scholarly journals The Protective Effect of Staphylococcus epidermidis Biofilm Matrix against Phage Predation

Viruses ◽  
2020 ◽  
Vol 12 (10) ◽  
pp. 1076
Author(s):  
Luís D. R. Melo ◽  
Graça Pinto ◽  
Fernando Oliveira ◽  
Diana Vilas-Boas ◽  
Carina Almeida ◽  
...  
Keyword(s):  
Protective Effect ◽  
Staphylococcus Epidermidis ◽  
Laser Scanning ◽  
3D Structure ◽  
Laser Scanning Microscopy ◽  
Confocal Laser ◽  
Planktonic Cells ◽  
The Matrix ◽  
The Impact ◽  
Biofilm Matrix

Staphylococcus epidermidis is a major causative agent of nosocomial infections, mainly associated with the use of indwelling devices, on which this bacterium forms structures known as biofilms. Due to biofilms’ high tolerance to antibiotics, virulent bacteriophages were previously tested as novel therapeutic agents. However, several staphylococcal bacteriophages were shown to be inefficient against biofilms. In this study, the previously characterized S. epidermidis-specific Sepunavirus phiIBB-SEP1 (SEP1), which has a broad spectrum and high activity against planktonic cells, was evaluated concerning its efficacy against S. epidermidis biofilms. The in vitro biofilm killing assays demonstrated a reduced activity of the phage. To understand the underlying factors impairing SEP1 inefficacy against biofilms, this phage was tested against distinct planktonic and biofilm-derived bacterial populations. Interestingly, SEP1 was able to lyse planktonic cells in different physiological states, suggesting that the inefficacy for biofilm control resulted from the biofilm 3D structure and the protective effect of the matrix. To assess the impact of the biofilm architecture on phage predation, SEP1 was tested in disrupted biofilms resulting in a 2 orders-of-magnitude reduction in the number of viable cells after 6 h of infection. The interaction between SEP1 and the biofilm matrix was further assessed by the addition of matrix to phage particles. Results showed that the matrix did not inactivate phages nor affected phage adsorption. Moreover, confocal laser scanning microscopy data demonstrated that phage infected cells were less predominant in the biofilm regions where the matrix was more abundant. Our results provide compelling evidence indicating that the biofilm matrix can work as a barrier, allowing the bacteria to be hindered from phage infection.

scholarly journals Hydrosol of Thymbra capitata Is a Highly Efficient Biocide against Salmonella enterica Serovar Typhimurium Biofilms

2016 ◽  
Vol 82 (17) ◽  
pp. 5309-5319 ◽  
Author(s):  
Foteini Karampoula ◽  
Efstathios Giaouris ◽  
Julien Deschamps ◽  
Agapi I. Doulgeraki ◽  
George-John E. Nychas ◽  
...  
Keyword(s):  
Salmonella Enterica ◽  
Laser Scanning ◽  
High Efficiency ◽  
3D Structure ◽  
Time Lapse ◽  
Laser Scanning Microscopy ◽  
Confocal Laser ◽  
Planktonic Cells ◽  
Content Type ◽  
Serovar Typhimurium

ABSTRACTSalmonellais recognized as one of the most significant enteric foodborne bacterial pathogens. In recent years, the resistance of pathogens to biocides and other environmental stresses, especially when they are embedded in biofilm structures, has led to the search for and development of novel antimicrobial strategies capable of displaying both high efficiency and safety. In this direction, the aims of the present work were to evaluate the antimicrobial activity of hydrosol of the Mediterranean spiceThymbracapitataagainst both planktonic and biofilm cells ofSalmonella entericaserovar Typhimurium and to compare its action with that of benzalkonium chloride (BC), a commonly used industrial biocide. In order to achieve this, the disinfectant activity following 6-min treatments was comparatively evaluated for both disinfectants by calculating the concentrations needed to achieve the same log reductions against both types of cells. Their bactericidal effect against biofilm cells was also comparatively determined byin situand real-time visualization of cell inactivation through the use of time-lapse confocal laser scanning microscopy (CLSM). Interestingly, results revealed that hydrosol was almost equally effective against biofilms and planktonic cells, whereas a 200-times-higher concentration of BC was needed to achieve the same effect against biofilm compared to planktonic cells. Similarly, time-lapse CLSM revealed the significant advantage of the hydrosol to easily penetrate within the biofilm structure and quickly kill the cells, despite the three-dimensional (3D) structure ofSalmonellabiofilm.IMPORTANCEThe results of this paper highlight the significant antimicrobial action of a natural compound, hydrosol ofThymbra capitata, against both planktonic and biofilm cells of a common foodborne pathogen. Hydrosol has numerous advantages as a disinfectant of food-contact surfaces. It is an aqueous solution which can easily be rinsed out from surfaces, it does not have the strong smell of the essential oil (EO) and it is a byproduct of the EO distillation procedure without any industrial application until now. Consequently, hydrosol obviously could be of great value to combat biofilms and thus to improve product safety not only for the food industries but probably also for many other industries which experience biofilm-related problems.

