title: Can genetically modified Escherichia coli with neutral buoyancy induced by gas vesicles be used as an alternative method to clinorotation for microgravity studies?
abstract: Space flight has been shown to affect various bacterial growth parameters. It is proposed that weightlessness allows the cells to remain evenly distributed, consequently altering the chemical makeup of their surrounding fluid, and hence indirectly affecting their physiological behaviour. In support of this argument, ground-based studies using clinostats to partially simulate the quiescent environment attained in microgravity have generally been successful in producing bacterial growth characteristics that mimic responses reported under actual space conditions. A novel approach for evaluating the effects of reduced cell sedimentation is presented here through use of Escherichia coli cultures genetically modified to be neutrally buoyant. Since clinorotation would not (or would only minimally) affect cell distribution of this already near-colloidal cell system, it was hypothesized that the effects on final population density would be eliminated relative to a static control. Gas-vesicle-producing E. coli cultures were grown under clinostat and static conditions and the culture densities at 60 h were compared. As a control, E. coli that do not produce gas vesicles, but were otherwise identical to the experimental strain, were also grown under clinostat and static conditions. As hypothesized, no significant difference was observed in cell populations at 60 h between the clinorotated and static gas-vesicle-producing E. coli cultures, while the cells that did not produce gas vesicles showed a mean increase in population density of 10.5 % (P = 0.001). These results further suggest that the lack of cumulative cell sedimentation is the dominant effect of space flight on non-stirred, in vitro E. coli cultures.
authors.affiliation: University of Colorado, Aerospace Engineering Sciences Department, BioServe Space Technologies, Boulder, CO 80309-0429, USA.
authors.forename: Michael
authors.initials: M
authors.lastname: Benoit
authors.affiliation: University of Colorado, Aerospace Engineering Sciences Department, BioServe Space Technologies, Boulder, CO 80309-0429, USA.
authors.forename: David
authors.initials: D
authors.lastname: Klaus
mesh_terms: D019149:Bioreactors; D002498:Centrifugation; D003470:Culture Media; D004926:Escherichia coli / Q000235:genetics / Q000254:growth & development / Q000502:physiology*; D030781:Organisms, Genetically Modified; D011506:Proteins / Q000235:genetics / Q000378:metabolism*; D012399:Rotation; D013026:Space Flight; D014893:Weightlessness; D018474:Weightlessness Simulation
chemical_list: D003470:Culture Media; D011506:Proteins; C038524:gas vesicle protein