Microbial contamination of human-tended spacecraft is unavoidable, making the study of microbial growth under space conditions essential for the preservation of astronauts' health and equipment integrity. Previous studies suggested that spaceflight conditions, such as microgravity, cause a range of physiological microbial alterations including increased growth yields and decreased antibiotic susceptibility. Because of its fast generation time, Vibrio natriegens could be used as a model organism for a variety of studies where generation time is a critical factor. In this study, V. natriegens was used as a tool to study growth characteristics by determining the viable cell number and antibiotic susceptibility under simulated microgravity using a 2-D clinostat (60 rpm) to establish a test system that resolves changes in microbial growth on a solid surface (agar) under microgravity. The data show that V. natriegens biomass increases significantly after 24 h at 37°C under simulated microgravity. The final cell population after cultivation under simulated microgravity was 60-fold greater than when cultivated under normal terrestrial gravity (1 × g). No change in susceptibility to the antibiotic rifampicin after cultivation under simulated microgravity or normal gravity was detected. These data show that V. natriegens is a new and innovative model organism for microbial microgravity research.
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References
1.
Albrecht-BuehlerG. (1992) The simulation of microgravity conditions on the ground. ASGSB Bull, 5:3–10.
2.
AnkenR. (2013) Simulation of microgravity for studies in gravitational biology: principles, devices and applications. Curr. Biotechnol. 2:192–200.
3.
AuninsT.R., EricksonK.E., PrasadN., LevyS.E., JonesA., ShresthaS., MastracchioR., StodieckL., KlausD., ZeaL., and ChatterjeeA. (2018) Spaceflight modifies Escherichia coli gene expression in response to antibiotic exposure and reveals role of oxidative stress response. Front Microbiol, 9, doi:10.3389/fmicb.2018.00310.
4.
AustinB., ZacharyA., and ColwellR.R. (1978) Recognition of Beneckea natriegens (Payne et al.) Baumann et al. as a member of the genus Vibrio, as previously proposed by Webb and Payne. Int J Syst Evol Microbiol, 28:315–317.
5.
BoulocP. and D'AriR. (1991) Escherichia coli metabolism in space. Microbiology, 137:2839–2843.
6.
BrieglebW. (1992) Some qualitative and quantitative aspects of the fast-rotating clinostat as a research tool. ASGSB Bull, 5:23–30.
7.
BrownR.B., KlausD., and ToddP. (2002) Effects of spaceflight, clinorotation, and centrifugation on the substrate utilization efficiency of E. coli. Microgravity Sci Technol, 13:24–29.
8.
BrungsS., EgliM., WuestS.L., ChristianenP.C., van LoonJ.J., AnhT.J.N., and HemmersbachR. (2016) Facilities for simulation of microgravity in the ESA ground-based facility programme. Microgravity Sci Technol, 28:191–203.
9.
CampbellE.A., KorzhevaN., MustaevA., MurakamiK., NairS., GoldfarbA., and DarstS.A. (2001) Structural mechanism for rifampicin inhibition of bacterial RNA polymerase. Cell, 104:901–912.
10.
CrucianB. and SamsC. (2009) Immune system dysregulation during spaceflight: clinical risk for exploration-class missions. J Leukoc Biol, 86:1017–1018.
11.
CrucianB., SimpsonR.J., MehtaS., StoweR., ChoukerA., HwangS.-A., ActorJ.K., SalamA.P., PiersonD., and SamsC. (2014) Terrestrial stress analogs for spaceflight associated immune system dysregulation. Brain Behav Immun, 39:23–32.
12.
DelpechR. (2001) Using Vibrio natriegens for studying bacterial population growth, artificial selection, and the effects of UV radiation and photo-reactivation. J Biol Educ, 35:93–97.
13.
EagonR.G. (1962) Pseudomonas natriegens, a marine bacterium with a generation time of less than 10 minutes. J Bacteriol, 83:736–737.
14.
Fajardo-CavazosP. and NicholsonW.L. (2016) Cultivation of Staphylococcus epidermidis in the human spaceflight environment leads to alterations in the frequency and spectrum of spontaneous rifampicin-resistance mutations in the rpoB gene. Front Microbiol, 7, doi:10.3389/fmicb.2016.00999.
