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Effects on microbial diversity of fermentation temperature (10℃ and 20℃), long-term storage at 5℃, and subsequent warming of corn silage

  • Received : 2018.10.23
  • Accepted : 2019.01.31
  • Published : 2019.10.01

Abstract

Objective: To evaluate the effects on microbial diversity and biochemical parameters of gradually increasing temperatures, from $5^{\circ}C$ to $25^{\circ}C$ on corn silage which was previously fermented at ambient or low temperature. Methods: Whole-plant corn silage was fermented in vacuum bag mini-silos at either $10^{\circ}C$ or $20^{\circ}C$ for two months and stored at $5^{\circ}C$ for two months. The mini-silos were then subjected to additional incubation from $5^{\circ}C$ to $25^{\circ}C$ in $5^{\circ}C$ increments. Bacterial and fungal diversity was assessed by polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) profiling and biochemical analysis from mini-silos collected at each temperature. Results: A temperature of $10^{\circ}C$ during fermentation restricted silage fermentation compared to fermentation temperature of $20^{\circ}C$. As storage temperature increased from $5^{\circ}C$ to $25^{\circ}C$, little changes occurred in silages fermented at $20^{\circ}C$, in terms of most biochemical parameters as well as bacterial and fungal populations. However, a high number of enterobacteria and yeasts (4 to $5\;log_{10}$ colony forming unit/g fresh materials) were detected at $15^{\circ}C$ and above. PCR-DGGE profile showed that Candida humilis predominated the fungi flora. For silage fermented at $10^{\circ}C$, no significant changes were observed in most silage characteristics when temperature was increased from $5^{\circ}C$ to $20^{\circ}C$. However, above $20^{\circ}C$, silage fermentation resumed as observed from the significantly increased number of lactic acid bacteria colonies, acetic acid content, and the rapid decline in pH and water-soluble carbohydrates concentration. DGGE results showed that Lactobacillus buchneri started to dominate the bacterial flora as temperature increased from $20^{\circ}C$ to $25^{\circ}C$. Conclusion: Temperature during fermentation as well as temperature during storage modulates microorganism population development and fermentation patterns. Silage fermented at $20^{\circ}C$ indicated that these silages should have lower aerobic stability at opening because of better survival of yeasts and enterobacteria.

