Identification and characterization of non-saccharomyces spoilage yeasts isolated from Brazilian wines

The industry of fine wines and also locally consumed table wines is emerging in Brazil with an increasing volume and economic impact. Enologists in this region currently lack information about the prevalence and characteristics of spoilage yeasts, which may contaminate and potentially undervalue Brazilian wines. Herein, we analyzed 50 local red wines including 27 fine wines (V. vinifera) and 23 table wines (V. labrusca). Presumptive spoilage yeasts were isolated on differential medium, and classified by RFLP-PCR and sequencing of the ITS1-5.8S-ITS2 and D1/D2 26S rDNA loci. The prevalence of spoilage yeasts in fine wines (11 %) was comparable to that reported in European and US wines, and significantly lower than that observed for local table wines (70 %). The majority of isolates belonged to Brettanomyces bruxelliensis, followed by Pichia guillermondii, and more rarely Candida wickerhamii and Trigonopsis cantarelli. The Brettanomyces isolates varied greatly in off-flavor production, displayed ethanol tolerance (>10 % by volume), tolerated sulfite (≥0.68 mg/l mSO₂), and 39 % of them grew on ethanol as sole carbon source. We discuss the causes and consequences of spoilage yeasts in relation to the Brazilian wine industry.     

Response of wheat seedlings to short-term drought stress with particular respect to nitrate utilization

Abstract. The effect of short-term changes in the water potential (from 0 to – 2.5 MPa) by addition of PEG 4000 to the nutrient solution was investigated with respect to nitrate uptake and reduction in 3-week-old wheat plants ( Triticum aestivum , cv Fidel). Plants were harvested at the end of 12-h treatments in the dark. The water potential of the mature leaves was similar to that of the medium down to – 0.8 MPa and was maintained at this level even though the external water potential was much lower. The medium water potential of 0.8 was a threshold level below which elongation of the youngest leaf was inhibited. Increase of the PEG concentration in the medium brought about a decrease of evapotranspiration and enhancement of nitrate uptake. No difference in the rate of nitrate reduction was observed, although the in vitro nitrate reductase activity was lowered. Nitrate accumulation in the shoot was ascribed both to the stimulation of net uptake from the medium, and to the mobilization and translocation of nitrate from the root. It is suggested that increase in the storage pool of nitrate in shoots was related to the role of NO₃⁻ as an osmoticum.     

A strategy for construction of industrial strains of distiller's yeast

A procedure was developed for construction of industrial strains of distiller's yeast (Saccharomyces cerevisiae). It includes several steps: construction of congenic genetically marked haploid strains of opposite mating types starting from an industrial strain of hybrid nature, integrative transformation of the above haploid strains with a DNA fragment containing an expression cassette responsible for new technological facilities, and hybridization of transformants and isolation of final industrial homozygous strains under experimental conditions simulating commercial fermentation processes. This strategy permits the generation of strains that have desirable characteristics of traditional races of distiller's yeast along with new technological facilities determined by the particular expression cassette. Using this procedure, we have constructed an industrial strain with improved amylolytic activity. (c) 1995 John Wiley & Sons, Inc.     

Individual anatomic variability of particular thalamic nuclei in children and adolescents with respect to stereotaxic coordinates

As demonstrate macro-microscopic and stereologic investigations performed in 120 preparations of children and adolescent brain, the anterior, medial and lateral nuclei in the thalamus have a complex geometric form which does not change with age. Their linear dimentions and stereotaxic coordinates increase somewhat. These changes are described by means of mathematical models as regression equations. Individual changeability of the nuclear groups, situating within the studied ones are evidently subjugated to the same regularities. The individual variants revealed are not connected with side of the brain, form of the skull and sex. The combination of the methods used gives reliable results.     

Ecovariant differences of Escherichia coli strains with respect to colicinogenicity and colicin resistance

The analysis of 173 Escherichia coli strains, isolated from different sources, for colicinogenicity and colicin resistance revealed that frequency of these signs increased in the following order: water in open reservoirs, intestine, extraintestinal localizations. In most cases resistance to 5 or more bacterial colicins was due to the absence of the corresponding receptors to colicins. Colicin resistance and colicinogenicity render E. coli selective advantages under the conditions of intestinal microbiocenosis.     

Selection and study of potent lactose-fermenting yeasts

Whey-fermenting Kluyveromyces cultures were revealed among 105 yeast strains assimilating lactose. Eighteen most potent strains isolated from milk products fermented galactose, sucrose, and raffinose, in addition to lactose. Many yeast strains fermented inulin. Most strains were resistant to cycloheximide and grew in medium containing glucose, NaCl, and ethanol at concentrations of up to 50, 11-12, and 10-12%, respectively (4 degrees C). Three strains had mycocinogenic activity. After fermentation of whey with selected yeast strains at 30 degrees C for 2-3 days, ethanol concentration was 4-5%.     

