Monitoring the succession of the biota grown on a selective medium for pseudomonads during storage of minced beef with molecular-based methods

Abstract

In the present study, the succession of the biota grown on a selective medium for pseudomonads (pseudomonas agar based medium - PAB) during the storage of meat under different conditions was monitored. Thus, minced beef was stored aerobically and under modified atmosphere packaging in the presence (MAP+) and absence (MAP-) of oregano essential oil at 0, 5, 10 and 15 °C. A total of 267 pure cultures were recovered from PAB throughout the storage period and subjected to PCR-Denaturing gradient gel electrophoresis (PCR-DGGE) for their differentiation. In parallel, the direct analysis of the whole cultivable community (WCC) from the same medium was applied. These two approaches were used in order to indicate the lack of selectivity. Fifteen different DGGE fingerprints were obtained after PCR - DGGE analysis of the isolates, which were assigned to Pseudomonas putida (3 fingerprints), Pseudomonas fragi and Pseudomonas fluorescens, Pseudomonas spp., Serratia liquefaciens (2), Citrobacter freundii, Serratia grimesii, Hafnia alvei (3), Rahnella spp. and Morganella morganii. Twelve of them occurred during the direct analysis of the WCC. The biota succession found to be affected from the different storage conditions. However, the outcome of the two strategies was quite different, which is leading to the use of different appropriated molecular approaches in order to widen the knowledge of bacterial succession of meat.