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Title: Noninvasive Quantitative Measurement of Bacterial Growth in Porous Media Under Unsaturated-Flow Conditions

Journal Article · · Applied and Environmental Microbiology

Glucose-dependent growth of the luxCDABE reporter bacterium Pseudomonas fluorescens HK44 was monitored noninvasively in quartz sand under unsaturated-flow conditions within a 45- by 56- by 1-cm two-dimensional light transmission chamber. The spatial and temporal development of growth were mapped daily over 7 days by quantifying salicylate-induced bioluminescence. A nonlinear model relating the rate of increase in light emission after salicylate exposure to microbial density successfully predicted growth over 4 orders of magnitude (r{sup 2}=0.95). Total model-predicted growth agreed with growth calculated from the mass balance of the system by using previously determined growth parameters of HK44 (predicted, 1.2 x 10{sup 12} cells; calculated, 1.7 x 10{sup 12} cells). Colonization expanded in all directions from the inoculation region, including upward migration against the liquid flow. Both the daily rate of expansion of the colonized zone and the population density of the first day's growth in each newly colonized region remained relatively constant throughout the experiment. Nonetheless, substantial growth continued to occur on subsequent days in the older regions of the colonized zone. The proportion of daily potential growth that remained within the chamber declined progressively between days 2 and 7 (from 97 to 13%). A densely populated, anoxic region developed in the interior of the colonized zone even though the sand was unsaturated and fresh growth medium continued to flow through the colonized zone. These data illustrate the potential of a light transmission chamber, bioluminescent bacteria, and sensitive digital camera technology to noninvasively study real-time hydrology-microbiology interactions associated with unsaturated flow in porous media.

Research Organization:
Pacific Northwest National Lab. (PNNL), Richland, WA (United States)
Sponsoring Organization:
US Department of Energy (US)
DOE Contract Number:
AC06-76RL01830
OSTI ID:
15001016
Report Number(s):
PNNL-SA-36835; KP1301020; TRN: US200323%%367
Journal Information:
Applied and Environmental Microbiology, Vol. 68, Issue 7; Other Information: PBD: 1 Jul 2002
Country of Publication:
United States
Language:
English