RESUMEN
AIMS: The yeast Saccharomyces boulardii is used as a probiotic for the prevention and treatment of diarrhoea. In this study, the quality of 15 probiotic products containing S. boulardii was verified. METHODS AND RESULTS: Using microsatellite typing, the identity of all Saccharomyces strains in the products was confirmed as S. boulardii. Additionally, solid-phase cytometry (SPC) and a plate method were used to enumerate S. boulardii cells. SPC was not only able to produce results more rapidly than plating (4h compared to 48h) but the cell counts obtained with SPC were significantly higher than the plate counts. Finally, we found that <1% of the S. boulardii cells survived 120min in gastric conditions and storage for 3months at 40°C with 75% relative humidity. CONCLUSIONS: We developed a SPC method for the quantification of viable S. boulardii cells in probiotics. Additionally, we demonstrated that gastric conditions and storage have a marked effect on the viability of the yeast cells. SIGNIFICANCE AND IMPACT OF THE STUDY: To our knowledge, this is the first time SPC is used for the quality control of probiotics with S. boulardii. Additionally, we demonstrated the need for gastric protection and accurate storage.
Asunto(s)
Probióticos/normas , Saccharomyces/fisiología , Recuento de Colonia Microbiana , Citometría de Imagen , Viabilidad Microbiana , Repeticiones de Microsatélite/genética , Control de Calidad , Saccharomyces/genética , Estómago/microbiología , Factores de TiempoRESUMEN
As part of studies on the spread of infections, risk factors and prevention, several typing methods were developed to investigate the epidemiology of Aspergillus fumigatus. In the present study, 52 clinical isolates of A. fumigatus from 12 airway specimens from patients with invasive aspergillosis (hospitalized in three different centres) were characterized by short tandem repeat (STR) typing and multilocus sequence typing (MLST). These isolates were previously typed by random amplified polymorphic DNA (RAPD), sequence-specific DNA polymorphism (SSDP), microsatellite polymorphism (MSP) and multilocus enzyme electrophoresis (MLEE). STR typing identified 30 genotypes and, for most patients, all isolates were grouped in one cluster of the unweighted pair group method with arithmetic mean dendrogram. Using MLST, 16 genotypes were identified among 50 isolates, while two isolates appeared untypeable. RAPD, MSP, SSDP and MLEE allowed identification of eight, 14, nine and eight genotypes, respectively. Combining the results of these methods led to the delineation of 25 genotypes and a similar clustering pattern as with STR typing. In general, STR typing led to similar results to the previous combination of RAPD, SSDP, MSP and MLEE, but had a higher resolution, whereas MLST was less discriminatory and resulted in a totally different clustering pattern. Therefore, this study suggests the use of STR typing for research concerning the local epidemiology of A. fumigatus, which requires a high discriminatory power.
Asunto(s)
Aspergilosis , Aspergillus fumigatus/clasificación , Proteínas Bacterianas/genética , Enfermedades Pulmonares Fúngicas , Repeticiones de Microsatélite/genética , Técnicas de Tipificación Micológica , Análisis de Secuencia de ADN/métodos , Aspergilosis/epidemiología , Aspergilosis/microbiología , Aspergillus fumigatus/genética , Aspergillus fumigatus/aislamiento & purificación , ADN de Hongos/análisis , ADN de Hongos/genética , Genotipo , Humanos , Enfermedades Pulmonares Fúngicas/epidemiología , Enfermedades Pulmonares Fúngicas/microbiología , Reproducibilidad de los Resultados , Especificidad de la EspecieRESUMEN
A series of 256 Aspergillus fumigatus isolates, recovered from eight patients with cystic fibrosis (CF), were genotyped using microsatellite-based typing. Only a limited number of genotypes were shared between patients and co-colonisation with multiple strains was indicated for all patients. Additionally, some genotypes were isolated recurrently, indicating that they are capable of prolonged colonisation.