R‐based method for quantitative analysis of biofilm thickness by using confocal laser scanning microscopy

Frühauf, Hanna Marianne; Stöckl, Markus; Holtmann, Dirk

doi:10.1002/elsc.202200008

“…All methods are described in detail in (Frühauf et al, 2022; Frühauf et al, 2022) and are only briefly described here. G. sulfurreducens strain PCA (DSM 12127) was obtained from DSMZ (German Collection of Microorganisms and Cell Cultures GmbH).…”

Section: Methodsmentioning

confidence: 99%

Geobacter sulfurreducens metabolism at different donor/acceptor ratios

Frühauf-Wyllie

¹

,

Holtmann

²

2022

MicrobiologyOpen

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Geobacter species have great application potential in remediation processes and electrobiotechnology. In all applications, understanding the metabolism will enable target-oriented optimization of the processes. The typical electron donor and carbon source of the Geobacter species is acetate, while fumarate is the usual electron acceptor. Here, we could show that depending on the donor/acceptor ratio in batch cultivation of Geobacter sulfurreducens different product patterns occur. With a donor/acceptor ratio of 1:2.5 malate accumulated as an intermediate product but was metabolized to succinate subsequently. At the end of the cultivation, the ratio of fumarate consumed and succinate produced was approximately 1:1. When fumarate was added in excess, malate accumulated in the fermentation broth without further metabolization. After the addition of acetate to stationary cells, malate concentration decreased immediately and additional succinate was synthesized. Finally, it was shown that also resting cells of G. sulfurreducens could efficiently convert fumarate to malate without an additional electron donor. Overall, it was demonstrated that by altering the donor/acceptor ratio, targeted optimization of the metabolite conversion by G. sulfurreducens can be realized.central metabolism, donor/acceptor ratio, Geobacter sulfurreducens, organic acid production | INTRODUCTIONGeobacter strains are deltaproteobacteria and can be found in different habitats, for example, aquatic systems, deep aquifer sediments, subsurface sediments, as well as several contaminated sites. Geobacter cells are Gram-negative, nonspore-forming, and obligate heterotrophs (Straub, 2011;Zhang et al., 2020). In general, Geobacter species grow only under strictly anoxic conditions and are unable to grow by fermentation (Straub, 2011). Geobacter species predominate under iron-reducing conditions (Straub, 2011). Acetate is a common electron and carbon donor among Geobacter species (Straub, 2011) and the majority of species could also metabolize alternative electron donors (e.g., ethanol, pyruvate, lactate, hydrogen, and formate). Geobacter species are using different electron acceptors, for example, Fe(III), nitrate, elemental sulfur (S 0 ), fumarate, malate, Mn(IV), U(VI), Co(III), and humic substances.Geobacter species appear to be the primary agents for coupling the oxidation of organic compounds to the reduction of insoluble Fe(III) and Mn(IV) oxides in many soils and sediments (Lovley et al., 2011).Different Geobacter species can oxidize aromatic hydrocarbons under anaerobic conditions. In addition, Geobacter species are important for aromatic hydrocarbon removal in contaminated

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“…The swift expansion of knowledge in the field of biofilms is currently propelled by advancements in microscopic imaging techniques, with Confocal Laser Scanning Microscopy (CLSM) standing out as the most commonly employed method for biofilm analysis (Gomes & Mergulhão, 2017). CLSM, a non-destructive and non-invasive approach, excels in providing threedimensional images of biofilms, offering valuable insights into their complexity and dynamics (Frühauf et al, 2022). Various optical technologies, including CLSM, are instrumental in visualizing the 3D structure and confirming the presence of biofilms (Kırmusaoğlu, 2019).…”

mentioning

confidence: 99%

Analysis of antibiofilm activity gold and silver nanoparticles against uropathogenic Escherichia coli using confocal laser scanning microscopy

