The biofilm architecture of sixty opportunistic pathogens deciphered using a high throughput CLSM method

Bridier, Arnaud; Dubois‐Brissonnet, Florence; Boubetra, A; Thomas, Vincent; Briandet, Romain

doi:10.1016/j.mimet.2010.04.006

Cited by 202 publications

(138 citation statements)

References 54 publications

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“…Evaluation of the time necessary for pioneer planktonic cells to reach the bottom of biofilms. Biofilms of B. thuringiensis 407 were grown in 96-well microscopic-grade microtiter plates (27). Bacterial suspensions were prepared as described above for flow biofilms.…”

Section: Methodsmentioning

confidence: 99%

“…Visualization of matrix strength evolution according to biofilm age. Twenty-fourhour and 72-h B. thuringiensis 407 biofilms grown in flow cells were perfused with 3 mL of an exponential phase culture of B. thuringiensis 407-GFP at a concentration of 10 8 cfu/mL The flow was stopped for 1 h. After this period, the flow was resumed at 15 mL/h, and after a 2-h incubation, fluorescent cells detected inside the biofilm were quantified by confocal microscopy and their biovolumes were estimated with the PHLIP MATLAB routine (27).…”

Section: Methodsmentioning

confidence: 99%

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Bacterial swimmers that infiltrate and take over the biofilm matrix

Houry

Gohar

Deschamps

et al. 2012

Proc. Natl. Acad. Sci. U.S.A.

176

148

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Bacteria grow in either planktonic form or as biofilms, which are attached to either inert or biological surfaces. Both growth forms are highly relevant states in nature and of paramount scientific focus. However, interchanges between bacteria in these two states have been little explored. We discovered that a subpopulation of planktonic bacilli is propelled by flagella to tunnel deep within a biofilm structure. Swimmers create transient pores that increase macromolecular transfer within the biofilm. Irrigation of the biofilm by swimmer bacteria may improve biofilm bacterial fitness by increasing nutrient flow in the matrix. However, we show that the opposite may also occur (i.e., swimmers can exacerbate killing of biofilm bacteria by facilitating penetration of toxic substances from the environment). We combined these observations with the fact that numerous bacteria produce antimicrobial substances in nature. We hypothesized and proved that motile bacilli expressing a bactericide can also kill a heterologous biofilm population, Staphylococcus aureus in this case, and then occupy the newly created space. These findings identify microbial motility as a determinant of the biofilm landscape and add motility to the complement of traits contributing to rapid alterations in biofilm populations.bacterial ecology | biofilm disruption | motile subpopulations | antimicrobials | time-lapse confocal imaging

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Bacterial swimmers that infiltrate and take over the biofilm matrix

Houry

Gohar

Deschamps

et al. 2012

Proc. Natl. Acad. Sci. U.S.A.

176

148

View full text Add to dashboard Cite

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“…CLSM markedly reduces the need for pretreatments such as disruption and fixation, which reduce or eliminate the evidence of microbial relationships, complex structures and biofilm organization, without the limitations encountered with scanning electron microscopes [21][22][23]. Bridier et al [24] proposed CLSM combined with the use of 96-well microtiter plates compatible with high resolution imaging for the study of biofilm formation and structure. The authors reported that the combined use of microplates and confocal imaging proved to be a good alternative to other high throughput methods commonly used since it permits the direct, in situ qualitative and quantitative characterization of biofilm architecture.…”

Section: Introductionmentioning

confidence: 99%

Comparison of methods for the detection of biofilm production in coagulase-negative staphylococci

Oliveira

Bonesso

Cunha

2010

Microorganisms in Industry and Environment

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Background: The ability of biofilm formation seems to play an essential role in the virulence of coagulase-negative staphylococci (CNS). The most clearly characterized component of staphylococcal biofilms is the polysaccharide intercellular adhesin (PIA) encoded by the icaADBC operon. Biofilm production was studied in 80 coagulasenegative staphylococci (CNS) strains isolated from clinical specimens of newborns with infection hospitalized at the Neonatal Unit of the University Hospital, Faculty of Medicine of Botucatu, and in 20 isolates obtained from the nares of healthy individuals without signs of infection. The objective was to compare three phenotypic methods with the detection of the icaA, icaD and icaC genes by PCR.

