Raymond C. Bartlett

Clinical, epidemiologic and bacteriologic observations of an outbreak of methicillin-resistant Staphylococcus aureus at a large community hospital.

Over a 12 month period, 61 isolates of methicillin-resistant Staphylococcus aureus (MR-SA) were obtained in 23 hospitalized patients. Eight-six per cent of the patients were over 50 years of age, and 91 per cent were in the postoperative period. In 10 patients (42 per cent), MR-SA was the major pathogen, producing either pneumonia, empyema, osteomyelitis, lung abscess, enterocolitis, wound infection or bacteremia with sepsis. Three patients in this group died despite therapy with antibiotics with in vitro activity against these organisms. All the patients probably acquired their MR-SA in the hospital, and five carriers of the organism were identified among hospital personnel. This outbreak demonstrates the ability of MR-SA not only to colonize many patients in a relatively brief period of time, but also to produce serious disease.

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 107 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.

Activity of Minocycline Against Acinetobacter calcoaceticus Var. anitratus (Syn. Herellea vaginicola) and Serratia Marcescens

The activity of minocycline and tetracycline against 23 isolates of Acinetobacter calcoaceticus var. anitratus (syn. Herellea vaginicola) and 178 strains of Serratia marcescens was determined by disk and microdilution methods. The results indicate that minocycline is highly active against this species of Acinetobacter, all but one strain being inhibited by 0.007 μg of the antibiotic per ml. Tetracycline was also active, though to a lesser degree, against A. calcoaceticus. Of the 178 strains of S. marcescens tested, only seven (3.9%) had a minimum inhibitory concentration of 2 μg or less of minocycline per ml. Tetracycline was less active than minocycline against S. marcescens; with 2 μg of tetracycline per ml, only 2 of 152 (1.3%) strains were inhibited. At concentrations of 8 and 16 μg of minocycline per ml, which can be achieved in the urine with the usual doses, 44.9 and 63.5% of S. marcescens strains were inhibited, which implies its possible usefulness for the therapy of urinary tract infection due to this organism.

Comparison of Microdilution and Broth Dilution Techniques for the Susceptibility Testing of Yeasts to 5-Fluorocytosine and Amphotericin B

Microdilution and broth dilution techniques were compared for the susceptibility testing of 50 clinical yeast isolates to 5-fluorocytosine and amphotericin B. Good correlation between methods was obtained with all isolates except Cryptococcus neoformans.

The comparative activity of fleroxacin, three other quinolones and eight unrelated antimicrobial agents.

The in vitro activity of fleroxacin, a new trifluorinated quinolone was evaluated against 432 bacterial isolates. Fleroxacin was 1- to 2-fold less active than ciprofloxacin and at least as active as ofloxacin and lomefloxacin against most members of the family Enterobacteriaceae. The MICs of fleroxacin for 90% of strains tested (MIC90) were < or = 0.25 micrograms/ml against all isolates of Enterobacteriaceae except Citrobacter freundii (MIC90, 4 micrograms/ml) and Serratia marcescens (MIC90, 2 micrograms/ml). Fleroxacin was as active as ciprofloxacin, ofloxacin and lomefloxacin against Pseudomonas spp, (MIC90 for all quinolones tested were > 8 micrograms/ml). Acinetobacter and Haemophilus influenzae were very susceptible to fleroxacin; however fleroxacin was 1-fold less active than lomefloxacin against Acinetobacter and at least 1-fold less active than ciprofloxacin or ofloxacin against H. influenzae. Methicillin-susceptible and -resistant strains of Staphylococcus epidermidis and methicillin-susceptible strains of S. aureus were very susceptible to fleroxacin, with an MIC90 < or = 1 microgram/ml (range 0.5-1 microgram/ml). Methicillin-resistant S. aureus and Staphylococcus spp. other than aureus and epidermidis were not susceptible to fleroxacin (MIC90 > 8 micrograms/ml). In addition, fleroxacin as well as ciprofloxacin, ofloxacin and lomefloxacin were inactive against Enterococcus spp. (MIC90 > 8 micrograms/ml). Streptococcus pneumoniae and S. pyogenes were resistant to both fleroxacin and lomefloxacin but were very susceptible to ciprofloxacin and ofloxacin. These results suggest that fleroxacin represents a valid therapeutic option in the treatment of infections caused by most Enterobacteriaceae and some species of staphylococcus.

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.

When We should be Testing, How often and what to Report

This chapter will offer practical suggestions for clinical microbiologists to help establish cost-effective policies for antimicrobial susceptibility testing of bacteria and efficient reporting of information to guide antimicrobial therapy. The cost of antimicrobic therapy greatly exceeds the cost of the bacteriology laboratory. Therefore, the combined efforts of the microbiologist, the pharmacist and the medical staff should be focused on producing information that reduces antimicrobic cost.1 Procedures will be recommended here to minimize the production of clinically unnecessary information and the reporting of excessive data that is potentially confusing to clinicians. The practices described are based on over 30 years of cooperative effort between the Microbiology Laboratory, Pharmacy Services and the Division of Infectious Diseases, Department of Medicine at Hartford Hospital. These efforts have reduced the percentage of antimicrobic cost in our pharmacy budget to about 15%. Nationally it is more typical for 30–50% of pharmacy budgets to be consumed by antimicrobic use.2,3 Further, about 20,000,000 bacterial isolates are subject to antimicrobial susceptibility testing in the nation’s laboratories annually.4 Compared to one other major center of comparable size and volume of testing we perform one fifth the number of antimicrobial susceptibility tests (Table 1). This results largely from controls that we have imposed on the quality of specimens and the extent of testing that is performed.5 Studies have shown that the cost for microbiologic processing in our laboratory is lower than that for conventional processing without controls.6