You are on page 1of 17

Journal of Microbiological Methods 83 (2010) 89–105

Contents lists available at ScienceDirect

Journal of Microbiological Methods
j o u r n a l h o m e p a g e : w w w. e l s ev i e r. c o m / l o c a t e / j m i c m e t h

Review

In vitro and in vivo model systems to study microbial biofilm formation
Tom Coenye ⁎, Hans J. Nelis
Laboratory of Pharmaceutical Microbiology, Ghent University, Gent, Belgium

a r t i c l e

i n f o

Article history:
Received 9 July 2010
Received in revised form 19 August 2010
Accepted 23 August 2010
Available online 8 September 2010
Keywords:
Biofilm
Model system
Animal model

a b s t r a c t
Biofilm formation is often considered the underlying reason why treatment with an antimicrobial agent fails
and as an estimated 65–80% of all human infections is thought to be biofilm-related, this presents a serious
challenge. Biofilm model systems are essential to gain a better understanding of the mechanisms involved in
biofilm formation and resistance. In this review a comprehensive overview of various in vitro and in vivo
systems is presented, and their advantages and disadvantages are discussed.
© 2010 Elsevier B.V. All rights reserved.

Contents
1.
2.

Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
In vitro biofilm model systems . . . . . . . . . . . . . . . . . . . . . .
2.1.
Microtiter plate-based model systems . . . . . . . . . . . . . . .
2.2.
Flow displacement biofilm model systems . . . . . . . . . . . . .
2.2.1.
Modified Robbins device (MRD) . . . . . . . . . . . . . .
2.2.2.
Other PFR systems . . . . . . . . . . . . . . . . . . . .
2.2.3.
Centers for Disease Control (CDC) biofilm reactor . . . . .
2.2.4.
Other flow displacement systems . . . . . . . . . . . . .
2.3.
Cell-culture-based model systems . . . . . . . . . . . . . . . . .
2.4.
Microfluidic devices . . . . . . . . . . . . . . . . . . . . . . . .
3.
In vivo biofilm model systems. . . . . . . . . . . . . . . . . . . . . . .
3.1.
Caenorhabditis elegans model . . . . . . . . . . . . . . . . . . . .
3.2.
Vertebrate animal models . . . . . . . . . . . . . . . . . . . . .
3.2.1.
Central venous catheter models . . . . . . . . . . . . . .
3.2.2.
Subcutaneous foreign body infection models . . . . . . . .
3.2.3.
Intraperitoneal foreign body infection models . . . . . . .
3.2.4.
Urinary tract infection models . . . . . . . . . . . . . . .
3.2.5.
Ear, nose and throat infection models . . . . . . . . . . .
3.2.6.
Respiratory tract infection models . . . . . . . . . . . . .
3.2.7.
Osteomyelitis infection models . . . . . . . . . . . . . .
3.2.8.
Other models . . . . . . . . . . . . . . . . . . . . . . .
4.
Wound biofilm models . . . . . . . . . . . . . . . . . . . . . . . . . .
4.1.
In vitro models . . . . . . . . . . . . . . . . . . . . . . . . . .
4.2.
In vivo models . . . . . . . . . . . . . . . . . . . . . . . . . .
5.
Quantification and visualisation of biofilms grown in various model systems
6.
Which model system to choose? . . . . . . . . . . . . . . . . . . . . .
Acknowledgements . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.

.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.

.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.

.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.

.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.

.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.

.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.

⁎ Corresponding author. Harelbekestraat 72, B-9000 Gent, Belgium. Tel.: + 32 9 2648141; fax: + 32 9 2648195.
E-mail address: Tom.Coenye@UGent.be (T. Coenye).
0167-7012/$ – see front matter © 2010 Elsevier B.V. All rights reserved.
doi:10.1016/j.mimet.2010.08.018

.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.

.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.

.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.

.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.

.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.

.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.

.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.

.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.

.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.

.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.

.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.

.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.

.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.

.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.

.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.

.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.

.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.

.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.

.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.

.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.

.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.

.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.

.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.

.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.

.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.

90
90
90
91
91
91
92
93
93
94
94
94
94
94
94
95
96
96
96
97
97
97
97
97
98
98
99
99

Firstly. 2003). which contains various (antibiotic) solutions.. 2008). Microtiter plate-based model systems Microtiter plate (MTP)-based systems are among the mostfrequently-used biofilm model systems (see for example Cerca et al. Flemming. 2003. This rapid and miniaturized biofilm assay is mostly applied to evaluate the effects of various antimicrobial agents on biofilm eradication (see for example Aaron et al. pegs are attached to the top lid of a microtiter plate and by closing the microtiter plate.. Coenye et al. Harrison et al. 1996). Coetser and Cloete. 2002. Peeters et al.. 1996. 1999. Coenye. 2007). Biofilms are defined as consortia of microorganisms that are attached to a biotic or abiotic surface. fermentation (Kunduru and Pometto. 2001. The focus of this review is on tools to study medically-relevant biofilms. a profound examination of the effects of modification. However. 2004). Juhna et al. 2008b.. Introduction Since 1943. 2005. Biofouling is especially problematic in systems in which materials come into contact with water.. 2008. Biofilm formation is a multistage process in which microbial cells adhere to the surface (initial reversible attachment). A magnet is used to collect the non-immobilised beads into a single spot which is then quantified through specialised image algorithms... it then took more than 30 years (and the paradigm-changing work of Bill Costerton and colleagues) to accept that for microorganisms (both bacteria and fungi) the biofilm mode of life is the rule rather than the exception (Costerton et al... 2010. 2005). Toté et al. 2007. 2003. Miyake et al. 2003). to determine the influence of matrix components on Leuconostoc mesenteroides biofilm formation (Badel et al. 2004.. 2009).. 2010). ship hulls and (marine) fish cages (Braithwaite and McEvoy. In this system. 2007. 2008c. In addition.. 2001. H. 2009). coating or impregnation of materials on various stages of biofilm development can easily be performed in microtiter plate model systems (Chandra et al. they may also pose a health risk to the users. 2007. 2009. Krom et al. the environment in the well of a MTP will change during the experiment (e.1. 1999.... 2001. De Prijck et al. proteins and DNA) results in a firmer attachment (Sauer. Biofilm formation is often considered the underlying reason why treatment with an antimicrobial agent fails and as an estimated 65–80% of all infections is thought to be biofilm-related. Finelli et al. Shakeri et al.b. Arias-Moliz et al. De Prijck et al. 2004). the lid can be transferred to a second plate. Pettit et al. this system also allows researchers to easily vary multiple parameters including the composition of growth media. Stepanovic et al.. presence or absence of shear stress and O2 and CO2 concentrations (Krom et al. In this review we present an overview of in vitro and in vivo model systems and discuss their advantages and disadvantages. 2005). De Kievit et al. 2007.. 2007. On the other hand. Ceri et al. Nelis / Journal of Microbiological Methods 83 (2010) 89–105 1.. Melchior et al. while the number of attached microorganisms increases) (ZoBell. Of particular relevance to human health is biofilm formation in drinking water reservoirs and distribution systems as these biofilms hinder the efficient operation of these systems. The multitude of advantages offered by these straightforward and (generally) user-friendly systems explains their widespread use. Following biofilm growth.. nutrients become depleted... 2007. Pitts et al. biofiltration (Cohen. when marine microbiologist Claude ZoBell described the so-called “bottle effect” (referring to the phenomenon that the number of free-living microorganisms in fresh sea water gradually declines when the water is kept in a glass bottle. 2. Staphylococcus carnosus and Staphylococcus xylosus (Chavant et al. 1999. 2007). 1999.. this presents a serious challenge (Costerton et al. but it has also been used to assess the influence of quorum sensing on biofilm formation (Tomlin et al..J. Pitts et al. 2010.. the top lid can either be transferred to a new microtiter plate containing media to allow regrowth. Parsek and Singh.90 T. Bardouniotis et al. Vandenbosch et al. 2005. 1999). 2010b. 2007). Ceri et al. 2007).c. Mah and O'Toole. signalling molecules accumulate. 2001. Wolfaardt et al. Imamura et al. Mowat et al... 2002.. Thirdly. MTP-based systems are closed (batch reactor-like) systems (Fig. Walker and Sedlacek. 2002. 2005. 2000.. Christensen et al. 1998) and to screen for the antimicrobial and anti-biofilm effects of various antibiotics. Sessile (biofilmassociated) cells are phenotypically and physiologically different from non-adhered (planktonic) cells and one of the typical properties of sessile cells is their increased resistance to antimicrobial agents (Donlan and Costerton. 2006).. 1). 2008. Uppuluri et al. humidity. Stewart and Costerton... 2007.. (1999) developed a variation of the traditional MTP model system. 2009b. wastewater treatment (Nicolella et al... 2000).. while the subsequent production of an extracellular matrix (containing polysaccharides. Peeters et al.. Ramage et al. Brackman et al. Amorena et al. Hall-Stoodley et al. in processes as diverse as bioremediation (Singh et al. 2006. 2007).. Hill et al. production of fine chemicals (Li et al. providing a habitat for pathogenic miroorganisms like Legionella pneumophila and Escherichia coli (Flemming. these pegs will be immersed in the media present in the wells of the 96-well MTP. 2007.... 2002). including heat exchangers... In vitro biofilm model systems 2. 2008. 2001). Another MTP-based commercially available method is the Biofilm Ring Test (BioFilm Control SAS) (Chavant et al. 2001. 1943) we have been aware of the fact that microorganisms are capable of living their life attached to a surface. O'Toole and Kolter. 12 or 24-well plate). 2007). Quave et al. Hamilton et al. 2009) and generation of electricity in microbial fuel cells (Rabaey et al. 2002. there are many (potential) applications of microbial biofilms. Stoodley et al. This technology has been used to study the kinetics of biofilm formation of Listeria monocytogenes. they provide the opportunity to perform a large number of tests simultaneously and this system is ideal for screening purposes (Niu and Gilbert. 1978... 2007)... 1992. 2003). Stepanovic et al.. coli. The “Calgary Biofilm Device” was introduced as a rapid technology to determine the antibiotic susceptibility of biofilms and it has been commercialized as the MBEC Assay (“Minimal biofilm eradication concentration” assay) by Innovotech. Secondly. 2003. 2005. 2001). 2006). Cells embedded in this matrix communicate with each other and show a coordinated group behaviour mediated by a process called quorum sensing (QS) (Zhang and Dong. As a consequence.g. MTP-based assays are fairly cheap as only small volumes of reagents are required.. 1985.. E. 2008a. 2003. biofuel production (Wang and Chen.. With this technology. In order to increase our knowledge concerning biofilm biology. to confirm that AI-2 based quorum sensing affects biofilm formation in Streptococcus mutans (Huang et al. 2005. Silva et al. to evaluate the effect of co-administration of antibiotics on Pseudomonas aeruginosa biofilms (Tré-Hardy et al. Shakeri et al.. 2003. etc). Biofilm formation can also have detrimental effects in industrial systems. the immobilisation of inert paramagnetic beads included in the culture medium during the formation of the biofilm is measured. In these systems.. unless the fluid is regularly replaced. but many of the models can of course also be used to mimick biofilm formation in other settings.. biofilm model systems to be used for the study of the often complex communities under controlled conditions are indispensable (Doyle. disinfectants. 2001. 2002).. to compare biofilm formation between . or the pegs can be clipped from the top lid and the biofilm biomass or the number of sessile cells present in the biofilm can be quantified using traditional viable plate counting or microscopic techniques. Ramage et al. incubation temperatures. Gabrielson et al. MTP-based model systems have been used to distinguish biofilm-deficient mutants from biofilmforming wild type strains (Heilmann et al. chemicals (including quorum sensing inhibitors) and plant extracts (Ali et al. in which there is no flow into or out of the reactor during the experiment (Heersink and Goeres. After the treatment. biofilms are either grown on the bottom and the walls of the microtiter plate (most commonly a 96-well plate) or they are grown on the surface of a coupon placed in the wells of the microtiter plate (most commonly a 6.

De Prijck et al. without changes in concentration over time. 2007). 2.. Many of these devices are custom-made and typically consist of a pump required to circulate the growth medium and/or cell suspension. Flow cells were first used in the 1990s to study environmental biofilm communities capable of biodegrading various recalcitrant xenobiotics (Wolfaardt et al. 2007) and P. aeruginosa biofilm formation (Nagant et al. Moller et al.. 2010). either following the “continuous flow stirred tank reactor” (CFSTR) approach or the “plug flow reactor” (PFR) approach (Heersink and Goeres. Krom et al.g. In addition.g. Nickel et al. the tubing at the inlet and outlet side is clamped off and the remaining cell suspension in the tubing at the inlet side is flushed out through the bypass.. the rate at which growth medium is added to the reactor) is identical to the effluent removal rate. 2003... 1995).. 2010. 2003. Once the devices are filled with the inoculum suspensions. the MRD has found application to assess the potential of antibiotic lock therapy for biofilm removal from colonised surfaces (Curtin et al. 2004. but many research groups use homemade MRDs. 2010) and to study early phases of P.e. aeruginosa (Haagensen et al. 2005. 2001) and has extensively been used as an “artificial throat” to study biofilm formation on laryngeal shunt prostheses (Leunisse et al. 2010a. pneumophila (Mampel et al. Uppuluri et al. the devices are flipped back.. 1997) and have since been used to study a wide range of microorganisms.. Percival et al.. A special type of plug flow reactor systems is the so-called “flow cell”. this is not the case for PFR systems. After the adhesion phase. Variations in this setup are possible.. a vessel with fresh growth medium connected by tubing to a chamber containing the material on which the biofilms are formed.2. 2. 1999.. While in CFSTR systems conditions are identical throughout the reactor..e.. 2007) and to study oral biofilms (see for example Foster and Kolenbrander. Ruiz et al.. 1). In PFR systems. planktonic cells are washed out of the reactor and only the sessile cells attached to a surface will remain and will be able to multiply... 2005. 2. the MRD can be attached to the effluent of a chemostat (Jass et al. Oosterhof et al. 1981. H. 2006.. with mixing (through diffusion) only occurring in the radial direction. 2006. These systems can be subdivided into two broad groups. the influent moves as a single “plug” in the direction of the flow (axial direction). the clamps are loosened and the pump is started to allow a continuous flow of the growth medium and biofilm development on the discs. 1994. Each port accepts a press-fit plug holding a disc on which a biofilm is formed (Fig. In CFSTR systems there is perfect mixing and the “feed rate” (i. 2008. Seidler et al.. 1995) or following the initial adhesion biofilms are allowed to mature in the absence of flow and the fluid flow is only switched on once mature biofilms are formed (Krom et al. 2006). 2009). 2009). Campylobacter coli and Campylobacter jejuni (Sulaeman et al. Schwandt et al. Foster and Kolenbrander. “continuous flow stirred tank reactor” (CFSTR) systems and “plug flow reactor” (PFR) systems.2. 1). surface-modified materials or materials impregnated with antimicrobial agents) under flow conditions (Nava-Ortiz et al. 1985. Krom et al. 2006. 2004.c.. 2003.. Flow displacement biofilm model systems In contrast to MTP-based systems.. Stovall. Raad et al. 2009). BioSurface Technologies) with glass chambers are particularly well-suited for real-time non-destructive microscopic analyses of biofilms. Modified Robbins device (MRD) The modified Robbins device (MRD) was developed by Jim Robbins at the University of Calgary to allow the reproducible and simultaneous formation of biofilms exposed to a fluid flow (McCoy et al.2. L. Foster et al.. 2006).b.e. 1.J..e. 2003) (Fig. the MRD is filled with a suspension of microorganisms and is flipped over to improve the adhesion of the planktonic cells to the discs. i. 1995). The MRD was found particularly useful to evaluate the effect of modified materials (i. 2) (Honraet and Nelis. 2004 and Palmer and Caldwell.1. These commercially available devices (e. Other PFR systems PFR devices other than the MRD have been used in numerous studies on microbial biofilms. 2009a. 2008. Honraet and Nelis. Nelis / Journal of Microbiological Methods 83 (2010) 89–105 91 Fig. 2010). As such a CFSTR system operates at steady state. van de Belt et al. . 2003) (Fig. Important conceptual differences between batch systems.. as environmental conditions change progressively through the reactor (Heersink and Goeres. Jass et al. and a second vessel for waste collection (e. including clinically relevant organisms like Neisseria meningitidis (Lappann et al. Coenye...2.. Busscher and van der Mei. i. When the dilution rate is higher than the doubling time of the microorganism(s) present in the reactor. Coenye et al. 2006). A MRD can be constructed from plastic or stainless steel and contains a number of individual ports in a linear array along a channel of rectangular cross-section. It is commercially available from several companies (including BioSurface Technologies and Tyler). flow displacement systems are “open” systems in which growth medium with nutrients is (semi-) continuously added and waste-products are (semi-)continuously removed (Heersink and Goeres. In a typical experiment..T. Staphylococcus epidermidis (Qin et al.

H. 2007). Goeres et al. 2005) (Fig. Coenye. Top left: single MRD unit. 3. albicans biofilm obtained on PMMA in the MRD (stained with calcofluor white). albicans biofilm.3. Each polypropylene rod can hold three removable coupons on which biofilms can form (Donlan et al. Left: two reactor vessels coupled to a bottle with fresh medium. Centers for Disease Control (CDC) biofilm reactor The CDC biofilm reactor (commercially available from BioSurface Technologies) consists of a glass vessel with a polyethylene top supporting eight removable polypropylene rods.. Nelis / Journal of Microbiological Methods 83 (2010) 89–105 Fig. albicans biofilm.. covered with C. Middle: PMMA disc loaded in MRD (view of luminal side). 2.. Setup of the CDC biofilm reactor. Top right: poly methyl-methacrylate (PMMA) discs ready to be used in the MRD. Bottom right: image of a C.92 T.J. 2. Right: poypropylene rod holding three medical grade silicone discs covered with a C.2. In this reactor. Bottom left: PMMA disc in plug. 3) and is oriented in such a way that the coupon is perpendicular to the rotating baffle (Buckingham-Meyer et al. . 2004. Setup of the MRD. the magnetic stirrer in the center of the vessel provides a Fig.

