Imaging Techniques for Detecting Prokaryotic Viruses in Environmental Samples

Turzynski, Victoria; Monsees, Indra; Moraru, Cristina; Probst, Alexander J.

doi:10.3390/v13112126

Cited by 10 publications

(7 citation statements)

References 164 publications

(278 reference statements)

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“…Transduction should be detected not only by classical methods such as conventional plating using selectable genetic markers. State-of-the-art imaging techniques, such as epifluorescence microscopy or atomic force microscopy have been applied to detect and characterize uncultivated phages in environmental samples, as recently reviewed by Turzynski et al [264].…”

Section: Challenges Of Studying Transduction In Biofilmsmentioning

confidence: 99%

Horizontal Gene Transfer of Antibiotic Resistance Genes in Biofilms

2023

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Most bacteria attach to biotic or abiotic surfaces and are embedded in a complex matrix which is known as biofilm. Biofilm formation is especially worrisome in clinical settings as it hinders the treatment of infections with antibiotics due to the facilitated acquisition of antibiotic resistance genes (ARGs). Environmental settings are now considered as pivotal for driving biofilm formation, biofilm-mediated antibiotic resistance development and dissemination. Several studies have demonstrated that environmental biofilms can be hotspots for the dissemination of ARGs. These genes can be encoded on mobile genetic elements (MGEs) such as conjugative and mobilizable plasmids or integrative and conjugative elements (ICEs). ARGs can be rapidly transferred through horizontal gene transfer (HGT) which has been shown to occur more frequently in biofilms than in planktonic cultures. Biofilm models are promising tools to mimic natural biofilms to study the dissemination of ARGs via HGT. This review summarizes the state-of-the-art of biofilm studies and the techniques that visualize the three main HGT mechanisms in biofilms: transformation, transduction, and conjugation.

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Section: Challenges Of Studying Transduction In Biofilmsmentioning

confidence: 99%

Horizontal Gene Transfer of Antibiotic Resistance Genes in Biofilms

2023

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“…Important facets of this interaction are also unknown, such as the flagellin structural requirements for adsorption, the precise rate of phage translocation along the filament, the molecular mechanism of the tail fiber wrapping around the filament, the transition between attachment to the flagellum, and the interaction with a cell surface component for ejection of DNA into the host cell. Observing phage translocation is a challenge, as phages are generally too small to be seen with typical light microscopy [232]. The use of electron microscopy to see biological processes occurring in living systems in real time is a major challenge.…”

Section: Translocation To the Cell Surfacementioning

confidence: 99%

Flagellotropic Bacteriophages: Opportunities and Challenges for Antimicrobial Applications

Esteves

Scharf

2022

IJMS

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Bacteriophages (phages) are the most abundant biological entities in the biosphere. As viruses that solely infect bacteria, phages have myriad healthcare and agricultural applications including phage therapy and antibacterial treatments in the foodservice industry. Phage therapy has been explored since the turn of the twentieth century but was no longer prioritized following the invention of antibiotics. As we approach a post-antibiotic society, phage therapy research has experienced a significant resurgence for the use of phages against antibiotic-resistant bacteria, a growing concern in modern medicine. Phages are extraordinarily diverse, as are their host receptor targets. Flagellotropic (flagellum-dependent) phages begin their infection cycle by attaching to the flagellum of their motile host, although the later stages of the infection process of most of these phages remain elusive. Flagella are helical appendages required for swimming and swarming motility and are also of great importance for virulence in many pathogenic bacteria of clinical relevance. Not only is bacterial motility itself frequently important for virulence, as it allows pathogenic bacteria to move toward their host and find nutrients more effectively, but flagella can also serve additional functions including mediating bacterial adhesion to surfaces. Flagella are also a potent antigen recognized by the human immune system. Phages utilizing the flagellum for infections are of particular interest due to the unique evolutionary tradeoff they force upon their hosts: by downregulating or abolishing motility to escape infection by a flagellotropic phage, a pathogenic bacterium would also likely attenuate its virulence. This factor may lead to flagellotropic phages becoming especially potent antibacterial agents. This review outlines past, present, and future research of flagellotropic phages, including their molecular mechanisms of infection and potential future applications.

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“…However, these efforts are not meant to replace precise, fine-scale, and high-quality local sampling conducted during oceanographic cruises or as part of site-specific observation sampling projects. In addition, super-resolution fluorescence microscopy approaches are changing our ability to visualize viruses and their interactions (Castelletto and Boretti 2021), but conventional wide-field fluorescence microscopes are still used to determine the abundance of viruses from environmental and culture samples (e.g., Turzynski et al 2021 and sources within). Therefore, efforts are needed to continue integrating the visualization of microorganisms within discrete studies to gain comprehensive insight into how marine microbial communities are structured and their influence on marine ecosystem functions (Sebastian and Gasol 2019).…”

Section: Introductionmentioning

confidence: 99%

Flexible and open-source programs for quantitative image analysis in microbial ecology

Pasulka

Hood

Michels

et al. 2022

Preprint

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Epifluorescence microscopy is an essential tool for obtaining reliable estimates of the abundance of marine microorganisms including viruses. However, computational analysis is required to gain consistent and quantitative data from digital microscopy images. Many imaging programs are proprietary and cost-prohibitive. The currently available free imaging programs are often platform specific and/or lack the flexibility to analyze microscopy images from natural samples, such as the planktonic environment, which can contain challenges such as debris and high background signals. Here we describe two MATLAB-based open-source image analysis programs that work across computer platforms and provide the tools to analyze a range of image types and cell sizes with a user-friendly interface. The Microbial Image Analysis (MiA) program aims to provide flexibility for the selection, identification, and quantification of cells that vary in size and fluorescence intensity within natural microbial communities. The Viral Image Analysis (ViA) program aims to provide an effective means for quantifying viral abundances from epifluorescence images as well as enumerating the intensity of a primary and secondary stain. In this paper, we provide an overview of the functionality of the MiA and ViA programs and highlight specific program features through several microbial image case studies.

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Imaging Techniques for Detecting Prokaryotic Viruses in Environmental Samples

Cited by 10 publications

References 164 publications

Horizontal Gene Transfer of Antibiotic Resistance Genes in Biofilms

Horizontal Gene Transfer of Antibiotic Resistance Genes in Biofilms

Flagellotropic Bacteriophages: Opportunities and Challenges for Antimicrobial Applications

Flexible and open-source programs for quantitative image analysis in microbial ecology

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