ALBERT

Description: We have successfully flown the EcAMSat (Escherichia coli Antimicrobial Satellite) free-flyer mission. This was a 6U small satellite that autonomously conducted an experiment in low Earth orbit to explore the impact of the space environment on antibiotic resistance in uropathogenic E. coli (UPEC) and the role a particular sigma factor plays in the response. After being held in stasis during transport to orbit, two strains a wildtype UPEC and an isogenic mutant with a deleted gene that encodes a sigma factor were grown to stationary phase in a fluidic card inside EcAMSat's payload, then incubated with three concentrations of the antibiotic gentamicin. The payload then administered alamarBlue, a redox indicator, into all wells of the fluidic card. The cells were then incubated for 144 hours and metabolic activity was measured optically using the payloads' LED and detector system. Data were then telemetered to the ground and compared to a control experiment conducted in an identical satellite in a lab. The results of this experiment will help us better understand important therapeutic targets for treating bacterial infections on Earth and in space. Such targets are particularly relevant to deep-space and long-duration missions where crew may be more susceptible to infection and treatments for them may work differently.

Description: We have successfully flown the EcAMSat (Escherichia coli Antimicrobial Satellite) free-flyer mission. This was a 6U (six unit - CubeSat) small satellite that autonomously conducted an experiment in low Earth orbit to explore the impact of the space environment on antibiotic resistance in uropathogenic E. coli (UPEC) and the role a particular sigma factor plays in the response. After being held in stasis during transport to orbit, two strains - a wildtype UPEC and an isogenic mutant with a deleted gene that encodes a sigma factor - were grown to stationary phase in a fluidic card inside EcAMSat's payload, then incubated with three concentrations of the antibiotic gentamicin. The payload then administered alamarBlue (registered trademark), a redox indicator, into all wells of the fluidic card. The cells were then incubated for 144 hours and metabolic activity was measured optically using the payloads' LED (Light-Emitting Diode) and detector system. Data were then telemetered to the ground and compared to a control experiment conducted in an identical satellite in a lab. The results of this experiment will help us better understand important therapeutic targets for treating bacterial infections on Earth and in space. Such targets are particularly relevant to deep-space and long-duration missions where crew may be more susceptible to infection and treatments for them may work differently.

Description: Human immune response is compromised and bacteria can become more antibiotic resistant in space microgravity (MG). We report that under low-shear modeled microgravity (LSMMG) stationary-phase uropathogenic Escherichia coli (UPEC) become more resistant to gentamicin (Gm). UPEC causes urinary tract infections (UTIs), reported to afflict astronauts; Gm is a standard treatment, so these findings could impact astronaut health. Because LSMMG has been shown to differ from MG, we report here preparations to examine UPEC's Gm sensitivity during spaceflight using the E. coli Anti-Microbial Satellite (EcAMSat) on a free flying nanosatellite in low Earth orbit. Within EcAMSats payload, a 48-microwell fluidic card contains and supports study of bacterial cultures at constant temperature; optical absorbance changes in cell suspensions are made at three wavelengths for each microwell and a fluid-delivery system provides growth medium and predefined Gm concentrations. Performance characterization is reported for spaceflight prototypes of this payload system. Using conventional microtiter plates, we show that Alamar Blue (AB) absorbance changes due to cellular metabolism accurately reflect E. coli viability changes: measuring AB absorbance onboard EcAMSat will enable telemetry of spaceflight data to Earth. Laboratory results using payload prototypes are consistent with wellplate and flask findings of differential sensitivity of UPEC and its delta rpoS strain to Gm. Space MG studies using EcAMSat should clarify inconsistencies from previous space experiments on bacterial antibiotic sensitivity. Further, if sigma (sup s) plays the same role in space MG as in LSMMG and Earth gravity, EcAMSat results would facilitate utilizing our previously developed terrestrial UTI countermeasures in astronauts.