Exploration of Microbial Communities using the Thermo Scientific Varioskan LUX Multimode Reader and the Invitrogen EVOS FL Cell Imaging System

Hiltunen, Anna; Skogman, Malena; Vuorela, Pia; Fallarero, Adyary

doi:10.2144/000114613

Cited by 2 publications

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“…For quantifying matrix poly-N-acetyl-β-(1-6)-glucosamine (PNAG), S. aureus ATCC 25923 biofilms were grown in 96-WPs and on coupons in 12-WPs for 18, 42 and 66 h, as described above. The previously reported WGA staining protocol [54] was applied with two modifications: a lower concentration (2.5 µg•mL −1 ) of WGA conjugate was used based on [55] and 100% DMSO was used to replace 33% acetic acid, which is not compatible with acid-unbearable materials, such as HA. The selection of 100% DMSO was based upon initial tests where a lower concentration of acetic acid (10% acetic acid), 96% ethanol and 100% DMSO were tested (acquired assay quality parameters for DMSO: screening window coefficient Z' factor = 0.431; signal to noise (S/N) = 5.4; signal to background (S/B) = 9.2) First, the 96-WPs were washed once with 200 µL of PBS, while the coupons were briefly soaked once in PBS, to detach planktonic and loosely attached cells.…”

Section: Quantification Of Matrix-associated Poly-n-acetyl-β-(1-6)-glmentioning

confidence: 99%

Structural and Functional Dynamics of Staphylococcus aureus Biofilms and Biofilm Matrix Proteins on Different Clinical Materials

et al. 2019

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Medical device-associated staphylococcal infections are a common and challenging problem. However, detailed knowledge of staphylococcal biofilm dynamics on clinically relevant surfaces is still limited. In the present study, biofilm formation of the Staphylococcus aureus ATCC 25923 strain was studied on clinically relevant materials—borosilicate glass, plexiglass, hydroxyapatite, titanium and polystyrene—at 18, 42 and 66 h. Materials with the highest surface roughness and porosity (hydroxyapatite and plexiglass) did not promote biofilm formation as efficiently as some other selected materials. Matrix-associated poly-N-acetyl-β-(1-6)-glucosamine (PNAG) was considered important in young (18 h) biofilms, whereas proteins appeared to play a more important role at later stages of biofilm development. A total of 460 proteins were identified from biofilm matrices formed on the indicated materials and time points—from which, 66 proteins were proposed to form the core surfaceome. At 18 h, the appearance of several r-proteins and glycolytic adhesive moonlighters, possibly via an autolysin (AtlA)-mediated release, was demonstrated in all materials, whereas classical surface adhesins, resistance- and virulence-associated proteins displayed greater variation in their abundances depending on the used material. Hydroxyapatite-associated biofilms were more susceptible to antibiotics than biofilms formed on titanium, but no clear correlation between the tolerance and biofilm age was observed. Thus, other factors, possibly the adhesive moonlighters, could have contributed to the observed chemotolerant phenotype. In addition, a protein-dependent matrix network was observed to be already well-established at the 18 h time point. To the best of our knowledge, this is among the first studies shedding light into matrix-associated surfaceomes of S. aureus biofilms grown on different clinically relevant materials and at different time points.

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Section: Quantification Of Matrix-associated Poly-n-acetyl-β-(1-6)-glmentioning

confidence: 99%

Structural and Functional Dynamics of Staphylococcus aureus Biofilms and Biofilm Matrix Proteins on Different Clinical Materials

et al. 2019

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“…In measuring the anti-biofilm activities of natural products, viability and matrix biomass can be assessed, where resazurin and crystal violet staining are performed sequentially in the same plate. Wheat germ agglutinin-Alexa Fluor 488 fluorescent Natural Products as Antibiofilm Agents DOI: http://dx.doi.org/10.5772/intechopen.104434 conjugate is mainly used to stain the matrix, which is essential to measure the biofilm matrix, biomass, and viability to investigate the potencies of anti-biofilm effects of natural products [95,96].…”

Section: Methods Used To Determine Anti-biofilm Effects Of Natural Pr...mentioning

confidence: 99%

Natural Products as Antibiofilm Agents

Danquah¹,

Minkah²,

Agana³

et al. 2022

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Biofilms, are vastly structured surface-associated communities of microorganisms, enclosed within a self-produced extracellular matrix. Microorganisms, especially bacteria are able to form complex structures known as biofilms. The presence of biofilms especially in health care settings increases resistance to antimicrobial agents which poses a major health problem. This is because biofilm-associated persistent infections are difficult to treat due to the presence of multidrug-resistant microorganisms. This chapter will give an idea about documented agents including isolated compounds, crude extracts, decoctions, fractions, etc. obtained from natural sources such as plants, bacteria, fungi, sponge and algae with antibiofilm activities. Furthermore, we have done phylogenetic analysis to identify plant families most prolific in producing plant species and compounds with good antibiofilm properties so as to aid in prioritizing plant species to investigate in future studies. The data in this chapter will help serve as valuable information and guidance for future antimicrobial development.

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Exploration of Microbial Communities using the Thermo Scientific Varioskan LUX Multimode Reader and the Invitrogen EVOS FL Cell Imaging System

Cited by 2 publications

References 5 publications

Structural and Functional Dynamics of Staphylococcus aureus Biofilms and Biofilm Matrix Proteins on Different Clinical Materials

Structural and Functional Dynamics of Staphylococcus aureus Biofilms and Biofilm Matrix Proteins on Different Clinical Materials

Natural Products as Antibiofilm Agents

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