Nailis et al. but also to assess the damage inflicted upon the human cells by this process. 2009) and the simulation of encrustation of urinary catheters (Gilmore et al. 2004). 2004. 2007. the development of a biofilm on a surface is limited to a predetermined depth by mechanically removing excess biofilm.. Jayatilake et al. 2009). 1988. 1998. Hadi et al. 2006) and to evaluate the efficacy of various disinfectants (Stewart et al. epidermidis biofilm formation by bacteriophages (Curtin and Donlan. 2010). 2010). which allows rotation of the disc and consequently creates liquid surface shear across the coupons. 2007). 2009. 2004).. 24 identical biofilms can be formed simultaneously and as the reactor setup allows for the easy removal of discs during the experiment. 2010. albicans to adhere to the endothelium under conditions of flow. 2009.. 2005). Honraet et al. many in vitro cell-culture-based models have been described for the study of the interaction of human cells with bacterial biofilms. vancomycin and moxifloxacin (alone or in combination with other antibiotics) against S.g.g. 2010. Zakikhany et al. Thorough statistical analysis has indicated that the CDC biofilm reactor is a reliable experimental tool to study biofilm formation by a wide range of organisms (Goeres et al. 2000). albicans superficial tissue infection models have recently been reviewed by Jayatilake and Samaranayake (2010). Human cell lines that mimic the in vivo situation in vitro can be used for this purpose. Nevertheless. albicans suspension.. aeruginosa biofilms on airway epithelial cells (Woodworth et al.3.. aeruginosa biofilms (Cotter et al. 2003. Jayatilake et al.. indicating that these models can serve as in vitro biofilm models systems (Green et al. which is in contrast with observations obtained in previous assays under static conditions.. b) and Stenotrophomonas maltophilia biofilms formed on cystic fibrosis derived IB3-1 bronchial cells (Pompilio et al. 2007). 2005.. The relatively large reactor volume (500–1000 ml) makes the CDC reactor less-suitable to assess the effect of antimicrobial agents in the reactor itself. aeruginosa biofilms under high shear and continuous flow (American Society for Testing and Materials [ASTM] standard method E5262-07: Standard Test Method for Quantification of Pseudomonas aeruginosa Biofilm Grown with High Shear and Continuous Flow using CDC Biofilm Reactor). Well-studied examples of mucosal biofilms include those formed by Candida albicans on oral and vaginal tissues (Dongari-Bagtzoglou. albicans inoculated on RHE forms a biofilmlike structure on top of the epithelial layer.. Hentzer et al. based on reconstituted human epithelia (RHE) have been established and are commercially available (Schaller et al. this model system allows the formation of a large number of identical couponattached biofilms for subsequent testing of disinfection and cleaning procedures (“off-line testing”) (Buckingham-Meyer et al. These models not only allow to monitor microbiological biofilm formation. 2010). a situation mimicking the movement of the 93 tongue over the teeth (Peters and Wimpenny. as the commensal flora and normal humoral and cell-mediated immune responses are not included. Coenye. C. 2002.... These differences highlight the importance of selecting the most appropriate assay conditions. immortalized human microvascular endothelial cells (HMEC-1 cell line) are coated on glass slides mounted in a parallel plate flow chamber.. H. A flow displacement model often used in (but not limited to) the study of oral biofilms (especially dental plaque) is the constant depth film fermenter (CDFF). Zakikhany et al.. The CDFF has been used to study the effect of antimicrobial agents added to the reactor or released from the substrate of the biofilm (Lamfon et al. Xu et al.. Buckingham-Meyer et al..J.. Leung et al. 2007. biofilms of enterohemorrhagic E.. internalisation of microbial cells and damage to human cells can be determined by microscopy (e. In addition.. 2007). Rotating disc reactors have been used to study various aspects (including resistance) of Staphylococcus aureus and P. to assess reduction of S.4.. 2005) as well as to evaluate particular treatments for eradication of microbial biofilms (e. 2008). 2010). Examples are P. In this system.. The drip flow reactor is recognised as a suitable tool to grow P.. although this model system was recently used to evaluate the activity of high dose daptomycin. Pratten. 1998. The RHE model is very suitable to study the interaction between sessile C..g. 2005). 2008) and to investigate the abilities of the different morphological forms of C. 2002).. A motor drives the inner cylinder. interactions in multispecies biofilms (Komlos et al. The bottom of the rotating disc contains a magnetic stirring bar. an in vitro flow adhesion assay that mimics the conditions found within blood vessels was developed (Grubb et al. to evaluate the effect of powered brushing on the removal of biofilm plaque (Adams et al. In this assay. 2005)... In the CDC biofilm reactor. 2005. 2.. Teitzel and Parsek. . 2001. 2007).. Other C. Williams et al. providing liquid/surface shear.g. Nelis / Journal of Microbiological Methods 83 (2010) 89–105 continuous flow of nutrients (introduced in the reactor by means of a peristaltic pump) over the colonised surfaces (Goeres et al. aeruginosa biofilms and is included in ASTM standard method E2647-08 (Standard Test Method for Quantification of a Pseudomonas aeruginosa Biofilm Grown Using a Drip Flow Biofilm Reactor with Low Shear and Continuous Flow). 2009. Just like the drip flow reactor and the CDC biofilm reactor. This system consists of an inner rotating cylinder on which a number of slides are mounted. 2004. BuckinghamMeyer et al. 2005)... Dongari-Bagtzoglou et al. Other flow displacement systems In drip flow reactors. The CDC biofilm reactor is recognised as a suitable tool to grow P. 2009). 2007. 2005). Goeres et al. Lee et al. 2001). Pompilio et al. Hope and Wilson. Cell-culture-based model systems Biofilms not only are formed on abiotic surfaces but also on biotic surfaces. Nailis et al. 2001. biofilms are grown on angled slides continuously irrigated with small volumes of (inoculated) media (BuckinghamMeyer et al. aureus biofilms (ParraRuiz et al. Honraet et al. Using this model it was found that yeast cells adhered to endothelial cells in significantly higher numbers than did pseudohyphal and hyphal forms..7 mm diameter coupons. 2006). 2010a.... coli on HeLa cells (Kim et al.. 2009). Models of oral and vaginal candidiasis. The disc containing the coupons is placed in a reactor vessel and liquid growth medium (with or without antimicrobial agents) is circulated through the vessel while the disc is rotating (Buckingham-Meyer et al. 2009).. albicans can also cause more serious systemic infections (Grubb et al. Streptococcus gallolyticus biofilms grown on EA.g. 2003). Similarly. aeruginosa biofilms and is included in an ASTM standard method (E2196-07 Standard Test Method for Quantification of a Pseudomonas aeruginosa Biofilm Grown with Shear and Continuous Flow Using a Rotating Disc Reactor). This type of model system has mostly been used in studies pertaining to disinfection of drinking water systems (see for example Butterfield et al. as large amounts of reagents would be required to do so.... Drip flow reactor-grown biofilms have been used to study spatial heterogeneity in bacterial biofilms (Huang et al..T. Malic et al. 2. 2005... roughness) on biofilm formation (Morgan and Wilson.. this model system is particularly well-suited to study biofilm formation over time (e. For example. 2003). the rotating disc reactor is recognised as a suitable tool to grow P. thereby providing a low shear environment with dispersive mixing (Stewart et al. 2004. 2005. 2005) and to study the influence of surface characteristics (e... Other applications include the testing of materials coated with antimicrobial agents (Agostinho et al..hy926 endothelial cells and primary human umbilical vein endothelial cells (Vollmer et al. 2007. Szabo et al. Hu et al. albicans cells and epithelial tissue (Green et al. Jayatilake et al.. this model also has its limitations.. Declerck et al.2. 2004. 2007). Zakikhany et al. 2001. Schaller et al. subsequently the flow chamber is perfused with a C.. Other assays (including those assessing metabolic activity with XTT or measuring the release of lactate dehydrogenase) also allow to obtain a picture of the inflicted tissue damage (e.. The rotating disc reactor consists of a teflon disc designed to hold six 12. C. 2010). The final type of flow displacement biofilm model system to be mentioned here is the annular biofilm reactor (Lawrence et al.

2. Subcutaneous foreign body infection models Subcutaneous foreign body infection models have been developed in guinea pigs. Lazzell et al. a number of studies employing C. This model system has extensively been used to evaluate the effect of various antimicrobial agents against S. Eun and Weibel. In this model. Coenye. Caenorhabditis elegans model Studies using the C. 3. 2003). This system consists of microchannels (370 μm wide and 70 μm deep) integrated into a MTP. Lee et al. Silastic lumen-within-lumen catheters are inserted in the external jugular vein of rats and advanced into the superior vena cava. resulting in very low Reynolds numbers and highly laminar flows (De La Fuente et al.. albicans CVC infection.. the CVC are contaminated with bacteria. rabbits and rats (Table 1. by screening a transposon-insertion mutant bank it was shown that Y. 2009). H. Darby et al. aeruginosa) (Kadurugamuwa et al. 2010).J. 2010. the effect of liposomal amphotericin B lock therapy (Schinabeck et al.2. 2005. Fig. 2003.2. 2007). 2010. 2006). Vertebrate animal models 3.. 2006a.. although other approaches have also been reported (e. Kim et al. pestis genes involved in the synthesis of the polysaccharide matrix are required for this biofilm formation. Rupp et al. 2009).. Kokai-Kun et al. The C. Other microorganisms tested in the C. With this model it was possible to develop an E. 1999b). pestis.and N-linked glycoconjugates on the nematode's cell surface and consequently less bacteria will adhere to the surface of this mutant (Cipollo et al. This BioFlux system (commercially available through Fluxion Biosciences) allows the simultaneous analysis of 96 biofilms. albicans is also an important causative agent of CVCrelated infections (Donlan. while the length can vary from 5 to 40 mm.1.. Lorenz et al.. In vivo CVC models not only allow to study the efficacy of various antimicrobial agents (including antimicrobial lock therapy) and/or fundamental aspects of microbial biofilm formation in a realistic setting.. The mostfrequently-used models are the rabbit CVC model (Schinabeck et al. while the small size of the chambers allows the microscopic analysis of the biofilm at single-cell resolution.. ponies.. epidermidis and S. Alternative animal models to study bacterial adhesion to CVC include mouse models (S. including physiological flow velocities and low fluid-tocell volume ratios. Several animal models have been developed to study biofilm formation on CVC in vivo.. the use of mice also has considerable advantages. encoding a nucleotide sugar transporter of the Golgi apparatus. 2008).1. 2007. elegans came from the study by Darby et al. the environmental conditions can be controlled very precisely (De La Fuente et al. 2010). but is typically in the range of 50–500 μm wide and 30–250 μm deep. 2007) and also to evaluate the effect of the semisynthetic glycopeptide LY333328 against Enterococcus faecium biofilms (Rupp et al. Additional studies confirmed that the infection phenotype is the result of biofilm formation and identified genes involved in haemin storage and lipopolysaccharide biosynthesis as important for biofilm formation (Joshua et al. The first indication that bacteria can form biofilms in C. 2007. 2005. In addition. the RNAIII-inhibiting peptide and the cathelicidin BMAP-28) (Cirrioni et al. 2007. amphotericin B and chitosan) against C. A first animal model developed was a rat model designed to study biofilm formation by S. 4). This situation has some analogy with that in the proventriculus of fleas infected with Y. 2004).. mice. and helped to increase our knowledge regarding the role of specific adhesins in biofilm formation (Li et al.c. 2007).. These models have been used to study in vivo susceptibility and molecular response to fluconazole treatment (Andes et al. Richter et al. 2008).. coli biofilm on HeLa cells under conditions mimicking gastrointestinal tract infections. 2004). where the presence of a bacterial biofilm results in blockage of the gut and increased transmission of the pathogen (Darby. 3. 1999b). Following insertion.1.. 2004).. Ghiselli et al.94 T. Microfluidic devices The use of microfluidic devices to study biofilm formation and eradication is rapidly gaining interest.4. However.. Mukherjee et al. 2004) and the rat CVC model (Andes et al. Furthermore. albicans biofilms (Martinez et al. 2009. including the possibility to compare the data with data obtained in the widely used mouse model for haematogenously disseminated candidiasis (tail vein infections).. Ding et al. elegans have specifically dealt with microbial biofilm formation. 2007) and Xenorhabdus nematophila (Drace and Darby. epidermidis (Ulphani and Rupp. 2008).. elegans model system usually focus on virulence as such (i. aureus and S.. C.2.. 3. In this study it was shown that Y. Fabrication of microfluidic devices for biofilm formation often requires photolithography. 2007). 2003.. elegans was demonstrated. Nelis / Journal of Microbiological Methods 83 (2010) 89–105 2. In both studies the importance of the extracellular matrix for biofilm formation on and/or in C. 2008) and a rabbit model (S. disinctin and protegrin IB-367. Joshua et al. The deletion of srf-3 results in reduced levels of O. determining whether infection results in reduced survival of the worms) and/or on the effect of particular chemical compounds on this survival. Rupp et al.b. and the effect of various antifungal compounds (including caspofungin. aureus.b) recently reported on the development of a microfluidic co-culture model. albicans biofilms on CVC (Lazzell et al. hamsters.. In vivo biofilm model systems 3. 2004). aureus biofilms (including linezolid. but also to investigate the dissemination of the microorganism to various organs. An interesting C. Conant et al. (2009) developed a mouse model for C. elegans model include staphylococci (S. Central venous catheter models Microbial biofilm formation on central venous catheters (CVC) can result in considerable morbidity and mortality (Donlan. These devices offer the possibility of developing biofilms under physiologically relevant conditions. from an inlet well (containing fresh medium) to an outlet well (containing spent medium).. P.1–50 μl/min).. ciprofloxacin.. the antimicrobial peptides citropin 1.. Recently.. 1999. Flow rates are typically very low (0.e. (2010a. Richter et al. a process in which a pattern is transferred from a mask onto a thin layer of photosensitive polymer and then onto the surface of a substrate (Weibel et al. In addition. 2008) and several animal models were developed to study biofilm formation on CVC in vivo. 2009.. Shuford et al. and it was suggested that this matrix may play an immunoprotective role during infection (Begun et al..g.. albicans biofilm formation. 2001). 2010. 2007. Giacometti et al. This model has been used to demonstrate the efficacy of caspofungin both for the prevention and the eradication of C. elegans gene identified this way is srf-3. 2009. Although the catheterisation in these smaller animals is more challenging. elegans model system also allows to identify host genes required for bacterial adhesion. 2007) as well as to obtain a better picture of changes in gene expression in in vivo biofilms over time (Nett et al. they have been used to increase our understanding of the molecular basis of C. 2008. 2007).. aureus) (Fernández-Hidalgo et al. Pneumatic pressure pushes fresh medium through the microchannel. vancomycin. 2004. pestis biofilm formation in the mouth and on the head prevented C. aureus) (Begun et al. epidermidis (Li et al. 2009)... In . This rat model was also used to study factors important for in vivo biofilm formation and experiments with the rat CVC model confirmed the importance of polysaccharide intercellular adhesin/hemagglutinin and the ica locus in S. Höflich et al.. Recently a “well plate microfluidic” device was described that allows high-throughput screening of biofilms in a microfluidic device (Benoit et al. elegans feeding. (2002) with Yersinia pestis. in which a microbial biofilm can be developed in the presence of an epithelial cell monolayer. Janakiraman et al. The size of the channels in these devices are model-dependent...

epidermidis S. (1995) Christensen et al. 2006). Fluckiger et al. Yasuda et al. 2004). ica−mutant) S. ica−mutant) S. 2006) and hence these models do not mimic perioperative infections (Ricicová et al. Patrick et al. In the latter case. Voermans et al. epidermidis. in vivo gene expression (Goerke et al. S. aureus S.. P. 2005). 1992. Rupp et al.. 2009). particularly well-suited to study the effect of (modified) substrates on biofilm formation (Nakamoto et al. Nelis / Journal of Microbiological Methods 83 (2010) 89–105 95 Table 1 Selection of subcutaneous foreign body infection models... 2010) or the animals can be infected post-implantation (Table 1). polymethylmethacrylate. Fluckiger et al. (1993) Van Wijngaerden et al. Morikawa et al. (2005) Nair et al. dltA−mutant) S. 2004).. aureus biofilms (Morikawa et al. a foreign body is inserted in subcutaneous pockets and a biofilm is allowed to develop on the implanted material. 2008. The tissue cage model has been used to study immune responses from the host (Zimmerli et al. aureus E. 1991. 1993. Gagnon and Richards. 4. 1995. (1982) Fluckiger et al. 2008. 2004. 1993) as well as the effect of arbekacin (alone or in combination with fosfomycin) on S.. 1983. S.. Engelsman et al. 2005... H. Voermans et al. albicans (WT. 2003.. aureus (WT. An advantage of these model systems is that they allow to establish a chronic infection which can be followed for longer periods (up to 6 months) (Gallimore et al.2. 1999a. 2005. aureus S.. aureus A. 2006. (2010). 2010).. (2008) Ricicová et al.. aureus (WT.. aureus (WT. epidermidis. Following implantation. epidermidis S.. albicans) have also been included. Actinomyces radicidentis.. (1992) Roehrborn et al.b) Kristian et al. (2004).3.T.b. Intraperitoneal foreign body infection models have been used to study the effect of antibiotics against bacterial biofilms (S. (2009) Voermans et al. Coenye. In the carboxymethylcellulose (CMC) pouch infection model. these models. the efficacy of particular antimicrobial agents (Murillo et al. albicans according to the model described by Ricicová et al. aureus S. 2004.. followed by the simultaneous injection of CMC and infecting organisms (Morikawa et al. Yoshikawa et al. epidermidis (WT. Kucharikova et al. coli P. In these model systems. the animals are allowed to recover from the surgery and subsequently bacterial suspensions can be injected in the tissue cage. aureus (WT. aureus) may not require immunosuppression. 1993... 1991. albicans) and for these microorganisms the use of immunosuppressive drugs (e. aureus and/or S. aeruginosa and C. 1994. Subcutaneous foreign body infection model systems are Fig. and to determine the role of particular genes in establishing biofilm-associated infections (Kristian et al. Gagnon and Richards. but other microorganisms (including E. (1999) Yoshikawa et al..J. Kristian et al. animals are injected with microorganisms several days. Materials can either be contaminated prior to implantation (Chang and Merritt 1994. A major advantage of this model is that microbial cells can easily be recovered from the fluid inside the tissue cage without the need for explantation (Handke and Rupp. (1999) Yasuda et al.. 1992. aeruginosa) (Carsenti-Etesse et al. Intraperitoneal foreign body infection models In many in vivo studies. The tissue cage model is based on the subcutaneous implantation of so-called tissue cages. aureus S. Roehrborn et al. These perforated cylinders (often made out of teflon) contain glass beads or other materials to increase the available surface area for biofilm formation (Kristian et al. Animal Reference Implantation ofa Micro-organism Guinea pig Zimmerli et al. Yoshikawa et al. no immunosuppression was used unless otherwise mentioned. 1993).. While studies with relatively virulent organisms (including S. S. 1995) as well as to evaluate the importance of quorum sensing in P. epidermidis. the inflammatory response associated with surgery may inhibit biofilm formation in less virulent organisms (e. 1991). 1999). dexamethasone) may be recommended (Ricicová et al.... Gallimore et al. (2006) Rediske et al. (2004) Fluckiger et al. materials can be precolonised or microorganisms can be injected intraperitoneally following implantation (Buret et al. 3. S. aureus. (1999a. coli.. aeruginosa S.g.. aeruginosa biofilms (Yasuda et al. an air pouch is created on the back of an animal (typically a rat) by the subcutaneous injection of air. (2008) Engelsman et al. 1999. (2010). C. 2005. radicidentis C. 2006)... Ricicová et al. epidermidis.g. Van Wijngaerden et al... P. (2003) Kristian et al. 1995. 2003. Nair et al.. PIA/HA mutant) S. epidermidis Hamster Mouse Pony Rabbit Rat a Nakamoto et al. aeruginosa biofilm persistence (by using mutant strains and quorum sensing inhibitors) . (1983) Patrick et al. 1982)... epidermidis S.. ica−mutant) S. Van Wijngaerden et al. Subcutaneous implantation of polyurethane catheter segments colonised with C.. biofilms are formed on biomaterials inserted in the peritoneal cavity of rabbits or mice. Owusu-Ababio et al. 2001). weeks or even months after implantation of the foreign body (Christensen et al. (2010) S. (2005) Nejadnik et al. aureus S. This model has been used to study the effects of antibiotics (ofloxacin and clarithromycin) on P. (1995) Rupp et al.. (2005) Chang and Merritt (1994) Tissue cage (teflon) Tissue cage (teflon) Stainless steel. Espersen et al. titanium (both sterile and colonised) Steel wire Plastic catheter Teflon catheter Teflon tubes Teflon catheter Tissue cage (teflon) (immunosuppression) Tissue cage (teflon) Tissue cage (teflon) Colonised silicone discs Colonised surgical meshes Tissue cage Infected polyethylene discs Carboxymethyl cellulose pouch Infected polyurethane catheters Carboxymethyl cellulose pouch Tissue cage (teflon) Colonised polyurethane catheters (immunosuppression) S. 2004). epidermidis. 2010. Nejadnik et al.. various mutants) Inserted material is sterile prior to insertion unless otherwise mentioned. Most studies have been carried out with S.

