Biofilms on tracheoesophageal voice prostheses: a confocal laser scanning microscopy demonstration of mixed bacterial and yeast biofilms.
The aim of this study was to demonstrate the presence of yeast and bacterial biofilms on the surface of tracheoesophageal voice prostheses (TVPs) by a double-staining technique with confocal laser scanning microscopy (CLSM). Biofilms of 12 removed TVPs were visualized by scanning electron microscopy, then stained with ConA-FITC and propidium iodide for CLSM. Microbial identification was by partial 16S rRNA gene analysis and ITS-2 sequence analysis. Microbial biofilms on the TVPs consisted of bacteria and filamentous cells. Bacterial cells were attached to the filamentous and unicellular yeast cells, thus forming a network. Sequence analyses of six voice prostheses identified the presence of a variety of bacterial and yeast species. In vivo studies showed that Klebsiella oxytoca and Micrococcus luteus efficiently attached to Candida albicans. CLSM with double fluorescence staining can be used to demonstrate biofilm formations composed of a mixture of yeast and bacterial cells on the surface of TVPs.
Kania
Romain E
RE
Department of Oto-Rhino-Laryngology, Head & Neck Surgery, Leiden University Medical Center, Leiden, The Netherlands.
Lamers
Gerda E M
GE
van de Laar
Nicole
N
Dijkhuizen
Marloes
M
Lagendijk
Ellen
E
Huy
Patrice Tran Ba
PT
Herman
Philippe
P
Hiemstra
Pieter
P
Grote
Jan J
JJ
Frijns
Johan
J
Bloemberg
Guido V
GV
D001419:Bacteria / Q000145:classification / Q000235:genetics / Q000254:growth & development / Q000302:isolation & purification; D018441:Biofilms; D002176:Candida albicans / Q000145:classification / Q000235:genetics / Q000254:growth & development / Q000302:isolation & purification; D003208:Concanavalin A / Q000378:metabolism; D004269:DNA, Bacterial / Q000032:analysis / Q000302:isolation & purification; D004271:DNA, Fungal / Q000032:analysis / Q000302:isolation & purification; D016650:Fluorescein-5-isothiocyanate / Q000378:metabolism; D007831:Larynx, Artificial / Q000382:microbiology*; D016982:Micrococcus luteus / Q000145:classification / Q000235:genetics / Q000254:growth & development / Q000302:isolation & purification; D018613:Microscopy, Confocal / Q000379:methods*; D008855:Microscopy, Electron, Scanning; D016133:Polymerase Chain Reaction; D011419:Propidium / Q000378:metabolism; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D017422:Sequence Analysis, DNA; D015003:Yeasts / Q000145:classification / Q000235:genetics / Q000254:growth & development / Q000302:isolation & purification
D004269:DNA, Bacterial; D004271:DNA, Fungal; D012336:RNA, Ribosomal, 16S; D003208:Concanavalin A; D011419:Propidium; D016650:Fluorescein-5-isothiocyanate