Characterization of Lactic Acid-Producing Bacteria Isolated from Rumen: Growth, Acid and Bile Salt Tolerance, and Antimicrobial Function

Abstract

Lactic acid bacteria are some of the dominant bacteria in the rumen, and they have a high ability for lactic acid production. The present study aimed to screen and evaluate the performance of culturable rumen bacteria from Chinese Holstein dairy cows as a potential probiotic or inoculant for silage production, in order to isolate ruminal lactic acid bacteria and evaluate their potential as probiotics. Three strains of Enterococcus avium (E. avium, EA1-3); three strains of Streptococcus lutetiensis (S. lutetiensis, SL1-3); and six strains of Streptococcus equinus (S. equinus, SE1-6) were successfully identified from the rumen fluid using modified De Man Rogosa sharp medium supplemented with 0.325% lactic acid. E. avium, S. lutetiensis and S. equinus are clustered in the phylogenetic tree. All the 12 Gram-positive strains reached the plateau growth phase in 6–10 h, with an OD600 at about 1.8. Both gas and acid accumulation reached plateaus at about 10–12 h in all strains, and S. equinus showed the strongest capacity. The highest lactic acid accumulation was detected in S. equinus broth (up to 219.77 μmol/L). The growth of all isolates was inhibited at pH 4.0, and EA2, SL1, SL2, SL3 and SE2 were tolerant to 0.1%, 0.2% and 0.3% bile salt. In addition, the supernatants of the strains had inhibitory effects on Escherichia coli and Staphylococcus aureus. Specifically, the S. equinus strains exhibited the strongest inhibition of the pathogens. In conclusion, these 12 strains had good potential as silage inoculants or probiotics for edible animals, especially S. equinus.

Keywords

• <i>Enterococcus avium</i>
• <i>Streptococcus lutetiensis</i>
• <i>Streptococcus equinus</i>
• identification
• lactic acid