2009. Finally. 1993. A modified agarbead model was developed by Sawai et al. In the rabbit model of acute sinusitis (Johansson et al. 1979). Winnie et al. To study catheter-associated urinary tract infections.. or the experiments can be carried out . a catheterized rabbit model was developed in Bill Costerton's group and subsequently used to evaluate the effect of various antibiotics on E.. several animal models have been developed to study these biofilm-associated infections. 1987) and rhesus monkeys (Cheung et al. 2008). 1989). instillation of P. 1993.g. 2009). 1981... Coenye.. (2000) that P. nose and throat (ENT) infections (Vlastarakos et al. followed by inoculation of S.. A second biofilm-associated ENT disease is chronic (rhino-)sinusitis (Cohen et al. 2007). 2005. aeruginosa..2. 1999)... A first model was described by Satoh et al. Cash et al. This model was subsequently used to demonstrate the importance of biofilm formation and matrix production in the development of urinary tract infections (Nickel et al.. 2009). 2007). (1995) used a rat model of renal infection in which glass beads covered with a bacterial biofilm were inserted in the bladder. Haraoka et al. In this model. in which a polyethylene tube is placed in the bladder transurethrally (without surgical manipulation) (Kadurugamuwa et al. aeruginosa and subsequently inserted in the trachea (through the mouth) (Nagata et al. 2006. 2007) and the effect of an antimicrobial peptide derived from the innate immunity protein LL-37 (Chennupati et al.. Starke et al. This polethylene tube can be colonised with pathogens before implantation.. 1987. 2007) and to assess the efficacy of topical antimicrobial agents (Le et al. 2009. Several animal models for chronic respiratory tract infections have been developed. Reid et al. In this model.. While high concentrations of these compounds result in the eradication of P. aeruginosa respiratory tract infection. 1987). This system has been used to evaluate the efficacy of antibiotics (including linezolid.. coated (either with RIP alone or with RIP combined with teicoplanin) and uncoated stents were surgically implanted in the bladder of rats.. 2005.. 2001). aeruginosa in the maxillary sinus allowed for the visualisation of biofilms with a pronounced three-dimensional structure from the sinus epithelium (Perloff and Palmer. 2005). Vlastarakos et al. 2003). Grimwood et al. More recently a non-surgical approach in rats and mice was developed. aureus in the bladder (Cirioni et al. a piece of a plastic intravenous catheter) is precoated with P. aureus infections.. followed by clamping of the urethra.. BALB/c) or “CF mice” (CFTR−/−) can be used (Hoffmann et al. Palmer 2006). microcolonies embedded in matrix material) and that CF sputum contains P. This model has mainly been used to study microorganisms involved in respiratory tract infections in CF patients. 1988).. 2008). 2007. 1992. 1976.2. 2004.. Kurosaka et al.4. 2009). these infection models are particularly well-suited to study the effect of modified peritoneal dialysis catheters (e. 3. Olson et al.5. Kukavica-Ibrulj et al. 2006). Harvey and Lund. rabbits and mice. 2007).. Juhn et al. 2005. Cheung et al. 1992. 2009). 2003. use of the chinchilla model led to an unambiguous establishment of a causal relationship between otitis media and biofilm. 1997). 2007)... 2001). 3. inoculation through the nasal or intratracheal route often only results in a transient pulmonary colonisation in which bacteria are rapidly cleared (O'Reilly. Sokol. 2001. The same model was later also used to demonstrate the ability of Streptococcus pneumoniae to form nasopharyngeal and middle-ear mucosal biofilms following transbullar inoculation (Hoa et al. Results from this study clearly indicate that RIP increases the efficacy of teicoplanin against surface-associated S. 1979.. In this model. rats were intravenously injected with agar beads (appr. 2002. followed by inoculation of the bladder with a defined number of microorganisms (Kadurugamuwa et al. 1997. animals are infected intratracheally with a small volume of a planktonic culture of an alginate-hyperproducing P. In addition.. 1981). confirmed the hypothesis that respiratory tract infections in CF patients are biofilmrelated (Costerton et al. These agar beads were subsequently transported into the pulmonary circulation to become entrapped in the pulmonary microvasculature. cats (Winnie et al.96 T. followed by transvesical inoculation with Proteus mirabilis. 2000). bacteria embedded in agar or agarose beads are used to establish pulmonary infections in laboratory animals. 1993) were used. Dohar et al. Ear. levofloxacin and erythromycin) against in vivo P. 2006. 2006).. Nelis / Journal of Microbiological Methods 83 (2010) 89–105 (Christensen et al. Finelli et al. Data from this experiment showed that regular treatment with mupirocin resulted in maximal reduction in biofilm surface coverage with sustained effects during the 8 day follow-up period (Le et al... 2009). In contrast... aeruginosa cells recovered from sputum of cystic fibrosis (CF) patients are in a biofilmlike structure (i. aureus infections. To this end either wild type mice (e. developed to mimick infections observed in diffuse panbronchiolitis (Yanagihara et al. 2002.. infection is restricted to the lungs and this model has been used to study the efficacy of various (combinations of) antibiotics (including clarithromycin. In a first set of models. aeruginosa biofilms.. 1995....e. 2009. and these can roughly be divided into three groups. More recently a sheep model was developed to study the role of bacterial biofilms (S.. but in later studies guinea pigs (Pennington et al. 2008. H. Urinary tract infection models Both surgical and non-surgical urinary tract infection models have been developed in rats. Two animal models were used to study the involvement of bacterial biofilms in this disease. aeruginosa and/or Burkholderia cepacia complex organisms (Bernier et al. aeruginosa biofilms in this model. 2007).. 2002. coli biofilms developing on these catheters and on adjacent tissues (Morck et al.. infections are characterised by consistent numbers of organisms and a relatively constant immunological response (Sokol. by allowing direct visualisation of the biofilm on the middle-ear mucosa (using SEM and CLSM) following transbullar injection with Haemophilus influenzae (Ehrlich et al.. nose and throat infection models As biofilms are increasingly recognised as important in ear.. a plastic tube (e... Mikuniya et al.g.2. (1997). In vivo models have also been used to study the efficacy of other coated catheters in preventing urinary tract infections (see for example Darouiche et al. 1977). 2007). aeruginosa strain. Respiratory tract infection models The observations by Singh et al. 2008).g. 200 μm in diameter) containing various bacterial pathogens (including S.. Meers et al. Yanagihara et al. 2008. 2003.. A second type of model is a murine model of chronic P. 2008.. 3. Orlando et al. quinolones and novel carbapenems) (Kihara et al. This model has also been used to study the in vivo efficacy of topical tobramycin (Chiu et al. Kim et al. proinflammatory and ciliotoxic effects on sinus mucosa may limit their application (Chennupati et al. Nickel et al. in agar-bead based animal infection models. The use of these encased bacteria results in proliferation and allows for the establishment of a chronic infection with histological damage similar to that observed in patients with cystic fibrosis or chronic obstructive pulmonary diseases. Cieri et al. (2005) developed a similar model in cynomolgus monkeys for the study of biofilm formation by P. Most attention has been paid to otitis media (middle-ear infection) and the chinchilla has emerged as the animal of choice to investigate this biofilm-associated infection (Bakaletz.. 1994. aureus) in rhinosinusitis (Ha et al. 2005). or a sterile segment can be implanted. (1984) in which a zinc disc was implanted in the bladder of rats.. Yanagihara et al. Sokol. Hachem et al. With this approach.. 1989. To evaluate the effect of coating urinary stents with RNAIII-inhibiting peptide (RIP) against S.J. aureus). 1982).6. mice (Starke et al. This model was originally developed using rats (Cash et al.. 2008. A similar model was developed in rabbits (Fung et al... aeruginosa quorum sensing molecules in the same ratios found in in vitro grown biofilms.. and the majority of studies were carried out with P.. 1991. 1982). A final model is the pulmonary infection model without artificial embedding (Hoffmann et al. Developed in the mid 1970s (Giebink et al. 2008. Pennington et al.. Post.

However. Treatment of these infections is particularly difficult and surgical interventions are often required (Brady et al. albicans and after a certain period the vagina was excised and investigated. 2006). red blood cells and plasma) found in the wound bed (Sun et al. 2009).. The medium used in this model (Bolton broth with 50% heparinised bovine plasma and 5% freeze–thaw laked horse red blood cells) was selected to contain the major host factors (damaged tissue. Histopathological examination of the rat oral mucosa following infection indicated that the model accurately mimics acute human denture stomatitis (including the presence of fungal invasion and neutrophil infiltration). and this effect was even more pronounced when the treatment was combined with intraperitoneal injections with . Graftskin contains human neonatal dermal fibroblast in a collagen matrix over which a cornified epidermis (derived from neonatal keratocytes) is produced..1% peptone) and a collagen matrix. aureus resulted in the formation of biofilm-like structures 48 h postinoculation (Davis et al. albicans biofilms on vaginal epithelial cells (Carrara et al. (2010) used a constant depth film fermentor to form multispecies biofilms consisting of wound isolates. osteomyelitis is induced by instilling a bacterial suspension followed by implanting a foreign body in the bone or by the direct implantation of a contaminated foreign body. 3. Following inoculation. Carrara et al. aureus. and to mimic this in the animal models. (2010) recently described an in vivo C. Campoccia et al. albicans cells. 2007). 2005). an orthodontic wire was threaded across the hard palate of immunosupressed rats and then secured between the cheek teeth. Experimental inoculation of partial thickness wounds in pigs with S. 2009).. Models were developed in dogs (Fitzgerald. Costerton. Biofilms formed in this model had a structure very similar to biofilm structures observed in vivo. it also allows to study anaerobes (which can constitute a considerable part of the population in chronic wounds) (Sun et al. Although in se an aerobic model. Other in vitro models have also been reported.2. It has been hypothesised that these biofilms and the lack of their elimination by leukocytes are responsible for the chronic nature of the infection (Bjarnsholt et al. rabbits (Eerenberg et al.. The role of quorum sensing in S. In a first group of models.. Osteomyelitis is a chronic disease. Hill et al.b. This model allowed to rapidly (24 h) grow multispecies biofilms (containing the three bacteria typically observed together in chronic wounds: P. it should not come as a surpise that several in vivo wound models were developed. To study the efficacy of conventional antibiotic treatment as well as of the potential beneficial effect of antibiotics delivered in situ in high concentrations. acrylic denture material was applied over the palate and. (in press) developed a murine model for vaginal candidiasis: following administration of 17-β-estradiol (3 days prior to inoculation) mice were intravaginally inoculated with C.... 2010. Del Pozo et al. 2008). Coenye. (2008) created pressure-induced ischemic wounds in rats and inoculated these with P. James et al. (2010) used rats in which pseudoestrus was induced by estradiol hexa-hydrobenzoate (1 week before inoculation). aeruginosa. 2008. Two recent studies report on the development of animal models for vaginal infections. Enterococcus faecalis and S. aeruginosa or S. aureus wound infections was investigated in a mouse model (full-thickness wound) by treating infected wounds with locally administered RIP (either as such or in a soaked wound dressing).J. The microorganism included in the vast majority of these studies is S. 1993. Gracia et al.. 2008). 2008). This model has the advantage that it is histologically similar to human skin and provides a controlled environment similar to the one encountered in in vivo wounds (Charles et al. Wound biofilm models 4. rats (Gerhart et al. aureus biofilms were grown on cellulose support matrices. the hard palate was inoculated with 107 C. implants are left in place for longer periods (weeks to months) (Handke and Rupp. Shirtliff et al. aureus) in a compact and relatively inexpensive model. H.. after solidification of the device. In vivo models As host-derived factors play an important role in chronic wound infections. Microscopic analyses (electron microscopy and CLSM) clearly indicated the presence of C. Harriott et al.. This model has been used to evaluate the efficacy of the octadecapeptide novispirin G10 (Song et al. a tissue engineered skin equivalent (Charles et al. The most advanced in vitro wound model used to study biofilms is based on Graftskin. In vitro models Recent evidence has shown that also in chronic wounds (e. Treatment with mupirocin or a triple antibiotic ointment had limited effect on these biofilms. in press). aureus biofilms underneath various wound dressings to evaluate their effect. aeruginosa or S. 2009). 2008).. 2006. as the role of biofilms in these chronic infections has only recently begun to be understood (Bjarnsholt et al.8. Damke et al. 2008. 2001. In this model. (2007) and Thorn and Greenman (2009) described model systems (including a flat-bed perfusion growth chamber) in which P. Subsequently the authors were able to demonstrate the presence of quorum sensing molecules in the infected wounds 3 and 7 days post wounding.. Other models Nett et al.7. The model has been used to study the effect of treatment of wounds with antimicrobial agents and it was shown that microorganisms grown in this model are much more resistant than planktonic cells (Sun et al. Nakagami et al.. 2002a. aeruginosa. 1983.2.2. the presence of biofilms has only been confirmed in a few in vivo models. 4. the infection was monitored for up to 3 weeks. aureus or P. Petty et al. Brady et al. 4. pressure ulcers) microbial biofilms (often polymicrobial) can be found.. without implanting a foreign body (Mader and Wilson 1983.. indicating the presence of a “wound-like” environment... Harriott et al.1. 1998.g. aeruginosa virulence (Hoffmann et al.. 2005). Subsequently. Thorn et al. 2008). At present it is unclear whether these systems – in which a fluid flow is present and/or marked shear forces are applied – have an added value over more conventional in vitro wound biofilm models. (2010) who developed a model in which biofilms can develop in the absence of a solid surface but in the presence of simulated wound fluid (containing 50% fetal calf serum and 0. Monzón et al.. 2002). Nelis / Journal of Microbiological Methods 83 (2010) 89–105 using wild type rats (Song et al. 2005) as well as to demonstrate the potential usefulness of azithromycin to attenuate P. 2010. Treatment with RIP-soaked wound dressing significantly reduced the bacterial load in the wounds. 2008. several animal models were developed (Crémieux and Carbon 1997). 2009).. USA) and was aptly named the Lubbock chronic wound biofilm model (Sun et al. James et al.T. planktonic cells are administered in the intramedullar cavity of the tibia of rabbits. Lipp et al.. diabetic foot ulcers. A somewhat similar model was described by Werthén et al. albicans biofilm denture model. On the Graftskin specimens full-thickness wounds can be made and these can be inoculated with typical wound pathogens like S.. 3. Osteomyelitis infection models Biofilm infections associated with orthopedic prosthetic devices are a major complication of orthopedic surgery and have a considerable impact on patient morbidity and mortality as well as on cost (An et al. aeruginosa. 2008. 1994) and mice (Li et al. Dowd et al.. 2009).. 2006.. The first chronic wound biofilm model was 97 developed at the Medical Biofilm Research Institute in Lubbock (Texas. 2008). In a second group of osteomyelitis infection models. 1985). 2008). (2010) used a colonydrip flow reactor model to grow P.

by using low-melting agarose) prior to further investigations (Pittman et al. resazurin staining) or the amount of extracellular polymers in the biofilm matrix (e. aureus in an osteomyelitis infection model in mice (Li et al.g. 2006. Visualisation techniques and advanced imaging techniques have recently been surveyed by Neu et al. 2010. when biofilms are grown on discs or pegs placed in the wells of the microtiter plate) (Heersink. Coenye. immunological factors) are missing. Using a similar model. it has been shown that the passage of a liquid–air interface (e. we want to stress the importance of standardising the techniques used to recover biofilm-grown cells from the surface. the reader is referred to Sjollema et al. aeruginosa infected acute wounds.. In most flow displacement biofilm model systems biofilms are formed on multiple coupons in the same reactor vessel and as a consequence only a single organism (or a single community) can be tested per run. by S. 2005). conventional plating is labour-intensive and time-consuming and does not allow to recover viable but non-culturable organisms. Schierle et al. Kanno et al. 2001.g.g. crystal violet staining... (2010).. (2010) described a rat skin model (full-thickness wounds) for P. 5. epidermidis on subcutaneously implanted catheters in mice (Vuong et al. coli (in combination with non-luminescent Bacteroides fragilis) in localised bacterial peritonitis in rats (Sharma et al. are generally less labour-intensive. 2005). staining with dimethylmethylene blue). For example. 2000b). multiple organisms and/or treatments can be included in a single run) and as such are very well-suited for screening purposes (especially when they are combined with rapid methods to quantify biofilm formation).. The same is true for “online” treatments in the reactor vessel: only treatment with a single product or a single combination of products is possible and only a single type of modified material can be tested per run.. the extent of biofilm formation can be measured in a variety of ways. do not require specialised equipment and are cheaper. 2005). and the strain being studied.. Nelis / Journal of Microbiological Methods 83 (2010) 89–105 teicoplanin (Simonetti et al.g. The combined use of flow-cell technology and confocal laser scanning microscopy (CLSM) has been very useful to study various aspects of microbial biofilm formation. 2008). 2010) are likely to futher increase our knowledge of biofilm biology in various model systems. Ramanan et al. In vitro “flow” systems have a lower throughput. including techniques to determine the total biofilm biomass (i. An important conclusion from the latter study was that. the number of viable sessile cells only (e. It is obvious that MTP-based systems permit a higher throughput.e. the technical developments in and increased use of nondestructive chemical analytical techniques. particular attention has been paid to the development of non-destructive “real-time” monitoring of biofilm development in various biofilm model systems.. this may require stabilisation (e. by S.g. 6.. and that this is much more important than obtaining high efficiencies for control biofilms. Pitt et al. The development of strains engineered to be constitutively bioluminescent (mostly by insertion of the Photorhabdus luminescens luxCDABE operon). 2003). the suspended cells can be diluted and plated. and their advantages and disadvantages. (2009. (2010). substratum coverage and surface to volume ratio (Heydorn et al. 2008). sessile cells can be removed from the surface by scraping and/or sonication (e.. in order to allow the comparison of biofilms grown under different conditions and/or following different treatments.g. 1993..g. (2005). While biofilms could be observed from 8 h post wounding until 7 days.g. Cellculture-based model systems often offer an elegant solution to some of the shortcomings of the other in vitro systems. (2010) and Hannig et al. aureus mutants resulted in improved wound healing kinetics. but few of these standardised systems are commercially available and developing and maintaining these model systems is not always straightforward. highlighting the need for standardised procedures (Gomez-Suarez et al. matrix and both living and dead cells. Various alternative techniques have been developed for the quantification of biofilms grown in microtiter plates. However.e. aureus on subcutaneously implanted surgical meshes (Engelsman et al.. These systems allow “multiplexing” (i. it is critical that efficiencies of harvesting and disaggregation are similar for the different biofilms. allows for the direct continuous monitoring of biofilm infections in vivo. 2009. aeruginosa and S. aeruginosa and P.g.. b)) to quantify three-dimensional biofilm image stacks has been of tremendous value in quantitative biofilm research. 2010). roughness.. These non-culture-based methods have been discussed in detail by Honraet et al. aureus or S. In recent years. Sandt et al. 2010). (2009) demonstrated that treatment of S. e. aureus on subcutaneously implanted teflon catheters in mice (Kadurugamuwa et al. when biofilms are grown on the bottom and the walls of the microtiter plate) or they can be detached from the surface by applying cycles of sonication and vortexing (e. (2008a) and by Toté et al. When studying “fragile” biofilms that dissociate easily from the surface during handling. by P.. 2010). aureus on cardiac tissue in rats (Xiong et al. Hamilton et al. 2009). FTIR spectromicroscopy (Holman et al. However.98 T. e. Features calculated by COMSTAT include thickness distribution and mean thickness. 2001). Subsequently. infection did not appear to delay wound healing in this model. their potential applications.. Peeters et al. 2009) and magnetic resonance imaging (Neu et al. While these systems have the advantage of incorporating the important aspect of fluid flow in the setup. 2003). Before providing a brief overview of the different approaches available. The use of dedicated software (e.. development and resistance in vitro and was recently reviewed by Pamp et al. (2000a. (2009) recently described methods that can be used to check the validity of harvesting and disaggregating biofilm cells from surfaces. In addition. As the extent of the detachment depends on the surface. epidermidis infected wounds with RIP speeds up wound healing and restores reepithelialisation to levels observed in uninfected wounds.J. other aspects encountered in the in vivo situation (e. COMSTAT. Syto9 staining). the speed of the air bubble. Which model system to choose? The goal of this review was to present an overview of commonly used in vitro and in vivo biofilm model systems. combined with the development of highly sensitive imaging procedures. are more labour-intensive and require specialised equipment and technical skills. developed by Heydorn et al. For a recent review on this topic. mirabilis on polyethylene tubes implanted in the bladder of mice (Kadurugamuwa et al. H. “offline” visualisation is used in which substrates containing the biofilm are removed from the biofilm reactor before being studied under the microscope. Pätzold et al.. they often require larger volumes of media and/or other reagents. They also showed that wounds infected with biofilm-deficient S. including Raman spectroscopy (Ivleva et al. Finally. 2007). 2008) and by E. Sharma et al. 2008. (2009). the influence of detachment artifacts introduced by rinsing and dipping on the final result is unpredictable.. by P. Quantification and visualisation of biofilms grown in various model systems Following biofilm growth in an in vitro or in vivo model system. The number of culturable cells can be determined using conventional plate count methods and to this end. by S.g. In cases where real-time monitoring is not possible (either due to the incompatibility of the biofilm model system with the microscope and/or practical problems associated with the use of the visualisation equipment for longer periods of time). by S. . an air bubble) can result in considerable detachment and often-used procedures like dipping and rinsing to remove loosely-adhered microorganisms may consequently result in artifacts (Gomez-Suarez et al. they are closed systems and the conditions in which biofilms are formed and/or tested are more often than not very different from the in vivo situation.

