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In Vitro Phage-Biofilm Interaction Assays

Creative Biolabs offers sophisticated and innovative one-stop phage solutions. We will find a way to manage the entire project to meet your needs. When working with us, there is no need to change providers, which can be time-consuming and expensive. You can rely on our skilled and enthusiastic staff to find the most appropriate path and guide you through your phage research journey.

Introduction

Bacteriophages and bacterial biofilms are widely found in the natural environment. Phage-host interactions in biofilm communities are quite complex, in which phages not only act as predators but can also establish symbiotic relationships to induce and strengthen biofilms. Biofilms are important survival strategies for bacteria and are often reported as major virulence factors for pathogenic bacteria. Bacteriophages have been shown to successfully control biofilms. The growing interest in phages as biofilm control agents relies on the fact that lytic phages can penetrate the 3D structure of biofilms and kill biofilm-associated cells, which are difficult to target with conventional antibiotics. Combining in vitro, ex vivo, or in vivo biofilm infection assays with computational simulations can help us to uncover and better understand the phage-biofilm interactions.

In Vitro Phage-Biofilm Interaction Assays at Creative Biolabs

Phage-biofilm interactions can be studied by a set of approaches that assess biofilm biomass and/or cell viability. These methods can be classified as cultural, molecular, physical, chemical, microscopic, computational, and mathematical model-based.

  • Microbiological Phage-Biofilm Interaction Assays

Determination of colony-forming units (CFUs) is the most widely used technique for evaluating bacteriophage killing in biofilms. The technique is based on serial dilutions of bacterial suspensions, this is a simple and widely used method. Biofilms are composed of subsets of cells that are viable but not culturable and are not normally detected by CFUs.

  • Physical Phage-Biofilm Interaction Assays

The aforementioned limitation on biofilm cell counting accuracy can be overcome by using flow cytometry in combination with bacterial cell staining with viability fluorophore. This methodology has been suggested as a very promising approach to studying, in almost real-time, phage-biofilm interactions. Total biofilm biomass can be obtained from dry or wet weight measurements. Electrochemical impedance spectroscopy (ECIS) has been widely used in the study of microbial electrochemical systems and can be used to indirectly assess biofilm biomass. A physical method extensively used to measure biofilm thickness is based on ultrasonic time-domain reflectometry.

  • Chemical Phage-Biofilm Interaction Assays

Chemical methods utilize dyes or fluorescent dyes that can bind or adsorb on biofilm components. They are indirect methods that can be used to measure specific biofilm components. Crystal Violet (CV) staining for biofilm quantification remains the most commonly used quantitative technique for microtiter plate analysis.

Microbial cells in biofilms can be well detected by simulated nucleic acids (peptide nucleic acid (PNA) and locked nucleic acid (LNA)) combined with fluorescence in situ hybridization (FISH). Numerous microscopy-based imaging modalities are also available to analyze biofilms.

Representation of the methods used for biofilm formation and studying phage-biofilm interactions.Fig.1 Representation of the methods used for biofilm formation and studying phage-biofilm interactions. (Pires, 2021)

As a leading international biotechnology company, Creative Biolabs Laboratories is flexible to meet the unique needs of phage client projects, and we have the expertise to optimize each stage to ensure you get the results you want. If you are interested in our in vitro phage-biofilm interaction assays, please feel free to contact us for more.

Reference:

  1. Pires, D.P.; et al. Understanding the complex phage-host interactions in biofilm communities. Annual Review of Virology. 2021, 8: 73-94.
For Research Use Only. Do NOT use in humans.

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