue silage. Journal of Dairy Science 94, 39023912.CrossRefGoogle ScholarPubMed
Desta, ST, Yuan, XJ, Li, JF and Tao, S (2016) Ensiling characteristics, structural and nonstructural carbohydrate composition and enzymatic digestibility of Napier grass ensiled with additives. Bioresource Technology 221, 447454.CrossRefGoogle ScholarPubMed
Du, Z, Sun, L, Chen, C, Lin, J, Yang, F and Cai, Y (2021) Exploring microbial community structure and metabolic gene clusters during silage fermentation of paper mulberry, a high-protein woody plant. Animal Feed Science and Technology 275, 114766.CrossRefGoogle Scholar
Flythe, MD and Russell, JB (2004) The effect of pH and a bacteriocin (bovicin HC5) on Clostridium sporogenes MD1, a bacterium that has the ability to degrade amino acids in ensiled plant materials. FEMS Microbiology Ecology 47, 215222.CrossRefGoogle Scholar
Gharechahi, J, Kharazian, ZA, Sarikhan, S, Jouzani, GS, Agh Da Si, M and Salekdeh, GH (2017) The dynamics of the bacterial communities developed in maize silage. Microbial Biotechnology 10, 16631676.CrossRefGoogle ScholarPubMed
Hu, Z, Chang, J, Yu, J, Li, S and Niu, H (2018) Diversity of bacterial community during ensiling and subsequent exposure to air in whole-plant maize silage. Asian-Australasian Journal of Animal Sciences 31, 14641473.CrossRefGoogle ScholarPubMed
Hu, Z, Niu, H, Tong, Q, Chang, J and Ma, D (2020) The microbiota dynamics of alfalfa silage during ensiling and after air exposure, and the metabolomics after air exposure are affected by Lactobacillus casei and cellulase addition. Frontiers in Microbiology 11, 519121.CrossRefGoogle ScholarPubMed
Keshri, J, Chen, Y, Pinto, R, Kroupitski, Y, Weinberg, ZG and Sela Saldinger, S (2018) Microbiome dynamics during ensiling of corn with and without Lactobacillus plantarum inoculant. Applied Microbiology Biotechnology 102, 40254037.CrossRefGoogle ScholarPubMed
Kilstrup, M, Hammer, K, Jensen, PR and Martinussen, J (2005) Nucleotide metabolism and its control in lactic acid bacteria. FEMS Microbiology Reviews 29, 555590.CrossRefGoogle ScholarPubMed
Krishnamoorthy, U, Muscato, TV, Sniffen, CJ and Van Soest, PJ (1982) Nitrogen fractions in selected feedstuffs. Journal of Dairy Science 65, 217225.CrossRefGoogle Scholar
Li, D, Ni, K, Zhang, Y, Lin, Y and Yang, F (2019) Fermentation characteristics, chemical composition and microbial community of tropical forage silage under different temperatures. Asian-Australasian Journal of Animal Sciences 32, 665.CrossRefGoogle ScholarPubMed
Ma, S, Fang, C, Sun, X, Han, L, He, X and Huang, G (2018) Bacterial community succession during pig manure and wheat straw aerobic composting covered with a semi-permeable membrane under slight positive pressure. Bioresource Technology 259, 221227.CrossRefGoogle ScholarPubMed
McDonald, P, Henderson, AR and Heron, S (1991) The Biochemistry of Silage. Abersytwyth, UK: Chalcombe Publications.Google Scholar
Ni, K, Minh, TT, Tu, T, Tsuruta, T, Pang, H and Nishino, N (2017 a) Comparative microbiota assessment of wilted Italian ryegrass, whole crop corn, and wilted alfalfa silage using denaturing gradient gel electrophoresis and next-generation sequencing. Applied Microbiology Biotechnology 101, 13851394.CrossRefGoogle ScholarPubMed
Ni, K, Wang, F, Zhu, B, Yang, J, Zhou, G, Pan, Y and Zhong, J (2017 b) Effects of lactic acid bacteria and molasses additives on the microbial community and fermentation quality of soybean silage. Bioresource Technology 238, 706715.CrossRefGoogle ScholarPubMed
Nishino, N, Li, Y, Wang, C and Parvin, S (2012) Effects of wilting and molasses addition on fermentation and bacterial community in guinea grass silage. Letters in Applied Microbiology 54, 175181.CrossRefGoogle ScholarPubMed
