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Removing biofilms from endoscopes: the importance of the cleaning chemistry
Wafi Siala1,2, Françoise Van Bambeke1 , Thomas Vanzieleghem2
1 Louvain Drug Research Institute, Université catholique de Louvain, Brussels; 2 OneLIFE S.A., Louvain-la-Neuve - Belgium
Introduction
Several studies have associated the use of endoscopes with several
outbreaks of infection in various clinical settings, including
gastrointestinal, thoracic, urological, and other endoscopic procedures.
The ability of bacteria to form biofilms is a major cause for endoscopes
contamination. Staphylococcus aureus, Klebsiella pneumoniae,
Enterococcus faecalis, Escherichia coli, Pseudomonas aeruginosa are
the most likely germs to cause these contaminations. We compared the
biofilm dispersion activity, against bacterial species mentioned above,
of one enzymatic cleaner developed by OneLIFE Sa (enziQure®) and
10 cleaners commonly used for the manual cleaning of endoscopes.
Results
References
1- Siala, W. et al. The antifungal caspofungin
increases fluoroquinolone activity against
Staphylococcus aureus biofilms by inhibiting N-
acetylglucosamine transferase. Nat Commun 2016;
7:13286.
Materials and Methods
Conclusions
While most of the current cleaning solutions are poorly active on biofilms, the enzymatic formulation of enziQure® allows marked
(> 50 %) biofilm dispersion for 80% of tested strains. This effect nevertheless remains strain-dependent, probably related to the
nature of the produced biofilm matrix.
Acknowledgements
WS is post-doctoral fellow of the program first
enterprise from the Region Wallonne. This work
was also supported by OneLIFE SA.
This poster will be available after the meeting at http://www.facm.ucl.ac.be/posters
We used 5 reference strains and 10 clinical isolates (C1-C10; infections
on medical devices and tissues). Biofilms were grown for 24 h in 96-
wells plates. biofilms were exposed for 1h at 40°C to concentrations
recommended by manufactures (0.5%-1%) of cleaners prepared in
standardized water (WSH :1.25 mM MgCl2, 2.5 mM CaCl2, 3.33 mM
NaHCO3). The ability of cleaners to remove biofilm biomass was
determined using crystal violet assay as previously described (1).
Tridimensional images were performed using confocal laser scanning
microscopy (CLSM). Biofilms were grown on coverslips, washed with
distilled water; after which coverslips were transferred into a fresh well
and incubated during 30 min at room temperature with LIVE/DEAD kit
(1). Image stacks were acquired at a resolution of 700x500 pixels,
recorded using Z-Stack module for acquisition of image series from
different focus planes, and used to construct three-dimensional images
with AxioVision software.
Figure 1 – Percentage of biomass reduction for 15 biofilms (formed by 3
isolates from 5 different species) after exposure to enziQure® and 10 other
cleaners in the absence of mechanical action (* : no enzyme; # : germicidal
activity). The dotted line shows the 50 % limit. Dots show the data for
individual strains, with mean and 95% confidence intervals shown in black for
each cleaner. Means (% ± SD): enziQure®: 62 ± 16; A: 48 ± 20; B: 45.7 ± 31;
C: 38 ± 35; D: 18 ± 17; E: 16 ± 23; F: 12 ± 24; G: 12 ± 19; H: 11 ± 17; I:
12 ± 14; J: 8± 12. Statistical analysis: one-way ANOVA with Dunnett’s multiple
comparison test (cleaner vs enziQure®): *: p < 0.05; ***: p < 0.001
Figure 2– Tridimensional images in confocal
laser scanning microscopy of PAO1 and
clinical isolate (C4) biofilms. Biofilms were
incubated during 1h at 40°C with 1%
enziQure®; 0.5% cleaner A or 0.5% cleaner J
and labelled with live/dead staining (red: dead;
green: live). PAO1 biofilms are shown on right ,
C4 biofilms are shown on left.
