The bactericidal effect of isoascorbic acid combined with mild heat

The thermal inactivation of Salmonella thompson, Escherichia coli, Staphylococcus aureus, Clostridium perfringens, Candida zeylanoides, Enterococcus faecium and E. faecalis was accelerated by the addition of sodium isoascorbate (1 mmol/l) to phosphate-buffer heating medium but not to complex food mixtures. The lethal effect of isoascorbate was nullified by heating under anaerobic conditions or by the addition of catalase. The scavengers of hydroxyl radicals, mannitol and formate were not protective whereas histidine was. Histidine may have protected by slowing the rate of isoascorbate autoxidation, a property common to other amino acids tested. Superoxide dismutase was not protective. Dehydroascorbic acid also enhanced heat killing and its action was also reversed by catalase. The bactericidal effects of mild heat plus isoascorbate or dehydroascorbic acid both apparently depend on oxidative processes but their relative effectiveness was not related to their respective rates of oxygen consumption or peroxide production. We speculate that site-specific redox reactions, involving amino-carbonyl intermediates are involved in the inactivation mechanism.     

Conductance measurements of the lag phase of injured Salmonella typhimurium

The duration of the lag phase of Salmonella typhimurium injured by heating, freezing, acidification or drying was measured using the 'Malthus' conductance meter. Results confirmed those previously obtained by viable counting and additionally, revealed the very wide variation in lag between and within populations and the extreme length of lag that can occur in some severely injured cells. In an extreme example, the measured lag times of low numbers of bacteria taken from the same heat-injured population ranged from 16 h to 70 h. The implications for the detection of injured micro-organisms in food are discussed.     

A comparison of solid and liquid media for measuring the sensitivity of heat-injured Salmonella typhimurium to selenite and tetrathionate media, and the time needed to recover resistance

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.     

The use of ethidium bromide to assess a novel injury/recovery phenomenon in Listeria monocytogenes in inhibitory NaCl conditions

An injury and recovery phenomenon was observed in Listeria monocytogenes inoculated into a medium containing 2·2 mmol l⁻¹ NaCl, a concentration that was inhibitory to growth. The apparent loss then recovery of viability, as determined by plate counts, was compared with the uptake of ethidium bromide by the cells and found to be inversely related. Injury was caused not only by the initial osmotic up-shock but also by the subsequent down-shock involved in the spread plate protocol.     

How rapidly does the human mitochondrial genome evolve?

The results of an empirical nucleotide-sequencing approach indicate that the evolution of the human mitochondrial noncoding D-loop is both more rapid and more complex than is revealed by standard phylogenetic approaches. The nucleotide sequence of the D-loop region of the mitochondrial genome was determined for 45 members of a large matrilineal Leber hereditary optic neuropathy pedigree. Two germ-line mutations have arisen in members of one branch of the family, thereby leading to triplasmic descendants with three mitochondrial genotypes. Segregation toward the homoplasmic state can occur within a single generation in some of these descendants, a result that suggests rapid fixation of mitochondrial mutations as a result of developmental bottlenecking. However, slow segregation was observed in other offspring, and therefore no single or simple pattern of segregation can be generalized from the available data. Evidence for rare mtDNA recombination within the D-loop was obtained for one family member. In addition to these germ-line mutations, a somatic mutation was found in the D-loop of one family member. When this genealogical approach was applied to the nucleotide sequences of mitochondrial coding regions, the results again indicated a very rapid rate of evolution.     

Effect of centrifugation on the pressure resistance of exponential phase cells of Escherichia coli 8164

M.A. casadei and B.M. mackey. 1997. Exponential phase cells of Escherichia coli NCTC 8164 that were centrifuged at 2000g for 20 min at 4 °C were more resistant to subsequent pressure treatment than cells grown in trypticase soya broth (TSB) and treated without any centrifugation steps. The effects of mild pressure stress (200 kPa for 20 min) and temperature stress (a shift from 37 °C to 4 °C) involved in the centrifugation procedure were analysed separately. It appeared that the increase in pressure resistance obtained following centrifugation was mainly due to the gradual temperature decrease during centrifugation, while the mild pressure stress seemed to play a smaller role in the response.