Pahlow, G, Muck, RE, Driehuis, F, Oude Elferink, SJWH and Spoelstr, SF (2003) Microbiology of ensiling. In Buxton DR, Muck RE and Harrison JH (eds), Silage Science and Technology. Agronomy no. 42. Madison, WI, USA: ASCSSA-SSSA, pp. 3194.Google Scholar
Pessione, A, Lamberti, C and Pessione, E (2010) Proteomics as a tool for studying energy metabolism in lactic acid bacteria. Molecular Biosystems 6, 14191430.CrossRefGoogle ScholarPubMed
Playne, MJ and McDonald, P (1966) The buffering constituents of herbage and of silage. Journal of the Science of Food & Agriculture 17, 264268.CrossRefGoogle Scholar
Rooke, JA and Hatfield, RD (2003) Biochemistry of ensiling. In Buxton DR, Muck RE and Harrison JH (eds), Silage Science and Technology. Agronomy no. 42. Madison, WI, USA: ASCSSA-SSSA, pp. 95140.Google Scholar
Santos, AO, Ávila, CLS, Pinto, JC, Carvalho, BF, Dias, DR and Schwan, RF (2015) Fermentative profile and bacterial diversity of corn silages inoculated with new tropical lactic acid bacteria. Journal of Applied Microbiology 120, 266279.CrossRefGoogle Scholar
Shao, T, Zhang, ZX, Shimojo, M, Wang, T and Masuda, Y (2005) Comparison of fermentation characteristics of Italian ryegrass (Lolium multiflorum Lam.) and guineagrass (Panicum maximum Jacq.) during the early stage of ensiling. Asian-Australasian Journal of Animal Sciences 18, 17271734.CrossRefGoogle Scholar
Thomas, TA (1977) An automated procedure for the determination of soluble carbohydrates in herbage. Journal of the Science of Food & Agriculture 28, 639642.CrossRefGoogle Scholar
Van Soest, PJ, Robertson, JB and Lewis, BA (1991) Methods for dietary fiber, neutral detergent fiber, and nonstarch polysaccharides in relation to animal nutrition. Journal of Dairy Science 74, 35833597.CrossRefGoogle ScholarPubMed
Ventura, M, Canchaya, C, Tauch, A, Chandra, G, Fitzgerald, GF and Chater, KF (2007) Genomics of actinobacteria: tracing the evolutionary history of an ancient phylum. Microbiology and Molecular Biology Reviews 71, 495548.CrossRefGoogle ScholarPubMed
Wang, S, Li, J, Dong, Z, Chen, L and Shao, T (2018) Inclusion of alfalfa improves nutritive value and in vitro digestibility of various straw–grass mixed silages in Tibet. Grass and Forage Science 73, 694704.CrossRefGoogle Scholar
Wang, Y, He, L, Xing, Y, Zhou, W, Pian, R, Yang, F, Chen, X and Zhang, Q (2019) Bacterial diversity and fermentation quality of Moringa oleifera leaves silage prepared with lactic acid bacteria inoculants and stored at different temperatures. Bioresource Technology 284, 349358.CrossRefGoogle ScholarPubMed
Wang, S, Zhao, J, Dong, Z, Li, J and Shao, T (2020) Sequencing and microbiota transplantation to determine the role of microbiota on the fermentation type of oat silage. Bioresource Technology 309, 123371.CrossRefGoogle ScholarPubMed
Xu, D, Wang, N, Rinne, M, Ke, W, Weinberg, ZG, Da, M, Bai, J, Zhang, Y, Li, F and Guo, X (2021) The bacterial community and metabolome dynamics and their interactions modulate fermentation process of whole crop corn silage prepared with or without inoculants. Microbial Biotechnology 14, 561576.CrossRefGoogle ScholarPubMed
Yan, Y, Li, X, Guan, H, Huang, L, Ma, X, Peng, Y, Li, Z, Nie, G, Zhou, J, Yang, W, Cai, Y and Zhang, X (2019) Microbial community and fermentation characteristic of Italian ryegrass silage prepared with corn stover and lactic acid bacteria. Bioresource Technology 279, 166173.CrossRefGoogle ScholarPubMed
Zheng, ML, Niu, DZ, Jiang, D, Zuo, SS and Xu, CC (2017) Dynamics of microbial community during ensiling direct-cut alfalfa with and without LAB inoculant and sugar. Journal of Applied Microbiology 122, 14561470.CrossRefGoogle ScholarPubMed