This study investigated two foodborne outbreaks of gastroenteritis that occurred 10 days apart among individuals who had meals at the restaurant of a farm holiday resort. Mild gastrointestinal symptoms were reported and none of the patients needed hospitalization. Mean incubation times were 45 and 33 h, and the overall attack rates were 43.5 and 58.3 %, respectively. Stool sample examination was negative for common enteric pathogens in both outbreaks. Specimens from 13 people involved in the second outbreak and 3 restaurant staff were examined for diarrhoeagenic Escherichia coli. An enteroaggregative E. coli (EAEC) strain of serotype O92 : H33 was isolated from six participants and one member of staff. In particular, the EAEC strain was isolated from five of the six cases of diarrhoea examined. The strain showed an aggregative pattern of adherence to HEp-2 cells, did not produce a biofilm and possessed the virulence-related genes aat, aggR, aap and set1A, but not the astA gene. A retrospective cohort study indicated a pecorino cheese made with unpasteurized sheep milk as the possible source (P,0.001). Samples of the cheese had E. coli counts higher than 10 6 c.f.u. g "1 , but the outbreak EAEC strain was not isolated. This report confirms that EAEC infections are probably underdiagnosed because of the limited availability of laboratories capable of identifying this group of pathogenic E. coli. INTRODUCTIONEnteroaggregative Escherichia coli (EAEC) are diarrhoeagenic E. coli characterized by the ability to adhere to HEp-2 cells in a characteristic 'stacked-brick' pattern (Nataro & Kaper, 1998). This property is usually due to the presence of aggregative adherence fimbriae, whose expression is positively controlled by the aggR gene, located on a large plasmid termed pAA (Nataro, 2005). EAEC infections are usually associated with watery diarrhoea, which is often persistent (Nataro & Kaper, 1998). Illness results from a complex interaction between pathogen and host, which implicates the initial adherence of the bacteria to the epithelium of terminal ileum and colon, in the characteristic aggregative pattern, followed by damage and a subsequent inflammatory response of the intestinal mucosa (Nataro, 2005;Huang et al., 2006).EAEC are a major cause of protracted diarrhoea in children in developing countries (Nataro & Kaper, 1998;Huang et al., 2006), where they are also linked to diarrhoeal illness of travellers and human immunodeficiency virus-positive patients (Huang et al., 2006). However, they have also been found to be associated with diarrhoea in industrialized countries (Tompkins et al., 1999;Weintraub, 2007). EAEC belong to a wide range of serotypes (Wilson et al., 2001) and are considered as emerging pathogens, characterized by a varied and complex epidemiology (Huang et al., 2006). Cases of EAEC infection are reported to be sporadic, but some outbreaks, involving both children and adults, have been described. The sources of infection have rarely been identified in these episodes. In France, an outbreak due to a...

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Aims: Enumeration of resistant bacteria in ultra-high temperature (UHT) treated milk; morphological characterization and phenotyping of resistant strains by traditional and nontraditional methods and their identification by molecular biology. Methods and Results: Modified standard plate count agar (PCA) and modified brain-heart infusion (BHI) agar were used for colony counts. Physiological culture traits were determined as suggested by Bergey's Manual of Systematic Bacteriology or in modified J-broth or in modified BHI agar. Scanning electron microscope (SEM) was used for microscopic examination. Strain identification was carried out by polymerase chain reaction (PCR). A total of 125 (62AE81% of 199) samples were positive and the bacterial load was higher than 10 5 CFU ml )1 in 46 samples (28AE80% of 125). The 16S rRNA sequence of bacterial cultures obtained from UHT-treated milk was similar to that of Bacillus sporothermodurans M215 type strain (T) and different biotypes were found by analysis of colony appearance, cell morphology and physiological traits. Conclusions: Bacillus sporothermodurans was the predominant sporigenous micro-organisms in UHT milk. Significance and Impact of the Study: BHI agar is more suitable than PCA for quality control of milk after UHT treatment. Modified J-broth medium is useful to determine selected physiological traits of B. sporothermodurans. The strains characterized and identified as B. sporothermodurans were significantly different compared with the type strain.

Cities are becoming increasingly complex, both in terms of their social and cultural context, and in the technological solutions that are necessary to make them function. In parallel, we are observing a growing attention toward the public dimension of design, addressing societal challenges and opportunities at an urban scale. Conceptualizing, ideating, and framing design problems at a larger scale may still prove challenging, even as cities are becoming more and more relevant for all branches of design. In this chapter, we address the use of game mechanics to produce strong concepts for better understanding complex problems in citymaking and communal participation, capitalizing on the necessity to shift the attention from smart cities to smart citizens. Through several examples we will show that games and play have a special quality of social bonding, providing context and motivational aspects that can be used to improve the dynamics and solutions within city-making.

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