A combination of direct viable count and fluorescence in situ hibridization for specific enumeration of viable Lactobacillus delbrueckii subsp. bulgaricus and Streptococcus thermophilus

[EN] Aims: We have developed a direct viable count (DVC)-FISH procedure for quickly and easily discriminating between viable and nonviable cells of Lactobacillus delbrueckii subsp. bulgaricus and Streptococcus thermophilus strains, the traditional yogurt bacteria. Methods and Results: direct viable...

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Detalles Bibliográficos
Autores: García Hernández, Jorge|||0000-0003-1258-6128, Moreno Trigos, Mª Yolanda|||0000-0003-3688-5157, Hernández Pérez, Manuel|||0000-0003-1986-6501, Amorocho Cruz, Claudia Milena
Tipo de recurso: artículo
Fecha de publicación:2011
País:España
Institución:Universitat Politècnica de València (UPV)
Repositorio:RiuNet. Repositorio Institucional de la Universitat Politécnica de Valéncia
Idioma:inglés
OAI Identifier:oai:riunet.upv.es:10251/74689
Acceso en línea:https://riunet.upv.es/handle/10251/74689
Access Level:acceso abierto
Palabra clave:Direct viable count
FISH
Lactobacillus delbrueckii subsp
Bulgaricus
Rapid method
Streptococcus thermophilus
MICROBIOLOGIA
Descripción
Sumario:[EN] Aims: We have developed a direct viable count (DVC)-FISH procedure for quickly and easily discriminating between viable and nonviable cells of Lactobacillus delbrueckii subsp. bulgaricus and Streptococcus thermophilus strains, the traditional yogurt bacteria. Methods and Results: direct viable count method has been modified and adapted for Lact. delbrueckii subsp, bulgaricus and Strep. thermophilus analysis by testing different times of incubation and concentrations of DNA-gyrase inhibitors. DVC procedure has been combined with fluorescent in situ hybridization (FISH) for the specific detection of viable cells of both bacteria with specific rRNA oligonucleotide probes (DVC-FISH). Of the four antibiotics tested (novobiocin, nalidixic acid, pipemidic acid and ciprofloxacin), novobiocin was the most effective for DVC method and the optimum incubation time was 7 h for both bacteria. The number of viable cells was obtained by the enumeration of specific hybridized cells that were elongated at least twice their original length for Lactobacillus and twice their original size for Streptococcus. Conclusions: This technique was successfully applied to detect viable cells in inoculated faeces. Significance and Impact of the Study: Results showed that this DVC-FISH procedure is a quick and culture-independent useful method to specifically detect viable Lact, delbrueckii subsp, bulgaricus and Strep thermophilus in different samples, being applied for the first time to lactic acid bacteria.