Disinfection or sterilization treatment by heating, irradiation, or chemicals can cause injury to microorganisms at sublethal levels. Microbial injury is the inability to grow under conditions suitable for the uninjured microorganisms. This inability of injured microorganisms to grow is explained in terms of more complex or different nutritional requirements or in terms of increased sensitivity to environmental conditions such as incubation conditions (time or temperature) or to chemical agents such as halogen compounds. Injured microorganisms can be distinguished from those that are dead or mutated by their ability to regain normal physiological activity when placed in appropriate conditions for cultivation. The return to normal physiological function has been termed repair. The extent and severity of sublethal injury, the mechanisms of injury, and the mechanisms and degree of recovery vary with the sterilization procedures, the species, the strains, the condition of the microorganism, and the methods of repair. Injury to spore formers has been detected at different stages of the spore cycle. The sites of injury include damage to enzymes, membrane disruption, and/or damage to DNA or RNA. Information on the sublethal injury and recovery of microorganisms is very important in evaluating sterilization/disinfection procedures. This paper supplies academic as well as practical information dealing with the repair, and detection of injured microorganisms for performing reproducible sterilization validation.
In this study, in order to construct a model of leftover bath water, we analyzed one hundred samples of used bath water samples which were provided by twenty-eight volunteer families. It appeared that the number of detected bacteria from such bath water was correlated closely with the number of bathers. Moreover, the pH, acidity, chemical oxygen demand (COD), ion, protein content of the leftover bath water were measured. The number of bathers had no connection with the pH, acidity, COD, and ion content of the leftover bath water. However, the protein content of the bath water correlated with the number of detected bacteria. Based on these results, the model of leftover bath water was constructed. Achromobacter xylosoxidans, Staphylococcus aureus, Staphylococcus epidermidis, Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa were incubated with the model bath wateras indices of bath water contamination. The number of incubated viable cells in the model bath water increased with increasing concentrations of casamino acid. Consequently, it was suggested that varying the concentration of casamino acid based on family size or contamination would be necessary in the efficient use of the constructed model of leftover bath water for microbial testing.
Bis-quaternary ammonium compounds (bis-QACs) have the ability to cause a rapid and abundant leakage of the turbid materials from cells, and such a bacterioclastic ability leads to a potent bactericidal activity. In order to clarify the detailed mechanism of the bactericidal action of bis-QACs, the correlation between the bacterioclastic action of 4, 4'-(1, 6-hexamethylenedithio) bis (1-octylpyridinium bromide) (4DTBP-6, 8) and the leakage of outer membrane pore protein E (OmpE) was investigated. Using the antiserum against a fusion protein consisting of GST and the OmpE protein of Escherichia coil encoded by the ompE gene, it was seen that the leakage of OmpE from E. coil cells was caused by treatment with low concentrations (much lower than the critical vesiculation concentration) of 4DTBP-6, 8. Furthermore, it was confirmed that 4DTBP-6, 8 caused an increase in the turbidity of the cell suspension of Klebsiella pneumoniae, Salmonella typhimurium and Serratia marcescences, and led to the leakage of several proteins which have a high percentage of homology with OmpE of E. coll. By immunoelectron microscopy investigation, it was revealed that the vesiculation from E. coil treated with 4DTBP-6, 8 contains OmpE. In addition, the bacteriolytic action of 4DTBP-6, 8 was investigated. The results suggested that the lysis of cells by bis-QACs was not an enzymatic action such as that by autolysin but a physical bacterioclastic action. Judging from these results, it is suggested that the leakage of OmpE is one of the major bacterioclastic actions of bis-QACs, and deals the bacterial cells a fatal blow.
The antifungal activity of scallop-shell powder heated at 1000 °C for 1 h against Trichophyton was kinetically investigated and the possibility of applying the powder to the treatment of dermatophytosis was examined. The death rate of T. mentagrophytes NBRC5466 in the heated shell powder slurry increased with powder concentration, following first-order reaction kinetics. Elevated slurry temperatures increased both the apparent first-order death rate constant (k) and the dilution coefficient (n) representing the dependence of k on reagent concentration. The activation energy for the death of NBRC5466 was almost equal to that for bacteria, whereas the n value was much smaller than that for bacteria. In addition, the trial using heated shell powder treatment on feet showed the possibility of its application to treat dermatophytosis.
For rapid, accurate and sensitive detection of Listeria monocytogenes in food samples, colonies developed on the selective agar (Oxford agar) after immunomagnetic separation (IMS) were subjected to polymerase chain reaction (PCR) assay with the prf A1-2 primer pair. The proposed assay system was shown experimentally to be capable of specifically detecting the bacteria from food samples contaminated at more than 102 cfu/g. However, the enrichment culture after a short period of 16 h with the appropriate selective broth was needed before IMS-plating, because the bacterial contents in most actual food were as low as less than 102 cfu/g. However, even if the enrichment cultivation was employed before IMS, L. monocytogenes was detected within 3 days.
This article describes the circumstances involved in the standardization of the evaluation methods for antimicrobial products. The quantitative method of JIS L 1902 was adopted as the test method for textile products. For plastic, metal and ceramic products, examination of test methods proceeded through the investigation of references and reference to test methods used by various industrial organizations, and a new test method was developed based on the film covering method. An evaluation standard for antimicrobial efficacy was also set up for these test methods and was established as a JIS standard (JIS Z 2801: 2000).