Polymorphism analysis of Haemophilus influenzae strains isolated from healthy children with respect to amplification reaction

The random-amplified polymorphic DNA (RAPD) assay was used to generate DNA fingerprints for 44 isolates H. influenzae obtained from healthy children. Problems with reproducibility and discriminatory power, frequently cited in the literature, were overcome by optimization procedure allowing to achieve reliable conditions for H. influenzae analysed. Particular parameters of RAPD fingerprinting were evaluated with respect to selection of best working primer, DNA polymerase and DNA concentration for amplification pattern. This study proved high sensitivity and efficiency of optimized RAPD profiling applicable for searching the epidemiology traces.     

Selection of phage-displayed peptides that bind to a particular ligand-bound antibody

Phage-displayed peptides that selectively bind to aldolase catalytic antibody 93F3 when bound to a particular 1,3-diketone hapten derivative have been developed using designed selection strategies with libraries containing 7-12 randomized amino acid residues. These phage-displayed peptides discriminated the particular 93F3-diketone complex from ligand-free 93F3 and from 93F3 bound to other 1,3-diketone hapten derivatives. By altering the selection procedures, phage-displayed peptides that bind to antibody 93F3 in the absence of 1,3-diketone hapten derivatives have also been developed. With using these phage-displayed peptides, ligand-bound states of the antibody were distinguished from each other. A docking model of one of the peptides bound to the antibody 93F3-diketone complex was created using a sequential divide-and-conquer peptide docking strategy; the model suggests that the peptide interacts with both the antibody and the ligand through a delicate hydrogen bonding network.     

Screening of Saccharomyces cerevisiae strains with respect to anaerobic growth in non-detoxified lignocellulose hydrolysate

A microplate screening method was used to assess anaerobic growth of 12 Saccharomyces cerevisiae strains in barley straw, spruce and wheat straw hydrolysate. The assay demonstrated significant differences in inhibitor tolerance among the strains. In addition, growth inhibition by the three hydrolysates differed so that wheat hydrolysate supported growth up to 70%, while barley hydrolysate only supported growth up to 50%, with dilute-acid spruce hydrolysate taking an intermediate position.     

Use of factor analysis to classify strains of yeasts: Application to genus torulopsis berlese

Summary A factor analytical approach, along with other statistical methods, was used to effect an orderly or scientific classification of 130 strains of yeasts designated as the genus Torulopsis, These strains were studied by means of 64 morphological, physiological and serological characteristics. From the correlations among these variables, factor analysis produced 12-dimensional profiles for the strains, which were then grouped according to these profiles. Initially 37 groups resulted, but these were classified again into 9 second-order groups or clusters. Discriminant analysis showed that the separations of the 9 clusters were statistically significant. Finally, the classifications of the strains were compared with the grouping into 9 phenons of 121 of these strains, reported in a previous paper (Kocková-Kratochvílová, 1973). These results corroborated the previous findings of agreement with published results of GC content in DNA.Laboratory measurements obtained at Institute of Chemistry of the Slovak Academy of Sciences, Bratislava, CSSR. Computer work at Educational Testing Service, Princeton, N. J., U.S.A.     

Selection and fermentation properties of cryophilic wine yeasts

Two cryophilic strains, YM-84 and YM-126, were selected by a double-layer agar fermenting technique from 100 strains of the wine yeast, Saccharomyces cerevisiae. The viability (specific growth rate) and fermentability of the two selected strains at low temperatures (7 and 13°C) were superior to those of wine yeast strains W3 and OC-2, indicating the usefulness of the two strains as cryophilic wine yeasts. Experiments using the two selected strains at intermediate temperatures (22 and 30°C) showed that their fermentation ceased prematurely and their ethanol yields were reduced.     

Antigenic analysis of Trypanosoma cruzi strains by crossed immunoelectrophoresis: demonstration and isolation of antigens particular to some strains

Antigenic differences among soluble extracts of Y, CL, SF, and Colombian strain epimastigotes of Trypanosoma cruzi have been demonstrated by crossed immunoelectrophoresis with an intermediate gel containing the heterologous antiserum. Using crossed immunoelectrophoresis with the homologous antiserum, over 30 precipitin lines could be demonstrated for each strain and, even though marked differences were observed in experimental infections, the strains shared a significant number of antigens. In addition, some strain-particular antigens were isolated using affinity chromatography. These antigens could be valuable in the study of biological, immunological, and pathological characteristics of experimental and natural T. cruzi infections.