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Varying ensiling conditions affect the fermentation quality and abundance of bacterial key players in lucerne silages

Published online by Cambridge University Press:  11 August 2020

T. Hartinger ,
K. Kube ,
N. Gresner  and
K.-H. Südekum Open the ORCID record for K.-H. Südekum [Opens in a new window]
T. Hartinger
Affiliation:
Institute of Animal Science, University of Bonn, Endenicher Allee 15, 53115Bonn, Germany
K. Kube
Affiliation:
Institute of Animal Science, University of Bonn, Endenicher Allee 15, 53115Bonn, Germany
N. Gresner
Affiliation:
Institute of Animal Science, University of Bonn, Endenicher Allee 15, 53115Bonn, Germany
K.-H. Südekum*
Affiliation:
Institute of Animal Science, University of Bonn, Endenicher Allee 15, 53115Bonn, Germany
*
Author for correspondence: K.-H. Südekum, E-mail: [email protected]
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Abstract

The successful ensiling of lucerne (Medicago sativa L.) depends on a rapid acidification in the silo and consequently relies on a sufficient proliferation of, particularly homofermentative, lactic acid bacteria. Similarly, growth of spoilage bacteria, such as enterobacteria and clostridia, must be suppressed and silage additives are therefore frequently applied to promote favourable conditions during ensiling. Three silage additives or soil were applied during lucerne ensiling and investigated for their effects on silage quality characteristics and abundances of total bacteria as well as the bacterial key players Lactobacillus spp., homofermentative Lact. plantarum, heterofermentative Lact. buchneri, Clostridium spp. and Enterobacteriaceae after 30 days of storage. Inoculation with viable Lact. plantarum resulted in highest concentration of this species and excellent silage quality, i.e. high lactic acid concentration coupled with low acetic acid and ammonia-nitrogen concentrations. A sodium nitrite and hexamine-based additive did not support growth of lactic acid bacteria, which was also apparent by higher pH and low lactic acid concentration. No effect of treatments was found on spoilage-related enterobacteria and clostridia, even not when adding soil to lucerne to increase initial clostridial contamination. However, soil treatment resulted in increased ammonia-nitrogen and acetic acid concentrations. Consequently, among the bacterial key players, lactic acid bacteria concentrations were related to silage quality. Regarding spoilage bacteria, however, alterations in silage quality characteristics were not reflected in the abundances of enterobacteria and clostridia. Future investigations should underpin the present findings and help to understand how silage additives affect microbial key players and silage fermentation.

Keywords

FermentationLactobacillussilage microbiologyspoilage bacteria
Type
Animal Research Paper
Information
The Journal of Agricultural Science , Volume 158 , Issue 4 , May 2020 , pp. 297 - 303
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Copyright
Copyright © The Author(s), 2020. Published by Cambridge University Press

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Footnotes

*

Present Address: Institute of Animal Nutrition and Functional Plant Compounds, Department for Farm Animals and Veterinary Public Health, University of Veterinary Medicine, Vienna, Austria

†

Present Address: Institute of Animal Nutrition, Livestock Products and Nutrition Physiology (TTE), University of Natural Resources and Life Sciences (BOKU), Vienna, Austria