. P.. the use of larger animals (sheep. J. M. Overall. 2002.. Heersink. 6023–6031. Which biofilm model system is selected not only depends on the preferences of the investigators and the resources available. pp. Appl. extracellular lipase activity and expression levels of genes encoding factors involved in adhesion and virulence (Fig. Wilkoff.. D. 1063–1082. 2008.. Hernandez-Yago. 2007). while the highest reductions were obtained in the static biofilm model. 2003. Clin. Antibiotic susceptibility assay for Staphylococcus aureus in biofilms developed in vitro. rats. Olson. Single and combination antibiotic susceptibilities of planktonic. UK. 2. Oteiza. 2006). 72. Bioresour. H. S. Lett. 2004. 2001. 1999. 263–267.. Winston. (Eds. 5. left axis) and fraction of filaments (red line. Sifri. . the expression of other virulence genes was strongly influenced by the biofilm model system (Fig. Vaccines 8. (2007). B. J. 59–85... Technol. Infect. but it is more distantly related to the natural pathology (O'Callaghan and Vergunst. aeruginosa and S. Heleen Nailis and Elke Peeters for valuable contributions.. Chinchilla as a robust.D. M.. Ferris. Cambridge.. Devine. goats.. B. Agostinho.J. Ceri. Sci. Khambaty. M. Immun. 5). 44. right axis) in 48 h old C... There are only a handful of studies in which a meaningful number of biofilm model systems were systematically compared..). M. D.. were compared.D. Wazni. Kristof De Prijck. M. Antimicrob.T. While the overall number of culturable cells recovered from these biofilms was similar. Mack. PLoS Pathog. J. M. aureus biofilms grown either in the CDC biofilm reactor... Gardarin. O...E. 2009. An. H.A. J. F. Friedman. 2010). M. R. small animals which are easy to infect and are relatively economical. J. but most importantly on the questions being addressed. K. J. James. C. J. C.. J. FEMS Microbiol. González-Rodríguez.. Endod. Ali. throughput and ethics. Marchillo. Reductions observed for biofilms grown in the drip flow reactor were intermediate.. P. The authors strongly recommend that efficacy of disinfectants against biofilms should be tested with a laboratory method producing a biofilm under conditions similar to the environment where the disinfectant will be applied. L.. Am..J. Eradication of Enterococcus faecalis biofilms by cetrimide and chlorhexidine. reductions were lowest in the CDC biofilm reactor. A. Citron. Medical Implications of Biofilms.H. W. especially when a relatively high number of animals is required (An et al.. Monzon. 1887–1893. G. Interestingly. 2007. Ramotar. 2003). Number of colony forming units (blue bars. K. a drip flow reactor or in a static biofilm. Ferrer-Luque. Coenye.. Albrecht. Fig. Costerton. 2009.. Calderwood. Leiva.. 364–370. Alabart... Investigating the suitability of the Calgary Biofilm Device for assessing the antimicrobial efficacy of new agents. R. References Aaron.J. 2003. In: Wilson. Stoodley. T... 2010. P. Shown are the fold changes compared to planktonic cultures. 2006. it is clear that vertebrate animal models should only be used if in vitro models or in vivo models with lower animals are not suitable to address the specific research question. Y. New method showing the influence of matrix components in Leuconostoc mesenteroides biofilm formation. SCR: subcutaneous rat model. on RHE cells and on polyurethane catheter segments implanted subcutaneously in rats were compared. Cambridge University Press. while the expression of some virulence genes appeared largely model system independent.. S. C.. Ausubel.. Staphylococcal biofilm exopolysaccharide protects against Caenorhabditis elegans immune defenses. Rohde.. 193–198. Inhibition of Staphylococcus aureus biofilms by a novel antibacterial envelope for use with implantable cardiac devices. Arias-Moliz. J.. K. Trans. 36. but these studies clearly indicate that the selection of the model system can have a profound influence on the results. 203. Handke and Rupp. J.. In this study. A... Dent. Hill. C. E. J. albicans biofilms grown in different model systems. J. Changes in gene expression in 48 h old C. P... Begun.. K. However. elegans (Mylonakis et al. Laroche.O.. albicans cells grown on silicone discs in MTPs and in a CDC biofilm reactor. the number of animals to be used should be kept to a minimum and considerable attention should be paid to minimize potential pain by using alternatives to painful procedures and/or appropriate anaesthesia (An et al. Handke and Rupp. G. Data are from Nailis et al. 2002. Microbiol. W. 4172–4179. Saginur. there were considerable differences in filamentation patterns. Characterization of biofilm growth and biocide susceptibility testing of Mycobacterium phlei using the MBEC assay system. A. Baca. and biofilmgrown Pseudomonas aeruginosa isolates cultured from sputa of adults with cystic fibrosis. Development of a laboratory model to assess the removal of biofilm from interproximal spaces by powered tooth brushing. albicans biofilms grown in different model systems. Vandemheen.. For that reason. rabbits.M. Chan. 15. D. This biofilm model offers considerable advantages in terms of logistics.T. Bardouniotis.L. (2010). Biochem. Development and characterization of an in vivo central venous catheter Candida albicans biofilm model. F. Biotechnol. B. 97. Andes.. Buckingham-Meyer. Valderrama. Huddleston. adherent. S. 40. A first example is the comparative evaluation Acknowledgements We wish to thank BOF-UGent and the Fund for Scientific ResearchFlanders for financial support and Drs.T.. Diachenko. R.. RHE: reconstituted human epithelial cell model.. Perez. CDC: CDC biofilm reactor. These examples clearly illustrate that data obtained in one model system cannot be extrapolated to another one and highlight the importance of the selection of an appropriate model system that most closely resembles the real-life situation the researcher wants to mimick. Gaiani. Nett. phenotypic and genotypic characteristics of sessile C. Adams. C. e57... reproducible and polymicrobial model of otitis media and its prevention. Non-mammalian animal models are increasingly used to study various aspects of infectious diseases and an animal frequently used for this purpose is the nematode C. 12B–17B.W. Chemother... Michaud. A second example is the study by Nailis et al.. dogs) is associated with high costs.. budget. Pitula. 151.. H. Furthermore. Animal models of orthopaedic implant infection. Amorena. Expert. F.. Clin..D.B. Oschel.M.M. Badel. Bernardi. guinea pigs and hamsters are preferred in most studies (An et al.M. Rev. C. M. 5). 3. H. E. Gracia.P. Bakaletz.. (2010) in which morphological.. M. the efficacy of three disinfectants against P. vertebrate in vivo biofilm models better take into account the host immune system and are indispensable for a better understanding of medical-device related infections. Nelis / Journal of Microbiological Methods 83 (2010) 89–105 99 of biofilm disinfectant tests reported by Buckingham-Meyer et al. MTP: microtiter plate. 43–55. 2006). While better mimicking the actual situation in a human host.. including mice.. When compared to non-mammalian in vivo models. 2003.. A. 87–90.

(Eds. M. Comparative assessment of antibiotic susceptibility of coagulase-negative staphylococci in biofilm versus planktonic cultures as assessed by bacterial enumeration or rapid XTT colorimetry. Infect. J. Immun.. A. Allergy 23. 50. Bjarnsholt. N. 73. Cheung. Rasmussen. Clin.. Ellis. 52893–52903. H.. 2003.S.W. Zanetti. Adherence of coagulase-negative staphylococci to plastic tissue culture plates: a quantitative model for the adherence of staphylococci to medical devices. Jefferson. Giacometti.100 T.. 3145–3148. Coenye. van der Mei. K. L.A.. Med. Mycopathologia. 865–874. 221–230.. F. Hoyer. 37. 407–410. 70. Ghiselli.. 1081–1086.. Daniels. Høiby. M. J. U.V.. 337–345.. A.. G... Gillis.J. Politz. Saba. 1210–1216.W. Chiu.. 127–141. V. Infect.M.. 3. Bernardi. 279. B. E.G.W. Beachey. 16. Batista. T. Skerlavaj. Goeres. S. K. Am. S.A.. Osteomyelitis and the role of biofilms in chronic infection.K. 2004.. 2007. W. 1985..M.. 331–336.. P. Donlan. M. Alternative biofilm growth reactors. Peptides 27. H. Licci.W. D. K. C.. Hoch. Braithwaite. Use of the modified Robbins device to study the in vitro biofilm removal efficacy of NitrAdine. Awad. Palmer. N. McCormick.. Experimental models of bone and prosthetic joint infections.. T. De Prijck. Costerton. R. 75. E. Microanal... Agents Chemother. R. a mutant of Caenorhabditis elegans. S. 51. Ward.. W. 31–51. K. G. Biomed.. R.B. R.A.K.. 158–164.P. The Biofilm Laboratory: Step-by-step Protocols for Experimental Design.. N. 2009. N. Carrara. 1268–1275. The use of quorum sensing inhibitors to interfere with biofilm formation and development in Burkholderia multivorans and Burkholderia cenocepacia. Moss.D. J. vancomycin.... 2007.J.H.. E. 47. J. 255–260. K..C.. 257–274. Costerton. R. . Res.. T. 2006. Silvestri. Chennupati. 2312–2320.M. 52. Costerton. 102–106. 405–411. C.B... Chakraborti. Merritt.. A new model of vaginal infection by Candida albicans in rats.N. M. Storey... Costerton. 16.. M.W.. E. N. Microbiol. Microbial adhesion in flow displacement systems.O. Methods 68. S. A. 77. Campoccia. L. J. J. Mocchegiani. Woods.L.A. 605–612. S. F. Coenye. Biol..G. T. Appl. Kofonow. Tam. 7–11. Res. Silvestri. Heersink.. A. M. Davis. Cirioni. Cirioni. Werner. Environ. Wound Repair Regen. Marine biofouling on fish farms and its remediation. 1761–1770. Montanes. Ricotti...A.. Bozeman. Morck. D. 2009.. Why chronic wounds will not heal: a novel hypothesis.. Charles. J. and Data Interpretation. Infection at the site of implanted materials with and without preadhered bacteria. Darouiche. 1978. 2010. E. Svidzinski. M... Entenza. E. Agents Chemother... V.. Kofonow. A. H. H. Christophersen. 1994.. Cohen. J.I. C. Methods 70. Orlando.. 16. Giacometti.C. Cohen. D. V. Tan. Mocchegiani.. Coetser. Caenorhabditis elegans mutants resistant to attachment of Yersinia biofilms. 651–657. Giacometti. 215–252. Res. G....). J.. J. 105.. Mycopathologia 169. Cirioni. Pradier. 257–262.. 453–459. 2008.. H. Efficacy of combination of chlorhexidine and protamine sulphate against device-associated pathogens.. Melton. 386–392. Microbiol.. Mocchegiani. Chang. C. C.. Schwarz. T. 733–740. C. T.. 31.J. J..A.. Consolaro.E. V. J. Ionescu-Zanetti. 1334–1341. Darby. Coenye..T. Ricotti. Drugs Exp. Ohman. Poole. F. Orlando. M.. K. R. Cohen. 46–51... M.. J. 2–10.. Biomaterials 27. Carrara.D. Palmer.. The Calgary Biofilm Device: new technology for rapid determination of antibiotic susceptibilities of bacterial biofilms. Immun. F. 2331–2339. Bleier. De La Fuente. 1771–1776. Muzzonigro. is deficient in glycoconjugates.A... 2007.G. Water Res. Madsen. Madhyastha. Costerton.D. T.. Genetics 176. 2010a.D..G. Impact of Pseudomonas aeruginosa quorum sensing on biofilm persistence in an in vivo intraperitoneal foreign-body infection model. Microbiol. Kurland. 5306–5313. Buckingham-Meyer. Biofiltration—the treatment of fluids by microorganisms immobilized into the filter bedding material: a review. M. T.. B. Antimicrob. A. J. H.. Agents Chemother. E. P. Scalise. Experimental foreign body infections in mice challenged with slime-producing Staphylococcus epidermidis..E.. 261–264. P.C.. 22.R. Calhoun. T.A. A. 2002. 2006.E.. Chemother. RNAIII-inhibiting peptide significantly reduces bacterial load and enhances the effect of antibiotics in the treatment of central venous catheter-associated Staphylococcus aureus infections. A. Saba. Multidrug efflux pumps: expression patterns and contribution to antibiotic resistance in Pseudomonas aeruginosa biofilms. C. pp. J. Kuhn. 12. Scalise. K...L.. D. L..... C. 2008... 2007. Mater.. Silvestri.I. Givskov. 16. O.. T... S. Linezolid compared with eperezolid. 183. Chemother. Mertz. C. The significance of infection related to orthopedic devices and issues of antibiotic resistance. Jones. Moss.... Orthop. M. Goeres. 19. 2005. P.. Christensen. L. A.. J.. 25. Chronic Pseudomonas aeruginosa endobronchitis in rhesus monkeys: I..P.H. Phipps. Biofouling 23.. G. M.. J. N. Peeters Nelis. Burr. Kjelleberg.. Curtin.. 2003. Crit. J.Ø. Kirsner.M.. N. Mayer-Hamblett.. Krogfelt. C. Surg.F.. Rhinol. Buret.. Jensen. Mansouri... 2001.. M.. Saba.. Gunther... Am. Martins. Curtin. B. Cirioni. N. Orlando. O. Nayak. Immun. Science 284. Dellamonica.. M. 1999. Casey. Darby. Leid. T. 2010. Clin. Cerca. G. J. Microbiol... Silvestri. Microbiol. McEvoy.G. Christensen. 996–1006..1-treated central venous catheters improve the efficacy of hydrophobic antibiotics in the treatment of experimental staphylococcal catheter-related infection. B. 59. Cormican.R. Bernard. Licci. Bisno.G. M. Ultrastructural imaging of Candida albicans adhesion to rat genital epithelium through scanning and transmission electron microscopy. Banks. Rayner. K. Geesey. J.S..J. W. 2007. 2001. Shirtliff. J. M.. 415–422. Infect. W..H.B.. Microbiol. Davis. Azeredo.. R. Dermatol. D. L. E. Cohen. A. Microbiol. 437.E. Scalise. Conant.. Chavant.... J. Consolaro.A. O... Colleran. G. 1991. Oliveira. C. M. M.. Relat. 1983.. Antimicrob..... M. H. F. Brackman..K..G.V. McCullough.. D. Cieri.. Mar..E. E. An in vivo model to study the pathobiology of infectious biofilms on biomaterial surfaces. Appl. R. T. A rat model of chronic respiratory infection with Pseudomonas aeruginosa.J.... Cohen. L.E.. M. 2009. K.. M.. E. Burns. De Kievit. Use of tissueengineered skin to study in vitro biofilm development.. Chiodi.. J. Antimicrob. doi:10. Cash. Cazzaniga. Bernier. Brady. Givskov. H... J.L.. Welsh.. Meng. A.. J. Effects of pentoxifylline on neutrophil influx. Chandra. J. S. Efficacy of silver-releasing rubber for the prevention of Pseudomonas aeruginosa biofilm formation in water. H. D. R. Primatol. RNAIII-inhibiting peptide affects biofilm formation in a rat model of staphylococcal ureteral stent infection..R. S. M.. Primatol. Patussi. R. J. Donatti.B.F. Baddour. L. Chiu. Jensen. K. Hillaert...R.. P. Res.. N.L. C. Biofilms in chronic rhinosinusitis: a review. G. Respir. L.A... F. C. J. Licci. M. Agents Chemother. Trends Microbiol. Bacteriol. Matin. A.. A. Appl.. Bergnach.B.L.P. Caenorhabditis elegans: plague bacteria biofilm blocks food intake. Oxygen-mediated regulation of biofilm development is controlled by the alternative sigma factor sigma(B) in Staphylococcus epidermidis. Honraet. G. A. W. J.G. Am. Cotter. R. 160. Appl..B. Adv. 4518–4520. D. 2008. Giacometti. B. Kirketerp-Møller. Van Calenbergh. Pre-treatment of central venous catheters with the cathelicidin BMAP-28 enhances the efficacy of antistaphylococcal agents in the treatment of experimental catheter-related infection.. Saba.. Biofilm formation by Propionibacterium acnes is associated with increased resistance against antimicrobial agents and increased production of putative virulence factors.... Microbiol. Crémieux. J. Biomol.B. Olson. C.B. S.W. Chem. Keelehan. W.. P. Clin. 2001. S. D.M. Science 2381. 4391–4405. A... Gawande. Wound Repair Regen.G. Arciola.. E. R. Busscher. FEMS Immunol. 23–29. Bjarnsholt.. S. J.. K.. 1992.V.A. De Prijck. Hébraud. 1979.. Screen. R..A. Kamysz. Antimicrob. P. F.J.. W. 243–244..... C....D. 22. C. 21.. 2008.. 2007.. W. Damke. Kumar. T. Tamashiro. M. J. V.H.. Antimicrob. Gaillard-Martinie. Balaban. L. Assessing adhesion forces of type I and type IV pili of Xylella fastidiosa bacteria by use of a microfluidic flow chamber.R. Well plate-coupled microfluidic devices designed for facile image-based cell adhesion and transmigration assays. Hirschberg.H. Novick Jr. De Prijck. C..A. New device for high-throughput viability screening of flow biofilms. 4136–4142. Cohen. S.E. N. Environ. 2005.. 1992. J.E.. Sokol.J. Iglewski. P. Hamilton.A.A. De Smet.. Ghori. Dis. J. J... J. D. Donatti... Balaban. 2010..E. Talon. P. Butterfield.. Dell'Acqua. De Wever.. T. 2006a. Buckingham-Meyer. 1999.G.S. Effects of an LL-37-derived antimicrobial peptide in an animal model of biofilm Pseudomonas sinusitis.. F.D.. a novel disinfecting formula for the maintenance of oral medical devices.. Bioresour. Rhinol.. 180–186. Comparative evaluation of biofilm disinfectant efficacy tests. Infect. 35. Nature 417. Leong. Pitts. Antunes.. M. 2009. Infect. G. 2002. 1318–1322. D. 15. P.A. Falkow. Environ..M.C. K. Nelis. Efficacy of subinhibitory concentration of pefloxacin in preventing experimental Staphylococcus aureus foreign body infection in mice.. Moser.. Darby.. C.M.. F. Svidizinski. G. 1997.B. Høiby. 144–151.M.. architecture and drug resistance. Camper. Cerca. J. C. Srikumar. J.. 213–232. P. Leong. 47... Woods. 2006c.A.. Coenye. Inhibition of Candida albicans biofilm formation by antimycotics released from modified polydimethyl siloxane. Drace. 86–95. R. Leid.N. Clin.. 2003.J. Damke. A new device for rapid evaluation of biofilm formation potential by bacteria. Mondain. K. Rev. J. Microbiol. 2002. Nelis.. Biofouling and biocorrosion in industrial water systems. N. 2005.. Microsc. 56. Scalise.. B. A...J. Nelis / Journal of Microbiological Methods 83 (2010) 89–105 Benoit. 193.Ø. M. Peptides 27. 36..A. 45. Comparative analysis of plant and animal models for characterization of Burkholderia cepacia virulence. Pier. H.. Schwartz. J. J.C.R. J.. Novick Jr. Nelis.J. Mack. 167–174. A. R. Cloete. 2009. 526–531. Bacterial biofilms: a common cause of persistent infections. Med. Microbiol. C. Stremick.. G. H. Younger...C. J. Ionescu-Zanetti.. L. In: Hamilton. Cytergy Publishing. Chiu... Silo-Suh. F.M. A. T... Christiaen.. 119. M.. Res. Wu. G. 5385–5394. N.. R. Biofilm theory can guide the treatment of device-related orthopaedic infections. E.. Simpson. Beachey. L.E. 13–22. J. Correlation between an in vitro invasion assay and a murine model of Burkholderia cepacia lung infection. Olson. Griffith.E.J..J. 18.... C....E. Dis. A. Bass... Microbiology 153. Greenberg. Batista.. Buret. C.. Microbiol... Costello. J. 2006b. Clin. Chronic Pseudomonas aeruginosa endobronchitis in rhesus monkeys: II. Minardi.. Christensen. Simpson.. Orlando. O. 1993. Citropin 1. resistant to bacterial infection and to biofilm binding. Using bacteriophages to reduce formation of catheterassociated biofilms by Staphylococcus epidermidis. Schacht. Analysis.H. K... 71. Conant..V. Biofilm formation by the fungal pathogen Candida albicans: development. C. 2690–2696. A.T. 357–362. 2007..M.. Stewart. O'Gara.. Storti-Filho. Uniquely insidious: Yersinia pestis biofilms. Coenye. D. Carbon. .M.. J. 25. and gentamicin in an in vitro model of antimicrobial lock therapy for Staphylococcus epidermidis central venous catheter-related biofilm infections. 2006. Ghiselli. Orthop. Biol. A histopathologic analysis. 76.. Y. A... 2008. C. G. Fleming.G. Allergy 231. Chlorination of model drinking water biofilm: implications for growth and organic carbon removal. 2007.... Dis. Heersink.. Mocchegiani. Ceri. 2104–2110. Rev. Res. Li..C. B. 1295–1302.1007/s11046-010-9326-1 Carsenti-Etesse.. Ghiselli. Barrett. S. Med. Antimicrob. J.. Read. 236–244. Hsu. J. Ghannoum. Ghiselli. Lukasiak. Microbiol. E. B. 158. Irie. Microbiol. Cipollo. Nelis.W.. Microscopic and physiologic evidence for biofilm-associated wound colonization in vivo. K. R. K. 2010. 40. Y.... How bacteria stick. Durant.. Evaluation of the in vivo efficacy of topical tobramycin against Pseudomonas sinonasal biofilms. Eaglstein. Parkins. E. A. A. Mukherjee. Buckingham-Meyer.... Antimicrob..K. M.. 2005. J. Mertz.. srf-3. Chemother. Montanaro.. Y. Ceri.. Coenye.. L. Clin. 2007... A. H. H. Rev... Johanson Jr. L.J. C. Cheung. E. 2008. A. Cheng..A. 1130–1134.. Technol.

