Quantitation of the Population Size and Metabolic Activity of a Resin Acid Degrading Bacterium in Activated Sludge Using Slot-Blot Hybridization to Measure the rRNA:rDNA Ratio
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- Muttray, A. & Mohn, W. Microb Ecol (1999) 38: 348. doi:10.1007/s002489901005
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The 16S rRNA:rDNA ratio is a useful parameter for measuring metabolic activity of a selected member of a complex microbial community, as in pulp effluent activated sludge systems. The RNA:DNA ratio of Sphingomonas sp. DhA-33, previously isolated from a sequencing batch reactor treating pulp mill effluent, is positively correlated with its growth rate (μ) under steady-state conditions. DhA-33 was grown in a chemostat with growth rates ranging from 0.04 to 0.15 cell divisions per hour. DhA-33 was also able to degrade dehydroabietic acid in bleached kraft mill effluent (BKME) plus mineral medium in batch culture. Slot-blot hybridization with radioactively labeled species-specific oligonucleotide probes for 16S rRNA and 16S rDNA was used to measure rRNA, rDNA, and the RNA:DNA ratio of this strain when in a mixed sludge community. An increase in DhA-33 rDNA indicated growth of DhA-33 within the community. The RNA:DNA ratio of DhA-33 increased sharply during exponential growth and declined as cells entered stationary phase. The RNA:DNA ratio decreased earlier and faster in DhA- 33/sludge co-cultures than in DhA-33 pure cultures, presumably due to an earlier depletion of nutrients. The species-specific quantification of the RNA:DNA ratio makes it possible to estimate the metabolic activity of selected members of a microbial community in situ.