ANALYSIS OF COMPLEX MICROBIAL ECOSYSTEMS WITH IN SITU HYBRIDIZATION AND EPIFLUORESCENT MICROSCOPY

Abstract

In situ hybridization with fluorescent oligonucleotide probes combined with epifluorescent microscopy was used to detect and localize microorganisms in rumen samples from a black and white Friesian cow. Bacterial cells were hybridized with 16S rRNA targeted oligonucleotide probes specific for the phylogenetic group Cytophaga-Flexibacter-Bacteroides (CFB) (BACPRE) and for species Prevotella bryantii B₁4 (PBB₁4) and P.ruminicola 23^T (PR23). The oligonucleotide probes were labelled with fluorescine isothiocyanate (FITC) or tetramethylrodamine isothiocyanate (TRITC). Both species specific probes proved to be highly specific and gave strong and clear signal.