Enhanced Bactericidal Efficacy of NaOCl at pH 12 Followed by Acidified NaOCl at pH 6.5 on Enterococcus faecalis Biofilm

ABSTRACT The association of Cryptosporidium oocysts with biofilm communities can influence the propagation of this pathogen through both environmental systems and water treatment systems. We observed the capture and retention of C. parvum oocysts in Pseudomonas aeruginosa biofilms using laboratory flow cells. Biofilms were developed in two different growth media using two different strains of P. aeruginosa, a wild-type strain (PAO1) and a strain that overproduces the exopolysaccharide alginate (PDO300). Confocal laser-scanning microscopy was used in conjunction with image analysis to assess the structure of the biofilms prior to introducing oocysts into the flow cells. More oocysts were captured by the biofilm-coated surfaces than the abiotic glass surface in both media. There was no significant difference in capture across the two strains of P. aeruginosa biofilm, but the fraction of oocysts captured was positively related to biofilm roughness and surface-area-to-volume ratio. Once captured, oocysts were retained in the biofilm for more than 24 h and were not released after a 40-fold increase in the system flow rate. We believe the capture and retention of oocysts by biofilm communities can impact the environmental transmission of C. parvum, and this interaction should be taken into consideration when predicting the migration of pathogens in the environment.

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Use of confocal laser scanning microscopy and a computer model to understand ink cavitation and filamentation

TAPPI Journal ◽

10.32964/tj9.10.7 ◽

2010 ◽

Vol 9 (10) ◽

pp. 7-15

Author(s):

HANNA KOIVULA ◽

DOUGLAS BOUSFIELD ◽

MARTTI TOIVAKKA

Keyword(s):

Laser Scanning ◽

Confocal Laser Scanning Microscope ◽

Laser Scanning Microscopy ◽

Confocal Laser Scanning ◽

Confocal Laser ◽

Print Quality ◽

Length Scale ◽

Offset Printing ◽

Significant Difference ◽

Printing Speed

In the offset printing process, ink film splitting has an important impact on formation of ink filaments. The filament size and its distribution influence the leveling of ink and hence affect ink setting and the print quality. However, ink filaments are difficult to image due to their short lifetime and fine length scale. Due to this difficulty, limited work has been reported on the parameters that influence filament size and methods to characterize it. We imaged ink filament remains and quantified some of their characteristics by changing printing speed, ink amount, and fountain solution type. Printed samples were prepared using a laboratory printability tester with varying ink levels and operating settings. Rhodamine B dye was incorporated into fountain solutions to aid in the detection of the filaments. The prints were then imaged with a confocal laser scanning microscope (CLSM) and images were further analyzed for their surface topography. Modeling of the pressure pulses in the printing nip was included to better understand the mechanism of filament formation and the origin of filament length scale. Printing speed and ink amount changed the size distribution of the observed filament remains. There was no significant difference between fountain solutions with or without isopropyl alcohol on the observed patterns of the filament remains.

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Bactericidal compounds controlling growth of the plant pathogen pseudomonas syringae pv. actinidiae, which forms biofilms composed of a novel exopolysaccharide

10.26686/wgtn.12597797 ◽

2020 ◽

Author(s):

S Ghods ◽

Ian Sims ◽

MF Moradali ◽

BHA Rehma

Keyword(s):

