Skip to main content Accessibility help
×
Home

A molecular method to assess bioburden embedded within silicon-based resins used on modern spacecraft materials

  • Christina N. Stam (a1), James Bruckner (a1), J. Andy Spry (a1), Kasthuri Venkateswaran (a1) and Myron T. La Duc (a1)...

Abstract

Current assessments of bioburden embedded in spacecraft materials are based on work performed in the Viking era (1970s), and the ability to culture organisms extracted from such materials. To circumvent the limitations of such approaches, DNA-based techniques were evaluated alongside established culturing techniques to determine the recovery and survival of bacterial spores encapsulated in spacecraft-qualified polymer materials. Varying concentrations of Bacillus pumilus SAFR-032 spores were completely embedded in silicone epoxy. An organic dimethylacetamide-based solvent was used to digest the epoxy and spore recovery was evaluated via gyrB-targeted qPCR, direct agar plating, most probably number analysis, and microscopy. Although full-strength solvent was shown to inhibit the germination and/or outgrowth of spores, dilution in excess of 100-fold allowed recovery with no significant decrease in cultivability. Similarly, qPCR (quantitative PCR) detection sensitivities as low as ∼103 CFU ml−1 were achieved upon removal of inhibitory substances associated with the epoxy and/or solvent. These detection and enumeration methods show promise for use in assessing the embedded bioburden of spacecraft hardware.

Copyright

Corresponding author

References

Hide All
Baker, S.S., Rugh, C.L. & Kamalay, J.C. (1990). Biotechniques 9, 268272.
Berhards, J., Weitzel, B., Werner, M., Rimpler, M. & Georgii, A. (1992). Histochem. 98, 145154.
Bizzini, A., Jaton, K., Romo, D., Bille, J., Prod'hom, G. & Greub, G. (2010). J. Clin. Microbiol. 48, 15491554.
Brewer, W.A., Paik, W.W., Smith, C.D., Robillard, C.L. & Green, R.H. (1972). Orig. Life Evol. Biosph. 3, 198205.
Cano, R., Poinar, H. & Poinar, G.O. Jr. (1992). Med. Sci. Res. 20, 249251.
Dickinson, D.N., La Duc, M.T., Haskins, W.E., Gornushkin, I., Winefordner, J.D., Powell, D.H. & Venkateswaran, K. (2004). Appl. Environ. Microbiol. 70, 475482.
Driks, A. (1999). Microbiol. Mol. Biol. Rev. 63, 120.
Gilbert, M.T., Cuccui, J., White, W., Lynnerup, N., Titball, R.W., Cooper, A. & Prentice, M.B. (2004). Microbiology 150, 341354.
Kempf, M.J., Chen, F., Kern, R. & Venkateswaran, K. (2005). Astrobiology 5, 391405.
La Duc, M.T., Nicholson, W., Kern, R. & Venkateswaran, K. (2003). Environ. Microbiol. 5, 977985.
La Duc, M.T., Osman, S. & Venkateswaran, K. (2009b). J. Rapid Meth. Auto. Microbiol. 17, 350368.
La Duc, M.T., Osman, S., Vaishampayan, P., Piceno, Y., Andersen, G., Spry, J.A. & Venkateswaran, K. (2009a). Appl. Environ. Microbiol. 75, 65596567.
Lampel, K.A., Orlandi, P.A. & Kornegay, L. (2000). Appl. Environ. Microbiol. 66, 45394542.
Muller, S. & Nebe-von-Caron, G. (2010). FEMS Microbiol. Rev. 34, 554587.
Nicholson, W. & Setlow, P. (1990). in Molecular Biological Methods for Bacillus, pp. 391450. John Wiley and Sons Inc., New York, NY.
NRC. (2006). Preventing the Forward Contamination of Mars, Committee on Preventing the Forward Contamination of Mars, National Research Council, National Academies Press, Washington, DC.
Olsson-Francis, K., de la Torre, R. & Cockell, C.S. (2010). Appl. Environ. Microbiol. 76, 21152121.
Puleo, J.R., Fields, N.D., Bergstrom, S.L., Oxborrow, G.S., Stabekis, P.D. & Koukol, R. (1977). Appl. Environ. Microbiol. 33, 379384.
Raghunathan, A., Ferguson, H.R. Jr., Bornarth, C.J., Song, W., Driscoll, M. & Lasken, R.S. (2005). Appl Environ Microbiol. 71, 33423347.
Rowe, R., Todd, R. & Waide, J. (1977). Appl. Environ. Microbiol. 33, 675680.
Satomi, M., La Duc, M.T. & Venkateswaran, K. (2006). Int. J. Syst. Evol. Microbiol. 56, 17351740.
Schaeffer, P., Millet, J. & Aubert, J. (1965). Proc. Natl. Acad. Sci. U.S.A 54, 704711.
Setlow, P. (2007). Trends Microbiol. 15, 172180.
Vasin, V.B. & Trofimov, V.I. (1995). Adv. Space Res. 15, 273276.
Venkateswaran, K., Chung, S., Allton, J. & Kern, R. (2004). Astrobiology 4, 377390.
Venkateswaran, K., Hattori, N., La Duc, M.T. & Kern, R. (2003). J. Microbiol. Methods 52, 367377.
Venkateswaran, K., Satomi, M., Chung, S., Kern, R.G., Koukol, R., Basic, C. & White, D. (2001). Syst. Appl. Microbiol. 24, 311320.
Wang, H. & Edwards, K.J. (2009). Geomicrobiol. J. 26, 463469.
Wang, Y., Hammes, F., De Roy, K., Verstraete, W. & Boon, N. (2010). Trends Biotechnol. 28, 416424.
Ward, D.M., Weller, R. & Bateson, M.M. (1990). Nature 345, 6365.
Zeitlin, C., Guetersloh, S.B., Heilbronn, L.H. & Miller, J. (2006). Nucl. Instrum. Methods Phys. Res. B, Beam Interact. Mat. At. (Netherlands) 252, 308318.
Zitterkopf, N. L. (2008). LabMedicine 39, 623625.

Keywords

A molecular method to assess bioburden embedded within silicon-based resins used on modern spacecraft materials

  • Christina N. Stam (a1), James Bruckner (a1), J. Andy Spry (a1), Kasthuri Venkateswaran (a1) and Myron T. La Duc (a1)...

Metrics

Altmetric attention score

Full text views

Total number of HTML views: 0
Total number of PDF views: 0 *
Loading metrics...

Abstract views

Total abstract views: 0 *
Loading metrics...

* Views captured on Cambridge Core between <date>. This data will be updated every 24 hours.

Usage data cannot currently be displayed