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Applied and Environmental Microbiology, April 2006, p. 2322-2330, Vol. 72, No. 4
0099-2240/06/$08.00+0     doi:10.1128/AEM.72.4.2322-2330.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

rRNA and Poly-ß-Hydroxybutyrate Dynamics in Bioreactors Subjected to Feast and Famine Cycles

Dominic Frigon,^1, Gerard Muyzer,² Mark van Loosdrecht,² and Lutgarde Raskin^1*

Department of Civil and Environmental Engineering, University of Illinois at Urbana-Champaign, Urbana, Illinois,¹ Environmental Biotechnology Group, Department of Biotechnology, Delft University of Technology, Delft, The Netherlands²

Received 15 April 2005/ Accepted 13 January 2006

Feast and famine cycles are common in activated sludge wastewater treatment systems, and they select for bacteria that accumulate storage compounds, such as poly-ß-hydroxybutyrate (PHB). Previous studies have shown that variations in influent substrate concentrations force bacteria to accumulate high levels of rRNA compared to the levels in bacteria grown in chemostats. Therefore, it can be hypothesized that bacteria accumulate more rRNA when they are subjected to feast and famine cycles. However, PHB-accumulating bacteria can form biomass (grow) throughout a feast and famine cycle and thus have a lower peak biomass formation rate during the cycle. Consequently, PHB-accumulating bacteria may accumulate less rRNA when they are subjected to feast and famine cycles than bacteria that are not capable of PHB accumulation. These hypotheses were tested with Wautersia eutropha H16 (wild type) and W. eutropha PHB-4 (a mutant not capable of accumulating PHB) grown in chemostat and semibatch reactors. For both strains, the cellular RNA level was higher when the organism was grown in semibatch reactors than when it was grown in chemostats, and the specific biomass formation rates during the feast phase were linearly related to the cellular RNA levels for cultures. Although the two strains exhibited maximum uptake rates when they were grown in semibatch reactors, the wild-type strain responded much more rapidly to the addition of fresh medium than the mutant responded. Furthermore, the chemostat-grown mutant culture was unable to exhibit maximum substrate uptake rates when it was subjected to pulse-wise addition of fresh medium. These data show that the ability to accumulate PHB does not prevent bacteria from accumulating high levels of rRNA when they are subjected to feast and famine cycles. Our results also demonstrate that the ability to accumulate PHB makes the bacteria more responsive to sudden increases in substrate concentrations, which explains their ecological advantage.

Present address: Department of Microbiology and Immunology, University of British Columbia, Vancouver, BC, Canada.