Differential recovery of phylogenetically disparate microbes from spacecraft-qualified metal surfaces

• Published in: Astrobiology Vol. 13; no. 2; p. 189
• Main Authors: Bargoma, E, La Duc, M T, Kwan, K, Vaishampayan, P, Venkateswaran, K
• Format: Journal Article
• Language: English
• Published: United States 01-02-2013
• Subjects: Bacteria - classification; Bacteria - genetics; Bacteria - isolation & purification; DNA, Bacterial - analysis; DNA, Ribosomal - analysis; DNA, Ribosomal - isolation & purification; Equipment Contamination; Metals; Models, Biological; Phylogeny; Polymerase Chain Reaction; RNA, Ribosomal, 16S - genetics; Spacecraft - instrumentation
• ISSN: 1557-8070
• DOI: 10.1089/ast.2012.0917

Universal and species-specific quantitative polymerase chain reaction-based methods were employed to compare the effectiveness of four distinct materials used to collect biological samples from metal surfaces. Known cell densities of a model microbial community (MMC) were deposited onto metal surfaces and subsequently collected with cotton and nylon-flocked swabs for small surface areas and biological sampling kits (BiSKits) and polyester wipes for large surface areas. Ribosomal RNA gene-based quantitative PCR (qPCR) analyses revealed that cotton swabs were superior to nylon-flocked swabs for recovering nucleic acids (i.e., DNA) from small surface areas. Similarly, BiSKits outperformed polyester wipes for sampling large surface areas. Species-specific qPCR results show a differential recovery of rRNA genes of the various MMC constituents, seemingly dependent on the type of sampling device employed. Both cotton swabs and BiSKits recovered the rDNA of all nine of the MMC constituent microbes assayed, whereas nylon-flocked swabs and polyester wipes recovered the rDNA of only six and four of these MMC strains, respectively. The findings of this study demonstrate the importance and proficiency of molecular techniques in gauging the effectiveness and efficiency of various modes of biological sample collection from metal surfaces.