Design and evaluation of 16S rRNA sequence based oligonucleotide probes for the detection and quantification of Comamonas testosteroni in mixed microbial communities.
BACKGROUND
The beta-proteobacterial species Comamonas testosteroni is capable of biotransformation and also biodegradation of a range of chemical compounds and thus potentially useful in chemical manufacturing and bioremediation. The ability to detect and quantify members of this species in mixed microbial communities thus may be desirable.


RESULTS
We have designed an oligonucleotide probe for use in fluorescent in situ hybridization (FISH) and two pairs of PCR primers targeting a C. testosteroni subgroup. The FISH probe and one of the PCR primer pairs are suitable for quantification of C. testosteroni in mixed microbial communities using FISH followed by quantitative image analysis or real-time quantitative PCR, respectively. This has been shown by analysis of samples from an enrichment of activated sludge on testosterone resulting in an increase in abundance and finally isolation of C. testosteroni. Additionally, we have successfully used quantitative PCR to follow the C. testosteroni abundance during a laboratory scale wastewater bioaugmentation experiment.


CONCLUSIONS
The oligonucleotides presented here provide a useful tool to study C. testosteroni population dynamics in mixed microbial communities.
Bathe
Stephan
S
Institute of Water Quality Control and Waste Management, Technical University of Munich, Am Coulombwall, 85748 Garching, Germany. steph-b@gmx.de
Hausner
Martina
M
D015169:Colony Count, Microbial; D020604:Comamonas testosteroni / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D017404:In Situ Hybridization, Fluorescence / Q000379:methods*; D008969:Molecular Sequence Data; D015345:Oligonucleotide Probes; D010802:Phylogeny; D016133:Polymerase Chain Reaction / Q000379:methods*; D012329:RNA, Bacterial / Q000235:genetics; D012336:RNA, Ribosomal, 16S / Q000032:analysis / Q000235:genetics*; D012722:Sewage / Q000382:microbiology; D013739:Testosterone / Q000378:metabolism
D015345:Oligonucleotide Probes; D012329:RNA, Bacterial; D012336:RNA, Ribosomal, 16S; D012722:Sewage; D013739:Testosterone