Pseudomonas Syringae ◽

Chlorine Dioxide ◽

Laser Scanning ◽

Extracellular Polymeric Substances ◽

Virulent Strain ◽

Bactericidal Effect ◽

Cell System ◽

Laser Scanning Microscopy ◽

Base Layer ◽

Confocal Laser

© 2015, American Society for Microbiology. Pseudomonas syringae pv. actinidiae is the major cause of bacterial canker and is a severe threat to kiwifruit production worldwide. Many aspects of the disease caused by P. syringae pv. actinidiae, such as the pathogenicity-relevant formation of a biofilm composed of extracellular polymeric substances (EPSs), are still unknown. Here, a highly virulent strain of P. syringae pv. actinidiae, NZ V-13, was studied with respect to biofilm formation and architecture using a flow cell system combined with confocal laser scanning microscopy. The biofilm formed by P. syringae pv. actinidiae NZ V-13 was heterogeneous, consisting of a thin cellular base layer 5 μm thick and microcolonies with irregular structures. The major component of the EPSs produced by P. syringae pv. actinidiae NZ V-13 bacteria was isolated and identified to be an exopolysaccharide. Extensive compositional and structural analysis showed that rhamnose, fucose, and glucose were the major constituents, present at a ratio of 5:1.5:2. Experimental evidence that P. syringae pv. actinidiae NZ V-13 produces two polysaccharides, a branched α-D-rhamnan with side chains of terminal α-D-Fucf and an α-D-1,4-linked glucan, was obtained. The susceptibility of the cells in biofilms to kasugamycin and chlorine dioxide was assessed. About 64 and 73% of P. syringae pv. actinidiae NZ V-13 cells in biofilms were killed when kasugamycin and chlorine dioxide were used at 5 and 10 ppm, respectively. Kasugamycin inhibited the attachment of P. syringae pv. actinidiae NZ V-13 to solid surfaces at concentrations of 80 and 100 ppm. Kasugamycin was bacteriostatic against P. syringae pv. actinidiae NZ V-13 growth in the planktonic mode, with the MIC being 40 to 60 ppm and a bactericidal effect being found at 100 ppm. Here we studied the formation, architecture, and composition of P. syringae pv. actinidiae biofilms as well as used the biofilm as a model to assess the efficacies of bactericidal compounds.

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Micromorphological analysis and bond strength comparison of two adhesives for different degrees of dental fluorosis

10.21203/rs.3.rs-16637/v3 ◽

2020 ◽

Author(s):

Shuangfeng Liu ◽

Yanxia Zhu ◽

Tana Gegen

Keyword(s):

Shear Strength ◽

Bond Strength ◽

Bonding Strength ◽

Laser Scanning ◽

Dental Fluorosis ◽

Laser Scanning Microscopy ◽

Confocal Laser ◽

Cohesive Failure ◽

Bonding System ◽

Significant Difference

Abstract The objective of this study was to analyze morphologically the all-etching bonding system and self-etching bonding system for enamel with different degrees of fluorosis and evaluate the bond strength of each system. Teeth that were indicated for extraction owing to orthodontic or periodontal problems were selected. According to Dean’s index and the Thylstrup-Fejerskov index, 180 extracted teeth were divided into three groups of mild, moderate, and severe dental fluorosis (DF), with 60 teeth in each group. The teeth in each group were randomly divided into two subgroups (n = 30), which were then subjected to the all-etching bonding system (Prime & Bond NT) and self-etching bonding system (SE-Bond). Each group of adhesives was used to bond Z350 universal resin (3M) to the etched dental enamel. Tensile and shear tests were conducted to determine the bond strength. Subsequently, the fractured specimens were investigated using scanning electron microscopy (SEM) and confocal laser scanning microscopy (CLSM). The Prime & Bond NT was statistically significant for the tensile and shear strength of enamel with mild fluorosis (P < 0.05) but did not exhibit a significant difference for moderate and severe DF (P > 0.05). The SE-Bond was not statistically significant for the tensile and shear strength of mild, moderate, or severe DF (P > 0.05). The SEM and CLSM results reveal that the mild fluorosis enamel crystals were relatively dense, and a small amount of resin remained. The moderate fluorosis enamel crystals were loosely arranged, and the gaps were widened. The severe fluorosis enamel crystals were irregularly arranged. The disorder was aggravated, and the dentinal orifice was exposed by partial enamel exfoliation. The bonding strength of mild fluorosis enamel with the Prime & Bond NT was better than that with the SE-Bond, and cohesive failure was the most common mode of failure. Because there was no difference in the bonding strength of the SE-Bond for different degrees of DF, we recommend the use of the all-etching adhesive system in the clinical treatment of teeth with mild fluorosis.

