Comparison of dichloran 18% glycerol (DG18) agar with general purpose mycological media for enumerating food spoilage yeasts.

Dichloran 18% glycerol (DG18) agar was originally developed to enumerate xerophilic foodborne moulds. However, some laboratories are using DG18 agar as a general medium to enumerate foodborne moulds and yeasts. A collaborative study, with the participation of seven laboratories, was undertaken to compare DG18 agar with dichloran rose bengal chloramphenicol (DRBC) agar, tryptone glucose yeast extract chloramphenicol (TGYC) agar, and plate count agar supplemented with chloramphenicol (PCAC) for enumerating 14 species of common food spoilage yeasts. Comparison of the mean values of populations of all yeasts recovered on each medium revealed no significant differences among DRBC agar, PCAC, and TGYC agar, while each of these media supported the development of significantly (P < or = 0.05) higher numbers of colonies than DG18 agar. However, differences were only 0.08 to 0.10 log10 cfu/ml, making the practical significance questionable. The overall coefficient of variation (CV) for within laboratory repeatability was 1.71%, while the CV for reproducibility of counts obtained among laboratories was 6.96%. Compared to DRBC agar, TGYC agar, and PCAC, yeast colonies were smaller on DG18 agar. Growth of Brettanomyces anomalus, Cryptococcus albidus, and Rhodotorula mucilaginosa was particularly retarded or inhibited on DG18 agar. Based on the performance of media in supporting colony development and ease of counting colonies, the use of DG18 agar as a general enumeration medium for foodborne yeasts cannot be recommended.

Performance of mycological media in enumerating desiccated food spoilage yeasts: an interlaboratory study.

Dichloran 18% glycerol agar (DG18) was originally formulated to enumerate nonfastidious xerophilic moulds in foods containing rapidly growing Eurotium species. Some laboratories are now using DG18 as a general purpose medium for enumerating yeasts and moulds, although its performance in recovering yeasts from dry foods has not been evaluated. An interlaboratory study compared DG18 with dichloran rose bengal chloramphenicol agar (DRBC), plate count agar supplemented with chloramphenicol (PCAC), tryptone glucose yeast extract chloramphenicol agar (TGYC), acidified potato dextrose agar (APDA), and orange serum agar (OSA) for their suitability to enumerate 14 species of lyophilized yeasts. The coefficient of variation for among-laboratories repeatability within yeast was 1.39% and reproducibility of counts among laboratories was 7.1%. The order of performance of media for recovering yeasts was TGYC > PCAC = OSA > APDA > DRBC > DG 18. A second study was done to determine the combined effects of storage time and temperature on viability of yeasts and suitability of media for recovery. Higher viability was retained at -18 degrees C than at 5 degrees C or 25 degrees C for up to 42 weeks, although the difference in mean counts of yeasts stored at -18 degrees C and 25 degrees C was only 0.78 log10 cfu/ml of rehydrated suspension. TGYC was equal to PCAC and superior to the other four media in recovering yeasts stored at -18 degrees C, 5 degrees C, or 25 degrees C for up to 42 weeks. Results from both the interlaboratory study and the storage study support the use of TGYC for enumerating desiccated yeasts. DG18 is not recommended as a general purpose medium for recovering yeasts from a desiccated condition.

Pichia sporocuriosa sp. nov., a new yeast isolated from rambutan.

A strain of a hitherto undescribed yeast species with a unique ascospore morphology was isolated from rambutan (Nephelium lappaceum). A description of the new species, Pichia sporocuriosa, is given.

Delimination of brewing yeast strains using different molecular techniques.

In general, the genetic characteristics, the phenotype and the microbial purity of the production brewing yeast strains are among the most important factors in maintaining a consistently good quality of products. Analysis of restriction fragment length polymorphism (RFLP) patterns of 18S rRNA-coding DNA was investigated to group ale and lager strains. All production brewing yeast strains showed the same RFLP pattern as the type strain and synonym type strains of S. cerevisiae, and were quite different from the type and synonym type strains of S. pastorianus. Based on these data, all production brewing yeast strains investigated in this study appeared to belong to S. cerevisiae. Electrophoretic karyotyping and random amplified polymorphic DNA (RAPD) analysis appeared to be suitable methods for distinguishing not only the type and synonym type strain of S. cerevisiae and S. pastorianus, but also the ale and the lager strains.

Restriction enzyme analysis of PCR amplified rDNA as a taxonomic tool in yeast identification.

A method has been developed to simplify the identification of yeast strains. We used the restriction fragment patterns of PCR-amplified 18S rRNA-coding DNA with the neighbouring ITS1 region for differentiation and identification of 169 yeast strains representing 128 species associated mainly with food, wine, beer, and soft drinks. The amplicons were digested with four different four-base-cutting restriction enzymes. To construct a database of restriction fragment patterns, the gels have been scanned and analyzed using the Molecular Analyst Fingerprint 2.0 software. The use of four enzymes proved to be sufficient for strain identification.

Candida novakii, sp. nov. a new anamorphic yeast species of ascomycetous affinity.

Two strains of an undescribed species of the genus Candida were isolated from decaying wood of Quercus sp. A description of the new species Candida novakii is given.

An opportunity to distinguish species of Saccharomyces sensu stricto by electrophoretic separation of the larger chromosomes.

Intact chromosomal DNA of the type strains of Saccharomyces sensu stricto species and some of their synonyms and 38 Hungarian wine and 14 brewing yeast strains were separated by rotating field gel electrophoresis (RFE). The applied electrophoretic conditions enabled us to distinguish strains of S. bayanus, S. pastorianus and S. cerevisiae from each other, for strains of the three species showed clear differences in their electrophoretic patterns at heavy chromosomes (> 1300 kb).

Some remarks on "a taxonomic key for the genus Saccharomyces" (Vaughan Martini and Martini 1993).

The practicability of Vaughan Martini and Martini's "Taxonomic Key for the Genus Saccharomyces" for the separation of S. bayanus from other Saccharomyces sensu stricto species was studied. It was concluded that the ability to grow in vitamin free medium was not a suitable character for this purpose. A new wild S. bayanus strain, isolated from exudate of Carpinus betulus was also included in this study. This appears to be the third documented strain of that species isolated outside of an artificial fermentation environment.