15.
Fajardo-CavazosP., NarvelR., and NicholsonW.L. (2014) Differing responses in growth and spontaneous mutation to antibiotic resistance in Bacillus subtilis and Staphylococcus epidermidis cells exposed to simulated microgravity. Gravitational and Space Research, 4:34–45.
16.
FuchsF.M., DriksA., SetlowP., and MoellerR. (2017) An improved protocol for harvesting Bacillus subtilis colony biofilms. J Microbiol Methods, 134:7–13.
17.
GassetG., TixadorR., EcheB., LapchineL., MoattiN., TooropP., and WoldringhC. (1994) Growth and division of Escherichia coli under microgravity conditions. Res Microbiol, 145:111–120.
18.
GueguinouN., Huin-SchohnC., BascoveM., BuebJ.L., TschirhartE., Legrand-FrossiC., and FrippiatJ.P. (2009) Could spaceflight-associated immune system weakening preclude the expansion of human presence beyond Earth's orbit?. J Leukoc Biol, 86:1027–1038.
19.
HammondT.G., StodieckL., BirdsallH.H., KoenigP., HammondJ.S., GunterM., and AllenP.L. (2016) Effects of microgravity and clinorotation on the virulence of Klebsiella, Streptococcus, Proteus, and Pseudomonas. Gravitational and Space Research, 4:39–50.
20.
HasensteinK.H., van LoonJ.J.W.A., and BeysensD.A. (2015) Clinostats and other rotating systems—design, function, and limitations. In Generation and Applications of Extra-Terrestrial Environments on Earth, edited by BeysensD.A. and van LoonJ.J.W.A., River Publishers, Aalborg, Denmark, pp 147–156.
21.
HemmersbachR., StrauchS.M., SeibtD., and SchuberM. (2006) Comparative studies on gravisensitive protists on ground (2D and 3D clinostats) and in microgravity. Microgravity Sci Technol, 18:257–259.
22.
HerranzR., AnkenR., BoonstraJ., BraunM., ChristianenP.C., de GeestM., HauslageJ., HilbigR., HillR.J., LebertM., MedinaF.J., VagtN., UllrichO., van LoonJ.J., and HemmersbachR. (2013) Ground-based facilities for simulation of microgravity: organism-specific recommendations for their use, and recommended terminology. Astrobiology, 13:1–17.
23.
HorneckG., KlausD.M., and MancinelliR.L. (2010) Space microbiology. Microbiol Mol Biol Rev, 74:121–156.
24.
HuangB., LiD.G., HuangY., and LiuC.T. (2018) Effects of spaceflight and simulated microgravity on microbial growth and secondary metabolism. Mil Med Res, 5, doi:10.1186/s40779-018-0162-9.
25.
KacenaM.A. and ToddP. (1999) Gentamicin: effect on E. coli in space. Microgravity Sci Technol, 12:135–137.
26.
KacenaM.A., MerrellG.A., ManfrediB., SmithE.E., KlausD.M., and ToddP. (1999) Bacterial growth in space flight: logistic growth curve parameters for Escherichia coli and Bacillus subtilis. Appl Microbiol Biotechnol, 51:229–234.
27.
KalpanaD., ImC., and LeeY.S. (2016) Comparative growth, cross stress resistance, transcriptomics of Streptococcus pyogenes cultured under low shear modeled microgravity and normal gravity. Saudi J Biol Sci, 23:24–33.
28.
KlausD., SimskeS., ToddP., and StodieckL. (1997) Investigation of spaceflight effects on Escherichia coli and a proposed model of underlying physical mechanisms. Microbiology, 143:449–455.
29.
KlausD.M. and HowardH.N. (2006) Antibiotic efficacy and microbial virulence during spaceflight. Trends Biotechnol, 24:131–136.
30.
MaidaI., BosiE., PerrinE., PapaleoM.C., OrlandiniV., FondiM., FaniR., WiegelJ., BianconiG., and CanganellaF. (2013) Draft genome sequence of the fast-growing bacterium Vibrio natriegens strain DSMZ 759. Genome Announc, 1, doi:10.1128/genomeA.00648-13.
