PMID- 30269994
OWN - NLM
STAT- MEDLINE
DCOM- 20190318
LR  - 20190318
IS  - 1618-0984 (Electronic)
IS  - 0723-2020 (Linking)
VI  - 42
IP  - 2
DP  - 2019 Mar
TI  - Autofluorescent characteristics of Candidatus Brocadia fulgida and the 
      consequences for FISH and microscopic detection.
PG  - 135-144
LID - S0723-2020(18)30249-2 [pii]
LID - 10.1016/j.syapm.2018.09.002 [doi]
AB  - An enrichment culture of Candidatus Brocadia fulgida was identified by three 
      independent methods: analysis of autofluorescence using different microscope 
      filter blocks and a fluorescence spectrometer, fluorescence in situ hybridization 
      (FISH) with anammox-specific probes and partial sequencing of the 16S rDNA, 
      hydrazine synthase hzsA and hydrazine oxidoreductase hzo. The filter block BV-2A 
      (400-440, 470 LP, Nikon) was suitable for preliminary detection of Ca. B. 
      fulgida. An excitation-emission matrix revealed three pairs of 
      excitation-emission maxima: 288-330 nm, 288-478 nm and 417-478 nm. Several 
      autofluorescent cell clusters could not be stained with DAPI or by FISH, 
      suggesting empty but intact cells (ghost cells) or inhibited permeability. 
      Successful staining of autofluorescent cells with the FISH probes Ban162 and 
      Bfu613, even at higher formamide concentrations, suggested insufficient 
      specificity of Ban162. Under certain conditions, Ca. B. fulgida lost its 
      autofluorescence, which reduced the reliability of autofluorescence for 
      identification and detection. Non-fluorescent Ca. Brocadia cells could not be 
      stained with Ban162, but with Bfu613 at higher formamide concentrations, 
      suggesting a dependency between both parameters. The phylogenetic analysis showed 
      only good taxonomical clustering of the 16S rDNA and hzsA. In conclusion, careful 
      consideration of autofluorescent characteristics is recommended when analysing 
      and presenting FISH observations of Ca. B. fulgida to avoid misinterpretations 
      and misidentifications.
CI  - Copyright (c) 2018 Elsevier GmbH. All rights reserved.
FAU - Bollmann, Jorg
AU  - Bollmann J
AD  - Department of Biotechnology for Water Treatment, BTU-Cottbus-Senftenberg, 
      Siemens-Halske-Ring 8, 03046 Cottbus, Germany. Electronic address: 
      boellman@b-tu.de.
FAU - Engelbrecht, Steffen
AU  - Engelbrecht S
AD  - Department of Biotechnology for Water Treatment, BTU-Cottbus-Senftenberg, 
      Siemens-Halske-Ring 8, 03046 Cottbus, Germany.
FAU - Martienssen, Marion
AU  - Martienssen M
AD  - Department of Biotechnology for Water Treatment, BTU-Cottbus-Senftenberg, 
      Siemens-Halske-Ring 8, 03046 Cottbus, Germany.
LA  - eng
PT  - Journal Article
DEP - 20180920
PL  - Germany
TA  - Syst Appl Microbiol
JT  - Systematic and applied microbiology
JID - 8306133
RN  - 0 (DNA, Bacterial)
RN  - 0 (RNA, Ribosomal, 16S)
RN  - 7664-41-7 (Ammonia)
SB  - IM
MH  - Ammonia
MH  - Bacteria/*classification/isolation & purification
MH  - Bacterial Typing Techniques
MH  - Bioreactors/microbiology
MH  - DNA, Bacterial/genetics
MH  - *Fluorescence
MH  - Genes, Bacterial
MH  - In Situ Hybridization, Fluorescence
MH  - Microscopy, Fluorescence
MH  - Phylogeny
MH  - RNA, Ribosomal, 16S/genetics
MH  - Sequence Analysis, DNA
OTO - NOTNLM
OT  - 16S rDNA
OT  - Candidatus Brocadia fulgida
OT  - Candidatus Kuenenia stuttgartiensis
OT  - FISH
OT  - Hydrazine oxidase hzo
OT  - Hydrazine synthase hzsA
EDAT- 2018/10/03 06:00
MHDA- 2019/03/19 06:00
CRDT- 2018/10/02 06:00
PHST- 2018/06/19 00:00 [received]
PHST- 2018/08/10 00:00 [revised]
PHST- 2018/09/13 00:00 [accepted]
PHST- 2018/10/03 06:00 [pubmed]
PHST- 2019/03/19 06:00 [medline]
PHST- 2018/10/02 06:00 [entrez]
AID - S0723-2020(18)30249-2 [pii]
AID - 10.1016/j.syapm.2018.09.002 [doi]
PST - ppublish
SO  - Syst Appl Microbiol. 2019 Mar;42(2):135-144. doi: 10.1016/j.syapm.2018.09.002. 
      Epub 2018 Sep 20.