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

Published online by Cambridge University Press:  16 February 2012

Christina N. Stam ,
James Bruckner ,
J. Andy Spry ,
Kasthuri Venkateswaran  and
Myron T. La Duc
Christina N. Stam*
Affiliation:
Jet Propulsion Laboratory, California Institute of Technology, Pasadena, CA 91109, USA
James Bruckner
Affiliation:
Jet Propulsion Laboratory, California Institute of Technology, Pasadena, CA 91109, USA
J. Andy Spry
Affiliation:
Jet Propulsion Laboratory, California Institute of Technology, Pasadena, CA 91109, USA
Kasthuri Venkateswaran
Affiliation:
Jet Propulsion Laboratory, California Institute of Technology, Pasadena, CA 91109, USA
Myron T. La Duc
Affiliation:
Jet Propulsion Laboratory, California Institute of Technology, Pasadena, CA 91109, USA
*
e-mail: Christina.N.Stam@jpl.nasa.gov
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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.

Keywords

DNAepoxyPCRspore
Type
Research Article
Information
International Journal of Astrobiology , Volume 11 , Issue 3 , July 2012 , pp. 141 - 145
Copyright
Copyright © Cambridge University Press 2012

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