J. Donlan.. Awad. L. Dial. Steinhuber. De Prijck. H. 6..P. 2005. Biomed. J... Saba. 4042–4045..J. 53. Ther.. Burrows. Characterization of mucosal Candida albicans biofilms.. Methods in Enzymology Volume 310: Biofilms..I. Kamysz.L. 5.M. Montana.. H. 1976. 2002.A. 2047–2053. 1811–1819.A. Tan. Dongari-Bagtzoglou. 2004. P. Microbiol. M... Tinkey... Experimental otitis media due to Streptococcus pneumoniae: immunopathogenic response in the chinchilla.. Microbiol. E.M.E.. California.C. Urol.. Goeres. J.E. MA. J... R. Murga. C. M. 2009. Raad.. Antimicrob. Fluckiger. Pretreatment with the protegrin IB-367 affects Gram-positive biofilm and enhances the therapeutic efficacy of linezolid in animal models of central venous catheter infection. M. Del Pozo.. Uthamanthil. Y.. Lorenz. Ghannoum.. A novel rabbit model for the evaluation of biomaterial associated urinary tract infection. Gagnon. Gorman.. 789–790.. Microbiol. In vivo models for the study of biomaterial-associated infection by biofilm-forming Staphylococci. J.. Fidel. 2009.L...S. 757–762. Statistical assessment of a laboratory method for growing biofilms. Noverr.. R. Loetterle.. N. Bozeman. 31... 1294–1297.F. R.D..D.J.. 2007.J.. Biofilms. C... Chemother. A. Effects of biofilm treatments on the multi-species Lubbock chronic wound biofilm model. W. Harrison.. R. 164. K. B. Development of a multispecies oral bacterial community in a saliva-conditioned flow cell..C.J. Ollevier. Appl.. I. 74.. Ehrlich. Miller. Monzón. G. T. V... In: Pace.S. M. Grubb. Matsumoto. Weibel. M. 70.. Thornhill.A.. van der Mei. Onrubia. R. K.. Busscher. De Keersmaecker. K. P..C. K. R. Pahissa. 2531–2537. T. E. Appl. Van Driessche.. J.S. Application of a rat osteomyelitis model to compare in vivo and in vitro the antibiotic efficacy against bacteria with high capacity to form biofilms. Saba. Gracia.P. Subang. Payne. Microbiology 151. P... Hellwig. Reitzel. E. Hamilton. .. Folkesson. U. Ghiselli. Biofilms on central venous catheters: is eradication possible? Curr.. Corneliussen. Grubb. L. 453–465. 1755–1762. G. Deva. 2009.G. 22. The Biofilm Laboratory.. Res. 65. Dwivedi. C. 7.. Biofilms: survival mechanisms of clinically relevant microorganisms. 2003. Doyle. Skinhøj. 59. (Eds. G. Haraoka.. R. M. Lopez-Ribot. Murdoch.. 269–275.. 1998. van Dam. Differentiation and distribution of colistin. Antimicrob.. San Diego. Stewart.. Biomed. 629–640. R. L.. S. A... C. Langmuir 25. Buckingham-Meyer. D. D. Ehrlich. Res. G. El-Sayed.E. 2010. Mol. G... 101 Gerhart. Comparative efficacies of quinupristin–dalfopristin. Eun. 2005. Saba.. 15. P.. 2001. De Smet. Green. Fluckiger. A.M. 1993.. J.. J. Jones. Handke. Natural history of chronic Staphylococcus epidermidis foreign body infection in a mouse model. 133–161.. Novel antiseptic urinary catheters for prevention of urinary tract infections: correlation of in vivo and in vitro test results. Shear-enhanced oral microbial adhesion.E. U..M. 5–12. 2005. Chandra.M.L.. DiTizio. Biofouling in water systems—cases. Biomater. 53. Experimental osteomyelitis — description of a canine model and the role of depot administration of antibiotics in the prevention and treatment of sepsis. J.. Finelli. S. H.T.. Nelis. Environ... Takahashi. K. Kirby. Academic Press.E. C. 233–239. H.. Mucosal biofilm formation on middle-ear mucosa in a nonhuman primate model of chronic suppurative otitis media. Foster. M. Rabin. Richards.M. R.. Perit. Orthop. 76.. G. M. J. sarA and sae of Staphylococcus aureus on the induction of alphatoxin during device-related infection resolved by direct quantitative transcript analysis. A 81. P. Silvestri.E.C.. Appl. J. Gomis..T. Res. De Fusco.. Mittelman.. 22... Patka. Appl. Hebda.E. M.W. I. Y. Dongari-Bagtzoglou.. Adhesion of Candida albicans to endothelial cells under physiological conditions of flow. Wang. Surg. Coenye. F.. Rev. Renzone. 59.. M. Tanaka.. 267–275. and polysaccharide intercellular adhesin synthesis by staphylococci in a device-related infection model. Am. A new model for posttraumatic osteomyelitis in rabbits.J. Gilmore. 2010c. Palmer. Follo. 383–387. Effects of antimicrobial agents on oral biofilms in a saliva-conditioned flowcell.J. S. F. Hanff. Parenter. 1439–1447. Busscher.. J. 1994.. 2002.P.. J. Surg. J. Nat. Kang.D.C. Schacht. J... Mater. Oreopoulos. Evaluation of redox indicators and the use of digital scanners and spectrophotometer for quantification of microbial growth in microplates. Veeh.E.. 2010. M. L. Evaluation of linezolid. F. Giacometti. Microbiol.. Cytergy Publishing. Rymarczyk-Machal. R. Dis. 2009. U. S. Hosp.. R. P. Cisar. Silvestri.. A. Lilly.. 63–73.V. Candida albicans biofilm formation on peptide functionalized polydimethylsiloxane. E. Colonization-resistant antimicrobial-coated peritoneal dialysis catheters: evaluation in a newly developed rat model of persistent Pseudomonas aeruginosa peritonitis. El-Sayed. Landmann. Thorner. Fung. A... Woods. Microbiol.. R. Appl. P. K. Bacteriol. Antimicrob. Checking the validity of the harvesting and disaggregating steps in laboratory tests of surface disinfectants. Gavaldà.. Microbiol. Med. F. Bone Joint Surg. Piede. F.. A. 2009. Visualization of adherent microorganisms using different techniques. vancomycin..... H. Walker. 3872–3878. vancomycin. 24. A method for growing a biofilm under low shear at the air–liquid interface using the drip flow biofilm reactor. Kashleva. 79. Khoury. Microbiol. L.. 5145–5149. A..O. Heersink. S. Ceri.. Hilyard. Fernández-Hidalgo.. Urol. Microbiol. C. Provinciali. W. Costerton. Wormald. 1995. Fabrication of microbial biofilm arrays by geometric control of cell adhesion. A. S.. Med. 4509–4515. Daigle. F..... C.. Stoodley. B. 167–193. J AOAC Int 92. C. Model system for growing and quantifying Streptococcus pneumoniae biofilms in situ and in real time. Effect of prednisolone on ascending renal infection due to biofilm disease and lower urinary tract obstruction in rats. 1–7. Hamilton..S. Environ. C.J. M. 160–167. A.K... Am... 93. Infect. Post. D. 38.. 1220–1223. D. Foster. J....J...G.. 2700–2710. H.. J. Expert. J. R. 322. 53.A. J. Mucosal biofilms: challenges and future directions.. R. Laryngoscope 115.A. Schacht.. Donlan. Sudbery.. 49. Kennedy. A. Environ. Buckingham-Meyer... Licci..F.. R. Chandra.. Haarman. Infections and Antimicrobial Therapy. Jiang. G. 2010b. Microbiol. Ploeg...W. 77. J.H. H.N. High-throughput metal susceptibility testing of microbial biofilms. Juhn. F. Steckelberg. Renshaw.. Richards. K. Kühn. J.E. A. 27–31. P. 708. Mater. Cirioni. D. G. Protoc..J.. K. T.. Sanii. E. G... Coenye. Costerton. Goeres. G.. Infect. Thornhill. Infect. Wolcott. 2003.. 2009. Silvestri. O. 250–255.. Mandrekar. Orlando. D. M. 1994. G.L... J..T. P. R. Wolz. Immun. 4980–4988.. Biofilm removal by medical device cleaners: comparison of two bioreactor detection assays.. 2002. Gomez-Suarez. Sinnadurai. Hayes. Cheng. N. 2009..L. gentamicin and ciprofloxacin in a rabbit model of antibiotic-lock technique for Staphylococcus aureus catheterrelated infection.. 73. J. Diaz. 2009. Almirante. Int. A. Candida albicans forms biofilms on the vaginal mucosa. C.C. A... G. Donlan. Beck. M....S..E.. Engelsman. Leiva. Int.A.. N. Licci. Della Vittoria. Hayes. Quie.. Darby. Bacterial biofilms: from the natural environment to infectious diseases. Giacometti. C. Cirioni.. A... Hayes. Ding.. Impact of the regulatory loci agr. 287.. M. D. Rouse... Res.E. R. Biofilms 1. J. Kolenbrander.K. Antimicrob. Scaloni. 4. 189. Mocchegiani.. 213–221. Analysis of bacterial detachment from substratum surfaces by the passage of air–liquid interfaces.S. Devreese. G. Donlan. Biomed.L. L.. Clin. I. Bacteriol. Buckingham-Meyer. P. E. A.M.. Environ.. Biofilm formation. Rosdahl. Hamill. 2005. B Appl. M.P.. R.. R. P. Flemming. RTPCR detection of Candida albicans ALS gene expression in the reconstituted human epithelium (RHE) model of oral candidiasis and in model biofilms. M. 1999. Euba. J. J. Res. M. 74. Real time noninvasive monitoring of contaminating bacteria in a soft tissue implant infection model. P. Mater. P. 2004. G. H. DiCosmo. Immun. Rupp.H. P. Scalise. E. A.. M. O. Patel. Cirioni. Chemother.. A. Wound Care 18 (508). A. E.. M. 593–603. Ghiselli. A. J. Immun. J. J. Tolker-Nielsen.M. Ghiselli. Mucosal biofilm formation on middle-ear mucosa in the chinchilla model of otitis media. linezolid.. Kolenbrander... 525–530. 2009.1099/ mic. Dis. Infect. Turner. 937–945. 783–788. 2007–2012. Appl. J.. C. 128–140. 4064–4068. N. J.M. Organs 16. Hachem.. Burrows.. 2002. A. J. N. Edmonds.W. Lopez-Ribot..J. T.. Costerton. L. Replication of Legionella pneumophila in biofilms of water distribution pipes.J. 4340–4348.L. Goeres. Antimicrob. C. Jarelöv. Invest.. 1989. J. Candida albicans–endothelial cell interactions: a key step in the pathogenesis of systemic candidiasis. F. J.N. 2003. T.. 2009.. B. Kubo. J.. P. 10. Vickery. and ciprofloxacin in treatment.G. Artif. I. J. 21. Biofouling 26.. K. P.....E. Hamilton.. Biomater.. D.039354-0.R. Goeres..E.. Frimodt-Møller.F. Goerke. Nat. Haagensen. Al-Ahmad. Orlando.. 2007... J. Am. Agents Chemother. Borne. 595–604.. 2010.0. Validation of the CDC biofilm reactor as a dynamic model for assessment of encrustation formation on urological device materials.. Roux. W. 2010. 28–37.. Can. Infect. Rhinol.F. Enteral Nutr. Francis. A sheep model for the study of biofilms in rhinosinusitis. A.M.J. P.. J... of experimental catheter-related infection due to Staphylococcus aureus. using the antibiotic-lock technique. T. Goerke.. F. Martín. A. DOI:10.. 2008. R. Mills.. S. J. M. C. B. M. Agents Chemother. 1710–1715.A. Gallimore. Grimwood. Khoury. Ha.. J.R.. D. Nadolski. R. B Appl. A mouse model of implant-associated infection. 2004. Fitzgerald.... Hoyer. H. M. Behets.. S.L.. M. 7967..L. Biotechnol. Murga. Oteiza. P.. Res. Eerenberg.. R.I. Hall-Stoodley.... H.. Klausen. 339–345. 2010. 1993. 1991. Dwars. Prevention of Candida albicans biofilm formation by covalently bound dimethylaminoethylmethacrylate and polyethylenimine. R. CRC Press. Ghannoum. Rupp. Microbiol. L. J. 123–129. Vasilakos... Microbiol.. Finch. To. T. Harriott. Boca Raton.L. Saville.A. 2004... PLoS ONE 4. Distinctin improves the efficacies of glycopeptides and betalactams against staphylococcal biofilm in an experimental model of central venous catheter infection.P.. H.D.M.. A. Gabrielson. S. Mycopathologia 170. Jarvi..B. S. A. A. D.. Method 50.. Use of in-biofilm expression technology to identify genes involved in Pseudomonas aeruginosa biofilm development.. 2005. Möllby.. Smith. 2. in press.J.L. 1983..... Infect. 2008.S...D. J. 4370–4377... Microbiol. Res. 141–144.R.. R. Loetterle. Horowitz. Finelli. L. 88. 2004..D. V. M.. Pan.. X. Rodriguez. Dowd. P. L. L.B. Gallant. Riber. Hart..E. Charlton.M. R.. S... Hadi. Hamilton. Testa. J.. Heyes. Agents Chemother. C. Giebink.. The hmsHFRS operon of Xenorhabdus nematophila is required for biofilm attachment to Caenorhabditis elegans. Laclériga. Nelis. 463–468. Kolenbrander. Wolz.. Rev. 2007.. 95–108. J. JPEN J.A. Immunol. J. 2006.. G.A. 134. P. D. A. 2009.P.E.C... K. V... Nelis / Journal of Microbiological Methods 83 (2010) 89–105 De Prijck. 510–512. BMC Microbiol.A.. Orlando. 371–380.A. X. K. V. Scalise.W. 76. B. A. Amorena. J. P. Scalise. M. Saville. The electricidal effect is active in an experimental model of Staphylococcus epidermidis chronic foreign body osteomyelitis..E.. P. Declerck.. Coenye. J. C. Effect of treatment with methicillin and gentamicin in a new experimental mouse model of foreign body infection. H. Ulrich. Hannig.W. Mukherjee. J. Espersen..J.. Assoc. Margineanu. P..). R. 4643–4654. Mocchegiani. pp. 2007. Veeh.H. 2008. Ehrlich.K. Anti. Steinhuber. Environ.A. Della Vittoria. R. van Hoef. T.. Jones.. Drace. C. B. Psaltis.W. A.E. V. Microbiology 150. D. 65. 164.C. 67. Łukasiak. Y.Z...K. Kumazawa. R... Top. McKenzie.. V. J.D. R.. Microbiol. Antibiotic-loaded biodegradable bone cement for prophylaxis and treatment of experimental osteomyelitis in rats. Giacometti.R. K. 1469–1472. Molin.. Sudbery. 11. Agents Chemother. 146–153.K. Ernst. O.. 40.. van der Mei. Antimicrob. Gagnon. Mack. J. B.. icaADBC transcription. causes and countermeasures.and sodium dodecyl sulfate-tolerant cells in Pseudomonas aeruginosa biofilms. Costerton. P. Infect.. F. Mocchegiani. L..J. J.. Microbiology.. Subinhibitory antibiotics reduce Pseudomonas aeruginosa tissue injury in the rat lung model. 2001.. Döring... Hu.. De Smet. R. C... Rev.. M. B. J. M.D. 185.. J.. Sun.J. Dohar. 289–303. Zimmerli. J. 2002. Murdoch..

