A fluorescence assay to determine the viable biomass of microcosm dental plaque biofilms

Filoche, Sara; Coleman, Michele; Angker, Linny; Sissons, C.H.

doi:10.1016/j.mimet.2007.02.015

Cited by 26 publications

(43 citation statements)

References 33 publications

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“…These limitations of plate counting techniques prompted the use of molecular techniques and other alternative methods (Vitali et al, 2003). New methods include molecular based techniques such as quantitative real-time polymerase chain reaction (PCR), fluorescent in situ hybridization (FISH) (Boulos et al, 1999;Veal et al, 2000), confocal scanning laser microscopy (CSLM) (Auty et al, 2001;Gardiner et al, 2000;Palencia et al, 2008), flow cytometry (Alakomi et al, 2005) and microplate scale fluorochrome staining assay (Filoche et al, 2007;Mättö et al, 2006). Flow cytometry is a rapid and sensitive technique that measures physiological characteristics such as membrane integrity, enzyme activity, respiration, membrane potential and intracellular pH (Bunthof et al, 2001) of each cell individually (Bunthoff & Abee, 2002).…”

Section: Methods For Quantification Of Probiotic Culturesmentioning

confidence: 99%

See 1 more Smart Citation

Probiotics - What They Are, Their Benefits and Challenges

Thantsha¹,

Mamvura²,

Booyens³

2012

New Advances in the Basic and Clinical Gastroenterology

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Section: Methods For Quantification Of Probiotic Culturesmentioning

confidence: 99%

“…Microplate scale fluorochrome staining assay is appropriate for assessing viability of fresh, freeze-dried and stressed cells. It can detect changes in the condition of probiotic cells earlier than can be done with conventional cultivation methods (Filoche et al, 2007;Mättö et al, 2006).…”

Section: Methods For Quantification Of Probiotic Culturesmentioning

confidence: 99%

Probiotics - What They Are, Their Benefits and Challenges

Thantsha¹,

Mamvura²,

Booyens³

2012

New Advances in the Basic and Clinical Gastroenterology

View full text Add to dashboard Cite

“…Cell damage started from the periphery of bacterial aggregates and slowly continued into the depths. Other authors have reported the inability of CHX to completely kill all bacteria in different in vitro biofilm systems (15,19,32) and in vivo studies (48,55) when a normally used clinical concentration was applied at usual exposure times. The killing efficacy was dependent on the age of the biofilm and thus on its thickness and composition.…”

mentioning

confidence: 99%

Real-Time Microsensor Measurement of Local Metabolic Activities in Ex Vivo Dental Biofilms Exposed to Sucrose and Treated with Chlorhexidine

Ohle¹,

Gieseke

Nistico

et al. 2010

Appl Environ Microbiol

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Dental biofilms are characterized by structural and functional heterogeneity. Due to bacterial metabolism, gradients develop and diverse ecological microniches exist. The aims of this study were (i) to determine the metabolic activity of microorganisms in naturally grown dental biofilms ex vivo by measuring dissolved oxygen (DO) and pH profiles with microelectrodes with high spatial resolution and (ii) to analyze the impact of an antimicrobial chlorhexidine (CHX) treatment on microbial physiology during stimulation by sucrose in real time. Biofilms were cultivated on standardized human enamel surfaces in vivo. DO and pH profiles were measured in a flow cell system in sterile human saliva, after sucrose addition (10%), again after alternative treatment of the sucrose exposed biofilms with CHX (0.2%) for 1 or 10 min or after being killed with paraformaldehyde (4%). Biofilm structure was visualized by vitality staining with confocal microscopy. With saliva as the sole nutrient source oxygen consumption was high within the superficial biofilm layers rendering deeper layers (>220 m) anoxic. Sucrose addition induced the thickness of the anaerobic zone to increase with a concurrent decrease in pH (7.1 to 4.4). CHX exposure reduced metabolic activity and microbial viability at the biofilm surface and drove metabolic activity deeper into the biofilm. CHX treatment led to a reduced viability at the biofilm surface with minor influence on overall biofilm physiology after 1 min; even after 10 min there was measurable respiration and fermentation inside the biofilm. However, the local microenvironment was more aerated, less acidogenic, and presumably less pathogenic.

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“…The microcosm technique or "artificial mouth" developed by Wong and Sissons 15 has already been explored for the assessment of antimicrobial substances and in the field of cariology [16][17][18][24][25][26][27][28] . However, few studies exist on adhesion and biofilm formation on titanium surfaces and on peri-implantitis 29 .…”

Section: Resultsmentioning

confidence: 99%

“…It was stimulated with parafilm (Parafilm "M"  , American National Can TM, Chicago, IL, USA) and collected 70 mL of saliva that was placed in a sterile graded collector and transported in a foam ice chest to the local Laboratory of Microbiology. The saliva was filtered in sterile glass wool and homogenized in a vortex mixer 17,18 . An aliquot of such saliva was used for bacterial quantification (CFU/mL).…”

Section: Saliva Collection and Processingmentioning

confidence: 99%

Microcosm Biofilm Formation on Titanium Surfaces

et al. 2015

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Risk factors associated with peri-implantitis are related to the biofilm composition around the implant and the ability of bacterial adhesion. This study evaluated the biofilm formation on different surfaces of commercially pure titanium (CP Ti) grade 4 after 12, 24, 48 and 168 hours using the microcosm technique and scanning electron microscopy (SEM). The following surface conditions were examined: S (control)-smooth and plain; B-sand-blasted with aluminum oxide; E-etched using nitric acid; and BE-the combination of J and A treatments. Statistical differences on biofilm formation (CFU/ mg) were found between the control (S) and B surfaces at the first 12 hours, which are related to the lowest (R a = 0.21 µm) and highest (R a = 0.62 µm) mean roughness values. At 168-hour all surfaces showed similar biofilm formation. Yet, microbial growth occurred on all surfaces, regardless of the surface treatment.

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A fluorescence assay to determine the viable biomass of microcosm dental plaque biofilms

Cited by 26 publications

References 33 publications

Probiotics - What They Are, Their Benefits and Challenges

Probiotics - What They Are, Their Benefits and Challenges

Real-Time Microsensor Measurement of Local Metabolic Activities in Ex Vivo Dental Biofilms Exposed to Sucrose and Treated with Chlorhexidine

Microcosm Biofilm Formation on Titanium Surfaces

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