Outside of the laboratory, bacterial cells are constantly exposed to stressful conditions, and an ability to resist those stresses is essential to their survival. However, the degree of stress required to bring about cell death varies with growth phase, amongst other parameters. Exponential phase cells are significantly more sensitive to stress than stationary phase ones, and a novel hypothesis has recently been advanced to explain this difference in sensitivity, the suicide response. Essentially, the suicide response predicts that rapidly growing and respiring bacterial cells will suffer growth arrest when subjected to relatively mild stresses, but their metabolism will continue: a burst of free-radical production results from this uncoupling of growth from metabolism, and it is this free-radical burst that is lethal to the cells, rather than the stress per se. The suicide response hypothesis unifies a variety of previously unrelated empirical observations, for instance induction of superoxide dismutase by heat shock, alkyl-hydroperoxide reductase by osmotic shock and catalase by ethanol shock. The suicide response also has major implications for current [food] processing methods.

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Listeria monocytogenes is a food-borne pathogen capable of adhering to a range of surfaces utilized within the food industry, including stainless steel. The factors required for the attachment of this ubiquitous organism to abiotic surfaces are still relatively unknown. In silico analysis of the L. monocytogenes EGD genome identified a putative cell wall-anchored protein (Lmo0435 [BapL]), which had similarity to proteins involved in biofilm formation by staphylococci. An insertion mutation was constructed in L. monocytogenes to determine the influence of this protein on attachment to abiotic surfaces. The results show that the protein may contribute to the surface adherence of strains that possess BapL, but it is not an essential requirement for all L. monocytogenes strains. Several BapL-negative field isolates demonstrated an ability to adhere to abiotic surfaces equivalent to that of BapL-positive strains. BapL is not required for the virulence of L. monocytogenes in mice.Listeria monocytogenes is a food-borne pathogen that causes serious illness, including meningitis, septicemia, and stillbirth, with a mortality rate of up to 30% (37). More recently, there have been reports of listerial gastroenteritis following the consumption of several different food types (16,34). A number of studies have demonstrated that this organism is able to persist in the food-processing environment for several months and even up to 10 years (23, 29). One of the major causes for concern about L. monocytogenes in these environments is its ability to attach to many different surfaces (2). Indeed, there is recent evidence to show that listerial biofilms formed inside the lumens of stainless steel tubes are able to withstand the shears generated by high-Reynolds-number flows (31). Biofilms, including those produced by L. monocytogenes, are more resistant to detergents and disinfectants (33) and also are a potential source of contamination within food-processing plants; hence, they pose a risk to the maintenance of product safety (27). Consequently, there is considerable interest in determining the mechanisms of attachment and biofilm formation.Our in silico analysis of the genome sequence of L. monocytogenes identified an open reading frame (lmo0435) for a protein with similarity to biofilm-associated proteins (Bap) believed to be important for the binding of staphylococci to abiotic surfaces (10). This Bap protein also has been implicated in the virulence of Staphylococcus aureus (10, 11). Thus, the aim of the current study was to establish if this protein (Lmo0435 [BapL]) of L. monocytogenes influenced biofilm formation and virulence and to determine the prevalence of the lmo0435 (bapL) gene within a selection of field isolates. MATERIALS AND METHODS Bacterial strains and plasmids.A list of the L. monocytogenes isolates and plasmids used in this study is given in Table 1. The strains were cultured in tryptone soya broth (TSB; Oxoid) or brain heart infusion agar (Oxoid) with shaking at 37°C unless otherwise stated. Escherichia coli JM...

The presence of a viable competitive microflora at cell densities of 108 CFU ml−1 protects an underlying population of 105 CFU of Salmonella typhimuriumml−1 against freeze injury. The mechanism of enhanced resistance was initially postulated to be via an RpoS-mediated adaptive response. By using an spvRA::luxCDABEreporter we have shown that although the onset of RpoS-mediated gene expression was brought forward by the addition of a competitive microflora, the time taken for induction was measured in hours. Since the protective effect of a competitive microflora is essentially instantaneous, the stationary-phase adaptive response is excluded as the physiological mechanism. The only instantaneous effect of the competitive microflora was a reduction in the percent saturation of oxygen from 100% to less than 10%. For both mild heat treatment (55°C) and freeze injury this change in oxygen tension affordsSalmonella a substantive (2 orders of magnitude) enhancement in survival. By reducing the levels of dissolved oxygen through active respiration, a competitive microflora reduces oxidative damage to exponential-phase cells irrespective of the inimical treatment. These results have led us to propose a suicide hypothesis for the destruction of rapidly growing cells by inimical processes. In essence, the suicide hypothesis proposes that a mild inimical process leads to the growth arrest of exponential-phase cells and to the decoupling of anabolic and catabolic metabolism. The result of this is a free radical burst which is lethal to unadapted cells.

Aims:The cross-contamination events within a commercial pork processing line were examined by a combination of ERIC-PCR DNA fingerprinting of Escherichia coli and plate counts. Methods and Results: Sponge sampling of environmental surfaces and carcasses was performed over an 8-h processing period. Prior to the start of processing the scraper and dry polisher blades were found to harbour substantial Enterobacteriaceae and Escherichia coli populations. From plate count data the key cross-contamination site for the transfer of bacteria between carcasses occurred during evisceration. However, DNA fingerprints of representative E. coli isolates identified that genotypes initially present on the scraper/dry polisher became distributed on wet polisher blades, band-saw and butcher's hands despite a singeing step being performed post dry polishing. A high proportion of E. coli on post-eviscerated carcasses could be traced to down-stream (pre-singe) environmental contact surfaces. Conclusions: DNA fingerprinting has demonstrated that E. coli and potential enteric pathogens can be transferred between pork carcasses throughout the processing line. In this respect scalding and singeing cannot be relied upon to control cross-contamination of enteric bacteria between carcasses. Significance and Impact of the Study: Sole reliance on indicator organism counts to identify cross-contamination events as currently advocated is limited.