References

Bartosch, S, Fite, A, Macfarlane, GT and McMurdo, MET (2004) Characterization of bacterial communities in feces from healthy elderly volunteers and hospitalized elderly patients by using real-time PCR and effects of antibiotic treatment on the fecal microbiota. Applied and Environmental Microbiology 70, 3575–3581.CrossRefGoogle ScholarPubMed
Bundesarbeitskreis Futterkonservierung (2011) Praxishandbuch Futter- und Substratkonservierung. Frankfurt am Main: DLG-Verlag GmbH.Google Scholar
Bustin, SA, Benes, V, Garson, JA, Hellemans, J, Huggett, J, Kubista, M, Mueller, R, Nolan, T, Pfaffl, MW, Shipley, GL, Vandesompele, J and Wittwer, CT (2009) The MIQE Guidelines: Minimum information for publication of quantitative real-time PCR experiments. Clinical Chemistry 55, 611–622.CrossRefGoogle ScholarPubMed
Conaghan, P, O'Kiely, P and O'Mara, FP (2010) Conservation characteristics of wilted perennial ryegrass silage made using biological or chemical additives. Journal of Dairy Science 93, 628–643.CrossRefGoogle ScholarPubMed
del Mar Lleo’, M, Tafi, MC and Canepari, P (1998) Nonculturable Enterococcus faecalis Cells are metabolically active and capable of resuming active growth. Systematic and Applied Microbiology 21, 333–339.CrossRefGoogle Scholar
Driehuis, F and Oude Elferink, SJ (2000) The impact of the quality of silage on animal health and food safety: a review. Veterinary Quarterly 22, 212–216.CrossRefGoogle ScholarPubMed
Eikmeyer, FG, Köfinger, P, Poschenel, A, Jünemann, S, Zakrzewski, M, Heinl, S, Mayrhuber, E, Grabherr, R, Pühler, A, Schwab, H and Schlüter, A (2013) Metagenome analyses reveal the influence of the inoculant Lactobacillus buchneri CD034 On the microbial community involved in grass ensiling. Journal of Biotechnology 167, 334–343.CrossRefGoogle ScholarPubMed
Fuller, Z, Louis, P, Mihajlovski, A, Rungapamestry, V, Ratcliffe, B and Duncan, AJ (2007) Influence of cabbage processing methods and prebiotic manipulation of colonic microflora on glucosinolate breakdown in man. British Journal of Nutrition 98, 364–372.CrossRefGoogle ScholarPubMed
Guo, XS, Ke, WC, Ding, WR, Ding, LM, Xu, DM, Wang, WW, Zhang, P and Yang, FY (2018) Profiling of metabolome and bacterial community dynamics in ensiled Medicago sativa Inoculated without or with Lactobacillus plantarum Or Lactobacillus buchneri. Scientific Reports 8, 357.CrossRefGoogle ScholarPubMed
Hartinger, T, Gresner, N and Südekum, K-H (2019) Effect of wilting intensity, dry matter content and sugar addition on nitrogen fractions in lucerne silages. Agriculture 9, 11.CrossRefGoogle Scholar
Hinds, AA and Lowe, LE (1980) Application of the Berthelot reaction to the determination of ammonium-N in soil extracts and soil digests. Communications in Soil Science and Plant Analysis 11, 469–475.CrossRefGoogle Scholar
Hoedtke, S, Gabel, M and Zeyner, A (2010) Der Proteinabbau im Futter während der Silierung und Veränderungen in der Zusammensetzung der Rohproteinfraktion. Übersichten zur Tierernährung 38, 157–179.Google Scholar
Kaewtapee, C, Burbach, K, Tomforde, G, Hartinger, T, Camarinha-Silva, A, Heinritz, S, Seifert, J, Wiltafsky, M, Mosenthin, R and Rosenfelder-Kuon, P (2017) Effect of Bacillus subtilis And Bacillus licheniformis Supplementation in diets with low- and high-protein content on ileal crude protein and amino acid digestibility and intestinal microbiota composition of growing pigs. Journal of Animal Science and Biotechnology 8, 9.CrossRefGoogle ScholarPubMed
Klocke, M, Mundt, K, Idler, C, McEniry, J, O'Kiely, P and Barth, S (2006) Monitoring Lactobacillus plantarum In grass silages with the aid of 16S rDNA-based quantitative real-time PCR assays. Systematic and Applied Microbiology 29, 49–58.CrossRefGoogle ScholarPubMed
Kung, L and Shaver, R (2001) Interpretation and use of silage fermentation analysis reports. Focus on Forage 3, 1–5.Google Scholar
Kung, L, Shaver, RD, Grant, RJ and Schmidt, RJ (2018) Silage review: interpretation of chemical, microbial, and organoleptic components of silages. Journal of Dairy Science 101, 4020–4033.CrossRefGoogle ScholarPubMed
Li, M, Penner, GB, Hernandez-Sanabria, E, Oba, M and Guan, LL (2009) Effects of sampling location and time, and host animal on assessment of bacterial diversity and fermentation parameters in the bovine rumen. Journal of Applied Microbiology 107, 1924–1934.CrossRefGoogle ScholarPubMed
Lingvall, P and Lättemäe, P (1999) Influence of hexamine and sodium nitrite in combination with sodium benzoate and sodium propionate on fermentation and hygienic quality of wilted and long cut grass silage. Journal of the Science of Food and Agriculture 79, 257–264.3.0.CO;2-J>CrossRefGoogle Scholar
McAllister, TA, Dunière, L, Drouin, P, Xu, S, Wang, Y, Munns, K and Zaheer, R (2018) Silage review: using molecular approaches to define the microbial ecology of silage. Journal of Dairy Science 101, 4060–4074.CrossRefGoogle ScholarPubMed
McDonald, P, Henderson, N and Heron, S (1991) The Biochemistry of Silage. Marlow, UK: Chalcombe.Google Scholar
