Performance of New Lactic Acid Bacteria Strains as Inoculants on the Microorganism Composition during Fermentation of Alfalfa Silage Containing Different Dry Matter Content

Abstract

Alfalfa is the most planted perennial legume in the world due to its high nutritive value, protein content, productivity, and digestibility in addition to high vitamin and mineral content. It is also one of the hardest plants to ensile owing to its low reducible sugar and dry matter (DM) contents and high buffering capacity. In this study, the effects of inoculation with Lactobacillus bifermentans which is homofermentative and Lactobacillus brevis which is heterofermentative on the silage fermentation of different DM containing alfalfa forage. Alfalfa forages were unwilted, or wilted for 9 or 24 hours in order to achieve low (L), moderate (M) and high (H) DM contents. As a result of the research, it was determined that wilting improved the fermentation properties, decreased the pH value, and increased dry matter recovery. Microbial inoculation decreased the pH value, increased the dry matter recovery, and decreased the number of undesirable enterobacteria in silage. As a result of the research, it was determined that the wilting and inoculation should be applied for successful fermentation of alfalfa silage. It was also determined that inoculation with L. bifermentans gave the highest crude protein (CP) content while L. brevis gave the highest dry matter recovery (DMR).

Keywords

• Alfalfa
• L. bifermantans
• L. brevis
• Lucerna
• Inoculant
• Wilting

Supporting Institution

TUBITAK

Project Number

110O694

Thanks

The microbial materials used in this study were isolated within the scope of the 110O694 project supported by TUBITAK.

References

1. Agarussi MCN, Pereira OG, da Silva VP, Leandro ES, Ribeiro KG, Santos SA. 2019. Fermentative profile and lactic acid bacterial dynamics in non-wilted and wilted alfalfa silage in tropical conditions. Molec Biol Rep, 46(1): 451-460.
2. AOAC 1990. Official Method of Analysis. 15th. edn. Association of Official Analytical Chemist, Washington, DC. US.
3. Arndt C, Powell JM, Aguerre MJ, Wattiaux MA. 2015. Performance, digestion, nitrogen balance, and emission of manure ammonia, enteric methane, and carbon dioxide in lactating cows fed diets with varying Alfalfa silage to corn silage ratios. J Dairy Sci, 98: 418-430.
4. Blajman JE, Signorini ML, Vinderola G, Lingua MS, Romero LA, Páez RB, Gaggiotti MC. 2022. Impact of native spray‐dried lactic acid bacteria, packing density and wilting time on fermentation characteristics of experimental maize and lucerne silages. Grass Forage Sci, 77(1): 66-78.
5. Bolsen KK, Ashbell G, Weinberg ZG. 1996. Silage fermentation and silage additives-review. Asian-Australasian J Anim Sci, 9(5): 483-494.
6. Canbolat Ö. 2012. Comparison of in vitro gas production, organic matter digestibility, relative feed value and metabolizable energy contents of some cereal forages. J Fac Vet Medic, 18(4): 571-577.
7. Coblentz WK, Muck RE. 2012. Effects of natural and simulated rainfall on indicators of ensilability and nutritive value for wilting alfalfa forages sampled before preservation as silage. J Dairy Sci, 95(11): 6635-6653.
8. Davies DR, Theodorou MK, Kingston-Smith AH, Merry RJ. 2005. Advances in silage quality. 14th International Silage Conference, July 10-12, 2005, Belfast, Northern Ireland. pp: 121-133.

