Science Progress

Weak organic acids have been used for centuries to preserve foods, but only recently has the possible mechanism for bacterial growth inhibition been investigated. Although the lowering of internal pH was favored as the cause of growth inhibition, the emphasis has shifted to the anion and its specificity. There are a number of applications of weak organic acids to foods and in the food industry be they pre-or postharvest, However, there is concern that the ability of foodborne pathogens to adapt to these acids may allow longer survival in these commodities and also to better survive transit through the gastric acid barrier of the stomach. Genomic and proteomic approaches have been applied to the identification of genes and proteins that may allow prokaryotes to cope with organic acid stress. These technologies in combination with genetic approaches may provide better identification of genes essential for survival to organic acids. These acids may have other roles: they can induce phenotypic antibiotic resistance, and the high concentrations of these acids in the colon may signal a relationship to diet, colonic microflora, and human health.

Studies of bacterial adaptation to alkaline pH have been less extensive to date compared with those of acidic pH. Recent development of novel methods for global analysis of gene expression under various conditions revealed that many genes were induced at high pH. These data led us to question why so many genes are required for adaptation to alkaline pH. The internal pH of bacteria growing at extremely high pH remains unclear because the methods for measuring interior acidic ΔpH developed to date are not so accurate, but it is generally accepted that cytoplasmic pH increases with medium alkalization, although the increase is lower than that of the change in medium pH. Therefore, activities of enzymes working in neutral cytoplasm may decrease with cytoplasmic alkalization under extreme alkaline conditions. Based on these findings, we propose in this article that genes whose products have an optimum activity at high pH are induced under alkaline stress to compensate for the decrease in activities of systems functioning at neutral pH.

In the environment, bacteria and other microorganisms are subjected to a variety of constantly changing chemical and physical agencies. Chemical ones include antimicrobial compounds (both biocides and antibiotics), pollutants, drugs, cosmetic and pharmaceutical ingredients and pesticides. The physical agents include desiccation and drying, osmotic pressure, hydrostatic pressure, temperature and pH changes and radiations (ultraviolet, sunlight, ionizing). Bacteria must thus adapt to survive these inimicable conditions. Organisms such as bacterial spores usually survive, whereas other types of microorganisms may be much more susceptible.

Depending on the type of organism, the bacterial cell wall, outer membrane or the spore outer layers may act as permeability barriers to the intracellular uptake of antibiotics and biocides. Some antibacterial agents interact with, and damage or modify, the outer components. Physical agencies are known to damage the cytoplasmic membrane or to produce alterations in DNA or proteins or enzymes. Nevertheless, significant damage to the cell wall or outer membrane may also occur.

Four types of organisms are considered: cocci, mycobactria, Gram-negative bacteria and bacterial spores. The nature of the damage inflicted on, or in some cases prevented by, their outer cell layers is discussed for each type of organism.

The ability of enterobacteria to become UV-tolerant is important because such tolerance may enable organisms to resist irradiation in the environment, in water treatment, in shell-fish, in stages of food processing, and at locations in the domestic, commercial and hospital environment The mechanism for regulation of tolerance induction and SOS response induction has been studied for many years, and is well understood, except for the early stages of induction. Such early stages, namely sensing of the stimulus (UV irradiation) and the way in which such sensing leads to signal production, have until now been poorly understood. The claim has been made that DNA is the sensor and that either damage to DNA or production of SS regions in DNA (following interaction of UV with DNA) triggers the signal that sets in train RecA activation and other stages of tolerance induction. This claimed induction mechanism is a “classical” one in the sense that it involves intracellular sensing (by DNA) of the stressing stimulus (UV), and production of an intracellular signalling molecule. It is not, however, firmly established as the mechanism for initiation of UV tolerance induction and SOS response induction. The results reviewed here give firm evidence for a different and unique mechanism for sensing of UV and production of the signal. These results establish without doubt that, for UV tolerance induction, the UV sensor is an extracellular protein, which is a UV tolerance-specific extracellular sensing component (ESC). This component is formed by unstressed cells and on interacting with the stimulus (UV) in the medium, is converted to the tolerance induction signalling molecule, which is a UV tolerance-specific extracellular induction component (EIC). It is this extracellular signal which interacts with the sensitive organisms and triggers tolerance induction. This pair of extracellular components (ECs) may offer the only means of switching-on such tolerance induction; certainly they offer the only known way for early warning to be given of impending UV challenge. Thus, the EIC can diffuse from a region of UV stress to a stress-free region and there warn organisms of impending stress and prepare them to resist it. As indicated here, UV irradiation not only induces UV tolerance, but also switches-on acid tolerance, alkali tolerance and thermotolerance responses. The fact that all three responses involve ESC/EIC pairs strongly supports the view that functioning of such EC pairs form the major, if not the only, means for UV tolerance induction. The UV tolerance-specific ESC can detect other stresses and becomes activated, leading to cross-tolerance responses. Of particular interest, this ESC acts as a biological thermometer, detecting increases in temperature, such increases leading to gradually increasing formation of the EIC and, accordingly, gradual increases in UV tolerance. This UV tolerance-specific ESC can also detect other stresses e.g acting as a pH sensor. In all cases, on activation, the EIC formed (from this specific ESC) only induces UV tolerance. It is proposed that the interaction of EICs with stress-sensitive organisms should be examined, and it is suggested that such EICs may, directly or indirectly, interact with and activate the same stress response regulators as are used to detect internal stressors and which, on activation, also trigger the switching-on of stress responses. For example, EICs either a in a protonated or oxidised state (formed by activation of ESCs by H⁺ or H₂O₂) or b produced by irradiation, may lead to protonation or oxidation or other forms of activation of the appropriate regulator (e.g. Fur or OxyR or RecA etc), leading to response induction.