Effects of Lacticaseibacillus paracaseiand Enterococcus faecium on the microbial community and metabolic potential of legume-cereal silage

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Silage is used as a major part of fodder all over the world. Lactic acid bacteria (LAB) play a critical role in silage production from legumes, grains, and other plants, lowering the pH by producing different kinds of organic acids during ensiling, which inhibits undesirable bacteria that decrease the quality and shelf life of the inoculated product. The aim of this study was to determine the effects of Lacticaseibacillus paracasei and Enterococcus faecium inoculated together and separately on the bacterial community, fermentation quality and metabolic pathways of legume-cereal silage after 30 days. We conducted a model ensiling of legume-cereal crops of the first harvest cut in the Leningrad region at the booting stage. Three experimental groups with different bacterial strains were added during ensiling (LE: L. paracasei and E. faecium; LP: L. paracasei; and EF: E. faecium). The microorganisms in the silage were counted at 0 and 30 days of ensiling, using Illumina MiSeq high-throughput sequencing to determine their composition. The DNA of the silage microbial community was isolated, amplified, sequenced, and processed, then used to predict functional metabolic pathway abundances based on 16S rRNA marker gene sequences using the database MetaCyc. LAB genera were presented as minor taxa pre-ensiling, while Weissella, Lactobacillus, and Pediococcus were the dominant genera after 30 days of silage. Overall, LE had the most desirable bacteria and metabolic pathways for silage. LP also performed favorably. EF had undesirable bacteria and metabolites, showing the worst fermentation quality. Our results found important differences in predicted carbohydrate and biogenic amine metabolism between the different groups as well. To our knowledge, little information is available regarding the dynamics of bacterial community succession and metabolic pathways of silage prepared with L. paracasei and E. faecium.

About the authors

Sophia J. Gallegos

ITMO University

Author for correspondence.
Email: gallegos_sophia@outlook.com
ORCID iD: 0009-0004-9981-3016

Bachelor’s student, Faculty of Biotechnology

49A Kronverksky ave., Saint Petersburg, 197101, Russian Federation

Ekaterina S. Ponomareva

ITMO University

Email: ololoven@mail.ru
ORCID iD: 0000-0002-4336-8273
SPIN-code: 4260-6755

Engineer, Advanced Engineering School of Interdisciplinary Engineering

49A Kronverksky ave., Saint Petersburg, 197101, Russian Federation

Larisa A. Ilina

Saint Petersburg State Agrarian University

Email: ilina@biotrof.ru
ORCID iD: 0000-0003-2789-4844
SPIN-code: 5826-7525

Doctor of Biological Sciences, Professor, Department of Large Livestock Farming

2A Peterburgskoe highway, Pushkin, Saint Petersburg, 196601, Russian Federation

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Copyright (c) 2026 Gallegos S.J., Ponomareva E.S., Ilina L.A.

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