RESUMEN
AIMS: The aim of the work was to apply PCR-temperature gradient gel electrophoresis (PCR-TGGE) and restriction enzyme analysis (RE) assays to identify commercially available starters of Saccharomyces cerevisiae sensu stricto complex. METHODS AND RESULTS: To characterize an analysed pool of 62 active dry yeasts of different brands used in wine fermentation practices, classical microbiological tests were also performed as well as evaluation of contamination with lactic acid bacteria and non-Saccharomyces yeasts. PCR-TGGE and RE were used in order to provide fast and reliable methods to identify and differentiate enological yeasts. Proposed molecular methods enabled to identify particular strains within 36 h after colony isolation and directly from dry yeast suspension. CONCLUSIONS: The methods are highly recommended to obtain reliable results on yeast strain differentiation in a significantly shorter time if compared to classical fermentation tests. SIGNIFICANCE AND IMPACT OF THE STUDY: The obtaining of yeast strain differentiation in a short time and without plating is a good tool for a rapid discrimination among enological strains used as starters in enological practices.
Asunto(s)
Saccharomyces cerevisiae/aislamiento & purificación , Vino/microbiología , Recuento de Colonia Microbiana/métodos , Medios de Cultivo , ADN de Hongos/análisis , Electroforesis en Gel de Poliacrilamida/métodos , Fermentación , Galactosa/metabolismo , Lactobacillus/aislamiento & purificación , Reacción en Cadena de la Polimerasa/métodos , Mapeo Restrictivo/métodos , Saccharomyces/aislamiento & purificaciónRESUMEN
AIMS: The aim of this work was to develop specific primers which are able to detect Bacillus cereus in a coffee concentrate sample. METHODS AND RESULTS: A pre-PCR step to clean the DNA, used for PCR, was developed to avoid PCR inhibition by Maillard products. The combination of centrifugation and washing the pellet, employing EDTA and water, before DNA extraction improved the detection of low numbers of B. cereus cells (10 cells ml-1). The development of specific primers enabled to detect low numbers of B. cereus without the need of a pre-enrichment step. CONCLUSIONS: The data obtained demonstrated the specificity and the sensitivity of the primers that could be used to check the presence of B. cereus in different food products, avoiding the need for labourious and time-consuming culture-based techniques. SIGNIFICANCE AND IMPACT OF THE STUDY: The method could help food microbiologists to check food samples quickly for the presence of B. cereus.