The effects of cinnamic, propionic, benzoic and sorbic acids on the growth and intracellular pH of Escherichia coli were investigated. The data suggest that the potency of weak acids as food preservatives is related to their capacity to reduce specifically the intracellular pH. The data also suggest that although both the undissociated forms of the acid cause the intracellular pH to fall, growth inhibition is due predominantly to the undissociated acid.

The primary habitat of the intracellular pathogen Listeria monocytogenes is considered to be soil and decaying vegetation. As an opportunistic pathogen it must be able to recognize its entry into host tissue and, in response, co-ordinately induce the expression of virulence factors. No signature molecule, which facilitates this regulation, has been identified for any human pathogen. Our studies have demonstrated for the first time that the expression of major virulence determinants in L. monocytogenes can be repressed by an environmentally ubiquitous molecule. Transcriptional hlyA and plcA fusions to luxAB were used to monitor virulent gene expression in the presence of various disaccharides. These studies revealed that the expression of listeriolysin O and phosphatidylinositol-specific phospholipase C is repressed specifically by the plant-derived disaccharide, cellobiose.

The role of K+ transport in the generation of a pH gradient in Escherichia coli has been investigated. In K+-depleted cells, net K+ uptake dissipated delta psi (membrane potential) and led to an increase in delta pH (pH gradient). The magnitude of the delta pH formed bore a simple relationship to the net K+ uptake and was substantially independent of the respiratory rate. In K+-replete cells, generation of a pH gradient was again K+-dependent, although no net uptake of this cation occurred. The results are discussed in terms of K+ cycling, and it is suggested that delta pH is in part a function of the rate of cycling and independent of the respiratory rate.

Growth of Listeria monocytogenes at pH 5·0 did not increase growth of the organism at pH 7·0 after exposure to low pH (3·0, 3·5), compared with cells initially grown at pH 7·0. However, growth at pH 5·0 significantly increased the survival of cells at low pH as determined by plate counts compared with cells grown at neutral pH. Thus, pH adaptation not only occurs in enteric bacteria but also in this Gram‐positive organism. Alterations in the cytoplasmic membrane could be responsible.

The capacity of E. coli cells to regulate intracellular pH (pHi) during net potassium uptake has been investigated. The data show: (a) that cells sense their intracellular pH; (b) that the pH gradient (delta pH) exerts a feedback regulation on pHi; (c) that a mechanism of regulation of pHi exists which may be independent of Na+ [Zilberstein, Agmon, Schuldiner & Padan (1982) J. Biol. Chem. 257, 3687-3691]; and (d) that cells have a limited capacity to raise their intracellular pH in the absence of net K+ transport.

SummaryListeria monocytogenessurvived and, under most conditions, multiplied when inoculated directly into the cheese milk of laboratory made Camembert cheeses. The rate and extent of growth was reduced at lower storage temperatures. Significantly higher rates of growth occurred at the surface compared with the centre of the cheeses, and these were probably associated with increased pH and proteolysis at the cheese surface due to the mould ripening process. Similar results were obtained with Camembert cheeses surface inoculated after manufacture. There was also temperature-dependent growth of List, monocytogenes on a range of inoculated commercially manufactured soft cheeses. Significant growth occurred in Cambazola, French and English Brie, blue and white Lymeswold, French Camembert and Brie with garlic. Little if any growth occurred in blue and white Stilton, Mycella, Chaume and full fat soft cheese with garlic and herbs at the temperatures examined.

Lectins from Helix pomatia, Canavalia ensiformis, Agaricus bisporus and Triticum vulgaris agglutinated cultures of Staphylococcus aureus, Escherichia coli, Listeria and Salmonella spp. This agglutination was specific as it was inhibited (except with A. bisporus lectin) by the competing sugar substrates. The ability of three of these lectins, immobilized on a variety of supports, to separate these micro-organisms from pure cultures was investigated. Immobilization of the lectins on magnetic microspheres was the most effective method. Immobilized T. vulgaris lectin bound 87-100% of cells from cultures of L. monocytogenes, 80-100% of Staph. aureus, 33-45% of Salmonella spp. and 42-77% of E. coli. The A. bisporus lectin bound 31-63% of cells in cultures of L. monocytogenes, 83% of Staph. aureus but only 3-5% of the salmonella cells. Similarly H. pomatia lectin bound greater than 92% of Staph. aureus and 64% of L. monocytogenes cells but was poor at binding the Gram-negative organisms. This preference for binding Gram-positive organisms was confirmed when mixed cultures were studied. The T. vulgaris lectin was effective in removing L. monocytogenes (43%) and Staph. aureus (26%) from diluted milk and Salmonella (31-54%) from raw egg. Agaricus bisporus lectin removed L. monocytogenes from undiluted milk (10-47%) or ground beef (32-50%).

Two methods for the successful extraction of DNA from foods are described. The rapid lysis method uses a proteinase K buffer system to lyse cells and solubilize food samples. DNA is then precipitated using isopropanol. The second method achieves cell lysis using toluene and mutanolysin, and solubilization using guanidium thiocyanate. Following protein removal with organic solvents DNA is precipitated with isopropanol. Both methods enabled the polymerase chain reaction to be applied directly to DNA extracted from samples of cheese, coleslaw and raw chicken and allowed the direct rapid, sensitive and specific detection of Yersinia enterocolitica, Aerococcus viridans and Listeria monocytogenes in these foods.