PMID- 24814284
OWN - NLM
STAT- MEDLINE
DCOM- 20141017
LR  - 20140815
IS  - 1472-765X (Electronic)
IS  - 0266-8254 (Linking)
VI  - 59
IP  - 3
DP  - 2014 Sep
TI  - 16S rRNA-targeted oligonucleotide probes for direct detection of 
      Propionibacterium freudenreichii in presence of Lactococcus lactis with 
      multicolour fluorescence in situ hybridization.
PG  - 320-7
LID - 10.1111/lam.12278 [doi]
AB  - Multicolour fluorescence in situ hybridization (FISH) has been applied to detect 
      Lactococcus lactis and Propionibacterium freudenreichii cells in mixed 
      populations in medium and skimmed milk, using epifluorescent microscopy. The 16S 
      rRNA-targeted 18-mer oligonucleotide probes, specific for P. freudenreichii were 
      designed and evaluated. Based on multiple alignments of designed sequences, eight 
      16S rRNA probes were selected for in vitro studies. The permeabilization protocol 
      was optimized for simultaneous hybridization of propionibacteria and lactococci 
      cells. The probes GLO62 (62-80), PEU64 (64-82) and PFX311 (311-329) were found 
      specific for P. freudenreichii in analysis in vitro. Lactococcus lactis cells 
      were labelled with LactV5 (822-840), (S-S-L.lact-0821-a-A-18) probe. The 
      following combinations of oligonucleotide probes: LactV5/GLO62 , LactV5/PEU64 and 
      LactV5/PFX311 , enabled differentiation of Lc. lactis and P. freudenreichii cells 
      in culture media and in skimmed milk. SIGNIFICANCE AND IMPACT OF THE STUDY: 
      Results showed that three newly designed 16S rRNA-targeted oligonucleotide probes 
      specific for Propionibacterium freudenreichii in combination with a probe 
      specific for Lactococcus lactis can be used to differentiate lactic and propionic 
      acid bacteria in mixed communities using multicolour fluorescence in situ 
      hybridization (FISH), without applying permeabilization step. This is a first 
      study on simultaneous detection of both bacterial species with FISH, which was 
      found as rapid, useful and culture-independent tool for direct visualization of 
      bacteria on single cell level. It might be applied in monitoring of mixed starter 
      cultures in dairy industry.
CI  - (c) 2014 The Society for Applied Microbiology.
FAU - Miks-Krajnik, M
AU  - Miks-Krajnik M
AD  - Chair of Industrial and Food Microbiology, Faculty of Food Science, University of 
      Warmia and Mazury in Olsztyn, Olsztyn, Poland; Food Science and Technology 
      Programme, c/o Department of Chemistry, National University of Singapore, 3 
      Science Drive 3, Singapore, 117543, Singapore.
FAU - Babuchowski, A
AU  - Babuchowski A
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20140602
PL  - England
TA  - Lett Appl Microbiol
JT  - Letters in applied microbiology
JID - 8510094
RN  - 0 (Oligonucleotide Probes)
RN  - 0 (RNA, Bacterial)
RN  - 0 (RNA, Ribosomal, 16S)
SB  - IM
MH  - Base Sequence
MH  - In Situ Hybridization, Fluorescence/methods
MH  - Lactococcus lactis/*genetics
MH  - Microscopy, Fluorescence
MH  - Oligonucleotide Probes
MH  - Permeability
MH  - Propionibacterium/*genetics
MH  - RNA, Bacterial/genetics
MH  - RNA, Ribosomal, 16S/*genetics
MH  - Sensitivity and Specificity
OTO - NOTNLM
OT  - 16S rRNA oligonucleotide probes
OT  - Lactococcus lactis
OT  - Propionibacterium freudenreichii
OT  - fluorescent in situ hybridization
OT  - milk
EDAT- 2014/05/13 06:00
MHDA- 2014/10/18 06:00
CRDT- 2014/05/13 06:00
PHST- 2014/02/22 00:00 [received]
PHST- 2014/05/05 00:00 [revised]
PHST- 2014/05/05 00:00 [accepted]
PHST- 2014/05/13 06:00 [entrez]
PHST- 2014/05/13 06:00 [pubmed]
PHST- 2014/10/18 06:00 [medline]
AID - 10.1111/lam.12278 [doi]
PST - ppublish
SO  - Lett Appl Microbiol. 2014 Sep;59(3):320-7. doi: 10.1111/lam.12278. Epub 2014 Jun 
      2.