scholarly journals A Reproducible Method for Growing Biofilms on Polystyrene Surfaces: Biomass and Bacterial Viability Evolution of Pseudomonas fluorescens and Staphylococcus epidermidis

2020 ◽  
Vol 10 (13) ◽  
pp. 4544
Author(s):  
Valeria Angarano ◽  
Cindy Smet ◽  
Simen Akkermans ◽  
Theodora Akritidou ◽  
Bart Huyck ◽  
...  
Keyword(s):  
Pseudomonas Fluorescens ◽  
Staphylococcus Epidermidis ◽  
Laser Scanning ◽  
Growth Parameters ◽  
3D Structure ◽  
Laser Scanning Microscopy ◽  
Confocal Laser ◽  
Total Biomass ◽  
Final Population ◽  
Biofilm Development

Since biofilm development represents a crucial issue within industrial, clinical and domestic sectors, innovative technologies/approaches (e.g., light technology for inactivation, antibiofilm coatings) are required to eradicate them. In this multidisciplinary scenario, protocols for the development of biofilms are necessary, particularly, in laboratories (not specialised in biofilm science) lacking in sophisticated devices for their growth. A protocol was developed for growing Pseudomonas fluorescens (Gram-negative) biofilms on wide, flat, polystyrene surfaces within 24 h. Several factors, such as inoculum level, area size and growth medium concentration, were investigated. Biofilm development was studied via viable cells and biomass quantification. A comparative analysis between kinetics and growth parameters, estimated using the Baranyi and Roberts model, was conducted at different inoculum levels (104 and 107 CFU/mL). The inoculum levels did not influence the final population within the 24-h-grown biofilms, but they influenced the total biomass development, which followed different kinetics. Confocal laser scanning microscopy confirmed that overnight growth allowed for development of a densely packed biofilm with its 3D structure. The developed protocol was validated for Staphylococcus epidermidis (Gram-positive). The present work is the first study to develop an easy-to-use protocol to obtain highly reproducible biofilms, on flat polystyrene surfaces, with no need for sophisticated technologies.

Confocal and SEM imaging to demonstrate food pathogen- biofilm biocontrol by pyocyanin from Pseudomonas aeruginosa BTRY1

2016 ◽  
Vol 6 (01) ◽  
pp. 5218
Author(s):  
Laxmi Mohandas ◽  
Anju T. R. ◽  
Sarita G. Bhat*
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Biofilm Formation ◽  
Laser Scanning ◽  
Laser Scanning Microscopy ◽  
Confocal Laser ◽  
Antibiofilm Activity ◽  
Opportunistic Pathogens ◽  
Redox Active ◽  
Sem Imaging ◽  
The Impact

An assortment of redox-active phenazine compounds like pyocyanin with their characteristic blue-green colour are synthesized by Pseudomonas aeruginosa, Gram-negative opportunistic pathogens, which are also considered one of the most commercially valuable microorganisms. In this study, pyocyanin from Pseudomonas aeruginosa BTRY1 from food sample was assessed for its antibiofilm activity by micro titer plate assay against strong biofilm producers belonging to the genera Bacillus, Staphylococcus, Brevibacterium and Micrococcus. Pyocyanin inhibited biofilm activity in very minute concentrations. This was also confirmed by Scanning Electron Microscopy (SEM) and Confocal Laser Scanning Microscopy (CLSM). Both SEM and CLSM helped to visualize the biocontrol of biofilm formation by eight pathogens. The imaging and quantification by CLSM also established the impact of pyocyanin on biofilm-biocontrol mainly in the food industry.

