Abstract
The objectives of this study were to evaluate the contribution of urea on the nutritional quality and microbial community of ensiled alfalfa (Medicago sativa L.). Alfalfa silage was control group without urea (AL), supplementation with 0.5% urea (AU1), or supplementation with 1% urea (AU2). The silage tanks were opened and sampled after silage at 0, 15, 30, and 60 d. Results showed that AU2 had higher pH, ratio of (ammonia–N)/(total nitrogen) (NH3–N/TN) and crude protein (CP) content than those in AL and AU1, while AU1 had higher acid detergent fiber (ADF) than that in AL and AU2 after 15 d silage. Richness and diversity indices of microbial communities in silage were no significant differences among AL, AU1 and AU2 group. Proteobacteria (58.23%) and Firmicutes (40.95%) were the predominant phylum in three groups during the silage process. The percent of community abundances on genera level of Enterobacteriaceae (37.61%) and Klebsiella (41.78%) in AL were a little higher than those in AU1 (30.39%, 25.02%) and AU2 (33.48%, 26.92%). These results showed that silage with urea alone could not improve the quality of alfalfa.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
Data availability
The data sets generated during and/or analyzed during the current study are available from the corresponding author on reasonable request.
References
Allen B, Kon M, Bar-Yam Y (2009) A new phylogenetic diversity measure generalizing the Shannon index and its application to phyllostomid bats. Am Nat 174:236–243. https://doi.org/10.1086/600101
Dunière L, Sindou J, Chaucheyras-Durand F, Chevallier I, Thévenot-Sergentet D (2013) Silage processing and strategies to prevent persistence of undesirable microorganisms. Anim Feed Sci Technol 182:1–15. https://doi.org/10.1016/j.anifeedsci.2013.04.006
Eikmeyer FG, Köfinger P, Poschenel A, Jünemann S, Zakrzewski M, Heinl S, Mayrhuber E, Grabherr R, Pühler A, Schwab H (2013) Metagenome analyses reveal the influence of the inoculant Lactobacillus buchneri CD034 on the microbial community involved in grass ensiling. J Biotechnol 167:334–343. https://doi.org/10.1016/j.anifeedsci.2013.04.006
Hu Z, Chang J, Yu J, Li S, Niu H (2018) Diversity of bacterial community during ensiling and subsequent exposure to air in whole-plant maize silage. Asian-Australas J Anim Sci 31:1464–1473. https://doi.org/10.5713/ajas.17.0860
Keshri J, Chen Y, Pinto R, Kroupitski Y, Weinberg ZG, Sela S (2018) Microbiome dynamics during ensiling of corn with and without Lactobacillus plantarum inoculant. Appl Microbiol Biotechnol 102:4025–4037. https://doi.org/10.1007/s00253-018-8903-y
Lin C, Bolsen KK, Brent BE, Fung DYC (1992) Epiphytic lactic acid bacteria succession during the pre-ensiling and ensiling periods of alfalfa and maize. J Appl Bacteriol 73:375–387. https://doi.org/10.1111/j.1365-2672.1992.tb04992.x
Lin H, Lin S, Awasthi MK, Wang Y, Xu P (2021) Exploring the bacterial community and fermentation characteristics during silage fermentation of abandoned fresh tea leaves. Chemosphere 283:131234. https://doi.org/10.1016/j.chemosphere.2021.131234
Liu JY, Yu M, Wang AK, Huang BZ, Li RS, Zhang MQ, Sun WT (2010) Influences of the Quality of Sugarcane Tip Silage by Adding Urea. Chin Cattle Sci 36(4):22–25. https://doi.org/10.3969/j.issn.1001-9111.2010.04.006
Magoč T, Salzberg SL (2011) FLASH: fast length adjustment of short reads to improve genome assemblies. Bioinformatics 27(21):2957–2963. https://doi.org/10.1093/bioinformatics/btr507
