Community analysis of biofilters using fluorescence in situ hybridization including a new probe for the <i>Xanthomonas</i> branch of the class <i>Proteobacteria</i>

Abstract

Domain-, class-, and subclass-specific rRNA-targeted probes were applied to investigate the microbial communities of three industrial and three laboratory-scale biofilters, The set of probes also included a new probe (named XAN818) specific for the Xanthomonas branch of the class Proteobacteria; this probe is described in this study, The members of the Xanthomonas branch do not hybridize with previously developed rRNA-targeted oligonucleotide probes for the alpha-, beta-, and gamma-Proteobacteria, Bacteria of the Xanthomonas branch accounted for up; to 4.5% of total direct counts obtained with 4',6-diamidino-2-phenylindole. In biofilter samples, the relative abundance of these bacteria was similar to that of the gamma-Proteobacteria, Actinobacteria (gram-positive bacteria with a high G+C DNA content) and alpha-Proteobacteria were the most dominant groups. Detection rates obtained with probe EUB338 varied between about 40 and 70%, For samples with high contents of gram-positive bacteria, these percentages were substantially improved when the calculations were corrected for the reduced permeability of gram-positive bacteria when formaldehyde was used as a fixative, The set of applied bacterial class- and subclass-specific probes yielded, on average, 58.5% (+/- a standard deviation of 23.0%) of the corrected eubacterial detection rates, thus indicating the necessity of additional probes for studies of biofilter communities. The Xanthomonas-specific probe presented here may serve as an efficient tool for identifying potential phytopathogens, In situ hybridization proved to be a practical tool for microbiological studies of biofiltration systems.