The volatile metabolites of three strains of Pseudomonas aeruginosa and one strain each of Pseudomonas cepacia, Pseudomonas maltophilia, Pseudomonas fluorescens, and Pseudomonas putida were analyzed using an automated headspace concentrator incorporating a gas chromatograph. The procedure does not require sample preparation and automates the entire analytical sequence to yield reproducible profiles of volatile constituents. Gas chromatographic profiles of the volatile metabolites of each species were obtained using a 20-min concentration period and two fused silica capillary columns of different polarities. The production of headspace metabolites from trypticase soy broth was studied in relationship to culture incubation time and initial cell concentration. The volatiles identified after 24 h incubation consisted of 1-butanol, isopentanol, toluene, 1-undecene, 2-butanone, 2-heptanone, 2-nonanone, and 2-undecanone. Sufficient amounts of specific metabolites were produced after 5 h incubation to provide information of possible diagnostic value. In particular, all P. aeruginosa strains produced a distinctive series of 1-undecene and methyl ketones after 5 h incubation of media inoculated to provide 2 X 10(6) cells/mL. The results indicate that when growth and analytical conditions are held constant, P. aeruginosa and related species produce characteristic profiles of headspace metabolites. Since conventional bacteriological tests require 24 h or more for the identification of these pseudomonads, automated volatile analysis could provide an alternative means for the rapid detection of these bacteria.
The cutaneous distribution of lipophilic diphtheroids was determined in normal human volunteers. The organisms were found to be plentiful in moist regions (scalp, nares, axilla, groin, and toe web) and scarce in dry and purely oily regions. The lipid requirement, cellular fatty acids, mycolic acid and cell wall diaminopimelic acid content of these lipophilic diphtheroids was compared to those of strains of Corynebacterium bovis, C. xerosis, C. diphtheriae, and C. minutissimum. Only lipophilic diphtheroids and C. bovis strains were found to have a strict lipid requirement. Lipophilic diphtheroids were found to have meso-diaminopimelic acid and corynemycolic acid in their cell walls, consistent with membership in the genus Corynebacterium. Lipophilic diphtheroids were also found to comprise a homogeneous group which was distinct from the speciated strains on the basis of cellular fatty acids and mycolic acids.
When incubated in natural (nonsterilized) soil, Pseudomonas aeruginosa died initially at a rate which approximated the rate for starvation of a pure culture in buffer. Predation by other soil microbes or phage did not appear to be involved, and pyocyanin either was not produced or was ineffective. The initial rate of death was followed by a second, considerably slower rate. Cells initially added in low numbers to soil also underwent biphasic death as above. Slow drying of the soil caused a period of rapid soil death of P. aeruginosa, but this then slowed to give residual numbers and a death rate similar to the second death rate noted for soil not allowed to dry. The cells in the dry soil had not changed genetically to a desiccation-resistant form. Pseudomonas aeruginosa died out completely in a relatively short time when the soil was first quickly dried to a water content similar to that obtained initially through slow drying and then further allowed to dry slowly. These observations appear to point to a dormant form, in some ways resembling a cyst, for P. aeruginosa in soil.
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