Keywords

References

  1. Allen MS, Coors JG, Roth GW. Corn silage. In: Buxton DR, Muck RE, Harrison JH, editors. Silage science and technology. Madison, WI, USA: ASA, CSSA, SSSA; 2003.
  2. Borreani G, Tabacco E, Schmidt RJ, Holmes BJ, Muck RE. Silage review: factors affecting dry matter and quality losses in silages. J Dairy Sci 2018;101:3952-79. https://doi.org/10.3168/jds.2017-13837
  3. Wilkinson JM, Davies DR. The aerobic stability of silage: key findings and recent developments. Grass Forage Sci 2013;68:1-19. https://doi.org/10.1111/j.1365-2494.2012.00891.x
  4. Moon NJ. Inhibition of the growth of acid tolerant yeasts by acetate, lactate and propionate and their synergistic mixtures. J Appl Microbiol 1983;55:453-60. https://doi.org/10.1111/j.1365-2672.1983.tb01685.x
  5. Weinberg ZG, Ashbell G, Hen Y, Azrieli A. The effect of applying lactic acid bacteria at ensiling on the aerobic stability of silages. J Appl Bacteriol 1993;75:512-8. https://doi.org/10.1111/j.1365-2672.1993.tb01588.x
  6. Muck RE, Moser LE, Pitt RE. Postharvest factors affecting ensiling. In. Buxton DR, Muck RE, Harrison JH, editors. Silage science and technology. Madison, WI, USA: ASA, CSSA, SSSA;2003.
  7. Bernardes TF, Daniel JLP, Adesogan AT, et al. Unique challenges of silages made in hot and cold regions. J Dairy Sci 2018;101:4001-19. https://doi.org/10.3168/jds.2017-13703
  8. Zhou Y, Drouin P, Lafreniere C. Effect of temperature (5-$25^{\circ}C$) on epiphytic lactic acid bacteria populations and fermentation of whole-plant corn silage. J Appl Microbiol 2016;121:657-71. https://doi.org/10.1111/jam.13198
  9. Hoedtke S, Zeyner A. Comparative evaluation of laboratoryscale silages using standard glass jar silages or vacuum-packed model silages. J Sci Food Agric 2011;91:841-9. https://doi.org/10.1002/jsfa.4255
  10. Johnson HE, Merry RJ, Davies DR, Kell DB, Theodorou MK, Griffith GW. Vacuum packing: a model system for laboratoryscale silage fermentations. J Appl Microbiol 2005;98:105-13. https://doi.org/10.1111/j.1365-2672.2004.02444.x
  11. Dubois M, Gilles KA, Hamilton JK, Rebers PA, Smith F. Colorimetric method for determination of sugars and related substances. Anal Chem 1956;28:350-6. https://doi.org/10.1021/ac60111a017
  12. Fussell RJ, McCalley DV. Determination of volatile fatty acids (C2-C5) and lactic acid in silage by gas chromatography. Analyst 1987;112:1213-6. https://doi.org/10.1039/an9871201213
  13. Taylor KACC. A simple colorimetric assay for muramic acid and lactic acid. Appl Biochem Biotechnol A 1996;56:49-58. https://doi.org/10.1007/BF02787869
  14. Flipot P, Mowat DN, Parkins JJ, Buchanan-Smith JG. Ensiling characteristics of silage treated with sodium hydroxyde. Can J Plant Sci 1976;56:935-40. https://doi.org/10.4141/cjps76-151
  15. Jonsson A. Enumeration and confirmation of Clostridium tyrobutyricum in silages using Neutral Red, D-cycloserine, and lactate dehydrogenase activity. J Dairy Sci 1990;73:719-25. https://doi.org/10.3168/jds.S0022-0302(90)78725-5
  16. Gao L, Yang H, Wang X, et al. Rice straw fermentation using lactic acid bacteria. Bioresour Technol 2008;99:2742-8. https://doi.org/10.1016/j.biortech.2007.07.001
  17. Kebli H, Drouin P, Brais S, Kernaghan G. Species composition of saproxylic fungal communities on decaying logs in the Boreal forest. Microb Ecol 2011;61:898-910. https://doi.org/10.1007/s00248-010-9790-7
  18. May LA, Smiley B, Schmidt MG. Comparative denaturing gradient gel electrophoresis analysis of fungal communities associated with whole plant corn silage. Can J Microbiol 2001;47:829-41. https://doi.org/10.1139/w01-086
  19. Stringini M, Comitini F, Taccari M, Ciani M. Yeast diversity in crop-growing environments in Cameroon. Int J Food Microbiol 2008;127:184-9. https://doi.org/10.1016/j.ijfoodmicro.2008.07.017
  20. Thanh VN, Mai LT, Tuan DA. Microbial diversity of traditional Vietnamese alcohol fermentation starters (banh men) as determined by PCR-mediated DGGE. Int J Food Microbiol 2008;128:268-73. https://doi.org/10.1016/j.ijfoodmicro.2008.08.020
  21. Bonito G, Isikhuemhen OS, Vilgalys R. Identification of fungi associated with municipal compost using DNA-based techniques. Bioresour Technol 2010;101:1021-7. https://doi.org/10.1016/j.biortech.2009.08.109