scholarly journals Influence of Aging on Corrosion Behaviour of the 6061 Cast Aluminium Alloy

Materials ◽  
2021 ◽  
Vol 14 (8) ◽  
pp. 1821
Author(s):  
Ting He ◽  
Wei Shi ◽  
Song Xiang ◽  
Chaowen Huang ◽  
Ronald G. Ballinger
Keyword(s):  
Aluminium Alloy ◽  
Laser Scanning ◽  
Corrosion Behaviour ◽  
Laser Scanning Microscopy ◽  
Confocal Laser ◽  
Kelvin Probe ◽  
Force Microscopy ◽  
The Matrix ◽  
6061 Aluminium Alloy

The influence of AlFeSi and Mg2Si phases on corrosion behaviour of the cast 6061 aluminium alloy was investigated. Scanning Kelvin probe force microscopy (SKPFM), electron probe microanalysis (EPMA), and in situ observations by confocal laser scanning microscopy (CLSM) were used. It was found that Mg2Si phases were anodic relative to the matrix and dissolved preferentially without significantly affecting corrosion propagation. The AlFeSi phases’ influence on 6061 aluminium alloy local corrosion was greater than that of the Mg2Si phases. The corroded region width reached five times that of the AlFeSi phase, and the accelerating effect was terminated as the AlFeSi dissolved.

scholarly journals The effect of the dispersion of microfibrillated cellulose on the mechanical properties of melt-compounded polypropylene–polyethylene copolymer

Cellulose ◽  
2019 ◽  
Vol 26 (18) ◽  
pp. 9645-9659 ◽  
Author(s):  
Caterina Palange ◽  
Marcus A. Johns ◽  
David J. Scurr ◽  
Jonathan S. Phipps ◽  
Stephen J. Eichhorn
Keyword(s):  
Tannic Acid ◽  
Laser Scanning ◽  
Secondary Ion Mass Spectrometry ◽  
High Surface ◽  
Microfibrillated Cellulose ◽  
Laser Scanning Microscopy ◽  
Confocal Laser ◽  
Cellulose Content ◽  
Reinforced Composites ◽  
The Impact

Abstract Microfibrillated cellulose (MFC) is a highly expanded, high surface area networked form of cellulose-based reinforcement. Due to the poor compatibility of cellulose with most common apolar thermoplastic matrices, the production of cellulose-reinforced composites in industry is currently limited to polar materials. In this study, a facile water-based chemistry, based on the reaction of MFC with tannic acid and subsequent functionalisation with an alkyl amine, is used to render the surface of the MFC fibrils hydrophobic and enhance the dispersion of the cellulose-based filler into an apolar thermoplastic matrix. The level of dispersion of the compatibilized MFC reinforced composites was evaluated using Time of Flight Secondary Ion Mass Spectrometry and multi-channel Spectral Confocal Laser Scanning Microscopy. The agglomeration of cellulosic filler within the composites was reduced by functionalising the surface of the MFC fibrils with tannic acid and octadecylamine. The resulting composites exhibited an increase in modulus at a high cellulose content. Despite the dispersion of a large portion of the functionalised filler, the presence of some remaining aggregates affected the impact properties of the composites produced.

scholarly journals Mosaic-CLSM Assessment of Bacterial Spatial Distribution in Cosmetic Matrices According to Matrix Viscosity and Bacterial Hydrophobicity

Cosmetics ◽  
2020 ◽  
Vol 7 (2) ◽  
pp. 32
Author(s):  
Samia Almoughrabie ◽  
Chrisse Ngari ◽  
Romain Briandet ◽  
Valérie Poulet ◽  
Florence Dubois-Brissonnet
Keyword(s):  
Spatial Distribution ◽  
Laser Scanning ◽  
Rapid Assessment ◽  
Challenge Test ◽  
Surface Hydrophobicity ◽  
Laser Scanning Microscopy ◽  
Confocal Laser ◽  
Bacterial Surface ◽  
Bacterial Hydrophobicity ◽  
The Impact