McEniry, J, O'Kiely, P, Clipson, NWJ, Forristal, PD and Doyle, EM (2007) Manipulating the ensilage of wilted, unchopped grass through the use of additive treatments. Irish Journal of Agricultural and Food Research 46, 77–91.Google Scholar
Muck, RE (2010) Silage microbiology and its control through additives. Revista Brasileira de Zootecnia 39, 183–191.CrossRefGoogle Scholar
Mullins, CR, Mamedova, LK, Carpenter, AJ, Ying, Y, Allen, MS, Yoon, I and Bradford, BJ (2013) Analysis of rumen microbial populations in lactating dairy cattle fed diets varying in carbohydrate profiles and Saccharomyces cerevisiae Fermentation product. Journal of Dairy Science 96, 5872–5881.CrossRefGoogle ScholarPubMed
Oude Elferink, SJWH, Krooneman, J, Gottschal, JC, Spoelstra, SF, Faber, F and Driehuis, F (2001) Anaerobic conversion of lactic acid to acetic acid and 1,2-propanediol by Lactobacillus buchneri. Applied and Environmental Microbiology 67, 125–132.CrossRefGoogle ScholarPubMed
Pahlow, G, Muck, R E, Driehuis, F, Oude Elferink, SJWH and Spoelstra, S F (2003) Microbiology of ensiling. In Buxton, DR, Muck, RE and Harrison, JH (eds), Silage Science and Technology. Madison, WI, USA: American Society of Agronomy; Crop Science Society of America; Soil Science Society of America, pp. 31–93.Google Scholar
Pang, H, Qin, G, Tan, Z, Li, Z, Wang, Y and Cai, Y (2011) Natural populations of lactic acid bacteria associated with silage fermentation as determined by phenotype, 16S ribosomal RNA and recA Gene analysis. Systematic and Applied Microbiology 34, 235–241.CrossRefGoogle ScholarPubMed
Rebollar, EA, Antwis, RE, Becker, MH, Belden, LK, Bletz, MC, Brucker, RM, Harrison, XA, Hughey, MC, Kueneman, JG, Loudon, AH, McKenzie, V, Medina, D, Minbiole, KPC, Rollins-Smith, LA, Walke, JB, Weiss, S, Woodhams, DC and Harris, RN (2016) Using “omics” and integrated multi-omics approaches to guide probiotic selection to mitigate chytridiomycosis and other emerging infectious diseases. Frontiers in Microbiology 7, 68.CrossRefGoogle ScholarPubMed
Rinttilä, T, Kassinen, A, Malinen, E, Krogius, L and Palva, A (2004) Development of an extensive set of 16S rDNA-targeted primers for quantification of pathogenic and indigenous bacteria in faecal samples by real-time PCR. Journal of Applied Microbiology 97, 1166–1177.CrossRefGoogle ScholarPubMed
Rochelle, PA, Leon, RD, Stewart, MH and Wolfe, RL (1997) Comparison of primers and optimization of PCR conditions for detection of Cryptosporidium parvum And Giardia Lamblia In water. Applied and Environmental Microbiology 63, 106–114.CrossRefGoogle ScholarPubMed
Scherer, R, Gerlach, K, Taubert, J, Adolph, S, Weiß, K and Südekum, K-H (2019) Effect of forage species and ensiling conditions on silage composition and quality and the feed choice behaviour of goats. Grass and Forage Science 74, 297–313.CrossRefGoogle Scholar
Schmidt, RJ, Emara, MG and Kung, L (2008) The use of a quantitative real-time polymerase chain reaction assay for identification and enumeration of Lactobacillus buchneri In silage. Journal of Applied Microbiology 105, 920–929.CrossRefGoogle ScholarPubMed
Schmidt, RJ, Hu, W, Mills, JA and Kung, L (2009) The development of lactic acid bacteria and Lactobacillus buchneri And their effects on the fermentation of alfalfa silage. Journal of Dairy Science 92, 5005–5010.CrossRefGoogle ScholarPubMed
Seale, DR, Henderson, AR, Pettersson, KO and Lowe, JF (1986) The effect of addition of sugar and inoculation with two commercial inoculants on the fermentation of lucerne silage in laboratory silos. Grass and Forage Science 41, 61–70.CrossRefGoogle Scholar
Stevenson, DM, Muck, RE, Shinners, KJ and Weimer, PJ (2006) Use of real time PCR to determine population profiles of individual species of lactic acid bacteria in alfalfa silage and stored corn stover. Applied Microbiology and Biotechnology 71, 329–338.CrossRefGoogle ScholarPubMed
VDLUFA (2012) Handbuch der landwirtschaftlichen Versuchs- und Untersuchungsmethodik (VDLUFA-Methodenbuch). Band III. Die chemische Untersuchung von Futtermitteln. Darmstadt: VDLUFA-Verlag.Google Scholar
von Lengerken, J and Zimmermann, K (1991) Handbuch Futtermittelprüfung. Berlin: Deutscher Landwirtschaftsverlag.Google Scholar
Weissbach, F and Kuhla, S (1995) Stoffverluste bei der Bestimmung des Trockenmassegehaltes von Silagen und Grünfutter: Entstehende Fehler und Möglichkeiten der Korrektur. Übersichten zur Tierernährung 23, 189–214.Google Scholar
Wen, A, Yuan, X, Wang, J, Desta, ST and Shao, T (2017) Effects of four short-chain fatty acids or salts on dynamics of fermentation and microbial characteristics of alfalfa silage. Animal Feed Science and Technology 223, 141–148.CrossRefGoogle Scholar
Woolford, MK (1975) Microbiological screening of food preservatives, cold sterilants and specific antimicrobial agents as potential silage additives. Journal of the Science of Food and Agriculture 26, 229–237.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, 1456–1470.CrossRefGoogle ScholarPubMed