9. Dewhurst RJ, Fisher WJ, Tweed JKS, Wilkins RJ. 2003. Comparison of grass and legume silages for milk production. 1. Production responses with different levels of concentrate. J Dairy Sci, 86(8): 2598-2611.
10. Ertekin İ, Kızılşimşek M. 2020. Effects of lactic acid bacteria inoculation in pre-harvesting period on fermentation and feed quality properties of alfalfa silage. Asian-Australasian J Anim Sci, 33(2): 245.
11. Gomes FM, Ribeiro KG, de Souza IA, de Silva J, Agarussi MCN, da Silva VP, da Silva TC, Pereira OG. 2021. Chemical composition, fermentation profile, microbial population and dry matter recovery of silages from mixtures of palisade grass and forage peanut. Tropical Grasslands Forrajes Tropicales, 9(1): 34-42.
12. Gül ZD, Tan M, Kaynar DF, Kharazmi K. 2015. Effects of some additives, harvest stage and wilting on quality characteristics of alfalfa silage. Atatürk Univ., J Agril Fac, 46(2): 113-118.
13. Günaydın T, Akbay F, Arıkan S, Kızılşimşek M. 2023. Effects of different lactic acid bacteria inoculants on alfalfa silage fermentation and quality. J Agri Sci, 29(2): 555-560.
14. Hanagasaki T. 2020. Identification and characterization of lactic acid bacteria associated with tropical grass silage produced in Okinawa. Trop Grasslands, 8(3): 234-249.
15. Jatkauskas J, Vrotniakiene V. 2016. Effect of the inoculant on the fermentation, microbial population and aerobic stability of whole plant maize ensiled in large tubes. Žemdirbystė Agriculture. Kėdainiai: Lietuvos Žemdirbystės Institutas, 103(3): 305-310.
16. Keklik K. 2020. Using possibilities of newly isolated lactic acid bacteria strains as microbial inoculant on alfaalfa (Medicago sativa L.) silages. MSc Thesis. Kahramanmaraş Sütçü imam University, institute of Science, Department of Field Crops, Kahramanmaras, Turkiye, pp: 60.
17. Kızılşimşek M, Adem E, Dönmez R, Katrancı B. 2016a. Silaj mikro florasının birbirleri ile ilişkileri, silaj fermentasyonu ve kalitesi üzerine etkileri. KSÜ Doğa Bil Derg, 19(2): 136-140.
18. Kızılşimşek M, Keklik K, Günaydın T. 2020. Using possibilities of new lactic acid bacteria ısolates as microbial ınoculant on different dry matter containing alfalfa (Medicago sativa L.) silage. KSU J Agric Nat, 23(5): 1331-1339.
19. Kızılşimşek M, Küsek M, Gezginc Y, Erol A. 2016b. Isolation and identification of high lactic acid producer bacteria from forage and their silages grown in different ecologies. J Fac Vet Medic, 22: 291-296.
20. Lee SS, Choi JS, Paradhipta DHV, Joo YH, Lee HJ, Noh HT, Kim SC. 2021. Application of selected inoculant producing antifungal and fibrinolytic substances on rye silage with different wilting time. Processes, 9(5): 879.
21. Li XL, Wan LQ. 2005. Research progress on Medicago sativa silage technology. Acta Pratacult Sci, 14: 9-15.
22. Liu C, Lai Y, Lu X, Guo P, Luo H. 2016. Effect of lactic acid bacteria inoculants on alfalfa (Medicago sativa L.) silage quality: assessment of degradation (in situ) and gas production (in vitro). j Integrat Agri, 15(12): 2834-2841.
23. Liu C, Zhao GQ, Wei SN, Kim HJ, Li YF, Kim JG. 2021. Changes in fermentation pattern and quality of Italian ryegrass (Lolium multiflorum Lam.) silage by wilting and inoculant treatments. Anim Biosci, 34(1): 48.
24. McDonald P, Henderson SAR, Heron JE. 1991. The biochemistry of silage (2nd ed.). Challacombe Publ., Church Lane, Kingston, Canterbury, Kent, UK. ISBN: 9780948617225.