Martins SCSG, Carvalho GGP, Pires AJV, Silva RR, Leite LC, Pereira FM, Mota ADS, Nicory IMC, Cruz CHda (2015) Qualitative parameters of sugarcane silages treated with urea and calcium oxide. Semina: Ciências Agrárias 36:1135–1144. https://doi.org/10.5433/1679-0359.2015v36n2p1135
McDonald P, Henderson AR, Herson SJE (1991) The Biochemistry of Silage, 2nd edn. Chalcombe publication, Marlow
Mceniry J, O’Kiely P, Clipson NJW, Forristal PD, Doyle EM (2007) The relative impacts of wilting, chopping, compaction and air infiltration on the conservation characteristics of ensiled grass. Grass Forage Sci 62(4):470–484. https://doi.org/10.1111/j.1365-2494.2007.00602.x
McGarvey JA, Franco RB, Palumbo JD, Hnasko R, Stanker L, Mitloehner FM (2013) Bacterial population dynamics during the ensiling of Medicago sativa (alfalfa) and subsequent exposure to air. J Appl Microbiol 114:1661–1670. https://doi.org/10.1111/jam.12179
Mendez-Garcia C, Peláez AI, Mesa V, Sanchez J, Golyshina OV, Ferrer M (2015) Microbial diversity and metabolic networks in acid mine drainage habitats. Front Microbio 6:475. https://doi.org/10.3389/fmicb.2015.00475
Moosa V, Mohsen DM, AliReza V (2012) Fermentation characteristics, in situ rumen degradation and aerobic stability of whole crop barley ensiled with urea or aqueous ammonia. Am J Exp Agric 2(1):21–30. https://www.webofscience.com/wos/alldb/full-record/CABI:20123099856
Muck RE, Nadeau EMG, McAllister TA, Contreras-Govea FE, Santos MC, Kung L Jr (2018) Silage review: Recent advances and future uses of silage additives. J Dairy Sci 101(5):3980–4000. https://doi.org/10.3168/jds.2017-13839
Ni K, Minh TT, Tu TT, Tsuruta T, Pang H, Nishino N (2017) Comparative microbiota assessment of wilted Italian ryegrass, whole crop corn, and wilted alfalfa silage using denaturing gradient gel electrophoresis and next-generation sequencing. Appl Microbiol Biot 101:1385–1394. https://doi.org/10.1007/s00253-016-7900-2
Ni K, Zhao J, Zhu B, Su R, Pan Y, Ma J, Zhou G, Tao Y, Liu X, Zhong J (2018) Assessing the fermentation quality and microbial community of the mixed silage of forage soybean with crop corn or sorghum. Bioresource Technol 265:563–567. https://doi.org/10.1016/j.biortech.2018.05.097
Pang H, Qin G, Tan Z, Li Z, WangY CY (2011) Natural populations of lactic acid bacteria associated with silage fermentation as determined by phenotype, 16S ribosomal RNA and recA gene analysis. Syst Appl Microbiol 34:235–241. https://doi.org/10.1016/j.syapm.2010.10.003
Putnam DH, Orloff SB (2014) Forage crops. Encyclopedia Agric Food Syst 3:381–405. https://doi.org/10.1016/B978-0-444-52512-3.00142-X
Rognes T, Flouri T, Nichols B, Quince C, Mahé F (2016) VSEARCH: a versatile open source tool for 516 metagenomics. Peer J 4:e2584. https://doi.org/10.7717/peerj.2584
Santos MC, Kung L Jr (2016) Short communication: The effects of dry matter and length of storage on the composition and nutritive value of alfalfa silage. J Dairy Sci 99(7):5466–5469. https://doi.org/10.3168/jds.2016-10866
Santos AO, Avila CLS, Pinto JC, Carvalho BF, Dias DR, Schwan RF (2016) Fermentative profile and bacterial diversity of corn silages inoculated with new tropical lactic acid bacteria. J Appl Microbiol 120:266–279. https://doi.org/10.1111/jam.12980
Stamilla A, Russo N, Messina A, Spadaro C, Natalello A, Caggia C, Randazzo CL, Lanza M (2020) Effects of microencapsulated blend of organic acids and essential oils as a feed additive on quality of chicken breast meat. Animals (basel) 10(4):640. https://doi.org/10.3390/ani10040640
