Utilization of Fluorescent Microspheres and a Green Fluorescent Protein-Marked Strain for Assessment of Microbiological Contamination of Permafrost and Ground Ice Core Samples from the Canadian High Arctic

Fluorescent microspheres were applied in a novel fashion during subsurface drilling of permafrost and ground ice in the Canadian High Arctic to monitor the exogenous microbiological contamination of core samples obtained during the drilling process. Prior to each drill run, a concentrated fluorescen...

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Bibliographic Details
Published in:Applied and Environmental Microbiology 2005-02, Vol.71 (2), p.1035-1041
Main Authors: Juck, D. F, Whissell, G, Steven, B, Pollard, W, McKay, C. P, Greer, C. W, Whyte, L. G
Format: Article
Language:English
Subjects:
Arctic Regions
Bacteria - genetics
Bacteria - isolation & purification
Biological and medical sciences
Cold Climate
Culture Media
DNA - chemistry
Equipment Contamination
Fluorescence
Fundamental and applied biological sciences. Psychology
Green Fluorescent Proteins - genetics
Green Fluorescent Proteins - metabolism
Ice
Ice - analysis
Methods
Microbiology
Microscopy, Fluorescence
Microspheres
Polymerase Chain Reaction - methods
Proteins
Pseudomonas - genetics
Pseudomonas - metabolism
Soil - analysis
Soil Microbiology
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Summary:Fluorescent microspheres were applied in a novel fashion during subsurface drilling of permafrost and ground ice in the Canadian High Arctic to monitor the exogenous microbiological contamination of core samples obtained during the drilling process. Prior to each drill run, a concentrated fluorescent microsphere (0.5-[micro]m diameter) solution was applied to the interior surfaces of the drill bit, core catcher, and core tube and allowed to dry. Macroscopic examination in the field demonstrated reliable transfer of the microspheres to core samples, while detailed microscopic examination revealed penetration levels of less than 1 cm from the core exterior. To monitor for microbial contamination during downstream processing of the permafrost and ground ice cores, a Pseudomonas strain expressing the green fluorescent protein (GFP) was painted on the core exterior prior to processing. Contamination of the processed core interiors with the GFP-expressing strain was not detected by culturing the samples or by PCR to detect the gfp marker gene. These methodologies were quick, were easy to apply, and should help to monitor the exogenous microbiological contamination of pristine permafrost and ground ice samples for downstream culture-dependent and culture-independent microbial analyses.
ISSN:0099-2240
1098-5336
DOI:10.1128/AEM.71.2.1035-1041.2005