Fenugreek sprout extract as a natural probiotic enhancer and antimicrobial agent for sustainable reproductive health management

Reproductive pathologies such as mastitis and uterine infections remain major challenges in dairy production. with limited treatment success, they are difficult to treat, often recur, and lead to economic losses due to reduced fertility, antibiotic residues, withdrawal periods, and antimicrobial resistance. Probiotics, particularly lactic acid bacteria (LAB), are increasingly explored as sustainable alternatives; however, their effectiveness depends on growth, adhesion, and biofilm formation within host environments. This study evaluated the prebiotic and antimicrobial potential of germinated fenugreek seed extract (GSF), rich in polyphenols (170.3 ± 10.7 mg GAE/g) and flavonoids (79.8 ± 9.3 mg QE/g), on two LAB strains: Streptococcus thermophilus (S. thermophilus) and Enterococcus durans ( E. durans) . At optimal concentrations (0.8–0.9 mg/mL), GSF significantly enhanced LAB planktonic growth by 25%, autoaggregation (71.9% vs. 65.1% in E. durans ), and biofilm formation (30% in S. thermophilus ). GSF also modulated membrane permeability, with protein release increasing up to 85.4% in E. durans compared to 4.9% in S. thermophilus , supporting quorum-sensing activity. Importantly, the combination of LAB cell-free supernatants with GSF extract exhibited synergistic inhibitory effects, with inhibition zones against Escherichia coli reaching 2.35 cm and Staphylococcus aureus 1.20 cm. These findings highlight the dual role of fenugreek sprout extracts as probiotic enhancers and antimicrobial agents, supporting their potential use as natural, antibiotic-free interventions to improve reproductive health and productivity in livestock.