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Evaluation of four final irrigation protocols for cleaning root canal walls

International Journal of Oral Science ◽

10.1038/s41368-020-00091-4 ◽

2020 ◽

Vol 12 (1) ◽

Author(s):

Qiang Li ◽

Qian Zhang ◽

Xiaoying Zou ◽

Lin Yue

Keyword(s):

Root Canal ◽

Laser Scanning ◽

Laser Scanning Microscopy ◽

Smear Layer ◽

Confocal Laser ◽

Bacterial Survival ◽

Bacterial Inhibition ◽

Layer Removal ◽

Significant Difference ◽

Canal Systems

Abstract The aim of this study was to compare the efficiency of four final irrigation protocols in smear layer removal and bacterial inhibition in root canal systems. Thirty roots inoculated with Enterococcus faecalis were prepared with ProTaper Universal files. The teeth were disinfected by conventional needle irrigation, sonic agitation using the EndoActivator device, passive ultrasonic irrigation, or an M3 Max file. Teeth with no root canal preparation served as blank controls for the establishment of the infection baseline. Teeth with preparation but no final irrigation served as a post-instrumentation baseline. After the final irrigation, the teeth were sectioned in half. One half of each tooth was examined by scanning electron microscopy (SEM) to assess smear layer removal using a five-point scale. The other half was examined by confocal laser scanning microscopy (CLSM) using the LIVE/DEAD BackLight bacterial viability kit to evaluate the depth of bacterial survival in dentinal tubules. SEM analysis revealed no significant difference in smear layer removal throughout the whole canal among the EA, PUI, and M3 Max groups (P > 0.05). CLSM revealed that PUI achieved the greatest bacterial inhibition depth in the coronal ((174.27 ± 31.63) μm), middle ((160.94 ± 37.77) μm), and apical ((119.53 ± 28.49) μm) thirds of the canal (all P < 0.05 vs. other groups). According to this comprehensive SEM and CLSM evaluation, PUI appears to have the best infection control ability in root canal systems.

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Bacterial Colonization in the Marginal Region of Ceramic Restorations: Effects of Different Cement Removal Methods and Polishing

Operative Dentistry ◽

10.2341/15-206-l ◽

2016 ◽

Vol 41 (6) ◽

pp. 642-654 ◽

Cited By ~ 5

Author(s):

SMB Pereira ◽

LC Anami ◽

CA Pereira ◽

ROA Souza ◽

KZ Kantorski ◽

...

Keyword(s):