A... 2001. N. T.. Susceptibility of Candida albicans biofilms grown in a constant depth film fermentor to chlorhexidine. Clin.. Heersink. F... Lopez-Ribot. Alanylation of teichoic acids protects Staphylococcus aureus against Toll-like receptor 2-dependent host defense in a mouse tissue cage infection model.D. R. Perit. 22. Berninsone. Englert. In: Hamilton..A. Noninvasive biophotonic imaging for monitoring of catheter-associated urinary tract infections and therapy in mice... 183. W. Swerhone. 62. H. 499. H. Libby.S. Chem.. Bell. Johansen.. Nelis / Journal of Microbiological Methods 83 (2010) 89–105 Harvey. Jayatilake. Otorhinolaryngol. B. 2008. Ghiorse.R. J... Dis.W. Molin. 2504–2514. Rennison. 8538–8544. J. 95–96.. Jass. Heydorn. Antibiotic susceptibilities of Pseudomonas aeruginosa isolates derived from patients with cystic fibrosis under aerobic. 2003. Lattif. A nonsurgical rat model of foreign body-associated urinary tract infection with Pseudomonas aeruginosa.. Rasmussen. Samaranayake. 4474–4475.J.. Joshua. Wolcott. Drettner. Microbiol.. A.. Honraet.. Tachi. Neumeister.K. Secor. Hentzer. R..K. Kumon. A bacteriological and histological study. Birzniece.. 1526–1531. Samaranayake. G. Otol. Sharp.. Microb. Microbiol..B. A. Pierce.M. Landmann. J.. 43.. Molin. 8564–8570. Heersink. Swogger.J. N. McElhaney Feser.P. Molin. Lab Chip 10. 105. R. Holman. 560–567. Mukherjee. 2004. A. M.. 1206–1216. Pearlman. Modeling growth and quorum sensing in biofilms grown in microfluidic chambers.. Porter.. Zulenkovs..... Pulcini. Cihlar. 3878–3887. G. C. Lauth. Microbiol. Pratten. Otani.. Seki. Antimicrob. A. 64. J. Givskov. Use of the modified Robbins device and fluorescent staining to screen plant extracts for the inhibition of S.J... 73. S. 393. B. McKee. W. 2008. mutans biofilm formation. 421–423.. A.. D. 567–570.. 383–385.P. Molin. F. Bozeman. Co-culture of epithelial cells and bacteria for investigating host–pathogen interactions... Infect. Lee.. 2008. Mol.. M. Lund. 283–289. S. Larsson.A. M. J. The efficacy of topical antibiofilm agents in a sheep model of rhinosinusitis. Arch.P.J. 16. G.B. Immunol..S. B. S.. 1998. 414–423. P..J. Berk.. J. Azithromycin blocks quorum sensing and alginate polymer formation and increases the sensitivity to serum and stationary-growth-phase killing of Pseudomonas aeruginosa and attenuates chronic P.. BellingerKawahara.. K. D.. Baskaran.. 171–182.W... Kristian.. Hanson. Y. 2003.J. B.. Am. P. K. Albert. Haagensen. J. Yanagihara. Castagnet..L. Tang.. H. M. S. 1977.E.D. F. Kadurugamuwa.L.. J.. Microbiol. M.. H. Effect in a rat model of heparinized peritoneal dialysis catheters on bacterial colonization and the healing of the exit site. Methods 63.H. Kim. Goeres. Givskov. G. Exp. Biol. Höflich.. Goetz. P. Miles. 2009.K.B. P. Ivleva. Gerke. MénardSzczebara. Wagner. Nakamura.S.. L... J. M. 2000a. M. Honraet.102 T.. Costerton.. Oral Pathol. F.. M. U. Agents Chemother. 21–25. P. Appl. G. Yamamura. F.. D. Heilmann. Wozei. Antimicrob.. Antimicrob. Purchio. Hoffmann. Yu.. 2000b. Antimicrob.R. Jayaraman... Syamal. Vogel.. 2005.. In: Hamilton. Appl. A. T.. Teitzel.. van der Mei.. Lee.. C.. An in vitro model of chronic wound biofilms to test wound dressings and assess antimicrobial susceptibilities.. Larm. R... 2395–2407. M. Parsek.. Immun. H.Y. H. H.H.. Mond.. Microdevices 10...E. S. L.W.. Coticchia. J. Analysis.. 64.M. Chandra. 34. J..... R. Ersbøll.... development and phenotypes in a standardized continuous flow system.. 215–224. 64. Ind. B. Stewart. Mol. Yamamoto. Continuous ethanol production by Zymomonas mobilis and Saccharomyces cerevisiae in biofilm reactors. 197–206.A. Fusarium and Candida albicans biofilms on soft contact lenses: model development. Perdreau-Remington. Winstanley. K. M. Hope.. S. Methods 42. J. 7456–7564..A. Shuler. J. Y. A Caenorhabditis elegans model of Yersinia infection: biofilm formation on a biotic surface.. 2010..G. J.. 5395–5401.. Microbiol. 1996. 2007. Peschel.. T. Haisch. Hentzer. Antimicrob. A.J.K... Sestrich. Spatial patterns of alkaline phosphatase expression within bacterial colonies and biofilms in response to phosphate starvation.. Y. The ability of biofilm formation does not influence virulence of Staphylococcus aureus and host response in a mouse tissue cage infection model. 71... M. F. G. K. Y. F. Anderson... Microbiology 146. Stub. Sachdeva... J. 2003. J. Chanturiya. Laryngol.. H. Ghannoum.K. Kohno.G.P.. In situ surface-enhanced Raman scattering analysis of biofilm. 2004. 1996. Kaplan. J. Keith.D. Hegde. Hu. J. Høiby.. G.. K.... J. Buckingham-Meyer. S.. 190. A.T. 54.. T. 9–15.E. Kumlien. 15. Optimized candidal biofilm microtiter assay. luxS-based quorumsensing signaling affects biofilm formation in Streptococcus mutans.R... M. Agents Chemother..... Bacteriol. 73.. B. Thomas. 2003. T. A. C. Morinaga. H. S. Cho.E.. Juhn.W. J. Determination of spatial distributions of zinc and active biomass in microbial biofilms by two-photon laser scanning microscopy.P. Pajkos. Infect. Rhinology 45. Biomed. Francis. Levesque..... G. M. Pathog. O.. R.L. J. Anal.. Lee.W. V. T. Götz. Ishida. A. Horn..A. 285–295. N... B.M. 3221–3229. T. Parsek.. H. .. (Eds. Hill. Anal. 2804–2813.. Quantification of biofilm structures by the novel computer program COMSTAT. 882–890.. C. 80. D. Lion. M... Ann.. Lee. In vivo efficacy of anidulafungin against mature Candida albicans biofilms in a novel rat model of catheter-associated candidiasis. 188. A. Immun. Towards a nondestructive chemical characterization of biofilm matrix by Raman microscopy. 2010. Heersink. O. Krom... Antimicrob. J.. B.W. 240–246. S. Chem..M. Microbiol. J. 2010. M.. Juhna. Paparella. M. Goeres... M. Xu. Heydorn.. Holtappels.. 2003. S.. Lysostaphin eradicates established Staphylococcus aureus biofilms in jugular vein catheterized mice.. Heydorn. Janakiraman..L. M. M. Microbiology 149. J. Chemother. M. Basic biofilm analytical methods. Kamihira. Appl.T. T. M. Lazzell. 2005. Stewart. Paroni... Infect.. 2005.. 3677–3687. Neumeister. Microb.. C. a novel carbapenem. R.. Dermatol. Candida biofilm analysis in the artificial throat using FISH. Bacteriol. Kukavica-Ibrulj. 5085–5090. O'Toole. K.T. D.. Keevil. Pometto.. Kokai-Kun. Francis. Hoa.. X. McCullough. 2007. Hodgkin. Rubin. E. P. Cohen.. Photobiol. H. L. C. Ersbøll... 53.A. 4014–4021. Ecol. Costerton. 277–282. G. J. E. Hegde. C. K. C. Politz... Cho. Chemother. 215. Agents Chemother. 237–245.. K. K. Huang. S..Ø.A. A. The Biofilm Laboratory: Step-by-step Protocols for Experimental Design..J. Azevedo. and Data Interpretation. Chaturvedi. P. Y.. J. K...D. P.K. Jayaraman. Infect.. Methods Mol. Bergström. Yu. Hoffmann. H.. C. and Data Interpretation. C. 2004.. M. S. Baumeister. Aaron.. Comparison of three assays for the quantification of Candida biomass in suspension and CDC reactor grown biofilms.). Cytergy Publishing. Sin. Mycoses 53. Isherwood.M. Claus. Real-time chemical imaging of bacterial activity in biofilms using open-channel microfluidics and synchrotron FTIR spectromicroscopy.. Experimental acute sinusitis in rabbits. S357–S358. 1461–1468.. Nelis. Houston. 1292–1309.. V..L. Demonstration of nasopharyngeal and middle ear mucosal biofilms in an animal model of acute otitis media. J.).R. Hentzer. 17. J. D.. An ultrastructural and a cytochemical study of candidal invasion of reconstituted human oral epithelium.. Féliers.. Loss of srf-3-encoded nucleotide sugar transporter activity in Caenorhabditis elegans alters surface antigenicity and prevents bacterial adherence. Izumikawa. C. T. Kanno.K. Nielsen. M. Environ. Giebink. S. C.B.. Z. K. 2009. E. Ivleva. Cramton. Rhinol. Prasad.T. 16. J. Nizet... D. Johansson. Z.. in hematogenous murine bronchopneumonia caused by methicillin-resistant and vancomycin-intermediate Staphylococcus aureus.. C. 3–13. current concepts and directions for research. Lappin-Scott. Microbiol. Meningococcal biofilm formation: structure.... Z.W. and susceptibility to lens care solutions. Microbiol... 2163–2168. 64. Götz.. Elkin. Sanschagrin. Biotechnol. J.. Liu. J. 2005. S. Kurosaka. Heersink. 1749. Göbel. Horn.. Analysis of the effects of chlorhexidine on oral biofilm vitality and structure based on viability profiling and an indicator of membrane integrity. N.. 2001.. Wren. M.. Microbiology 146.. Contag. Neu. Treatment and prevention of Candida albicans biofilms with caspofungin in a novel central venous catheter murine model of candidiasis. delta-Aminolaevulinic acid mediated photodynamic antimicrobial chemotherapy on Pseudomonas aeruginosa planktonic and biofilm cultures.. S. Methods 64. H. G..J. The Biofilm Laboratory: Step-by-step Protocols for Experimental Design. Wound Repair Regen.. 2004. Samaranayake. 48.R.. B. Hay. Ma. Molin. Kunduru.. Bioanal..A. Johansen. Balzer.B. Huang. Araki. J.. Chemother. 13–15.L.J.. Niessner. Szczotka-Flynn.. Method 68. J.. Kucharikova. P... Lappann. Kadurugamuwa.L. 16–23.. Kim. E. D. C... J. Experimental superficial candidiasis on tissue models. P. V. Rose. Microbiol. Direct continuous method for monitoring biofilm infection in a mouse model. Givskov. 2009. 2010a. 2409–2415. influence of lens type. Psaltis. E. Landmann. N. Microbiol.. 279. J. Y. DeBoer.. J... Exp. M. Camper.. 38. B 75.. Environ. J. aeruginosa lung infection in Cftr(−/−) mice. Reactor design considerations. Kim. Biofilms and chronic rhinosinusitis: systematic review of evidence. 2005. pp.. S. Harbour. A. Agents Chemother... G.. Sharipo... Contag. 2004. Y. B. M.. P. J. Mathiesen. S.. Microbiol.C. 81. Agents Chemother 51. N. Dial.. J. Interaction of Klebsiella oxytoca and Burkholderia cepacia in dual-species batch cultures and biofilms as a function of growth rate and substrate concentration. 2009. 65.. C. 2008. A.P. H. 2008... Golda. L.B. 2009.E. Int. Lee...C. J. M.. J.. D.. N. R. 2000. Modi. T.P. 71. and biofilm conditions. E.R. S. C. B. M..C. H. Abruña. Chem. Smith. 2010b. Tan. Busscher.. 37–44. Toriyabe. 2009. J.. S. Hidalgo. Chen. Buijssen... Lagrou. Rasmussen. A. R.. Wilson. R. Z. Immun.. Givskov. M. Jayatilake. C. Parr.. Imai.. Eng..R.K. Sternberg. H. Z. 1195–1206. 45–54. 2007..F. Y. Haisch. J. M. M.. Biofilm formation on rat skin wounds by Pseudomonas aeruginosa carrying the green fluorescent protein gene... (Eds. anaerobic. A simple rotating annular reactor for replicated biofilm studies. L. 52. Hirschberg. H.. 2006.. Acta Otolaryngol. Hentzer. M. 12–19.. 114–125. Vis. D. S. A. Novel mouse model of chronic Pseudomonas aeruginosa lung infection mimicking cystic fibrosis. K.K.. L. Hao. Takase. D. R.R. J.. B. Lass.. 30440–30448. A. Lamfon. 2007. S. Coenye. In vivo growth of Pseudomonas aeruginosa strains PAO1 and PA14 and the hypervirulent strain LESB58 in a rat model of chronic lung infection.. Bozeman. Thompson. Imamura. Darby. 2005. Alginate overproduction affects Pseudomonas aeruginosa biofilm structure and function.Y.. Potency of SMP-601. C. J. N. 37. B.. Carlsöö. Infect. Nord.. R. J.A. Lejeune..F. J. 2010. Buckingham-Meyer. Anal. Macleod. L. pp. Kakeya. A.. Williams...H. Ind.. Biofilms in chronic wounds. 1988.. Jensen.. Huang. D. E. Antimicrob. 2009. Van Dijck. Lawrence. Rhinol.P.. Robinson.. Goetghebeur. 73. M.. H. 287–295.. 2008. R.F. G. 2009. Analysis. Kihara. M.. 2006. Wormald.. T. 45. E. 217–224. Experimentally induced acute otitis media—an animal model. Detection of Escherichia coli in biofilms from pipe samples and coupons in drinking water distribution networks. Gravato-Nobre. S. K.J. P.. 154–156. 1995.. Y.C. Microfluidic co-culture of epithelial cells and bacteria for investigating soluble signal-mediated interactions.J. J. M.. Microbiol. Song. I. Krom. Assessment of a chemostat-coupled modified Robbins device to study biofilms. K. Chemother.. Givskov. J.L. 36. K. R. H. S. Z. Ferracin. Bye. Characterization of Tn917 insertion mutants of Staphylococcus epidermidis affected in biofilm formation. Environ.W.. Wagner. 19. P.. A. Antimicrob. Jayaraman.. Cytergy Publishing.. Kristian.J. Komlos. C.B. Experimental reproducibility in flow-chamber biofilms. Microbiol.W. B. 49.. P. L. Ciofu. J. A. Immun. R. Microfluidic devices for studying growth and detachment of Staphylococcus epidermidis biofilms... Chem. T. J. Kim.G. C. McFeters.. 43–50. 489–498. M. 2001. fluconazole and miconazole: a longitudinal study. Biomed. Yang. James. Karlyshev.R. R.. Høiby. J. Biol. Antimicrob. Kim. 118. A... 2005.. Malic. J.. Hentzer.. Lindholm. Titball.. 94–100. Uppuluri. Goeres.. 2008.. L. Zhang.. 357–366. Med. K. Meric. Peschel. H. Niessner. 249–256.. Cunningham. Photochem. Tournu. Tsukamoto.K.. Ann. 52. O'Donnell.. J. C. C.W.V. 21. P.. Bragonzi. Nelis.. 292–298. Le.H.. C. M. Hill.L.R.

