No abstract

The degradation of lactic acid under anoxic conditions was studied in several strains of Lactobacillus buchneri and in close relatives such as Lactobacillus parabuchneri, Lactobacillus kefir, and Lactobacillus hilgardii. Of these lactobacilli, L. buchneri and L. parabuchneri were able to degrade lactic acid under anoxic conditions, without requiring an external electron acceptor. Each mole of lactic acid was converted into approximately 0.5 mol of acetic acid, 0.5 mol of 1,2-propanediol, and traces of ethanol. Based on stoichiometry studies and the high levels of NAD-linked 1, 2-propanediol-dependent oxidoreductase (530 to 790 nmol min(-1) mg of protein(-1)), a novel pathway for anaerobic lactic acid degradation is proposed. The anaerobic degradation of lactic acid by L. buchneri does not support cell growth and is pH dependent. Acidic conditions are needed to induce the lactic-acid-degrading capacity of the cells and to maintain the lactic-acid-degrading activity. At a pH above 5.8 hardly any lactic acid degradation was observed. The exact function of anaerobic lactic acid degradation by L. buchneri is not certain, but some results indicate that it plays a role in maintaining cell viability.

Aerobic deterioration of silages is initiated by (facultative) aerobic micro‐organisms, usually yeasts, that oxidize the preserving organic acids. In this study, a Lactobacillus buchneri strain isolated from maize silage was evaluated for its potential as a bacterial inoculant that enhances aerobic stability of silages. In four experiments, chopped whole crop maize (30–43% dry matter (DM)) was inoculated with Lact. buchneri and ensiled in laboratory silos. Uninoculated silages served as controls. Analysis of silages treated with Lact. buchneri at levels of 103−106 cfu g−1 after about 3 months of anaerobic storage showedthat acetic acid and 1‐propanol contents increased with inoculum levels above 104 cfu g−1,whereas lactic acid decreased. Propionic acid, silage pH and DM loss increased withinoculum levels above 105 cfu g−1. Time course experiments with maize inoculated with Lact. buchneri at 4 × 104−2 × 105 cfu g−1 showed that up to 7–14 d after ensiling, Lact. buchneri had no effect on silage characteristics. Thereafter, the lactic acid content of the inoculated silages declined and, simultaneously, acetic acid and, to a lesser extent, propionic acid and 1‐propanol, accumulated. Inoculation reduced survival of yeasts during the anaerobic storage phase and inhibited yeast growth when the silage was exposed to O2, resulting in a substantial improvement in aerobic stability. The results indicate that the use of Lact. buchneri as a silage inoculant can enhance aerobic stability by inhibition of yeasts. The ability of the organism to ferment lactic acid to acetic acid appears to be an important underlying principle of this effect.

Aerobic spoilage by yeasts and moulds is a major cause of reduced nutritional value of silage and increases the risk of potential pathogenic microorganisms. Recent studies have shown that inoculation with Lactobacillus buchneri inhibits yeast growth and reduces the susceptibility to aerobic spoilage of various ensiled forages. The aim of this study was to determine whether these effects are retained when L. buchneri is added in combination with homofermentative lactic acid bacteria. In three experiments, silages were produced from perennial ryegrass [240±421 g kg )1 dry matter (DM)] inoculated with L. buchneri or L. buchneri plus a mixture of Pediococcus pentosaceus and Lactobacillus plantarum (inoculant PL). Uninoculated silage and silage inoculated with PL alone served as controls. Silages were examined for pH and DM loss in the course of ensilage and chemical and microbiological composition and aerobic stability after 3±4 months. L. buchneri plus PL and PL alone increased the initial rate of pH decline. L. buchneri alone and L. buchneri plus PL enhanced aerobic stability and, in general, reduced yeast and mould counts. In addition, these inoculants increased the ®nal pH and DM loss and the concentrations of acetic acid and 1,2-propanediol (or propionic acid and 1-propanol instead of 1,2-propanediol), and decreased the concentration of lactic acid. The effects of L. buchneri on fermentation products increased with decreasing DM content. In silages of less than 270 g kg )1 DM, L. buchneri increased the ammonia-N concentration. It is suggested that this was associated with the relatively high ®nal pH resulting from the high metabolic activity of L. buchneri in these silages.

Inoculation of maize silage with Lactobacillus buchneri (5 x 10(5) c.f.u. g(-1) of maize silage) prior to ensiling results in the formation of aerobically stable silage. After 9 months, lactic acid bacterium counts are approximately 10(10) c.f.u. g(-1) in these treated silages. An important subpopulation (5.9 x 10(7) c.f.u. g(-1)) is able to degrade 1,2-propanediol, a fermentation product of L. buchneri, under anoxic conditions to 1-propanol and propionic acid. From this group of 1,2-propanediol-fermenting, facultatively anaerobic, heterofermentative lactobacilli, two rod-shaped isolates were purified and characterized. Comparative 16S rDNA sequence analysis revealed that the newly isolated bacteria have identical 16S rDNA sequences and belong phylogenetically to the L. buchneri group. DNA-DNA hybridizations, whole-cell protein fingerprinting and examination of phenotypic properties indicated that these two isolates represent a novel species, for which the name Lactobacillus diolivorans sp. nov. is proposed. The type strain is LMG 19667T (= DSM 14421T).