Purbowati,

Ama,

Tjandra

et al. 2024

Berkala Penelitian Hayati

0

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Infectious diseases, including urinary tract infections (UTIs), significantly impact global health, diminishing the quality of life and increasing clinical and economic burdens. Concerns arise due to the potential development of antimicrobial resistance (AMR) facilitated by biofilm formation. Nanotechnology, specifically silver and silver nanoparticles (1–100 nm), proves effective against bacterial strains, making them valuable in the biomedical field. Advances in microscopic imaging techniques, such as Confocal Laser Scanning Microscopy (CLSM), enhance our understanding of biofilms. This study aims to assess the effectiveness and optimal concentration of gold and silver nanoparticles in combating biofilms formed by uropathogenic Escherichia coli (UPEC) through CLSM analysis. The research comprises the following stages: rejuvenation of bacterial isolates and inoculum preparation, testing the anti-biofilm activity of gold and silver nanoparticles at concentrations of 25, 50, 75, and 100 ppm against UPEC using CLSM analysis. The findings reveal that both gold and silver nanoparticles exhibit anti-biofilm activity against UPEC at concentrations of 25, 50, 75, and 100 ppm, as confirmed by CLSM analysis. The optimal concentration for anti-biofilm activity of gold and silver nanoparticles against UPEC is identified as 100 ppm.

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“…The swift expansion of knowledge in the field of biofilms is currently propelled by advancements in microscopic imaging techniques, with Confocal Laser Scanning Microscopy (CLSM) standing out as the most commonly employed method for biofilm analysis (Gomes & Mergulhão, 2017). CLSM, a non-destructive and non-invasive approach, excels in providing threedimensional images of biofilms, offering valuable insights into their complexity and dynamics (Frühauf et al, 2022). Various optical technologies, including CLSM, are instrumental in visualizing the 3D structure and confirming the presence of biofilms (Kırmusaoğlu, 2019).…”

mentioning

confidence: 99%

Analysis of antibiofilm activity gold and silver nanoparticles against uropathogenic Escherichia coli using confocal laser scanning microscopy

Purbowati,

Ama,

Tjandra

et al. 2023

Berkala Penelitian Hayati

0

View full text Add to dashboard Cite

Infectious diseases, including urinary tract infections (UTIs), significantly impact global health, diminishing the quality of life and increasing clinical and economic burdens. Concerns arise due to the potential development of antimicrobial resistance (AMR) facilitated by biofilm formation. Nanotechnology, specifically silver and silver nanoparticles (1–100 nm), proves effective against bacterial strains, making them valuable in the biomedical field. Advances in microscopic imaging techniques, such as Confocal Laser Scanning Microscopy (CLSM), enhance our understanding of biofilms. This study aims to assess the effectiveness and optimal concentration of gold and silver nanoparticles in combating biofilms formed by uropathogenic Escherichia coli (UPEC) through CLSM analysis. The research comprises the following stages: rejuvenation of bacterial isolates and inoculum preparation, testing the anti-biofilm activity of gold and silver nanoparticles at concentrations of 25, 50, 75, and 100 ppm against UPEC using CLSM analysis. The findings reveal that both gold and silver nanoparticles exhibit anti-biofilm activity against UPEC at concentrations of 25, 50, 75, and 100 ppm, as confirmed by CLSM analysis. The optimal concentration for anti-biofilm activity of gold and silver nanoparticles against UPEC is identified as 100 ppm.

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R‐based method for quantitative analysis of biofilm thickness by using confocal laser scanning microscopy

Cited by 7 publications

References 26 publications

Geobacter sulfurreducens metabolism at different donor/acceptor ratios

Geobacter sulfurreducens metabolism at different donor/acceptor ratios

Analysis of antibiofilm activity gold and silver nanoparticles against uropathogenic Escherichia coli using confocal laser scanning microscopy

Analysis of antibiofilm activity gold and silver nanoparticles against uropathogenic Escherichia coli using confocal laser scanning microscopy

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