C. J. van der Mei. Tomono. C.C. 151. Observations of fouling biofilm formation. Saba.M.. Mowat. K. Therapeutic efficacy of fleroxacin for eliminating catheter-associated urinary tract infection in a rabbit model. J... Palmer Jr. C. Environ. Acta Otolaryngol. Weber. Med. Mukae. Infect.. J.A. D. Mackinson. G. Coenye...... S. J.... 47–53. T. H. B. J.M. K.. Y. Gracia. G. A. M.. J. N. 1639–1644... 65. Li.. J. 63... Shirai. 341–344.. L.J. Wagner. Biotechnol. 6951–6956. D. M. Scalise. D.. W. Devleeschouwer. Amdinocillin treatment of catheter-associated bacteriuria in rabbits. Mocchegiani.. A flowcell for the study of plaque removal and regrowth. Haaga. James. B. 26. Effect of erythromycin on chronic respiratory infection caused by Pseudomonas aeruginosa with biofilm formation in an experimental murine model. F.E. Karlsson. J.... Engelsman. C. Li.. Casadevall. 2885–2895. 2007. H.. M. Immun. Buijssen. G... 2004... Development of a simple model for studying the effects of antifungal agents on multicellular communities of Aspergillus fumigatus. L. 43.. Gromov. 1983. 1065–1072. J. Otto. Detection of Pseudomonas aeruginosa quorum sensing signals in an infected ischemic wound: an experimental study in rats... Monden. J. A simple infection model using pre-colonized implants to reproduce rat chronic Staphylococcus aureus osteomyelitis and study antibiotic treatment. S.Y.R.R. S. T. I.. 50.. Chemother. Giacometti. Li.. 2009.. H.J. J.D. H.. Spigel.E. Nickel.W. Crit.. B. D.E. Korber.. Tré-Hardy.. 91. Amorena. Kumon. 171–182.W.C. Torii.. e101.. A. R. T. Sardaryan. J.W... Exploiting amoeboid and nonvertebrate animal model systems to study the virulence of human pathogenic fungi. N. Environ. Vet. Bove. Eur. Lam.E.. Olson. M. 1–21. Molin.. Use of fibrinolytic agents to coat wire implants to decrease infection. Immunol. 2006. R. Enhanced benzaldehyde tolerance in Zymomonas mobilis biofilms and the potential of biofilm applications in finechemical production. D.. E. Study of the initial phase of biofilm formation using a biofomic approach. T.. Urol. Busscher.J.. Nonaka. C. 285–290.. Gao... Microbiol. Infect.. 2008. Non-mammalian animal models to study infectious disease: worms or fly fishing? Curr. osteolysis. Hacker.T.. Appl.C. 80. Lipp. Biomaterials 26. Lepak.. K. A. 21–30. Rowland.M. G. Fink-Gremmels.. Niu. 2002. J. Oral Surg. J. Saldarriaga Fernandez. García-Alvarez. Meth..R. Environ.. An animal model. Nickel.. H.R. 1981. 2008.. Peptides 29. Drissi.. Kolter. Antimicrob.. Agents 4. C. 19. J. 30–36.. S. Nelis. T.. Haagensen. Antimicrob. J. 2010. K.. D. B. K..W. K....D. Tomasinsig. Elving... 159. Matthijs. Merritt. 2010. Coenye. I. P. J. 2001. J..W. Spratt.. Colorimetric method for identifying plant essential oil components that affect biofilm formation and structure. Meers. Ziebuhr. A. and humoral immunity. Vergunst. 347–353. Nailis... Y. Grant. Nelis / Journal of Microbiological Methods 83 (2010) 89–105 Leung. Potts. K. 201. Marrie. 73. Mampel. Sjögren. J. M. 1985. J. Mader... 1993.K.. 61. J. Kurumunda.D. Invest. Friedman.W.. Leiva.P. Morck. Hill.. Oosterhof. J. J. 1118–11123. Microbiol. 910–917. Malinin.W. D.J. 217–223. Deforce. J. Long. 2005. Sugama. G. S. J. Yanagihara. Mylonakis. Lam.. M. Tar. R.. using an in vivo central venous catheter model.... H. Schäfer. Am. 2006. H.. Hayes. S. J.B. 9–17. 103 Murillo. composition and function in biofilm systems.C.G. A. Microbiol. J. G. Oosterhof. The alternative sigma factor sigma B of Staphylococcus aureus modulates virulence in experimental central venous catheter-related infections. Martinez. Caldwell...... 1995. Thiede...E. Ricicová... Suginaka.K. Microbiol. Conversion of Staphylococcus epidermidis strains from commensal to invasive by expression of the ica locus encoding production of biofilm exopolysaccharide. Cirioni. Mukherjee.W.. 2009. A.J. M. G. McCoy. M. Fujii. 2005. Costerton. S.A. Otol. Am. McLean. Orlando.. M. Bryers.. Can.. Respir. H.. Microbiol. H. Ghiselli. Flagellar and twitching motility are necessary for Pseudomonas aeruginosa biofilm development.. M. H. 295–304.R. Wang. F. The influence of antimicrobial peptides and mucolytics on the integrity of biofilms consisting of bacteria and yeasts as affecting voice prosthetic air flow resistances. Volke. Extended antimicrobial susceptibility assay for Staphylococcus aureus isolates from bovine mastitis growing in biofilms.. 2006.. Vuong. Costerton. Cleeland. Burillo.. F. Han. Ecol. 3673–3677. J. Environ. Y. M. U. Nickel. 72. Xu. 3188–3191. Hitchcock. J. Costerton. 2008. Nelis. Bucio..J. J. Concheiro. Andes.. S.. C.. Microbiol. K. Hüttinger..J. R. J. Sundqvist.. 1989. Dynes. K. R. Res. G. 1998. Mikuniya. 70. Costerton. J. Svarovsky.. T.G. Comparative evaluation of cefamandole and cephalothin in the treatment of experimental Staphylococcus aureus osteomyelitis in rabbits. Microbiol. Ann. F. Appl. Appl. Palecek. R.. Y. P. Gudiol. Dehaye. 79–85. Manz. 2007.. Eap1p.E. Development and validation of an in vivo Candida albicans biofilm denture model.. Biofilm penetration.. 13... P. M. A. Ausubel. 859–868.J.. Zanetti. F. J.J. Li. Agents 33. Morohoshi. Agents 25...M.C.J. 73.E. T. Planktonic replication is essential for biofilm formation by Legionella pneumophila in a complex medium under static and dynamic flow conditions. 96–105. Dis. Rosche. A. Comparative evaluation of fleroxacin. 1987... A.... 368–376.. Ida. C. Mol. Ariza. Acta Biomater. ampicillin.A.. E. K. T. Minardi. Bacteriologically stressed animal model of new closed catheter drainage system with microbicidal outlet tube. Butcher. Infect. van der Mei.. M. Oral Microbiol. J. 604–608. Mordan.J. M. McKay.W. A.E... J. Environ. and gentamicin as treatments of catheter-associated urinary tract infection in a rabbit model.. Antimicrob. Spirig... Webb. Antimicrob. Schwarz. Endod. Cleeland. Laclériga.... 30. 125. Grant. J. Nickel. 27. R. Kariyama. J.. Lorenz. Wastewater treatment with particulate biofilm reactors.. J.. S. O'Callaghan. Brundin. M. 2008. J. an adhesin that mediates Candida albicans biofilm formation in vitro and in vivo. Appl. 2010. R. 2007. A. Amphotericin B lipid complex is efficacious in the treatment of Candida albicans biofilms using a model of catheter-associated Candida biofilms. J. Evaluation of four experimental osteomyelitis infection models by using precolonized implants and bacterial suspensions. Caldwell. Olson. Nagata. D. E. J.. T. J.. Busscher... Williams.A. Nakagami. Trends Microbiol. Coenye. Busscher. V. Treatment of Pseudomonas aeruginosa biofilms with a combination of fluoroquinolones and fosfomycin in a rat urinary tract infection model. Marchillo. 10. Ohta. T. D.. Wilson.. Muzzonigro. W.. S.S.. Pratten. 16. Prosser. Am. 1995.. 2251–2259.A. J. S. Biofouling 19.. Nagant.R. Kadota. Int. Microbiol. Puzas. K. Lawrence..E.F. B. J..A.. Morck. Van Dijck.. Wound Care 19.. W... L. An ecological study of infected urinary stone genesis in an animal model.Z. Kato. 3. 507–513.. P. D. Microbiol. Testing wound dressings using an in vitro wound model. 1994... A.. 23S–30S.J. Monzón. 2008. Heijnen. Appl. 59. 2004. T. B. Med. T. K. Miyake. PLoS Pathog. D.. Microbiol. Perkins.. van der Mei. F... Oral Pathol.. J. Skerlavaj.. 48. Mechanisms of biofilm resistance to antimicrobial agents. Demonstration of antibiofilm and antifungal efficacy of chitosan against candidal biofilms. Int. 1991.A. O'Toole. D. Stewart. J.. 2009... Int. Leunisse. 56.F. Nejadnik. A. H.. H. J. K. A.J. Bacterial colonization of polymer brush-coated and pristine silicone rubber implanted in infected pockets in mice. Simple method for measuring the antibiotic concentration required to kill adherent bacteria. 94.J. C. Ohlsen..P. 2005. Robbins. P. B..E. J.. F..W. E. Fujiwara. Prosser. Microbiol. 2007. 72. Scotto.. F.. D.. Quantitative mouse model of implant-associated osteomyelitis and the kinetics of microbial growth. J. K. Chemother. G. Mycopathologia 167.V.. 4. Deforce. Costerton. 149–153... van Weissenbruch. C. H. Olson. . Nosanchuk. H. Building biofilms in vital host tissues: a survival strategy of Actinomyces radicidentis.. 7145–7153.. 595–603. Nett. X.... J. 2001.R. Appl. 10.F. Tilleman. 820–826. 286–290.J. Engelmann.. L.. James.. Ellis. 119.F. M.. Eukaryot. Laryngol. 2010.W..J. P... Cleeland. Morikawa.. 188–194.. 2007. R. 1992. van der Laan. Synergistic effect of fosfomycin and arbekacin on a methicillin-resistant Staphylococcus aureus-induced biofilm in a rat model.P.. A. Cyclodextrin-functionalized biomaterials loaded with miconazole prevent Candida albicans biofilm formation in vitro. 2010. Young. 6. M.J... E. Microbiol. S.. 2006. Kim. Wen. 72. Olson. S. S. Palmer. Wilson.. T.C. Marchillo. Molin. Radiol. Laclériga. F.. Garcia. 1997. J. T... T. D. 1995. G. O'Keefe. Doménech.. C. Real-time PCR expression profiling of genes encoding potential virulence factors in Candida albicans biofilms: identification of model-dependent and -independent gene expression. 2010. T. 106. 2008.T.J. Gulabivala. Infect. Antibiotic resistance of Pseudomonas aeruginosa colonizing a urinary catheter in vitro.. van Loosdrecht. Oral Med. Nett. Wolfaardt. Q. Agents Chemother. 62. Alvarez-Lorenzo.. Melchior. Morck. Microbes Infect. O'Reilly. C. Y. 11–19. Br. 24.. 200. Nicolella. Albers. G.J. Andes. J.. G. K. R. M. 9.P. Clin. Ruseska. Care Med. W. Oral Radiol.. Chemotherapy 38. K. G. Hilbi. W. 2000. Gilbert. Hecker. J.C. Scand.K. J.... Nakamoto.. S. A. 72. C.. R. Bone Joint Surg. 2001. Mihu.. 82. Malic.R.. T. Res.. O. Kjelleberg. Time course global gene expression analysis of an in vivo Candida biofilm. E. Stokroos. V.. Albers. Williams. 931–939. Orthop. 34–39. 220–226. G. Olson.. H. 1398–1404. Wound Repair Regen. trimethoprimsulfamethoxazole. Kuroki. M. H. 1–33.. T. J.. 22. H. Kirker.A. Mah.. Characterization of Candida albicans infection of an in vitro oral epithelial model using confocal laser scanning microscopy. O... van der Mei.W.D. Ghannoum. P. 2007. 35–39. Silvestri. M. Antimicrob.S.. Antimicrob. R... J. R. B. Friedman. A. Rhinol.. O'Toole.G.. Microbiol. Coenye... B.. 2006... Marchillo. 3650–3659.W.. C. Hayashi.. C. 2101–2107.W. 1999. McKay.. Oshel. M.. Efficacy of sustained release ciprofloxacin microspheres against device-associated Pseudomonas aeruginosa biofilm infection in a rabbit peritoneal model. Cell 6. 1323–1325.. 1436–1440. A. Chlorhexidine-releasing methacrylate dental composite materials. 2432–2438. Ikeda. Tubau. Dis.. Infect. Neville. The effects of surface roughness and type of denture acrylic on biofilm formation by Streptococcus oralis in a constant depth film fermentor. M. K. G. Lang. Khoury. Olson. 213–218. Lam. Bacterial biofilms in chronic rhinosinusitis. J. H. C.... Med. Chemother.. I. A. Nair. Effects of quaternary ammonium silane coatings on mixed fungal and bacterial biofilms on tracheoesophageal shunt prostheses.... Agostinho. H. Thomas. Advanced imaging techniques for assessment of structure. Amorena.W. D.. Dis. H. Vandenbroucke.. Søballe. H. Nava-Ortiz. Rogers. 2008.R. J. 243–248.J. 196. FEMS Microbiol..J. García-Alvarez. 1995. 2010. Morck. D.. 13.. J... Yoshikawa.. C. Amerongen. Kohno. Usui. The artificial throat: a new method for standardization of in vitro experiments with tracheo-oesophageal voice prostheses.. R.. J. Orthop. Nailis... M. Neu.C. H. 114. J. triggered release and in vivo activity of inhaled liposomal amikacin in chronic Pseudomonas aeruginosa lung infections... Costerton. van Weissenbruch.W. Antimicrob. J. I.. A.B.. J... 1205–1212. F.. Meth. D.C. S. J. Sanada. Impact of nutrient composition on a degradative biofilm community..... BMAP-28 improves the efficacy of vancomycin in rat models of grampositive cocci ureteral stent infection. Ralphs. K.. Relevance of animal models for chronic bacterial airway infections in humans..... Owusu-Ababio. J. 280–289.. 141–149. M. Acta Orthop. J. U. H. Maebashi. 2010.. Fisher. Oteiza. Immun. D. S21–S27. Opin. BMC Microbiol. Efficacy of high doses of levofloxacin in experimental foreign-body infection by methicillin-susceptible Staphylococcus aureus. R. Busscher. D. Ramage.... M. Cordero. Wright. Monzón. 307–313.N.E. Andes..J. K.E.. 30. K. S.. T.J.. Koji... Agents Chemother. A. F. A. Microbiol. Cabellos..S.... 4011–4017. Norde.. Riva.. 78. Costerton. M. Nelis. 2003. Whitfield. Infect. Gaastra.A. J. Awad.M.. Kucharíková. D..R.. Microbiol.. Urology 38.R. D. Morgan... 44–50. K. Moller. Monitoring ALS1 and ALS3 gene expression during in vitro Candida albicans biofilm formation under continuous flow conditions.

Microbiology 156. The genomics and proteomics of biofilm formation..R.. Mohamed. Williams.L.. Agents Chemother. P.M. Microbiol.D.V. 354–359. M. 67.A. 4.. Characterization of the importance of polysaccharide intercellular adhesin/hemagglutinin of Staphylococcus epidermidis in the pathogenesis of biomaterial-based infection in a mouse foreign body infection model... Antimicrob.. Rodríguez. K..1–1B. Stubblefield.L. Mukherjee. H. Burillo. Engels. Yoshida. Ultrasonic enhancement of antibiotic action on Escherichia coli biofilms: an in vivo model.. Eur. Blackwood. B. Antimicrob. Streptococcus pneumoniae forms surface-attached communities in the middle ear of experimentally infected chinchillas. Nat. Rediske. Piper. R. Hamm. Agents Chemother.. L.. Yang.. T.M.J. Molin... J..G. Guarnieri.L. Yang. FEMS Yeast Res. Vidaillac. Reinthaler. Rouse.. 677–701. Moghaddam.M. S. P. Staphylococcal biofilms impair wound healing by delaying reepithelialization in a murine cutaneous wound model. 1–6.A. Wymondham. W.A. Munakata. biofilm formation and adherence of methicillin-resistant Staphylococcus aureus. Agents Chemother. R.K. Alvarez-Lorenzo. Rose. Microbiology 150... 1988. J. (Eds. Bacterial biofilms: an emerging link to disease pathogenesis. Mak. T. P... Batstone. Environ.18. Naglik.F. Peters. Hansbrough. J. I.. J. M. 2008b. van Weissenbruch. Appl.. Nelson. 4329–4334. 1211–1214.M.. K. Takeuchi.J.. Antimicrob.. R.. J. Relat. 1247–1249.. RNAIII-inhibiting peptide enhances healing of wounds infected with methicillin-resistant Staphylococcus aureus.. A. Microplate alamar blue assay for Staphylococcus epidermidis biofilm susceptibility testing. Microbiol. Microbiol.. T. Characterization of Staphylococcus epidermidis polysaccharide intercellular adhesin/hemagglutinin in the pathogenesis of intravascular catheter-associated infection in a rat model.E. G. Koga. S. T. Riva. van der Mei.A. J. M... 78–83..J. Urol... R. Schaller. P. E.. F.. J. BMC Microbiol. Bartscht. Richter. Ruiz. J. 710–713. H. T..M. O. Candida albicans biofilm formation in a new in vivo rat model.. Appl.R.J.. Sharma... Reid. Piccolomini. Med..J. Ghiselli.. J.J. H. Mader. J. 67. 19. M. 2205–2211. F.... 2003. 239–247.. Rakhorst. Antimicrob. N. 10. Singh.. G.. P. Patrick. De Prijck. 2005.H. H. H. Microbial ecology meets electrochemistry: electricity-driven and driving communities. B.. Henriques. 2475–2479. Ertl. Cytom. 786–794... Osborn. Pantuso. Kermanshahi. 76. Yamamoto.C. P. Fey.D. M. Effect of LY333328 against vancomycinresistant Enterococcus faecium in a rat central venous catheter-associated infection model. 157–165... Azeredo. A microtiter-plate screening method for biofilm disinfection and removal.104 T... Robison. S. K.S. Dis.. H. K. J.A. G. Robbins. 263–270. Anal.. M. Bioeng.C. M.. . L.. J.... Rhinol. Satoh. Ethnopharmacol. Sjollema. Hickey. R.T.. Plaunt. H.D. Hachem. Active immunization with lipopolysaccharide Pseudomonas antigen for chronic Pseudomonas bronchopneumonia in guinea pigs. K. and 25percent ethanol. Singh. K. Tetrasodium EDTA as a novel central venous catheter lock solution against biofilm. J. 2010. D. Silva. 2002a. 32. Hoffmann. 2006. Protoc.. P. 1723–1731. Sharma. H.. D.K.. P. K. 418–428. Pettit.. 467–477. 90–103.R... W. J.N.. Direct evidence of bacterial biofilms in otitis media. Pitts. Gualdoni.J.. Effects of extracts from Italian medicinal plants on planktonic growth. Lagrou.. J. J.. Laryngoscope 111.. M. The influence of skeletal implants on incidence of infection.. De la Garza. 2007. Biofilms: implications in bioremediation. Genome Biol. E.. Chemother. Quorum-sensing signals indicate that cystic fibrosis lungs are infected with bacterial biofilms. Long. Offidani. L. Lab Chip 7. 89–97. Curr. Percival.L. 2010. Nature 407. C...L. 2007.. H. Agents Chemother. Am. L. Tomono.5.. 70. 471–477. Polypropylene grafted with smart polymers (PNIPAAm/PAAc) for loading and controlled release of vancomycin. J.. A. 1062–1065. Shuford.. 2007. Agents Chemother.E.E. 2807–2813. J. 9–18.J..H.. J. S. Polymorphonuclear leukocytes (PMNs) induce protective Th1-type cytokine epithelial responses in an in vitro model of oral candidosis. 2010. 51. J.. Evaluation of caspofungin and amphotericin B deoxycholate against Candida albicans biofilms in an experimental intravascular catheter infection model... van der Mei. Raad.. Rupp.J. 56. Horton. V. Hosp. P. 2010. 103. J. 1984. G.. Agents Chemother.. Hetherington.. 2005. 43. K. Arduino... T. 2612–2617. 49. 2006. Eastwood. Application of paramagnetically tagged molecules for magnetic resonance imaging of biofilm mass transport processes. Salvenmoser. Paul. T.... J. Quave. Schaefer.. Simonetti. S. J... J.. S. M. C. W. Evaluation of the efficacy of disinfection procedures against Burkholderia cenocepacia biofilms. Murga..W. Pink. Anders-von Ahlften.. Control Hosp.. Boeld.. Gilbert. 909–919. S. C. S.J.. C.. D. Oberbauer. 2009.R..W. Antimicrob. J.C.... T. A constant-depth laboratory model film fermentor. In: Coenye. Dew.. A. G. Hube. Mustoe. A 75...L. M. Van Dijck.S. Sloan. 2008.. Oliveira. Surgery 147. Fiscarelli. E. Wound Repair Regen. Phoenix. Weber. Hollingshead. Fey.L..M.. G. Nelis. G.. Optimal antimicrobial catheter lock solution.. M. 286–292. Agents Chemother.. P.. 132.. Palmer. Ploeg. B. Infect. T. Antimicrob. M. Pittman. T.S. K. Madanahally.... M.. Protoc. M. J.K. 61. Liposomal amphotericin B eradicates Candida albicans biofilm in a continuous catheter flow model.A.B. 1808–1820. Gibcus... Balaban.. Carr. R. Tolker-Nielsen. M.. Tournu.. 1236–1244. J. Biopharm. T. G.. G. 1363–1367.. Environ. Watt. Bioanal. Stepper... 2007. Invest.. J. Sharma. R... M. Coenye. D. Parsek. 2008c. Nelis. 361–368. L.C.. G. Concheiro.L. Pätzold. Head Neck 27.. Insight into the microbial multicellular lifestyle via flow-cell technology and confocal microscopy. Infect.E. Nelis.. O.. J. Nelis.. Immun. W. Antimicrob... Schaller... A new approach to nondestructive analysis of biofilms by confocal Raman microscopy. R..S. P. Microbiol. H. Qin. 101–107. Post. J. Immun..M..C. Greenberg. Tan. A.J. Savini... 194. 231–233. J. Microbiol. Clin. Ramage.L. Infect.E. Z. Appl. Welsh. O. 3668–3673. J. Pitt... 1997.R. E. 1. A.. Microbiol. E. Taboada. Sauer. M.. A. Influence of fluid shear and microbubbles on bacterial detachment from a surface.L.. Bujdáková.. Singh.. Draper. H. P. pp. Di Bonaventura. Rybak.O.K. Glover. Galiano. 1993. Rupp. van der Mei.J. 2005.J.F.. B. R. 466–471... Antimicrob.. Rev. Chandra. K. C. Holmes. Giacometti.T. Franks. J. Spanier. 1140–1148.. A. Lipid-based slow-release formulation of amikacin sulfate reduces foreign body-associated infections in mice.. Peeters.. N. M. Orthop.L. J. Y. Y. R.E. Cirioni. Winn.. K. S. J... rapidly eradicates organisms embedded in biofilm. Gatifloxacin efficacy in treatment of experimental methicillin-sensitive Staphylococcus aureus-induced osteomyelitis in rabbits. Albers. Vandamme.D. C... M.J. Coenye. Infect.. Pennington. D. A. D. P..M. S. Immun. Standardized method for in vitro antifungal susceptibility testing of Candida albicans biofilms. Ou. 2005.M..mc01b05s6. Agents Chemother. Brennan. Stewart. K.. Infect. Smith-Palmer. Spatiotemperal progression of localised bacterial peritonitis before and after open abdomen lavage monitored by in vivo bioluminescent imaging. Sandt. 2004. Gross.. L. B.R. Donlan. Pitt. 2007. Van Dam. M. M. Nelis / Journal of Microbiological Methods 83 (2010) 89–105 Pamp. Activities of high-dose daptomycin. Kite. Growing oral biofilms in a constant depth film fermentor (CDFF). T.E.T.. Schierle.. J. Emtiazi... A. Ricicová. 1999b... Peeters. M. Confalone. P. Model systems of Burkholderia cepacia complex infection.A.. C.M. ISME J. L. J. W.O.D. 4027–4036. B. Clin. 2007. 45. 70. G.. Tashiro. S.. 2009.C. Goteri. Chaiban. H.R. R. Schaalje. 2005. Dis... Orlando. Hanna. Ghannoum. B.... Kean. Barton. Am. 52. M. 605–608.. 54... Zelver. H. Biomaterials 31. Agents Chemother.C. Med. Effect of dairy products on the lifetime of Provox2 voice prostheses in vitro and in vivo. 269–276.R. M. Experimental osteomyelitis treatment with antibiotic-impregnated hydroxyapatite. R.C.O.. Jiang. Biomaterials 14..L. C. 68. F. Engelsman.R. Kohno. 2008.K. Pink. Van Eldere. McBride. Mack. Moninger.. 81.... Busscher. D.. Chemother. M. Korting. Roeder. E.. 199. 515–519. 60. R.. V. M. 2767–2773. 47. 762–764... G. Vande Walle. 2007. Annu. C. V.A. doi:10.1002/9780471729259. M. Sternberg..5. W. Methods 54.. Sokol. Yanagihara.A. 1981. G. J. Perloff.A. H. G. Immun. Qu. Nicoletti. 26. W. M.. M. 17. W. Chem. J. 79–86. Biofouling 23.M. 10. Busscher.. J. 389–397.B. Pratten.. 492–495. Calhoun. S. D. 46. 48. Pompilio. 118. 1995.E. R. Shuster. X.. 2001. U.K. Microbiol. 6:1B.. Methods 72. vancomycin and moxifloxacin alone or in combination with clarithromycin or rifampin in a novel in vitro model of Staphylococcus aureus biofilm. Mader. Plano.H.. 59.T. Saba.C. Rabbit model of Candida albicans biofilm infection: liposomal amphotericin B antifungal lock therapy. Pettit. Role of coagulase in a murine model of hematogenous pulmonary infection induced by intravenous injection of Staphylococcus aureus enmeshed in agar beads. Epidemiol. T. Hamilton. Rupp. 57.. 2006... 2003. Sagers. Wimpenny. M. K. Infect.. Petty. G. B.. May. EDTA..). Air–water interface displaces adsorbed bacteria.. Biotechnol. Res... Pang. Microbiol. 1.. Peeters. Scalise. Burkholderia: Molecular Microbiology and Genomics.E.D. Schinabeck. K. Van Oeveren. J. Pharm. Shirtliff.. C..S. K. J. Infect. S. C.. K. 1984–1995. 2004.A.. Silicone colonization by non-Candida albicans Candida species in the presence of urine. Jain. Dvorak.K. 2627–2632.. Y..A. E. Hong. M. Parra-Ruiz.. Infect. Parsek. Zakikhany.K. van der Mei. Verstraete. Brückl.. 1985. 83–93. K. Blackall.G. D. Kim. 1999. R.. Ulphani. B. 2010. Weindl... H.D. J. Kitoh.K. B. Coenye.....K.. Kucharíková. Keuntje..W. J... F. Meth.. L. M.. A. J. A. Shakeri. C. A. Robison. R. S. Lopez-Ribot. Bucio. A. 747–754. using different combinations of minocycline. 705–707. Role of the Staphylococcus epidermidis slime layer in experimental tunnel tract infections. Sawai. 65. 219. Coenye. D. J. 2008. 1752–1755. Kaku. Silvestri. Longo. 67.. R.E. Schwandt. Seidler. Petrucca. K. The potential of bio-optical imaging of biomaterialassociated infection in vivo.... van Dam. S.. Wickes. J. Keller.. H. Horizon Bioscience.J. Shirtliff. Antimicrob.. M. 1999a. E. 2000.K.. Assessment of biofilm cell removal and killing and biocide efficacy using the microtiter plate test.J.. Trends Microbiol. Müller. Heer.. 2006. Ramanan. Development of a microfluidic biochip for online monitoring of fungal biofilm dynamics.C. Calhoun... S. K. P. Silverthorne. J. 39. H. A.... Negri... 2656–2659. 2010.... A newly designed model for infection-induced bladder stone formation in the rat. Microbiol. Roehrborn. 2002b... Steckelberg.. Hirakata.. Arnaut. V. H.J. 2010.P. Nealson.. Resistance of planktonic and biofilm-grown Burkholderia cepacia complex against the transition metal gallium.F. H. Microbiol. Formation and properties of in vitro biofilms of ica-negative Staphylococcus epidermidis clinical isolates. Hendrix. 229–250. M. Patel... 2003.. Hossain. Busscher. 2006. W. Bennett. 2008a. Antimicrob. 2010. R. Busscher.J. Coenye.S. 386. 14. W. W. 2001. P.. Adhesion to and biofilm formation on IB3-1 bronchial cells by Stenotrophomonas maltophilia isolates from cystic fibrosis patients. Antimicrob. Ulphani. Confocal Raman microspectroscopy as a tool for studying the chemical heterogeneities of biofilms in situ. Mack. Scalise.. M. Brown.. Models of oral and vaginal candidiasis based on in vitro reconstituted human epithelia. A.. Hube. Bone Joint Surg. Evidence of bacterial biofilms in a rabbit model of sinusitis. H. 1992.. Experiments in a canine model. H. Kast.J. Fey. 1727–1732. 2010. P. 71. R. 2083–2094. 102. Rabaey.. Swords.J. 2001. J. Comparison of multiple methods for quantification of microbial biofilms grown in microtiter plates.. J.Q.. S.. R. Crocetta. P. A. R. 2009. Dijkstra.. Agarose stabilization of fragile biofilms for quantitative structure analysis...