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“…While the submerged model represents environmental conditions confronted by B. subtilis in its natural habitats, it has only recently been applied to the species (22,27), likely due to experimental limitations associated with the bacterium (17,27). In a previous study, we proposed a new methodology to visualize and quantify B. subtilis submerged biofilms using a microplate-based model combined with confocal laser scanning microscopy (CLSM) (27,28). This experimental system enabled the identification of a remarkable B. subtilis strain (NDmed), isolated from an endoscope washer-disinfector (29), capable of forming thick and protruding biofilms highly resistant to the biocides commonly used for endoscope disinfection in hospitals (30).…”

mentioning

confidence: 99%

Identification of ypqP as a New Bacillus subtilis Biofilm Determinant That Mediates the Protection of Staphylococcus aureus against Antimicrobial Agents in Mixed-Species Communities

Sanchez-Vizuete

Coq

Bridier³

et al. 2015

Appl Environ Microbiol

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In most habitats, microbial life is organized in biofilms, three-dimensional edifices sustained by extracellular polymeric substances that enable bacteria to resist harsh and changing environments. Under multispecies conditions, bacteria can benefit from the polymers produced by other species ("public goods"), thus improving their survival under toxic conditions. A recent study showed that a Bacillus subtilis hospital isolate (NDmed) was able to protect Staphylococcus aureus from biocide action in multispecies biofilms. In this work, we identified ypqP, a gene whose product is required in NDmed for thick-biofilm formation on submerged surfaces and for resistance to two biocides widely used in hospitals. NDmed and S. aureus formed mixed biofilms, and both their spatial arrangement and pathogen protection were mediated by YpqP. Functional ypqP is present in other natural B. subtilis biofilm-forming isolates. However, the gene is disrupted by the SP␤ prophage in the weak submerged-biofilm-forming strains NCIB3610 and 168, which are both less resistant than NDmed to the biocides tested. Furthermore, in a 168 laboratory strain cured of the SP␤ prophage, the reestablishment of a functional ypqP gene led to increased thickness and resistance to biocides of the associated biofilms. We therefore propose that YpqP is a new and important determinant of B. subtilis surface biofilm architecture, protection against exposure to toxic compounds, and social behavior in bacterial communities. Bacillus subtilis is a nonpathogenic Gram-positive bacterium that can be found in its natural habitats as free cells or associated with surfaces in biofilms. In the soil, B. subtilis strains have been shown to form surface-associated communities on plant tissues that protect them from infection by pathogens (1-3). Because of its ability to resist stress through biofilm or spore formation, the bacterium has been isolated from extreme environments, such as sand deserts, clouds, and the digestive tracts of animals (4 -6). As well as its ubiquitous presence in diverse habitats, B. subtilis has been used extensively in biotechnological applications, such as the production of natto, a traditional Japanese food made of fermented soybeans (7), and the production of industrial enzymes and pharmaceutical proteins (8, 9). As a generally recognized as safe (GRAS) organism, B. subtilis is also used as a biocontrol agent in agriculture and livestock buildings (10 -14) and as a probiotic agent to improve human and animal health by preventing gastrointestinal infections (15,16).In fundamental research, B. subtilis has emerged as the model organism for deciphering the complex genetic regulation involved in the biofilm mode of life of Gram-positive bacteria. The domesticated strain B. subtilis 168 has been widely used to dissect metabolic and cellular processes. However, this strain is not able to form robust pellicles and complex colonies like those of its parental strain, NCIB3610, a descendant of the original Marburg strain that was deposited in 1951 (17)....

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The biofilm architecture of sixty opportunistic pathogens deciphered using a high throughput CLSM method

Cited by 202 publications

References 54 publications

Bacterial swimmers that infiltrate and take over the biofilm matrix

Bacterial swimmers that infiltrate and take over the biofilm matrix

Comparison of methods for the detection of biofilm production in coagulase-negative staphylococci

Identification of ypqP as a New Bacillus subtilis Biofilm Determinant That Mediates the Protection of Staphylococcus aureus against Antimicrobial Agents in Mixed-Species Communities

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