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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).

A survey was conducted to determine the occurrence of mycotoxins in feedstuffs of dairy cows in the Netherlands and to estimate total dietary intakes of these compounds. Twenty-four dairy farms were visited twice and samples taken of all diet ingredients. Feed intake data were collected by means of questionnaires. A total of 169 feed samples were collected and analyzed for 20 mycotoxins using a liquid chromatography tandem mass spectrometry multimethod. Silage and compound feed were the main diet ingredients, representing on average 67 and 23% of dry matter intake, respectively. Deoxynivalenol (DON), zearalenone, roquefortine C, and mycophenolic acid were the mycotoxins with the highest incidence. The incidence of DON in silage, compound feed, and feed commodity samples was 38 to 54%. The incidence of zearalenone in silage, compound feed, and feed commodity samples was 17 to 38%. The DON and zearalenone had a low incidence in forage samples and were not detected in ensiled by-product samples. Roquefortine C and mycophenolic acid were only detected in silage and ensiled by-product samples (incidence 7 to 19%). Fumonisins B(1) and B(2) were detected in 2 compound feed samples and one feed commodity sample. Aflatoxins B(1), B(2), G(1), and G(2), ochratoxin A, T-2 and HT-2 toxin, 3-acetyl-DON, 15-acetyl-DON, diacetoxyscirpenol, sterigmatocystin, fusarenon-X, ergotamine, and penicillinic acid were not detected in any of the samples. Average concentrations of DON, zearalenone, roquefortine C, and mycophenolic acid in complete diets were 273, 28, 114, and 54 microg/kg, respectively. Maximum concentrations were 969, 203, 2,211, and 1,840 microg/kg, respectively. Calculated average daily intakes of these mycotoxins were 5.0, 0.5, 2.0, and 0.9 mg/animal, respectively, and maximum daily intakes 19.3, 3.5, 38.9, and 32.3 mg/animal, respectively. Corn silage was the major source of all 4 of these mycotoxins in the diet. Extremely high concentrations of roquefortine C and mycophenolic acid (up to 45 and 25 mg/kg, respectively) were detected in visibly molded areas in surface layers of corn silage. These areas appeared to be the main source of roquefortine C and mycophenolic acid in the diet. Because carry-over of DON, zearale-none, roquefortine C, and mycophenolic acid into milk is negligible, their occurrence in feedstuffs is not considered of significant concern with respect to the safety of dairy products for consumers. Potential implications for animal health are discussed.

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Germination and growth of spores of butyric acid bacteria (BAB) may cause severe defects in semihard cheeses. Silage is the main source of BAB spores in cheese milk. The objectives of the study were to determine the significance of grass silages and corn silages as sources of BAB spores and to investigate the relationships between high concentrations of BAB spores in corn silage and aerobic deterioration. In the first survey, samples were taken from various locations in silos containing grass and corn silages and from mixed silages in the ration offered to the cows on 21 farms. We demonstrated that the quantity of BAB spores consumed by cows was determined by a small fraction of silage with a high concentration of spores (above 5 log10 BAB/g). High concentrations were most often found in corn silage within areas with visible molds (69% of the samples). Areas with visible molds in grass silage and surface layers of corn silage contained, respectively, 21 and 19% of the cases of concentrations above 5 log10 BAB spores/g. Based on these results, we concluded that currently in the Netherlands, corn silage is a more important source of BAB than is grass silage. In a second survey, 8 corn silages were divided into 16 sections and each section was studied in detail. High concentrations of BAB spores were found in only the top 50 cm of these 8 silages. Elevated concentrations of BAB spores were associated with different signs of aerobic deterioration. In 13% of the sections in corn silage with more than 5 log10 yeasts and molds/g, more than 5 log10 BAB spores/g were found. Sections with a temperature of more than 5 degrees C above ambient temperature contained, in 21% of the cases, more than 5 log10 BAB spores/g. Concentrations above 5 log10 BAB spores/g were measured in 50% of the sections with a pH above 4.4. All sections with a pH above 4.4 also showed a temperature that was more than 5 degrees C above ambient temperature and a concentration of yeasts and molds above 5 log10 cfu/g. Based on these results, we postulated that high concentrations of BAB spores in corn silage are the result of oxygen penetration into the silage, resulting in aerobic deterioration and the formation of anaerobic niches with an increased pH just below the surface. Growth of BAB in these anaerobic niches with an increased pH caused the locally high concentrations of BAB in corn silage.