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The aim of the present study was to investigate the prevalence of enterovirulent Escherichia coli strains in cheese manufactured in the Italian Alpine region. Cheese samples obtained from raw and pasteurised milk were collected from cheese-making plants. Enumeration of E. coli and the presence of verocytotoxigenic (VTEC), enteropathogenic (EPEC) and enterotoxic (ETEC) E. coli were carried out. A multiplex-PCR for the amplification of the sequences of VT1, VT2, eae, ehxA, ST, LT genes was used to identify pathogenic E. coli strains. The presence of E. coli O157 was investigated using an immunomagnetic separation technique. E. coli O157 was not recovered in cheese made from raw or pasteurised milk. The multiplex PCR approach detected at least one virulence determinant in only ten cheese samples, in four of them, a verotoxin-producing gene (VT2 and/or VT1) was detected and in the remaining six only the attaching and effacing lesion genes (eae) were found. No ETEC strains were detected. The statistical analyses of the results demonstrated significant associations between the use of raw milk in cheese production and the presence of detectable virulence genes in cheese. Moreover, high E. coli counts were also associated with the use of raw milk in fresh and ripened cheese production. The results revealed the survival and recovery of pathogenic strains even after a 60-day ripening period, highlighting the importance of the hygienic and sanitary requirements for milk used in the production of both fresh and ripened cheese. [ABSTRACT FROM AUTHOR]

Background: Sustainability is a broad and debated subject, often difficult to be defined and applied into real projects, especially when dealing with a complex scenario as the one of healthcare. Many research studies and evaluation systems have handled this topic from different perspectives, but many limits and criticalities still have to be overcome to properly cope with actual needs., Methods: The Sustainable Healthcare project has been developed through three main phases: a deep study of the state of the art, unraveling pros and cons of available sustainability scoring systems; an accurate analysis of the stakeholders network and their needs; the realization of an objective evaluation framework, through scientific methods, as the ANP., Results: The newly developed evaluation system takes into consideration all the three pillars of sustainability, analyzing social, environmental and economic sustainability through a set of criteria, specified by measurable indicators. So the system identifies both global sustainability and specific critical areas, pointing out possible strategic solutions to improve sustainability. The evaluation is achieved through technical analyses and qualitative surveys, which eventually allow to quantitatively assess sustainability, through a sound scoring method., Conclusions: This study proposes an innovative evaluation method to determine the sustainability of a hospital, already existing or in the design phase, within the European context. The Sustainable Healthcare system overcomes some of the current evaluation systems' limits by establishing a multidisciplinary approach and being an easy-to-use tool. This protocol is intended to be of support in the identification of the main hospital's weaknesses and in setting priorities for implementation of the solutions.

Aims: To investigate the genetic relatedness between Lactococcus garvieae strains isolated from fish and dairy samples collected in northern Italy, using random-amplified polymorphic DNA (RAPD)-polymerase chain reaction (PCR), Sau-PCR and amplified fragment length polymorphism (AFLP)., Methods and Results: Eighty-one isolates from bovine and caprine dairy products (n = 53) and from diseased rainbow trouts and other fishes (n = 28) were examined. All methods showed a typeability of 100%, repeatability ranging from 84.4% to 97.5% and discriminatory powers from 0.798 to 0.986. Dairy and fish strains revealed a low genetic relatedness as they are often grouped into distinct clusters. RAPD analysis discriminated 52 genotypes when primer M13 was used, whereas with primer P5 only 27 genotypes were identified. When Sau-PCR was performed, 13 genotypes were detected while AFLP analysis allowed the differentiation of 32 genotypes., Conclusion: L. garvieae strains isolated from dairy samples are generally not related to those collected from fish lactococcosis outbreaks., Significance and Impact of the Study: L. garvieae strains exhibit a genetic diversity related to the specific animal host they colonize. RAPD M13 fingerprinting proved to be a molecular tool for comparing isolates, whereas Sau-PCR and AFLP analyses were useful techniques to investigate the distribution of L. garvieae populations in the environment.