The survival of Salmonella typhimurium after a standard heat challenge at 55 degrees C for 25 min increased by several orders of magnitude when cells grown at 37 degrees C were pre-incubated at 42 degrees, 45 degrees or 48 degrees C before heating at the higher temperature. Heat resistance increased rapidly after the temperature shift, reaching near maximum levels within 30 min. Elevated heat resistance persisted for at least 10 h. Pre-incubation of cells at 48 degrees C for 30 min increased their resistance to subsequent heating at 50 degrees, 52 degrees, 55 degrees, 57 degrees or 59 degrees C. Survival curves of resistant cells were curvilinear. Estimated times for a '7D' inactivation increased by 2.6- to 20-fold compared with cells not pre-incubated before heat challenge.
The heat resistance of Salmonella typhimurium, measured as survival following a standard heat challenge at 55°C for 25 min, increased progressively as cells were heated up at linearly rising temperatures. The amount by which heat resistance increased depended on the rate of temperature rise; the slower the temperature rise, the greater the increase in resistance.
The duration of the lag phase of Salmonella typhimurium surviving heat, freezing, drying and gamma‐radiation was used to indicate the time needed to repair sublethal injury. Following equivalent lethal treatments, heat and freeze‐injured cells needed longer to repair than those injured by drying or gamma‐radiation. Measurement of repair on membrane filters showed that in a heat‐injured population having a lag time of 9 h, some individual cells needed up to 14 h to recover maximum tolerance to 3% NaCl.
The resistance of Salmonella thompson to heating at 54° or 60°C in tryptone soya broth, liquid whole egg, 10% or 40% reconstituted dried milk or minced beef was increased if cells were held at 48°C for 30 min before heating at the higher temperatures. Induction of thermotolerance by mild heat shock is thus not confined to cells grown and heated in broth systems. The heat shock phenomenon may therefore have implications for the safety of foods given marginal heat treatment.
The resistance of stationary phase Salmonella typhimurium to heating at 55 degrees C was greater in cells grown in nutritionally rich than in minimal media, but in all media tested resistance was enhanced by exposing cells to a primary heat shock at 48 degrees C. Chloramphenicol reduced the acquisition of thermotolerance in all media but did not completely prevent it in any. The onset of thermotolerance was accompanied by increased synthesis of major heat shock proteins of molecular weight about 83, 72, 64 and 25 kDa. When cells were shifted from 48 degrees C to 37 degrees C, however, thermotolerance was rapidly lost with no corresponding decrease in the levels of these proteins. There is thus no direct relationship between thermotolerance and the cellular content of the major heat shock proteins. One minor protein of molecular weight about 34 kDa disappeared rapidly following a temperature down-shift. Its presence in the cell was thus correlated with the thermotolerant state.
Sensitivity of heat‐injured Salmonella typhimurium to selenite and tetrathionate media was measured by viable counts in liquid and on agar‐solidified versions of these media and on nutrient media. All solid media, including the supposedly non‐inhibitory nutrient agar, were more inhibitory to injured cells than the corresponding liquid media. Catalase or pyruvate increased counts on nutrient agar to the level obtained in nutrient broth. Therefore nutrient agar plus pyruvate was the most suitable reference medium against which to compare recoveries on other media. Although recoveries of injured cells varied widely depending on the composition and physical state of the medium, this had a minor effect on estimates of repair time because resistance to all selective media was regained by the end of the lag phase.
Cold-shocked Salmonella typhimurium displayed minimal medium recovery (MMR), viable counts on M9 minimal agar being much higher than those on tryptone soya yeast extract agar (TSYA). The addition of catalase to TSYA restored counts to the level found on M9 agar. Peroxide concentrations between 12 and 30 mumol/l were measured in TSYB but none was detected in M9 medium. Cold-shocked cells were sensitive to reagent hydrogen peroxide at a concentration similar to that found in TSYB. The minimal medium recovery phenomenon of cold-shocked cells is thus a manifestation of peroxide sensitivity. Changing the composition of growth media affected both cellular catalase activity and the magnitude of the MMR effect but the two properties were not directly related. Factors additional to cellular catalase activity must therefore affect susceptibility to peroxide following cold shock. Mutational loss of catalase, exonuclease III or recA-dependent DNA repair functions all increased the sensitivity of cold-shocked Escherichia coli to the inhibitory effects of peroxide present in rich medium. The peroxide resistant fraction of a cold-shocked population of Salm. typhimurium (i.e. those cells able to grow on TSYA) was more resistant to gamma radiation than the population as a whole. Cold shock thus sensitizes cells to more than one form of oxidative stress. Prior exposure of growing cells to 30 mumol/l hydrogen peroxide abolished their sensitivity to rich medium following cold shock implying that Salm. typhimurium contains an inducible system protecting against oxidative stress.
Possible routes by which bacteria might reach the deep tissues of carcasses were tested by placing genetically 'marked' strains of Escherichia coli, Clostridium pe$ringens or Bacillus thuringiensis on slaughter instruments before use and examining deep tissue samples for their presence post morteem. Bacteria present on the captive bolt pistol were recovered from the spleens of beef cattle and those placed on the pithing rod were found in both spleen and muscle of the flank and neck. Bacteria from the throat cutting ('stick') knife were isolated, on different occasions, from the heart, lung, spleen, liver and kidneys of sheep though rarely from their muscles. Orally administered bacteria were found in the spleen and lung, but not the musculature, of pigs.
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