Optimization of conditions for profiling bacterial populations in food by culture-independent methods
| Autor: | Ana M. Diez, Kalliopi Rantsiou, Luca Simone Cocolin, Rosalinda Urso, Claudia Dr. Beimfohr, Ingrid Bergmaier, Giuseppe Comi |
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| Jazyk: | angličtina |
| Rok vydání: | 2007 |
| Předmět: |
DNA
Bacterial Streptococcus thermophilus Meat Food industry Colony Count Microbial Food Contamination Biology Polymerase Chain Reaction Microbiology FISH Species Specificity Bacterial ecology Cheese Leuconostoc Food microbiology Food science DGGE In Situ Hybridization Fluorescence Phylogeny Culture-independent methods Bacteria business.industry Lactococcus lactis Gene Amplification food and beverages General Medicine biology.organism_classification Electrophoresis Gel Pulsed-Field Lactobacillus sakei Food Microbiology business Temperature gradient gel electrophoresis Food Science Food contaminant |
| Popis: | In this study we used culture-independent methods to profile bacterial populations in food products. Denaturing gradient gel electrophoresis (DGGE) and fluorescence in situ hybridization (FISH) were employed in order to identify bacterial species without the need of isolation and biochemical identification. The protocols used to extract the DNA, subsequently subjected to PCR amplification for DGGE, as well as the hybridization procedure for FISH, were optimised. Moreover, an extensive study on the primers and probes to be used for the direct detection and identification of microorganisms commonly found in food, was carried out. Meat and cheese samples, fresh or processed, were subjected to DGGE and FISH analysis and the results obtained highlighted how the processing in food industry is decreasing the bacterial biodiversity. Not only processed cheese or meat but also fermented products were dominated by only one or few species. Lactobacillus sakei, Lactobacillus curvatus and Brochothrix thermosphacta were the main species found in meat products, while in cheese(s) Lactococcus lactis, Streptococcus thermophilus and Leuconostoc spp. were repeatedly detected. The results obtained by the two culture-independent methods used always correlated well. |
| Databáze: | OpenAIRE |
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