Laser Scanning ◽

Bacterial Colonization ◽

Resin Cement ◽

Laser Scanning Microscopy ◽

Confocal Laser ◽

Significant Difference ◽

Ceramic Blocks ◽

Cement Removal ◽

Ceramic Restorations ◽

Marginal Region

SUMMARY This study evaluated the effects of excess cement removal techniques, with or without subsequent polishing, on biofilm formation and micromorphology in the marginal region of the tooth/restoration. From bovine teeth, 96 dentin blocks (4 × 8 × 2 mm) were produced, molded, and reproduced in type IV gypsum, on which 96 pressed ceramic blocks (Vita PM9, Vita Zahnfabrik; 4 × 8 × 2 mm) were produced via the lost wax technique. The dentin blocks and their respective ceramic blocks were cemented with a self-adhesive resin cement (RelyX U200, 3M ESPE), and cement excess was removed from the margin using four different techniques, followed or not by polishing with silicone rubber tips: MBr, removal with microbrush and photoactivation; MBr-Pol, MBr + polishing; Br, removal with brush and photoactivation; Br-Pol, Br + polishing; Photo-Expl, 5 seconds of initial photoactivation, removal with explorer, and final curing; Photo-Expl-Pol, Photo-Expl + polishing; Photo-SB, 5 seconds of initial photoactivation, removal with scalpel, and final curing; and Photo-SB-Pol, Photo-SB + polishing. After 24 hours, the roughness in the marginal region was analyzed using a profilometer (three measurements on each sample). Micromorphological analyses of the region were performed by stereomicroscope and scanning electron microscopy (SEM). Then the samples were contaminated with sucrose broth standardized suspension with Streptococcus mutans, Staphylococcus aureus, and Candida albicans and incubated for a period of 48 hours. The samples were quantitatively analyzed for bacterial adherence in the marginal region by confocal laser scanning microscopy and counting of colony-forming units (CFUs/mL) and qualitatively analyzed using SEM. Roughness data (Ra) were submitted to two-way analysis of variance, Tukey test at a confidence level of 95%, and Student t-tests. CFU, biomass, and biothickness data were analyzed by Kruskal-Wallis, Mann-Whitney, and Dunn tests. The removing technique statistically influenced Ra (MBr, p=0.0019; Br, p=0.002; Photo-Expl, p=0.0262; Photo-SB, p=0.0196) when comparing the polished and unpolished groups. The MBr and MBr-Pol technique differed significantly for CFU/mL values (p=0.010). There was no significant difference in the amounts of biomass and biothickness comparing polished and unpolished groups and when all groups were compared (p>0.05). Different morphological patterns were observed (more regular surface for polished groups). We conclude that margin polishing after cementation of feldspar/pressed ceramic restorations is decisive for achieving smoother surfaces, as the excess cement around the edges can increase the surface roughness in these areas, influencing bacterial adhesion.

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Influence of Light-curing Parameters on Biofilm Development and Flexural Strength of a Silorane-based Composite

Operative Dentistry ◽

10.2341/14-279-l ◽

2016 ◽

Vol 41 (2) ◽

pp. 219-227 ◽

Cited By ~ 3

Author(s):

E Brambilla ◽

A Ionescu ◽

G Cazzaniga ◽

M Ottobelli

Keyword(s):

Flexural Strength ◽

Laser Scanning ◽

Testing Machine ◽

Laser Scanning Microscopy ◽

Confocal Laser ◽

Curing Time ◽

Two Samples ◽

Significant Difference ◽

Light Curing ◽

Biofilm Development

SUMMARYObjectives: The aim of this study was to evaluate the differences in biological and mechanical performances of a silorane-based and a methacrylate-based composite. Another aim was to assess the influence of light-curing time and light-curing intensity on in vitro biofilm formation and flexural strength of the two tested composites.Methods: Experiment 1: 432 specimens obtained from a silorane-based composite and from a standard methacrylate-based composite were divided into six groups and light-cured for 10, 20, 30, 40, 60, or 80 seconds, using one of two light-curing intensities, 400 mW/cm2 or 800 mW/cm2. At 24 hours, a monospecific Streptococcus mutans biofilm adherent to the surfaces of the samples was obtained. Then, a colorimetric technique (MTT assay) was used to evaluate the adherent viable biomass. Two samples per group were observed using confocal laser scanning microscopy. Analysis of variance (ANOVA) and Tukey tests were used to analyze the results (p<0.05). Experiment 2: 192 bar-shaped specimens were obtained and light-cured as in the previous experiment. A three-point bend test using a universal testing machine was performed to obtain flexural strength values. ANOVA and Tukey tests were used to analyze the results (p<0.05).Results: In experiment 1, a highly significant difference (p<0.0001) in biofilm development was shown between silorane-based and methacrylate-based composites. In fact, the silorane-based composite exhibited better biological performance. Significant differences were also found between the two light-curing intensities (p<0.018) and for curing times (p<0.0001): silorane-based composite light-cured for 80 seconds at 800 mW/cm2 light-curing intensity showed the lowest biofilm development. In experiment 2, a significant difference in flexural strength (p<0.0318) was only found between the different composites. Nevertheless, both resin composites showed flexural strength values in accordance with International Organization for Standardization guidelines even after 10 seconds of light-curing time.Conclusions: Silorane-based composite was less prone to biofilm development compared with a methacrylate-based composite. Acceptable flexural strength values for both composites were obtained after 10 seconds of light-curing time.