Van Lierde. Appl. 155. K.. Maes.. Stepanovic.. Kaneko. Microbiol. Y. Vuong.. Appl. Malott. Garland. Microbiol. Persistence and decontamination of Bacillus atrophaeus subsp. Stewart. M. Agents Chemother. Environ. 2009. H. Horemans. C.. O...J. Clinical efficacy of intravenous administration of marbofloxacin in a Staphylococcus aureus infection in tissue cages in ponies.T. 2010.. 2003. Willard. Tamashiro.. H. 2009a.L. Kristensen. S. Mol.. Maragoudakis... Hinse. M. Chemother.. Szabo.. Y. Induction of phagocytic inhibitory activity in cats with chronic Pseudomonas aeruginosa pulmonary infection...A.. 5208–5218.S. Vanden Berghe. R. Wang. Thomas. Henriksson.. 2001. D. K. The effect of solid surfaces upon bacterial activity. Environ. Dong. Y. Greenman.. Y.W.. Quorum sensing and signal interference: diverse implications.. 2008. Kawada.W. Antimicrob. Peetermans. Zhang. as a model of wound infection.R. 2003. Stoodley. K. Antimicrob. 2009. Xia. F. Maes.. Development of realtime in vivo imaging of device-related Staphylococcus epidermidis infection in mice and influence of animal immune status on susceptibility to infection. Stepanovic.. An in vitro model of bacterial infections in wounds and other soft tissues. Sci. Microfabrication meets microbiology. Environ.M. R.. Zimmerli.. J. Nelson.. Antimicrob. Dowd.. Kakeya.G.). Yanagihara. pp. An in vitro biofilm model of subgingival plaque. D. T. Dowd.. 107. globigii spores on corroded iron in a model drinking water system. Tomono. G. Tzagaroulakis. architecture. P. Bayer....D. T. Washington. 487–497.. Appl.D.R. L. Meckes. H. N. 2005... Whitesides. Combination therapy for chronic Pseudomonas aeruginosa respiratory infection associated with biofilm formation. S. Van Wijngaerden.L. J. Efficacy of ME1036 against methicillin-resistant Staphylococcus aureus and vancomycininsensitive S. 2009. Kohno. Ajiki.. K. Yanagihara. 337–342. J. Izumikawa.. J. An in vitro model of Pseudomonas aeruginosa biofilms on viable airway epithelial cell monolayers..S. Sawai. J.. T. 1982. J. Yamamoto. Santo Domingo. Z.. S.. Holland.. Microbiol. Environ. M. Kakeya. A new colorimetric microtitre model for the detection of Staphylococcus aureus biofilms.. Huang. Cultivation of microbial consortia and communities. Agents Chemother. M.. J.. F.. V. J.. 78. D. M. 1999.. Vaudaux. 46. 53. J. Antimicrob.. Environ. M.N. S. J. Lipson. Appl.. Res.... Inhibitory effect of biocides on the viable masses and matrices of Staphylococcus aureus and Pseudomonas aeruginosa biofilms.. Biofouling 21. Givskov.. M. Chaturvedi.. Microbiol. Maes. A. J. 47. K. 1303–1312..Q. Pediatr.. Clin.. R. 606–615. A modified microtiter-plate test for quantification of staphylococcal biofilm formation. H.J.L. Baker..C. Thorn. Kadota.. 83. Robarts. J. Care Med. Cos.. S. C.W.. Chemother. Schenk. 1–18. Rees. In: Hurst.H. 9.D. S. Yu. J.L. Le Bihan. Ther.J. M. Dé...J.H. 71.A.. Microbiol. 2000. Mizuta. K.. Ceri.L. Bénéré. W. Araki. 1987.. Multicellular organization in a degradative biofilm community. Sonksen. Uppuluri. B. T. American Society for Microbiology Press. C. Levecque. S. S. 1997. Zobell. Potential of biofilm-based biofuel production. J. E. G.. B. Storey. T. S.. Mygind.. M. Macé....K.. 258–261. Microbiol. P. Vanden Berghe. K. K. Rayner. Y.. J. Hirakata. N.L.. S. Microbiol. T. a novel des-F(6)-quinolone... Yamada. 2007. Dis. Antimicrob.. 3135–3142. Morinaga.. Winnie. BMC Microbiol. Busscher. 2001. Y. Sawai. Agents 34. Lawrence. K.. L. Efficacy of linezolid against Panton-Valentine leukocidin (PVL)-positive methicillin-resistant Staphylococcus aureus (MRSA) in a mouse model of haematogenous pulmonary infection... Effect of clarithromycin on lymphocytes in chronic respiratory Pseudomonas aeruginosa infection. H. 1982. Anim... Microbiol. G. R. In vivo transcript profiling of Candida albicans identifies a gene essential for interepithelial dissemination.. Wound Repair Regen.. P. Yamada.. Antimicrob.E. S. 56. Z. Klinger. G. Agents 28. Roe. E. P. 2007.. C.. P. Tré-Hardy. Devleeschouwer. M. Tashiro. N.... T. K. Høiby. Costerton. Microbiol.L. M.. van Soest.. Sun. D. 187–209. 235–238. S.. Vandenbosch.. P. Higashiyama. Kurihara. Y.. Y. Lopez-Ribot.M. Edwards. 2000. Diluzio. 2007. Annu. 2010. 65..M.. 1994. Higashiyama. 1749–1755. Quorumsensing mutations affect attachment and stability of Burkholderia cenocepacia biofilms. Ensink. 16. P. V. A. Y. Kohno.. 434–446.R. G.. M. P. Cohen..P.. E. A....P. H. 1999.. Kihara. C. D. C. 2004. W. A novel in vitro flat-bed perfusion biofilm model for determining the potential antimicrobial efficacy of topical wound treatments.. Kadurugamuwa. Yanagihara. 156–164. Clin. Am. El Manssouri... Seki. 10. 2001.. 69. Izumikawa. 2010.... Dakic. 2938–2954. W. Int... Bacteriol. Costerton.. S. aureus in a model of haematogenous pulmonary infection. 4078–4083. Y. K. 105 Vollmer. Miyazaki. Kocianova. J. S. Waldvogel. Nelis. 401–404. M.. K. Biomaterials 22.E.. K.. R. Y. R. S. H.. J. 37.. Infect.M. 64.... A. P.... 2005. Cos..S. 29. Appl. and drug resistance. Respir... J.M. Infect.. 175–179. Smith.. D. Vanderbist.. Tresse.. Otto. A mouse model of chronic pulmonary infection with Pseudomonas aeruginosa and Pseudomonas cepacia. Yu. 1943. Effect of antibiotic co-administration on young and mature biofilms of cystic fibrosis clinical isolates: the importance of the biofilm model.. Appl. Seki. van Der Mei.. Yamamoto. Microbiol. Kohno. Sternberg. Yanagihara. K..G. J. Spatial physiological heterogeneity in Pseudomonas aeruginosa biofilm is determined by oxygen availability. Immunol. E. F. Design of a simple model of Candida albicans biofilms formed under conditions of flow: development. Manual of Environmental Microbiology. Werthén.R.. Torii.K. Neut. Yokota. D. J. R. Antimicrob. Agents Chemother. S.L. 51.. Djukic.. 669–674.. Interactions between endocarditisderived Streptococcus gallolyticus subsp. Agents Chemother. G. J..... 209–218. Parsek. C. Morinaga. K. Rice. Lawrence. Infect. H. Fungicidal activity of miconazole against Candida spp. S.. Toté. 2007. I.. H. Wolfaardt. Stetzenbach.D.A. Ohno.G. S. Int. K. van Horn.. J. J.. Tomlin. 2007. 212–216. 69–72. Microbiol. Djordjevic. Y. Pathogenesis of foreign body infection: description and characteristics of an animal model. D.A.. Kohno. 2007. J.E. Kadota. E.. 2313–2320. 2007. nose. 49. Uppuluri. Nelis / Journal of Microbiological Methods 83 (2010) 89–105 Song. Sulaeman. Chemother. J. Crawford. 10. C. 38. 380–387. 2010.B. Antimicrob. Antimicrob.. Antimicrob.A. S. Rossero. Immun. W. Zakikhany. .. Ferekidis. 39–56.. Population diversity in model potable water biofilms receiving chlorine or chloramine residual.. Voermans. van de Belt. Hirakata. E. Use of a bioluminescent Pseudomonas aeruginosa strain within an in vitro microbiological system. Rupp. W.. Bishop. Seki. 2008. Mycopathologia 168.. T.. J.. J. 2008.E. M. Starke. J.. J. 268–273.. Chaturvedi.W. Appl. Van Eldere. Nikolopoulos... J. K. Biotechnol.. 2009.. D. Yoshikawa. Greenman. K. Y. 2009. 955–958. 40–45..R. Microbiol.R.. Wu. 1088–1093. P. Potency of DX-619.. Caldwell. J. 22. Appl. M. Oral Microbiol..J. Wolfaardt. APMIS 118. 694–700.. Y. 3868–3874. 4035–4039. D. Antibiotic resistance of bacteria in biofilms. Y. 146. 44. Microbiol. Schmidt-Westhausen..M. In vitro multispecies Lubbock chronic wound biofilm model. Y.S. H. Int. Influence of the incubation atmosphere on the production of biofilm by staphylococci...R.. L.. G. Palmer. Rhinol.. Nonaka.E... D. 2004. P.. 477–481. N. and throat infections: how important are they? Laryngoscope 117. Ramage. L. Teitzel.. Yanagihara. Stewart. Smith. Francis.M.. 3 rd Ed. Coenye.. 279–288. Lab.. Microbiol.. Miyazaki. D. P. Coenye. Antimicrob. E.Ø. 9. K. Tomono.. Korber. J. Federighi. J. Woodworth. M... B. A. P. 2008. Y.. Y. M.. L..D. Y. A. Foreign body infection: a new rat model for prophylaxis and treatment. Koga. Hube. Appl.J. J.. 108. E... Ohnishi. Vandersmissen.C. D.L. P... Biofilms in ear. 426–431.M. Nakamura. Sherman. Kadurugamuwa. Ulphani. Svabic-Vlahovic.. Infect..T. S. Tsukamoto. van Duijkeren. J. H.A. G.M. in haematogenous murine bronchopneumonia caused by methicillin-resistant and vancomycin-intermediate Staphylococcus aureus. K. 22.. M. Lopez-Ribot.. 101–111. Langston. Effects of intratracheal administration of novispirin G10 on a rat model of mucoid Pseudomonas aeruginosa lung infection. 60. L.M.H... D. 698–702. to assess the antimicrobial efficacy of wound dressings by monitoring light production. Chemother. Methods 40.. J. Tsukamoto. Nat.. Wound Care 18.V. J.. T. J. H. Pharmacol..D. C. 2006. F.... 1993.. R. 198. 525–532. H. Morikawa.E. 1998. Bobbaers. Rhoads. Biofilm penetration and disinfection efficacy of alkaline hypochlorite and chlorosulfamates. K... Djukic. Vanden Berghe. Staphylococcus aureus biofilm formation on different gentamicin-loaded polymethylmethacrylate bone cements. 1607–1611.J. J. Walker. 2009. Tomono. Microbiol. Sun.. J. J. T. McFeters. Thorn. 555–560. Infect. Naglik... Bjarnsholt. Mills. J.. S. Davies. H. Kuroki. M. Comparison between the biofilm initiation of Campylobacter jejuni and Campylobacter coli strains to an inert surface using BioFilm Ring Test. G. 5.J. Kamihira. Sokol. Appl. Savic. 49. H. J. 152–161. 22.. K.. J.. Traore. Interaction between biofilms formed by Pseudomonas aeruginosa and clarithromycin. Characteristics of Candida albicans biofilms grown in a synthetic urine medium.. Toté. 2009b. Dinakaran. Jensen. Cos. M. Sedlacek.. 135–138.S. Appl.J. Crit. M... Environ. Stewart. K. 2008.. J. D. Vukovic.B.. 668–673. C. Xu. Lett.. J. Chen.. Wolcott.R. 283–287. biofilms. Kleesiek. Vet. Dis. E. Nydegger. Kamihira. Propagation of anaerobic bacteria within an aerobic multi-species chronic wound biofilm model. J. 2451–2457. 1563–1571. gallolyticus isolates and human endothelial cells. Bhargave. Microbiol. (Eds.. Toté.. Thomassen. Xiong.. Dreier. Lancet 358. 2006.. 2070–2079. 249–254. Vlastarakos. 3217–3224. Realtime in vivo bioluminescent imaging for evaluating the efficacy of antibiotics in a rat Staphylococcus aureus endocarditis model.W. 2010. I.. Heavy metal resistance of biofilm and planktonic Pseudomonas aeruginosa. J... Y.E. H. Kohno. 91.. Weibel. Braeckmans... D. Schaller.. P. U.. J. Y. Koga. 2005.M. Izumikawa. Sauer. J. P. T. Microbiol. Cell. 2002. Microbiol. Agents 32. 2005. Wolcott.S.. 73. Williams. D. 49...C. Model of Staphylococcus aureus central venous catheterassociated infection in rats. Agents 33. Raventos...P. 107. Effect of arbekacin on a methicillin-resistant Staphylococcus aureus-induced biofilm in a rat model. 101–109. H. K. Rev. Hirakata. Yasuda. Microbiol. Biofilms as complex differentiated communities. Am.M. J. S. Int... Maesaki.. J.. 46.. 805–813. Evaluation of hydrogen peroxide-based disinfectants in a new resazurin microplate method for rapid efficacy testing of biocides. Rev. J. Caldwell. A. Tashiro. Appl.. Y. 46. G... 76.