L. GRANCHI, M. BOSCO, A. MESSINI and M. VINCENZINI.1999.PCR–RFLP analysis of the rDNA–ITS (internal transcribed spacer) region was applied to 174 yeast strains belonging to 30 species of oenological significance and including 27 type strains in order to define a rapid identification protocol for yeast colonies. DraI‐or HaeIII‐PCR–RFLP patterns were species‐specific with the exception of teleomorphic and anamorphic forms. An improved protocol taking about 30 h was used for the detection and quantification of yeast species occurring in the course of a spontaneous wine fermentation at industrial level. Wine samples were taken and plated daily on an agar medium and the developed colonies were analysed by PCR–RFLP after 24 h of incubation. A representative sample of these colonies was also identified by traditional methods. Both procedures gave identical results. However, PCR–RFLP analysis allowed a more precise enumeration of the yeast populations, proving to be a reliable and simple method for monitoring the development of the yeast community throughout wine fermentation.

BackgroundStoned olive pomace (SOP), which represents approximately 50% of the conversion process of olives to olive oil, is largely not utilised and creates costs for its disposal and has negative environmental impacts. In vitro trial experiments were employed to study the effect of feeds integrated with this bio-waste, which is rich in polyphenols, on rumen biohydrogenation, using sheep rumen liquor as inoculum.ResultsFatty acid (FA) analysis and a polymerase chain reaction denaturing gradient gel electrophoresis (PCR-DGGE) approach aimed at characterising the microbial community indicated that including SOP in feeds at the level of 50 g/kg and 90 g/kg induced changes in the FA profile and microbial populations. The simultaneous decrease of Butyrivibrio proteoclasticus and accumulation of vaccenic acid was observed. A depression in the populations of Neisseria weaveri, Ruminobacter amylophilus and other unclassified bacteria related to members of the Lachnospiraceae and Pasteurellaceae families was detected, suggesting that these microbial groups may be involved in rumen biohydrogenation.ConclusionsSupplementation of feeds with SOP alters the rumen bacterial community, including bacteria responsible for the hydrogenation of vaccenic acid to stearic acid, thereby modifying the FA profile of the rumen liquor. Hence, a use of SOP aimed to produce meat or dairy products enriched in functional lipids can be hypothesised.

Quality changes and shelf-life in European sea bass fillets packed under 40% CO 2 : 60% N 2 (MAP) and air (AIR) or prepared from the whole ungutted fish stored in ice (ROUND) were compared. Raw and cooked sensory scores, pH, colour, expressible water, malonaldehyde, total viable count, Pseudomonas spp., H 2 S-producing bacteria, Lactobacillus spp., Streptococcus spp., Staphylococcus spp., coliforms and proteolytic counts, were determined until AIR and MAP raw fillet spoilage. Raw ROUND fillets had the best quality and shelf-life (10 vs. 8 vs. 7 days after slaughtering in ROUND, MAP and AIR, respectively), while the MAP fillets had better sensorial scores, lower pH values and better microbiological counts, but greater lightness values than the corresponding AIR fillets. MAP fillets also had the highest malonaldehyde levels. The higher correlation between Streptococcus spp. and odour scores (r ¼ )0.971, P < 0.01) compared to the other species could suggest that it is a specific spoilage organism in the MAP condition used.

Strains of Hanseniaspora osmophila and Kloeckera corticis, isolated from wines produced by spontaneous fermentations of normal and dried grapes, were characterized for their fermentation behavior with and without SO(2) at 25 degrees C. All isolates behaved as glucophilic yeasts and yielded ethanol at concentrations of about 9% (v/v); acetic acid, acetaldehyde, ethyl acetate and acetoin were always produced to high concentrations. SO(2) addition had no significant effect on growth yield and fermentation rate. These metabolic features were maintained in the presence of 400 g l(-1) of sugars and at 15 degrees C, and were quite similar to those shown by Saccharomycodes ludwigii. Therefore, H. osmophila and K. corticis should be considered detrimental yeast species, particularly in fermentations of musts from dried grapes.

The addition of polyphenol extracts in ruminant diets is an effective strategy to modulate rumen microflora. The aim of this in vitro trial was to study the effects of chestnut tannin extract (CHT), vescalagin (VES) and gallic acid (GAL) on dietary fibre degradability and on the dimethyl acetals (DMA) profile and microbial community composition of rumen liquor. Four diets (basal diet; basal diet plus CHT; basal diet plus VES; basal diet plus GAL) were fermented for 24 h using ewe rumen liquor. At the end of the fermentation, the microbial communities were characterized by sequencing the 16S rRNA gene. The DMA profile was analyzed by gas chromatography. Chestnut tannin extract did not affect fibre degradability, whereas VES and GAL showed a detrimental effect. The presence of CHT, VES and GAL influenced the concentration of several DMA (i.e., 12:0, 13:0, 14:0, 15:0, 18:0 and 18:1 trans-11), whereas the composition of the microbial community was marginally affected. The inclusion of CHT led to the enrichment of the genera Anaerovibrio, Bibersteinia, Escherichia/Shigella, Pseudobutyrivibrio and Streptococcus. The results of this study support the hypothesis that the activity of CHT is due to the synergistic effect of all components rather than the property of a single component.