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Individual or Combined Effects of Meropenem, Imipenem, Sulbactam, Colistin, and Tigecycline on Biofilm-Embedded Acinetobacter baumannii and Biofilm Architecture

Antimicrobial Agents and Chemotherapy ◽

10.1128/aac.00551-16 ◽

2016 ◽

Vol 60 (8) ◽

pp. 4670-4676 ◽

Cited By ~ 7

Author(s):

Yung-Chih Wang ◽

Shu-Chen Kuo ◽

Ya-Sung Yang ◽

Yi-Tzu Lee ◽

Chun-Hsiang Chiu ◽

...

Keyword(s):

Acinetobacter Baumannii ◽

Laser Scanning ◽

Bactericidal Effect ◽

Laser Scanning Microscopy ◽

Confocal Laser ◽

Roughness Coefficient ◽

Content Type ◽

Biofilm Thickness ◽

Carbapenem Resistant ◽

Biofilm Architecture

ABSTRACTAcinetobacter baumanniibiofilms are difficult to eradicate. We investigated the effects of meropenem (2 mg/liter), imipenem (2 mg/liter), sulbactam (4 mg/liter), colistin (2 mg/liter), and tigecycline (2 mg/liter), alone or in combination, on biofilm-embedded carbapenem-resistant and carbapenem-susceptibleA. baumannii(CRAb and CSAb, respectively) cells, as well as on the architecture of the biofilms.A. baumanniiATCC 15151 (Ab15151) and its OXA-82-overproducing transformant, along with two clinical CSAb and two clinical CRAb isolates of differing clonalities, were used. The minimal bactericidal concentrations for biofilm-embedded cells of the six tested isolates were >50-fold those of their planktonic cells. When used individually, meropenem exhibited a higher killing effect than the other four antimicrobials on biofilm-embedded CSAb cells in the colony biofilm assay. For two clinical CRAb isolates, meropenem plus sulbactam or sulbactam plus tigecycline showed >100-fold the bactericidal effect exhibited by these agents used alone after 48 h of treatment. The effect of antimicrobials on the architecture of Ab15151 biofilm emitting green fluorescence was determined by confocal laser scanning microscopy using COMSTAT software. Significant decreases in the maximum biofilm thickness were observed after exposure to meropenem and imipenem. Meropenem plus sulbactam significantly decreased the biomass and mean thickness and increased the roughness coefficient of biofilms, but sulbactam plus tigecycline only decreased the maximum and mean biofilm thickness compared to any of these agents used alone. Meropenem was active against biofilm-embedded CSAb, whereas meropenem plus sulbactam exhibited synergism against biofilm-embedded CRAb and caused significantly more damage to the biofilm architecture than did any of the agents used alone.

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Bactericidal compounds controlling growth of the plant pathogen pseudomonas syringae pv. actinidiae, which forms biofilms composed of a novel exopolysaccharide

10.26686/wgtn.12597797.v1 ◽

2020 ◽

Author(s):

S Ghods ◽

Ian Sims ◽

MF Moradali ◽

BHA Rehma

Keyword(s):

