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Biomedical subjects

F M MacKenzie

Publications and source records attributed to F M MacKenzie.

3 recordsLinked to original sources

Comparison of methodologies used in assessing the postantibiotic effect.

The postantibiotic effect (PAE) of the carbapenem antibiotic meropenem was determined for the reference strains of Escherichia coli NCTC 4174 and E. coli NCTC 12210. Regrowth of bacteria after antibiotic exposure was determined by viable counting and bioluminescence alone and in combination with an impedance technique and a morphological technique was also employed. Different methods of calculating the PAE were also used. After exposure of E. coli to 0.1-100 x MIC of meropenem for 2 h, concentration dependent differences in counts by bioluminescence, and viable counting were observed, the latter always being lower. The unexposed control of E. coli NCTC 4174 yielded counts of 1.1 x 10(6) +/- 1.1 x 10(5) and 1.3 x 10(6) +/- 4.7 x 10(5) by viable counting and bioluminescence respectively and E. coli NCTC 12210 gave counts of 4.2 x 10(6) +/- 1.8 x 10(6) and 1.1 x 10(7) +/- 4.3 x 10(6) by the same methods. After exposure to 100 x MIC of meropenem, NCTC 4174 yielded counts of 1.28 x 10(3) +/- 5.35 x 10(2) and 2.59 x 10(5) +/- 8.61 x 10(4) and NCTC 12210 gave counts of 5.22 x 10(3) +/- 9.74 x 10(2) and 5.21 x 10(6) +/- 1.45 x 10(6) by viable counting and bioluminescence, respectively. The discrepancies were due to the inability of the viable counting procedure to detect spheroplasts. Falsely low post exposure counts led to falsely low determinations of PAE by viable counting alone and in combination with the impedance technique.(ABSTRACT TRUNCATED AT 250 WORDS)

Anti-Bacterial Agents

Automatic procedures for measuring post-antibiotic effect and determining random errors.

Four methods are described for determining the post-antibiotic effect (PAE) automatically using the high resolution capabilities of the conductance measurement of bacterial growth. The time resolution of each method is derived by a statistical procedure for calculating random error from the component errors involved in each determination. The procedures are illustrated using data generated from the measurements of the growth of Escherichia coli NCTC 4174 after exposure of cultures to five antibiotic concentrations. Excellent agreement was found to exist between three of the four methods. However, that based on the determination of the time of peak growth rate is shown to provide better resolution than the other methods, i.e., those based on growth to 10(7) organisms/mL, the duration of the lag phase or the mean retardation of growth averaged over the entire period of observation. The results strongly suggest that PAE may be ascribed entirely to a prolongation of lag time.

Anti-Bacterial Agents

Postantibiotic effect of meropenem on members of the family Enterobacteriaceae determined by five methods.

The postantibiotic effect (PAE) of meropenem was determined for 11 strains, both clinical isolates and reference strains of members of the family Enterobacteriaceae. The study compares PAE results obtained by five methods used to monitor bacterial regrowth, including viable counting, alone and in combination with impedance; bioluminescence, alone and in combination with impedance; and a morphological technique. After exposure of the test organisms to meropenem (0.1 x to 100 x MIC) for 2 h, concentration-dependent differences in counts by bioluminescence and viable counts were observed, the latter always being lower. The differences varied with the test organism. For example, after exposure of Providentia stuartii NCTC 10318 to 0.1 x MIC, the counts were 5.5 x 10(5) and 2.0 x 10(5) whereas after exposure of Citrobacter freundii MR76 to 0.1 x MIC of meropenem the counts were 2.3 x 10(6) and 6.8 x 10(3) by bioluminescence and viable counting, respectively. The discrepancies were probably due to the relative inability of the viable counting procedure to detect fragile aberrant morphologies and resulted in differences in the calculated PAE values. With methods which do not detect fragile morphologies, the PAE may be underestimated. A general trend was observed for the order of magnitude of the PAEs by the following methods (in order of decreasing magnitude of PAE): (i) morphological technique, (ii) bioluminescence technique alone, (iii) bioluminescence in combination with impedance, (iv) viable counting in combination with impedance, and (v) viable counting alone. It is our opinion that of the methods examined in this study, bioluminescence in combination with impedance best reflects the true values for PAEs, and these results were examined more closely.

Bacteriological Techniques