Search

The effects of pH, temperature and initial yeast:bacteria ratio on Lactobacillus hilgardii and Saccharomyces florentinus cultivated either in pure or mixed culture were studied. Quadratic polynomial as a function of factors was proposed to express the lactic acid production at different sampling times, and the percentage increase in lactic acid production by Lact. hilgardii in mixed culture compared with pure culture. Temperature was the factor which had the main influence on lactic acid production in mixed culture, whereas stimulation of bacteria depended greatly on pH value. In the range 0.1-20%, the initial yeast-bacteria ratio had no effect on these responses, but presence of the yeast was absolutely necessary to obtain high production of lactic acid. Optimum culture conditions were determined to maximize these characteristics.

Growth, sugar and organic acid metabolism of Lactobacillus hilgardii X 1 B and Leuconostoc oenos X 2 L isolated from Argentinian wine were examined in pure and mixed cultures. In mixed culture Leuc. oenos X 2 L did not grow, no viable cells were detected after 24 h, but the consumption of glucose and fructose by Lact. hilgardii was higher. An increase of mannitol and acetic acid production was detected during the early stages of growth. Over 12 h, higher levels of D(-) lactic acid and slightly increased levels of L(+) lactic acid were observed. Citric and malic acid were simultaneously metabolized, but a slight increase in citric acid consumption was observed in mixed culture

Eighty‐two ethanol‐tolerant (≥ 15% v/v) isolates of Lactobacillus were obtained from Douro fortified wines and associated winery equipment. The diversity and specific identity of these isolates was studied with computer‐assisted comparison of the whole‐cell, SDS‐solubilized protein profiles (as generated by SDS‐PAGE); with this method four species were identified. The majority of isolates (89%) were identified as Lact. hilgardii, and three differentiable electrophoretic groups of this species were evident. The identification was confirmed by DNA‐DNA dot‐blot hybridization studies. Also identified were one strain each of Lact. fructivorans, Lact. collinoides and Lact. mali.

Stimulation of the growth of Lactobacillus hilgardii and production of lactic acid in the presence of Saccharomyces florentinus have already been observed in a poor glucose medium. In this study we showed that CO2, pyruvate, propionate, acetate and succinate excreted by the yeast were responsible for this phenomenon, whereas ethanol, fumarate and a cocktail of vitamins had no effect. Fermentation by the yeast did not enrich the total nitrogen or free amino acid content of the medium. The use of proteases confirmed that it was improbable that any nitrogenous compound could be responsible for the bacterial activation. Between values of 0.1 and 5%, the initial yeast: bacteria ratio had no effect on the stimulation of Lact. hilgardii, even though yeast fermentation was drastically affected by a low initial value. The interactions differed greatly according to the sugar source. With monosaccharides, stimulation began after 48 h of culture, and was higher on glucose than on fructose. With sucrose, stimulation appeared at the beginning of fermentation. Different mechanisms seemed to be involved.

Different techniques were tested for studying the synergism between the micro-organisms of sugary kefir grains. Agar cultures in Petri dishes did not give reproducible results. In sequential cultures, i.e. growing one organism, sterile filtering and then inoculating the other, 10 of 18 selected lactic acid bacteria/yeast pairs revealed stimulation of bacterial growth in a poor glucose medium. In mixed culture, Saccharomyces florentinus supported better survival of Lactobacillus hilgardii and a significant increase in lactic acid production; at the same time, the growth and alcoholic fermentation of S. florentinus were drastically reduced. The inter-relationships between these two strains were the same when immobilized in calcium alginate beads, even though total metabolite production was always lower than with free cells. The stimulation of Lact. hilgardii by Candida lambica in sequential culture was not confirmed in mixed culture, where the two organisms grew as in pure culture, and bacterial growth and lactic acid production were inhibited in the immobilized system.

Nineteen atypical heterofermentative strains of Lactobacillus spp. isolated from South African red wines were identified with the API 50 CHL system and by computerized comparison of their total soluble cell protein patterns. Eleven strains were identified as Lactobacillus hilgardii and eight strains as L. brevis. Three strains of L. hilgardii and three strains of L. brevis produced small amounts of H2S in peptone water supplemented with 001%l-cysteine. All strains produced mannitol from fructose. Proteolytic activity was detected in all except two strains of L. hilgardii. Strains capable of digesting gelatine, hydrolized casein, but not vice versa. The excessive production of sulphur-containing compounds, mannitol and acetaldehyde by lactic acid bacteria could have serious quality implications for the wine industry. Three strains of L. hilgardii and all strains of L. brevis decarboxylated l-malic acid to l-lactic acid.

SUMMARY: An improved growth medium for lactobacilli is described. It supports good growth of lactobacilli generally and also is particularly useful for a number of fastidious strains which grow only poorly in other general media. In addition, tomato juice, a highly variable material, is not required. In a slightly modified form, it can also be used as a basal medium for fermentation tests.