The reliability of the challenge test depends, among other parameters, on the spatial distribution of microorganisms in the matrix. The present study aims to quickly identify factors that are susceptible to impair a uniform distribution of inoculated bacteria in cosmetic matrices in this context. We used mosaic confocal laser scanning microscopy (M-CLSM) to obtain rapid assessment of the impact of the composition and viscosity of cosmetic matrices on S. aureus spatial distribution. Several models of cosmetic matrices were formulated with different concentrations of two thickeners and were inoculated with three S. aureus strains having different levels of hydrophobicity. The spatial distribution of S. aureus in each matrix was evaluated according to the frequency distribution of the fluorescence values of at least 1350 CLSM images. We showed that, whatever the thickener used, an increasingly concentration of thickener results in increasingly bacterial clustered distribution. Moreover, higher bacterial hydrophobicity also resulted in a more clustered spatial distribution. In conclusion, CLSM-based method allows a rapid characterization of bacterial spatial distribution in complex emulsified systems. Both matrix viscosity and bacterial surface hydrophobicity affect the bacterial spatial distribution which can have an impact on the reliability of bacterial enumeration during challenge test.

scholarly journals Enhancement of Vancomycin Activity against Biofilms by Using Ultrasound-Targeted Microbubble Destruction

2011 ◽  
Vol 55 (11) ◽  
pp. 5331-5337 ◽  
Author(s):  
Nianan He ◽  
Jian Hu ◽  
Huayong Liu ◽  
Tao Zhu ◽  
Beijian Huang ◽  
...  
Keyword(s):  
Staphylococcus Epidermidis ◽  
Laser Scanning ◽  
Rabbit Model ◽  
Antibiotic Activity ◽  
Laser Scanning Microscopy ◽  
Confocal Laser ◽  
Content Type ◽  
Implanted Medical Devices

ABSTRACTTreating biofilm infections on implanted medical devices is formidable, even with extensive antibiotic therapy. The aim of this study was to investigate whether ultrasound (US)-targeted microbubble (MB) destruction (UTMD) could enhance vancomycin activity againstStaphylococcus epidermidisRP62A biofilms. Twelve-hour biofilms were treated with vancomycin combined with UTMD. The vancomycin and MB (SonoVue) were used at concentrations of 100 μg/ml and 30% (vol/vol), respectively, in studiesin vitro. After US exposure (0.08 MHz, 1.0 W/cm2, 50% duty cycle, and 10-min duration), the biofilms were cultured at 37°C for another 12 h. The results showed that many micropores were found in biofilms treated with vancomycin combined with UTMD. Biofilm densities (A570values) and the viable counts ofS. epidermidisrecovered from the biofilm were significantly decreased compared with those of any other groups. Furthermore, the highest percentage of dead cells was found, using confocal laser scanning microscopy, in the biofilm treated with vancomycin combined with UTMD. The viable counts of bacteria in biofilms in anin vivorabbit model also confirmed the enhanced effect of vancomycin combined with UTMD. UTMD may have great potential for improving antibiotic activity against biofilm infections.

scholarly journals High Adhesion and Increased Cell Death Contribute to Strong Biofilm Formation in Klebsiella pneumoniae

Pathogens ◽  
2019 ◽  
Vol 8 (4) ◽  
pp. 277
Author(s):  
Siddhi Desai ◽  
Kinjal Sanghrajka ◽  
Devarshi Gajjar
Keyword(s):  
Cell Death ◽  
Klebsiella Pneumoniae ◽  
Biofilm Formation ◽  
Laser Scanning ◽  
Laser Scanning Microscopy ◽  
Extracellular Dna ◽  
Confocal Laser ◽  
Bacterial Cells ◽  
High Adhesion ◽  
Biofilm Matrix

Klebsiella pneumoniae (Kp), is a frequent cause of hospital and community-acquired infections and WHO had declared it as a “priority pathogen”. Biofilm is a major virulence factor of Kp and yet the mechanism of strong biofilm formation in Kp is unclear. A key objective of the present study is to investigate the differences between strong and weak biofilms formed by clinical isolates of Kp on various catheters and in different media conditions and to identify constituents contributing to strong biofilm formation. Quantification of matrix components (extracellular DNA (eDNA), protein, exopolysaccharides (EPS), and bacterial cells), confocal laser scanning microscopy (CLSM), field emission gun scanning electron microscopy (FEG-SEM) and flow-cytometry analysis were performed to compare strong and weak biofilm matrix. Our results suggest increased biofilm formation on latex catheters compared to silicone and silicone-coated latex catheters. Higher amounts of eDNA, protein, EPS, and dead cells were observed in the strong biofilm of Kp. High adhesion capacity and cell death seem to play a major role in formation of strong Kp biofilms. The enhanced eDNA, EPS, and protein in the biofilm matrix appear as a consequence of increased cell death.