25. Muck RE, Filya I, Contreras-Govea FE. 2007. Inoculant effects on alfalfa silage: in vitro gas and volatile fatty acid production. J Dairy Sci, 90: 5115-5125.
26. Muck RE, Kung L. 1997. Effects of Silage Additives on Ensiling. in Silage: Field to Feedbunk. NRAES-99. Northeast Regional Agricultural Engineering Service, Ithaca, US, pp: 187-199.
27. Muck RE, Nadeau EMG, McAllister TA, Contreras-Govea FE, Santos MC, Kung JrL. 2018. Silage review: Recent advances and future uses of silage additives. J Dairy Sci, 101(5): 3980-4000.
28. Muck RE. 1987. Dry matter level effect on alfalfa silage quality. I. Nitrogen Transformation. Transact American Soc Agri Eng, 30: 7-14.
29. Muck RE. 2010. Silage microbiology and its control through additives. Rev Bras Zootec, 39: 183-191.
30. NLO. 2010. National Lucerne Organization of South Africa, URL: www.lusern.org (accessed date: June 3, 2011).
31. Oliveira AS, Weinberg ZG, Ogunade IM, Cervantes AA, Arriola KG, Jiang Y, Adesogan AT. 2017. Meta-analysis of effects of inoculation with homofermentative and facultative heterofermentative lactic acid bacteria on silage fermentation, aerobic stability, and the performance of dairy cows. J Dairy Sci, 100(6): 4587-4603.
32. Rangrab LH, Mühlbach PRF, Berto JL. 2000. Alfalfa silage harvested at the beginning of flowering and subjected to wilting and the action of biological additives (in Portuguese). R Bras de Zootec, 29(2): 349-356.
33. Silva VP, Pereira OG, Leandro ES, Da Silva TC, Ribeiro KG, Mantovani HC, Santos SA. 2016. Effects of lactic acid bacteria with Bacteriocinogenic potential on the fermentation profile and chemical composition of alfalfa silage in Tropical conditions. J Dairy Sci, 99(3): 1895-1902.
34. Tao X, Ji C, Chen S, Zhao J, Wang S, Li J, Sun F, Shao T. 2021. Fermentation quality and aerobic stability of Napier grass ensiled with citric acid residue and lactic acid bacteria. Trop Grasslands, 9(1): 52-59.
35. Tharanathan RN, Mahadevamma S. 2003. Grain legumes–a boon to human nutrition. Trends Food Sci Tech, 14(1): 507‐518.
36. Tyrolová Y, Výborná A. 2011. The effects of wilting and biological and chemical additives on the fermentation process in field pea silage. Czech J Anim Sci, 56:427-432.
37. Uslu OS, Kızılşimşek M, Kamalak A, Erol A, Kızıl S, Aydemır IE, Yanar K. 2017. Effects of wilting on dry matter recovery and feed quality of ryegrass silage. 2nd Internatıonal Balkan Agriculture Congress, May 16-18, 2017, Tekirdağ, Türkiye, pp: 127.
38. Wan JC, Xie KY, Wang YX, Liu L, Yu Z, Wang B. 2021. Effects of wilting and additives on the ensiling quality and in vitro rumen fermentation characteristics of sudangrass silage. Anim Biosci, 34(1): 56.
39. Wilkinson JM, Davies DR. 2013. The aerobic stability of silage: key findings and recent developments. Grass Forage Sci, 68(1): 1-19.
40. Xiccato G, Cinetto M, Carazzolo A, Cossu ME. 1994. The effect of silo type and dry matter content on the maize silage fermentation process and ensiling loss. Anim Feed Sci Tech, 49(3-4): 311-323.
41. Yang YX, Wang CZ, Lian HX, Zhang CM, Hu XF 2004. Effect of alfalfa meal on production performance, egg quality and egg yolk color of layers. J Huazh Agric, 23: 314-319.
42. Zhang YC, Wang XK, Li DX, Lin YL, Yang FY, Ni KK. 2020. Impact of wilting and additives on fermentation quality and carbohydrate composition of mulberry silage. Asian-Australis J Anim Sci, 33: 254-263.