Development of an Antimicrobial Susceptibility Testing Method Suitable for Performing During Space Flight

Very little is known regarding the affects of the microgravity environment of space flight upon the action of antimicrobial agents on bacterial pathogens. This study was undertaken to develop a simple method for conducting antibacterial susceptibility tests during a Space Shuttle mission. Specially prepared susceptibility test research cards (bioMerieux Vitek, Hazelwood, MO) were designed to include 6-11 serial two-fold dilutions of 14 antimicrobial agents, including penicillins, cephalosporins, a Beta-lactamase inhibitor, vancomycin, erythromycin, tetracycline, gentamicin, ciprofloxacin, and trimethoprim/sulfamethoxazole. Minimal inhibitory concentrations (MICS) of the drugs were determined by visual reading of color endpoints in the Vitek research cards made possible by incorporation of a colorimetric growth indicator (alamarBlue(Trademark), Accumed International, Westlake, OH). This study has demonstrated reproducible susceptibility results when testing isolates of Staphylococcus aurezis, Group A Streptococcus, Enterococcusfaecalis, Escherichia coli (beta-lactamase positive and negative strains), Klebsiella pneumoniae, Enterobacter cloacae, and Pseudomoiias aeruginosa. In some instances, the MICs were comparable to those determined using a standard broth microdilution method, while in some cases the unique test media and format yielded slightly different values, that were themselves reproducible. The proposed in-flight experiment will include inoculation of the Vitek cards on the ground prior to launch of the Space Shuttle, storage of inoculated cards at refrigeration temperature aboard the Space Shuttle until experiment initiation, then incubation of the cards for 18-48 h prior to visual interpretation of MICs by the mission's astronauts. Ground-based studies have shown reproducible MICs following storage of inoculated cards for 7 days at 4-8 C to accommodate the mission's time schedule and the astronauts' activities. For comparison, ground-based control (normal gravity) MIC values will be generated by simultaneous inoculation and incubation of a second set of test cards in a laboratory at the launch site. This procedure can provide a safe and compact experiment that should yield new information on the affects of microgravity on the biological activities of various classes of antibiotics.