Autofluorescent characteristics of Candidatus Brocadia fulgida and the consequences for FISH and microscopic detection.

摘要: Abstract An enrichment culture of Candidatus Brocadia fulgida was identified by three independent methods: analysis autofluorescence using different microscope filter blocks and a fluorescence spectrometer, in situ hybridization (FISH) with anammox-specific probes partial sequencing the 16S rDNA, hydrazine synthase hzsA oxidoreductase hzo. The block BV-2A (400–440, 470 LP, Nikon) suitable for preliminary detection Ca. B. fulgida. excitation-emission matrix revealed pairs maxima: 288–330 nm, 288–478 nm 417–478 nm. Several autofluorescent cell clusters could not be stained DAPI or FISH, suggesting empty but intact cells (ghost cells) inhibited permeability. Successful staining FISH Ban162 Bfu613, even at higher formamide concentrations, suggested insufficient specificity Ban162. Under certain conditions, lost its autofluorescence, which reduced reliability identification detection. Non-fluorescent Ban162, Bfu613 dependency between both parameters. phylogenetic showed only good taxonomical clustering rDNA hzsA. In conclusion, careful consideration characteristics is recommended when analysing presenting observations to avoid misinterpretations misidentifications.