31.
MennigmannH.D. and LangeM. (1986) Growth and differentiation of Bacillus subtilis under microgravitiy. Naturwissenschaften, 73:415–417.
32.
MorrisonM.D., Fajardo-CavazosP., and NicholsonW.L. (2017) Cultivation in spaceflight produces minimal alterations in the susceptibility of Bacillus subtilis cells to 72 different antibiotics and growth-inhibiting compounds. Appl Environ Microbiol, 83, doi:10.1128/AEM.01584-17.
33.
NickersonC.A., OttC.M., WilsonJ.W., RamamurthyR., and PiersonD.L. (2004) Microbial responses to microgravity and other low-shear environments. Microbiol Mol Biol Rev, 68:345–361.
34.
PayneW.J. (1958) Studies on bacterial utilization of uronic acids III: induction of oxidative enzymes in a marine isolate. J Bacteriol, 76:301–307.
35.
PayneW.J., EagonR.G., and WilliamsA.K. (1961) Some observations on the physiology of Pseudomonas natriegens nov. spec. Antonie van Leeuwenhoek, 27:121–128.
36.
PolinskiE., SchuelerO., KrauseL., WimmerM.A., HemmersbachR., and GoldbachH.E. (2017) 2-D clinorotation alters the uptake of some nutrients in Arabidopsis thaliana. J Plant Physiol, 212:54–57.
37.
RosadoH., StapletonP.D., and TaylorP.W. (2006) Effect of simulated microgravity on the virulence properties of the opportunistic bacterial pathogen Staphylococcus aureus [IAC-06-A1.7/A2.7.06]. In Proceedings of the 57th International Astronautical Federation Congress, doi:10.2514/6.IAC-06-A1.7.06.
38.
RosenzweigJ.A., AhmedS., EunsonJ., and ChopraA.K. (2014) Low-shear force associated with modeled microgravity and spaceflight does not similarly impact the virulence of notable bacterial pathogens. Appl Microbiol Biotechnol, 98:8797–8807.
39.
SchiwonK., ArendsK., RogowskiK.M., FürchS., PreschaK., SakincT., Van HoudtR., WernerG., and GrohmannE. (2013) Comparison of antibiotic resistance, biofilm formation and conjugative transfer of Staphylococcus and Enterococcus isolates from International Space Station and Antarctic Research Station Concordia. Microb Ecol, 65:638–651.
40.
SenatoreG., MastroleoF., LeysN., and MaurielloG. (2018) Effect of microgravity and space radiation on microbes. Future Microbiol, 13:831–847.
TaylorP.W. (2015) Impact of spaceflight on bacterial virulence and antibiotic susceptibility. Infect Drug Resist, 8:249–262.
43.
ThévenetD., D'ariR., and BoulocP. (1996) The SIGNAL experiment in BIORACK: Escherichia coli in microgravity. J Biotechnol, 47:89–97.
44.
TirumalaiM.R., KarouiaF., TranQ., StepanovV.G., BruceR.J., OttC.M., PiersonD.L., and FoxG.E. (2017) The adaptation of Escherichia coli cells grown in simulated microgravity for an extended period is both phenotypic and genomic. NPJ Microgravity, 3, doi:10.1038/s41526-017-0020-1.
45.
VukantiR., ModelM.A., and LeffL.G. (2012) Effect of modeled reduced gravity conditions on bacterial morphology and physiology. BMC Microbiol, 12, doi:10.1186/1471-2180-12-4.
46.
WeinstockM.T., HesekE.D., WilsonC.M., and GibsonD.G. (2016) Vibrio natriegens as a fast-growing host for molecular biology. Nat Methods, 13:849–851.
47.
ZeaL., LarsenM., EstanteF., QvortrupK., MoellerR., Dias de OliveiraS., StodieckL., and KlausD. (2017) Phenotypic changes exhibited by E. coli cultured in space. Front Microbiol, 8, doi:10.3389/fmicb.2017.01598.
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