  22. Green SJ, Leigh MB, Neufeld JD. Denaturing gradient gel electrophoresis (DGGE) for microbial community analysis. In: Timmis KN, editors. Microbiology of hydrocarbons, oils, lipids and derived compounds. Heidelberg, Germany: Springer; 2017. https://doi.org/10.1007/978-3-662-52778-8
  23. Wieringa GW. Influence of moisture and nutrient content of forage plants on fermentation processes. 3rd General Meeting of the European Grassland Federation; 1969; Branshweig, Germany.
  24. Yang HY, Wang XF, Liu JB, et al. Effects of water-soluble carbohydrate content on silage fermentation of wheat straw. J Biosci Bioeng 2006;101:232-7. https://doi.org/10.1263/jbb.101.232
  25. Kim SC, Adesogan AT. Influence of ensiling temperature, simulated rainfall, and delayed sealing on fermentation characteristics and aerobic stability of corn silage. J Dairy Sci 2006;89:3122-32. https://doi.org/10.3168/jds.S0022-0302(06)72586-3
  26. Middelhoven WJ, Franzen MM. The yeast flora of ensiled whole-crop maize. J Sci Food Agric 1986;37:855-61. https//doi.org/10.1002/jsfa.2740370906.
  27. Santos MC, Golt C, Joerger RD, Mechor GD, Mourao GB, Kung LJ. Identification of the major yeasts isolated from high moisture corn and corn silages in the United States using genetic and biochemical methods. J Dairy Sci 2017;100:1151-60. https://doi.org/10.3168/jds.2016-11450
  28. Romero JJ, Joo Y, Park J, Tiezzi F, Gutierrez-Rodriguez E, Castillo MS. Bacterial and fungal communities, fermentation, and aerobic stability of conventional hybrids and brown midrib hybrids ensiled at low moisture with or without a homo- and heterofermentative inoculant. J Dairy Sci 2018;101:3057-76. https://doi.org/10.3168/jds.2017-13754
  29. Pahlow G, Muck RE, Driehuis F, Oude Elferink SJWH, Spoelstra SF. Microbiology of ensiling. In. Buxton DR, Muck RE, Harrison JH, editors. Silage science and technology. Madison, WI, USA: American Society of Agronomy; 2003.
  30. Pitt RE, Muck RE, Pickering NB. A model of aerobic fungal growth in silage. 2. Aerobic stability. Grass Forage Sci 1991; 46:301-12. https://doi.org/10.1111/j.1365-2494.1991.tb02235.x
  31. Oude Elferink SJWH, Krooneman J, Gottschal JC, Spoelstra SF, Faber F, Driehuis F. Anaerobic conversion of lactic acid to acetic acid and 1,2-propanediol by Lactobacillus buchneri. Appl Environ Microbiol 2001;67:125-32. https://doi.org/10.1128/AEM.67.1.125-132.2001
  32. Schmidt RJ, Hu W, Mills JA, Kung LJ. The development of lactic acid bacteria and Lactobacillus buchneri and their effects on the fermentation of alfalfa silage. J Dairy Sci 2009;92:5005-10. https://doi.org/10.3168/jds.2008-1701
  33. Tabacco E, Piano S, Revello-Chion A, Borreani G. Effect of Lactobacillus buchneri LN4637 and Lactobacillus buchneri LN40177 on the aerobic stability, fermentation products, and microbial populations of corn silage under farm conditions. J Dairy Sci 2011;94:5589-98. https://doi.org/10.3168/jds.2011-4286
  34. Wambacq E, Latre JP, Haesaert G. The effect of Lactobacillus buchneri inoculation on the aerobic stability and fermentation characteristics of alfalfa-ryegrass, red clover and maize silage. Agric Food Sci 2013;22:127-36. https://doi.org/10.23986/afsci.6711
  35. Rault A, Bouix M, Beal C. Fermentation pH influences the physiological-state dynamics of Lactobacillus bulgaricus CFL1 during pH-controlled culture. Appl Environ Microbiol 2009;75:4374-81. https://doi.org/10.1128/AEM.02725-08
  36. Eikmeyer F, Kofinger P, Poschenel A, et al. Metagenome analyses reveal the influence of the inoculant Lactobacillus buchneri CD034 on the microbial community involved in grass ensiling. J Biotechnol 2013;167:334-43. https://doi.org/10.1016/j.jbiotec.2013.07.021
  37. Ridwan R, Rusmana I, Widyastuti Y, et al. Fermentation characteristics and microbial diversity of tropical grass-legumes silages. Asian-Australas J Anim Sci 2015;28:511-8. https://doi.org/10.5713/ajas.14.0622
  38. Behrendt U, Ulrich A, Schumann P. Chryseobacterium gregarium sp. nov., isolated from decaying plant material. Int J Syst Evol Microbiol 2008;58:1069-74. https://dx.doi.org/10.1099/ijs.0.65544-0

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