0
10
20
30
40
50
60
70
80
90
100
enziQ
ure
A#
B
C
D
E#
F*,#
G#
H
*,#
I
J
0
10
20
30
40
50
60
70
80
90
100
ATCC33591
C1
C2
PAO1
C3
C4
ATCC700603
C5
C6
ATCC25922
C7
C8
ATCC29512
C9
C10
%reductionofbiomass
S. aureus P. aeruginosa K. pneumoniae E. coli E. faecalis
* *** *** *** *** *** *** ***
P1484
PAO1	UNTREATED	
PAO1	VS	CLEANER	A	
PAO1	VS	CLEANER	J	
C4	UNTREATED	
C4	VS	CLEANER	A	
C4	VS	CLEANER	J	
C4	VS	enziQure®	PAO1	VS	enziQure®

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Biofilm removal - the importance of chemistry

  • 1. Removing biofilms from endoscopes: the importance of the cleaning chemistry Wafi Siala1,2, Françoise Van Bambeke1 , Thomas Vanzieleghem2 1 Louvain Drug Research Institute, Université catholique de Louvain, Brussels; 2 OneLIFE S.A., Louvain-la-Neuve - Belgium Introduction Several studies have associated the use of endoscopes with several outbreaks of infection in various clinical settings, including gastrointestinal, thoracic, urological, and other endoscopic procedures. The ability of bacteria to form biofilms is a major cause for endoscopes contamination. Staphylococcus aureus, Klebsiella pneumoniae, Enterococcus faecalis, Escherichia coli, Pseudomonas aeruginosa are the most likely germs to cause these contaminations. We compared the biofilm dispersion activity, against bacterial species mentioned above, of one enzymatic cleaner developed by OneLIFE Sa (enziQure®) and 10 cleaners commonly used for the manual cleaning of endoscopes. Results References 1- Siala, W. et al. The antifungal caspofungin increases fluoroquinolone activity against Staphylococcus aureus biofilms by inhibiting N- acetylglucosamine transferase. Nat Commun 2016; 7:13286. Materials and Methods Conclusions While most of the current cleaning solutions are poorly active on biofilms, the enzymatic formulation of enziQure® allows marked (> 50 %) biofilm dispersion for 80% of tested strains. This effect nevertheless remains strain-dependent, probably related to the nature of the produced biofilm matrix. Acknowledgements WS is post-doctoral fellow of the program first enterprise from the Region Wallonne. This work was also supported by OneLIFE SA. This poster will be available after the meeting at http://www.facm.ucl.ac.be/posters We used 5 reference strains and 10 clinical isolates (C1-C10; infections on medical devices and tissues). Biofilms were grown for 24 h in 96- wells plates. biofilms were exposed for 1h at 40°C to concentrations recommended by manufactures (0.5%-1%) of cleaners prepared in standardized water (WSH :1.25 mM MgCl2, 2.5 mM CaCl2, 3.33 mM NaHCO3). The ability of cleaners to remove biofilm biomass was determined using crystal violet assay as previously described (1). Tridimensional images were performed using confocal laser scanning microscopy (CLSM). Biofilms were grown on coverslips, washed with distilled water; after which coverslips were transferred into a fresh well and incubated during 30 min at room temperature with LIVE/DEAD kit (1). Image stacks were acquired at a resolution of 700x500 pixels, recorded using Z-Stack module for acquisition of image series from different focus planes, and used to construct three-dimensional images with AxioVision software. Figure 1 – Percentage of biomass reduction for 15 biofilms (formed by 3 isolates from 5 different species) after exposure to enziQure® and 10 other cleaners in the absence of mechanical action (* : no enzyme; # : germicidal activity). The dotted line shows the 50 % limit. Dots show the data for individual strains, with mean and 95% confidence intervals shown in black for each cleaner. Means (% ± SD): enziQure®: 62 ± 16; A: 48 ± 20; B: 45.7 ± 31; C: 38 ± 35; D: 18 ± 17; E: 16 ± 23; F: 12 ± 24; G: 12 ± 19; H: 11 ± 17; I: 12 ± 14; J: 8± 12. Statistical analysis: one-way ANOVA with Dunnett’s multiple comparison test (cleaner vs enziQure®): *: p < 0.05; ***: p < 0.001 Figure 2– Tridimensional images in confocal laser scanning microscopy of PAO1 and clinical isolate (C4) biofilms. Biofilms were incubated during 1h at 40°C with 1% enziQure®; 0.5% cleaner A or 0.5% cleaner J and labelled with live/dead staining (red: dead; green: live). PAO1 biofilms are shown on right , C4 biofilms are shown on left. 0 10 20 30 40 50 60 70 80 90 100 enziQ ure A# B C D E# F*,# G# H *,# I J 0 10 20 30 40 50 60 70 80 90 100 ATCC33591 C1 C2 PAO1 C3 C4 ATCC700603 C5 C6 ATCC25922 C7 C8 ATCC29512 C9 C10 %reductionofbiomass S. aureus P. aeruginosa K. pneumoniae E. coli E. faecalis * *** *** *** *** *** *** *** P1484 PAO1 UNTREATED PAO1 VS CLEANER A PAO1 VS CLEANER J C4 UNTREATED C4 VS CLEANER A C4 VS CLEANER J C4 VS enziQure® PAO1 VS enziQure®