scholarly journals Electrochemical and surface analytical techniques applied to microbiologically influenced corrosion investigation

2018 ◽  
Vol 36 (4) ◽  
pp. 349-363 ◽  
Author(s):  
László Trif ◽  
Abdul Shaban ◽  
Judit Telegdi
Keyword(s):  
Photoelectron Spectroscopy ◽  
Laser Scanning ◽  
Electrochemical Impedance ◽  
Electrochemical Techniques ◽  
Analytical Techniques ◽  
Laser Scanning Microscopy ◽  
Microbiologically Influenced Corrosion ◽  
Microbial Consortia ◽  
Confocal Laser ◽  
The Impact

AbstractSuitable application of techniques for detection and monitoring of microbiologically influenced corrosion (MIC) is crucial for understanding the mechanisms of the interactions and for selecting inhibition and control approaches. This paper presents a review of the application of electrochemical and surface analytical techniques in studying the MIC process of metals and their alloys. Conventional electrochemical techniques, such as corrosion potential (Ecorr), redox potential, dual-cell technique, polarization curves, electrochemical impedance spectroscopy (EIS), electrochemical noise (EN) analysis, and microelectrode techniques, are discussed, with examples of their use in various MIC studies. Electrochemical quartz crystal microbalance, which is newly used in MIC study, is also discussed. Microscopic techniques [scanning electron microscopy (SEM), environmental SEM (ESEM), atomic force microscopy (AFM), confocal laser microscopy (CLM), confocal laser scanning microscopy (CLSM), confocal Raman microscopy] and spectroscopic analytical methods [Fourier transform infrared spectroscopy (FTIR) and X-ray photoelectron spectroscopy (XPS)] are also highlighted. This review highlights the heterogeneous characteristics of microbial consortia and use of special techniques to study their probable effects on the metal substrata. The aim of this review is to motivate using a combination of new procedures for research and practical measurement and calculation of the impact of MIC and biofilms on metals and their alloys.

Surface Functionalization-Dependent Physicochemical Interactions between Nanoparticles and the Biofilm EPS Matrix

2020 ◽  
Author(s):  
Dishon Hiebner ◽  
Caio Barros ◽  
Laura Quinn ◽  
Stefania Vitale ◽  
Eoin Casey
Keyword(s):  
Laser Scanning ◽  
Three Dimensional ◽  
Engineered Nanoparticles ◽  
Laser Scanning Microscopy ◽  
Confocal Laser ◽  
Innovative Strategy ◽  
Biofilm Control ◽  
Physicochemical Interactions ◽  
The Matrix ◽  
Underlying Mechanisms

<p>The contribution of the biofilm extracellular polymeric substance (EPS) matrix to reduced antimicrobial susceptibility in biofilms is widely recognised.  As such, directly targeting the EPS matrix is a promising biofilm control strategy that allows for efficient disruption of the matrix to allow an increase in susceptibility to antibiofilm agents. To this end, engineered nanoparticles (NPs) have received considerable attention. However, the fundamental understanding of the physicochemical interactions occurring between NPs and the EPS matrix has not yet been fully elucidated. An insight into the underlying mechanisms involved when a NP interacts with molecules in the EPS matrix will aid in the design of more efficient systems for biofilm control. The use of highly specific fluorescent probes in confocal laser scanning microscopy (CLSM) to illustrate the spatial distribution of EPS macromolecules within the biofilm is demonstrated. Three-dimensional (3D) colocalization analysis was used to assess the affinity of differently functionalized silica NPs (SiNPs) for specific EPS macromolecules from <em>Pseudomonas fluorescens</em> biofilms. Results show that both the charge and surface functional groups of SiNPs dramatically affect the extent to which SiNPs interact and localize with EPS macromolecules, including proteins, polysaccharides, and DNA. This research not only develops an innovative strategy for biofilm-nanoparticle interaction studies but also provides a platform on which to build more efficient NP systems for biofilm control.</p>

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