Endoscopic stenting is a palliative approach for the treatment of diseases involving biliary obstruction. Its major limitation is represented by stent occlusion, followed by life-threatening cholangitis, often requiring stent removal and replacement. Although it has been suggested that microbial colonization of biliary stents could play a role in the clogging process, the so far available data, particularly on the role of anaerobic bacteria, are not enough for a comprehensive description of this phenomenon. Our study was focused on the analysis of 28 explanted biliary stents by culturing, denaturing gradient gel electrophoresis and scanning electron microscopy to identify all the aerobic/anaerobic bacteria and fungi involved in the colonization of devices and to verify the ability of isolated anaerobic bacterial strains to form a biofilm in order to better understand the mechanisms of stent clogging.
Aims: To investigate the bacterial dynamics of a Caciotta cheese traditionally manufactured in the Montefeltro area (Central Italy) with raw cow’s milk and an aqueous extract of dried flowers from Cynara cardunculus as a coagulating agent.
Methods and Results: Conventional methods and a combined PCR‐DGGE approach, relying on culture‐dependent and ‐independent analyses, were used to investigate the cheese bacterial community, with a special focus on lactic acid bacteria. A heterogeneous population, including enterococci, lactococci, lactobacilli, food spoilage and other banal micro‐organisms, was found.
Significance and Impact of the Study: The study contributed to highlighting the influence of different technological parameters on bacterial dynamics of a raw milk Caciotta cheese coagulated with vegetable rennet.
Conclusions: None of the species found in the vegetable rennet became dominant during the cheese‐making and a prevailing role of the adventitions microbita coming from the raw milk and the dairy environment was highlighted.
An investigation aimed at assessing the microbiological quality of meals consumed at a university canteen after implementation of the HACCP system and personnel training was carried out. Cooked and warm-served products (74 samples), cooked and cold-served products (92 samples) and cold gastronomy products (63 samples) sampled from 2000 to 2007 underwent microbiological analyses. All the samples were tested for: Samonella spp., Listeria monocytogenes, total mesophilic aerobes, coliforms, Escherichia coli, Staphylococcus aureus, Bacillus cereus, and sulphite-reducing clostridia. The microbiological contamination of work surfaces (tables, tablewares, cutters, ladles, slicing machines, wash-basins, etc.), hands and white coats of members of the canteen staff was also assessed. The microbiological results clearly demonstrated the success of the HACCP plan implementation, through a general improvement of the hygiene conditions of both meals and work surfaces.
Polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) represents a still valid molecular tool for the profiling of complex microbial ecosystems, including cheeses. In the present study, a double PCR-DGGE approach has been applied to the investigation of the bacterial diversity of seven cheese models to objectively assess strengths and weaknesses of such an approach. To that end, the bacterial DNA was extracted directly from both the cheese replicates and the bulks of colonies harvested from the serial dilution agar plates of selective solid media used for the enumeration of presumptive lactobacilli, lactococci and thermophilic cocci, respectively. The results overall collected allowed the main bacterial taxa to be identified and roughly quantified. Rough quantification of the main cultivable species represents a strength of the PCR-DGGE approach applied, whereas its main weaknesses were represented by the low degree of selectivity of the conventional growth media used for cultivation of lactic acid bacteria and the underestimation of the effective microbial diversity occurring in the seven cheese models.
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