%0 Journal Article
%T Design and evaluation of 16S rRNA sequence based oligonucleotide probes for the detection and quantification of Comamonas testosteroni in mixed microbial communities
%A Stephan Bathe
%A Martina Hausner
%J BMC Microbiology
%D 2006
%I BioMed Central
%R 10.1186/1471-2180-6-54
%X 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.The oligonucleotides presented here provide a useful tool to study C. testosteroni population dynamics in mixed microbial communities.Comamonas testosteroni is a ubiquitously occuring ¦Â-proteobacterial species which has been isolated from aquatic as well as terrestrial environments. It has been shown to be capable of transformations of steroid compounds [1-4] and also of degradation of aromatic hydrocarbons [5-10]. C. testosteroni is thus of interest for potential biotechnological applications such as chemical transformations in fine chemical manufacturing [11] and bioremediation processes [7,12], as indicated by laboratory results.Due to the widespread environmental distribution and the potential relevance of C. testosteroni in biotransformation processes, it may be of interest to follow the population dynamics of this species over time in a mixed-species environment. Since culture-dependent methods are not sufficiently accurate to detect and quantify one particular species within a mixed bacterial community, methods targeting the 16S rRNA and/or its gene(s) need to be applied to such a problem, as has already been done in previous investigations [13-16]. Probes for species of the genera Comamonas and Delftia (COM1424 [17], and PPT [18]) as well as for s
%U http://www.biomedcentral.com/1471-2180/6/54