Bacteriocin-Like Activity of Butyrivibrio fibrisolvens JL5 and Its Effect on Other Ruminal Bacteria and Ammonia Production

Abstract

When ruminal bacteria from a cow fed hay were serially diluted into an anaerobic medium that had only peptides and amino acids as energy sources, little growth or ammonia production was detected at dilutions greater than 10 ؊6 . The 10 ؊8 and 10 ؊9 dilutions contained bacteria that fermented carbohydrates, and some of these bacteria inhibited Clostridium sticklandii SR, an obligate amino acid-fermenting bacterium. Phylogenetic analysis indicated that the most active isolate (JL5) was closely related to Butyrivibrio fibrisolvens B835. Strain JL5 inhibited B. fibrisolvens 49 and a variety of other gram-positive organisms, but it had little effect on most gramnegative ruminal bacteria. Strain JL5 did not produce a bacteriocin-like inhibitory substance (BLIS) until it reached the late log or stationary phase. The JL5 BLIS did not cause the lysis of B. fibrisolvens 49, but the intracellular potassium level, the ATP level, the electrical potential, and the viability decreased rapidly. The JL5 BLIS also caused marked decreases in the viability and cellular potassium level of C. sticklandii SR. The membrane potential and intracellular ATP level also declined. The BLIS was degraded very slowly by pronase E, but it could be precipitated with 60% ammonium sulfate and dialyzed (3,500-Da cutoff). The BLIS could be separated from other peptides by polyacrylamide gel electrophoresis, and C. sticklandii SR overlays indicated that the molecular size of this compound was approximately 3,600 Da. Based on these results, it appeared that the JL5 BLIS was a pore-forming peptide. Because carbohydrate-fermenting ruminal bacteria could inhibit the growth of obligate amino acid-fermenting bacteria, BLIS may play a role in regulating ammonia production in vivo.

Distribution and Genetic Diversity of Bacteriocin Gene Clusters in Rumen Microbial Genomes

“…Besides all the efforts to characterize bacteriocins produced by ruminal bacteria, until now only three lantibiotics (butyrivibriocin OR79A, butyrivibriocin AR10, bovicin HC5), four class II bacteriocins (lichenin, bovicin 255, enterocin BC25, enterocin CCM4231), and two class III bacteriocins (enterolysin, albusin B) have been identified or at least partially characterized at the biochemical and genetic levels among strains of ruminal bacteria (10,15,16,18,19,21,(35)(36)(37). Moreover, although the occurrence of c Frequency refers to the proportion of the time that clusters containing the specified protein with a particular domain were identified in this study.…”

The complex world of gastrointestinal bacteria

“…Antimicrobial peptide production has been detected from several GI bacteria, both gram-negative and gram-positive (Miranda et al 1993;Leer et al 1995;Kalmokoff and Teather 1997;Avelar et al 1999;Yildirim et al 1999;Gomez et al 2002;Rychlik and Russell 2002). This strongly suggests AMPs have a role in altering and coordinating the GI bacterial community, although the exact nature of this role is not yet known.…”

“…Also, the inhibitory effect of nisin on some GI bacteria (Rada and Dlabal 1998;Mantovani and Russell 2001) suggests it has an inhibitory role in the GI tract. Ruminal isolates of Streptococcus bovis are sensitive to a bacteriocin produced by Enterococcus (Morovskyl et al 1998), and many gram-positive bacteria are sensitive to a peptide produced by Butyrivibrio fibrisolvens (Rychlik and Russell 2002). Also, the addition of nisin to an artificial rumen altered the fermentation pattern, which included increasing propionate and decreasing butyrate production (Jalc and Lauková 2002).…”

Production and Characterization of a Bacteriocin from Ruminal BacteriumRuminococcus albus7

“…[3][4][5] In recent years, they have been isolated from the rumen bacteria Streptococcus bovis and Butyrivibrio fibrisolvens. [6][7][8][9] Both predominant cellulolytics and readily isolated from ruminants, Ruminococcus albus 8 has been shown to produce bateriocin to inhibit the growth of Ruminococcus flavefaciens when co-cultured on cellobiose. 10) Fatty acids and surfactants have enhanced the growth performance and cellulytic activity of Ruminococci.…”

“…10) Previous studies have suggested that the chemical character and inhibition mechanism of bacteriocin vary among different strains of the same species bacteria. 2,18) Although many studies of bacteriocins from rumen bacteria have focused on their effects on rumen fermentation, [6][7][8][9]18) much fewer data are available concerning the antimicrobial activity of rumen bacteriocin as to other gut flora. The purpose of this study was to investigate the characteristics of the bacteriocin from R. albus 7 in detail and to examine its potential use as an alternative to antibiotics for pathogenic bacteria.…”