Pseudomonas Syringae ◽

Chlorine Dioxide ◽

Laser Scanning ◽

Extracellular Polymeric Substances ◽

Virulent Strain ◽

Bactericidal Effect ◽

Cell System ◽

Laser Scanning Microscopy ◽

Base Layer ◽

Confocal Laser

© 2015, American Society for Microbiology. Pseudomonas syringae pv. actinidiae is the major cause of bacterial canker and is a severe threat to kiwifruit production worldwide. Many aspects of the disease caused by P. syringae pv. actinidiae, such as the pathogenicity-relevant formation of a biofilm composed of extracellular polymeric substances (EPSs), are still unknown. Here, a highly virulent strain of P. syringae pv. actinidiae, NZ V-13, was studied with respect to biofilm formation and architecture using a flow cell system combined with confocal laser scanning microscopy. The biofilm formed by P. syringae pv. actinidiae NZ V-13 was heterogeneous, consisting of a thin cellular base layer 5 μm thick and microcolonies with irregular structures. The major component of the EPSs produced by P. syringae pv. actinidiae NZ V-13 bacteria was isolated and identified to be an exopolysaccharide. Extensive compositional and structural analysis showed that rhamnose, fucose, and glucose were the major constituents, present at a ratio of 5:1.5:2. Experimental evidence that P. syringae pv. actinidiae NZ V-13 produces two polysaccharides, a branched α-D-rhamnan with side chains of terminal α-D-Fucf and an α-D-1,4-linked glucan, was obtained. The susceptibility of the cells in biofilms to kasugamycin and chlorine dioxide was assessed. About 64 and 73% of P. syringae pv. actinidiae NZ V-13 cells in biofilms were killed when kasugamycin and chlorine dioxide were used at 5 and 10 ppm, respectively. Kasugamycin inhibited the attachment of P. syringae pv. actinidiae NZ V-13 to solid surfaces at concentrations of 80 and 100 ppm. Kasugamycin was bacteriostatic against P. syringae pv. actinidiae NZ V-13 growth in the planktonic mode, with the MIC being 40 to 60 ppm and a bactericidal effect being found at 100 ppm. Here we studied the formation, architecture, and composition of P. syringae pv. actinidiae biofilms as well as used the biofilm as a model to assess the efficacies of bactericidal compounds.

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Correlation between Bond Strength to Dentin and Sealers Penetration by Push-Out Test and CLSM Analysis

Brazilian Dental Journal ◽

10.1590/0103-6440201902766 ◽

2019 ◽

Vol 30 (6) ◽

pp. 555-562

Author(s):

Maybell Tedesco ◽

Marcelo Carvalho Chain ◽

Wilson Tadeu Felippe ◽

Ana Maria Hecke Alves ◽

Lucas da Fonseca Roberti Garcia ◽

...

Keyword(s):

Bond Strength ◽

Laser Scanning ◽

Regional Analysis ◽

Kendall Test ◽

Laser Scanning Microscopy ◽

Confocal Laser ◽

Dentinal Tubules ◽

Significant Difference ◽

Ah Plus ◽

Push Out

Abstract This study correlated the bond strength (BS) and dentin penetration of different sealers by push-out test and Confocal Laser Scanning Microscopy (CLSM) analysis. Forty-five root canals were prepared according to the crown-down technique and filled with gutta-percha associated to the following sealers (n=15): Endofill, AH Plus and MTA Fillapex. Five canals of each group were filled with the sealers added with 0.1% Rhodamine B dye. Next, the specimens were transversely sectioned and submitted to the push-out test (n=10) and CLSM analysis (n=5). The BS data showed the following means (MPa) and standard deviation: AH Plus (4.17±1.86); MTA Fillapex (3.13±1.96) and Endofill (2.10±1.03). Statistical analysis (two-way ANOVA, α=0.05) showed significant difference among sealers (p<0.001) and root canal thirds (p<0.001). The BS results of Endofill and MTA Fillapex were statistically similar (p>0.05), however, they were statistically different from AH Plus (p<0.001). The regional analysis of BS showed similarity between middle and apical thirds (p>0.05), and both were different from coronal portion (p<0.001). CLSM analysis verified tags formation in all groups and higher penetration of the specimens filled with AH Plus (p<0.05). The Kendall test (correlation between BS to dentin and sealer penetration into dentinal tubules) and the Pearson test (between failures pattern and sealer penetration into dentinal tubules) did not show correlation between the variables evaluated for all the tested sealers (p>0.05). AH Plus group had higher BS to dentin, and deeper tags formation than the other sealers. There was no significant correlation between BS and intratubular penetration of the tested sealers.

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