Wanapat M, Kang S, Khejornsart P, Pilajun R (2013) Improvement of whole crop rice silage nutritive value and rumen degradability by molasses and urea supplementation. Trop Anim Health pro 45(8):1777–1781. https://doi.org/10.1007/s11250-013-0433-0
Wang D, Khurshid M, Sun ZM, Tang YX, Zhou ML, Wu YM (2016) Genetic engineering of Alfalfa (Medicago sativa L.). Protein Peptide Lett 23(5):495–502. https://doi.org/10.2174/0929866523666160314152618
Wang C, Pian R, Chen X, Lv H, Zhou W, Zhang Q (2020) Beneficial effects of tannic acid on the quality of bacterial communities present in high-moisture mulberry leaf and stylo silage. Front Microbio 11:586412. https://doi.org/10.3389/fmicb.2020.586412
Weatherburn MW (1967) Phenol-hypochlorite reaction for determination of ammonia. Anal Chem 39(8):971–974. https://doi.org/10.1021/ac60252a045
William H, George W (Eds) (2007) Official methods of analysis of AOAC International,18th edn, AOAC International, Gaithersburg
Wu JD, Chen YB, Sun W (2007) Influence of adding urea on the quality of silage corn straw. J West Anhui Univer 23(5):87–89. https://doi.org/10.3969/j.issn.1009-9735.2007.05.027
Yunus M, Ohba N, Tobisa M, Shimojo M, Masuda Y (2001) Effect of glucose and formic acid on the quality of napiergrass silage after treatment with urea. Asian Austral J Anim 14(2):211–215. https://doi.org/10.5713/ajas.2001.211
Yunus M, Ohba N, Shao T, Yano Y, Tobisa M, Shimojo M, Furuse M, Masuda Y, Shao T (2000) Effect of slower nitrogen releasing additive on the quality of napiergrass (Pennisetum purpureum Schumach.) silage. J Fac Agr Kyushu U 45(2):415–421.https://doi.org/10.5109/24390
Zhao LP, Ren LP, Zhou ZM, Meng QX, Hou YL, Wang F (2016) Improving ruminal degradability and energetic values of bamboo shoot shell using chemical treatments. Anim Sci J 87:896–903. https://doi.org/10.1111/asj.12512
Funding
This study was financially supported by Chinese Academy of Agricultural Sciences and Central Public-interest Scientific Institution Basal Research Fund by Chinese Academy of Agricultural Sciences (1,610,242,021,005) provided by Dr. Zheng, Natural Science Foundation of Hunan Province by Hunan Provincial Department of science and technology (2021JJ50028) provided by Dr. Hou, Agricultural Science and Technology Innovation Project Special Fund of Chinese Academy of Agricultural Sciences by Chinese Academy of Agricultural Sciences (ASTIP–IBFC) provided by Dr. Wu. We thank all present and past members of our laboratory who have contributed comments and ideas.
Author information
Authors and Affiliations
Contributions
The authors’ contributions are as follows: Conceptualization, Hou Z.P. and Wu D.Q.; Methodology, ZHENG.X. and Zheng X.L.; Analysis, Z.H. and X.Z.; Data curation, Wu D.Q; Writing & Editing—Hou Z.P. and Wu D.Q.
Corresponding author
Ethics declarations
Competing interests
The authors have no relevant financial or non-financial interests to disclose.
Additional information
Communicated by Erko Stackebrandt.
Publisher's Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Rights and permissions
About this article
Cite this article
Hou, Z., Zheng, X., Zhang, X. et al. Effects of urea supplementation on the nutritional quality and microbial community of alfalfa (Medicago sativa L.) silage. Arch Microbiol 204, 414 (2022). https://doi.org/10.1007/s00203-022-03046-x
Received:
Revised:
Accepted:
Published:
DOI: https://doi.org/10.1007/s00203-022-03046-x