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 fluorescent microsphere (0.5-microm 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.
Juck
D F
DF
Biotechnology Research Institute, National Research Council of Canada, Montreal, Quebec H4P 2R2, Canada. david.juck@cnrc-nrc.gc.ca
Whissell
G
G
Steven
B
B
Pollard
W
W
McKay
C P
CP
Greer
C W
CW
Whyte
L G
LG
D001110:Arctic Regions; D001419:Bacteria / Q000235:genetics / Q000302:isolation & purification; D003081:Cold Climate; D003470:Culture Media; D004247:DNA / Q000737:chemistry; D004866:Equipment Contamination; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism*; D007053:Ice / Q000032:analysis*; D008856:Microscopy, Fluorescence; D008863:Microspheres; D016133:Polymerase Chain Reaction / Q000379:methods; D011549:Pseudomonas / Q000235:genetics / Q000378:metabolism; D012987:Soil / Q000032:analysis; D012988:Soil Microbiology
D003470:Culture Media; D007053:Ice; D012987:Soil; D049452:Green Fluorescent Proteins; D004247:DNA