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Biomedical subjects

M F Mazens

Publications and source records attributed to M F Mazens.

8 recordsLinked to original sources

Differentiation of Pseudomonas aeruginosa and Enterobacteriaceae in direct smears based on measurements by scanning electron microscopy.

We believe that experienced observers can often distinguish between Enterobacteriaceae and Pseudomonas aeruginosa, by differences in dimensions in Gram-stained direct smears. Many microbiologists question whether this should be attempted because of overlap in dimensions. We have found that culture results confirm our observations about 80% of the time and that such reporting is helpful in diagnosis and treatment. We decided to try to verify the differences in dimensions objectively. Because of the limitations of making measurements by light microscopy, exudate from clinical specimens and from experimental mouse infections were examined by scanning electron microscopy. Discriminant function analysis was applied to the dimensions of length and width, and this showed that 83% of the organisms in both groups could be correctly classified on the basis of the dimensions. This supports our premise that experienced observers should be able to differentiate between these organisms in Gram-stained direct smears, using light microscopy with sufficient confidence to provide clinically useful information.

Animals↗

Antimicrobial synergy testing based on antibiotic levels, minimal bactericidal concentration, and serum bactericidal activity.

The ratio between the concentrations of different antimicrobial agents varies widely during therapy and often bears no resemblance to the ratios assessed by in vitro methods to evaluate the effectiveness of multiple antimicrobial therapy. The authors examined an alternative method using patient serum that reflects the actual antibiotic levels achieved in the patient. Previous investigators have shown that the ratio of the concentration of free drug (drug-f) to the minimal bactericidal concentration (MBC) in broth approximates the serum bactericidal titer (SBT) when pooled normal human serum is used as the diluent. Theoretically, when two antimicrobial agents are administered, the SBT should be equivalent to (drug-f A/MBC drug A) + (drug-f B/MBC drug B). SBT and drug level determinations were performed on peak and trough serum specimens from ten patients with endocarditis or osteomyelitis who were receiving multiple antimicrobial therapy. The serum dilution synergy method predicted additive interactions twice as often as the checkerboard and kill curve, and predicted synergy less frequently than the kill curve. The checkerboard predicted antagonism four times more often than the other methods and provided equivocal results in four of ten cases. The suggested method may offer an alternative procedure to assess antimicrobial interactions, which is based on antibiotic levels actually achieved in vivo instead of the arbitrary concentrations often used in in vitro tests.

Adult↗

Rapid antimicrobial susceptibility test using tetrazolium reduction.

A rapid method of antimicrobial susceptibility testing has been developed, which uses a modified microdilution procedure and an inoculum of 10(7) bacteria per ml. Results are determined within 4 h with an indicator consisting of 2(p-iodophenyl)-3(p-nitrophenyl)-5-phenyltetrazolium chloride. The precipitation of a red formazan by bacteria uninhibited by antimicrobials is accelerated by the addition of phenazine methosulfate. Isolates are classified as resistant, indeterminate, or susceptible, based on growth in up to two antimicrobial concentrations which conform closely to concentrations which correlate with the millimeter breakpoints used in the Bauer-Kirby method. Results of testing 10 antimicrobial agents against 1,126 isolates were compared with results obtained when the Bauer-Kirby method and the agar dilution procedure were used as reference methods. Enterococci were excluded because of false resistance. Discrepancies were classified as very major (false susceptibility), major (false resistance), and minor (combinations of susceptibility or resistance with indeterminate results). The rapid method versus the agar dilution method yielded 2.3% very major, 0.7% major, and 2.9% minor discrepancies, for a total of 6.0%. Of 58 organism-antimicrobial agent combinations tested, 23 displayed 1% very major discrepancies between the rapid method and the agar dilution method. Six were not therapeutically important. The remainder involved Staphylococcus aureus, Staphylococcus epidermidis, Acinetobacter sp., and most organisms tested with chloramphenicol. It is suggested that adjustments in antibiotic concentrations and/or inoculum size may eliminate these discrepancies. The rapid method appeared economical when compared with Autobac 1 and the Bauer-Kirby procedure.

Evaluation Studies as Topic↗

Effect of plate size and location of disc on zone diameter in the disc antimicrobial susceptibility test.

An established standard antimicrobial disc susceptibility test recommends the use of a 150-mm petri dish. Many workers substitute 100-mm plates and use various types of mechanical dispensers. A comparison of zone diameters was made by testing Staphylococcus aureus and Escherichia coli against penicillin and chloramphenicol with the use of these different sized plates and dispensers. Zone diameters were consistently smaller at the edges of plates than in the center (0.4 to 1.2 mm). Significant small differences of up to 1.2 mm or no differences were observed when the mean diameters from these various plates were compared. All of the zone diameter measurements fell within accepted values for analytical variability in this procedure. There is no evidence that any individual combination of plates, dispensers, or disc locations provides greater precision in zone diameter measurement. More difficulty was encountered in measuring zones at the periphery of small plates. This suggests that busy clinical laboratory workers might not produce work of comparable quality with the smaller plates.

Chloramphenicol↗