A Multiwell-Plate Caenorhabditis elegans Assay for Assessing the Therapeutic Potential of Bacteriophages against Clinical Pathogens

Manohar, Prasanth; Leptihn, Sebastian; Elangovan, Namasivayam; Loganathan, Archana; Nachimuthu, Ramesh

doi:10.1128/spectrum.01393-21

Cited by 10 publications

(11 citation statements)

References 47 publications

(58 reference statements)

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“…Another consideration is the use of "phage cocktails" consisting of multiple phages, because this vastly reduces the chance that the bacterium develops resistance to the treatment [211][212][213][214]. Including a flagellotropic phage in a phage cocktail could certainly be advantageous; however, before a flagellotropic phage could be included in an antimicrobial cocktail, more simple in vivo experimentation must be conducted on models such as mice [215], Caenorhabditis elegans [216], or Galleria mellonella [217,218].…”

Section: In Vivo Researchmentioning

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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Section: In Vivo Researchmentioning

confidence: 99%

Flagellotropic Bacteriophages: Opportunities and Challenges for Antimicrobial Applications

Esteves

Scharf

2022

IJMS

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“…To study the anti-biofilm activity of phages, 10 strong biofilm-forming (based on OD) P. aeruginosa isolates were chosen (P. aeruginosa strains 01, 08, 11,16,27,32,35,37,42,47). Anti-biofilm activity of the phages was determined after adding the phages (10 6 PFU/mL) to the biofilm.…”

Section: Biofilm Clearance Assaymentioning

confidence: 99%

“…A liquid-based assay to study the efficacy of phages against pathogenic bacteria was previously developed by our research team (42). To test the efficacy of Pseudomonas bacteriophage in recovering the nematodes from pseudomonal infection, two types of studies were performed i.e., therapeutic treatment and prophylactic treatment.…”

Section: Phage Efficacy Assaymentioning

confidence: 99%

“…To analyze the presence of phages inside the nematodes or uptake of phages by the nematodes (42), phage numbers were determined as follows: Briefly, after 4 days, 10 nematodes were removed from groups 5, 6 and 7, which are phage only, therapeutic treatment and prophylactic treatment, respectively, to determine the phage titer using the double agar overlay method. In brief, the nematodes were washed thrice with buffer, vortexed, ground (with mortar and pestle), and centrifuged at 10,000 ´ g for 5 min.…”

Section: Phage Efficacy Assaymentioning

confidence: 99%

“…In brief, the nematodes were washed thrice with buffer, vortexed, ground (with mortar and pestle), and centrifuged at 10,000 ´ g for 5 min. The supernatant was used to determine the phage titer (42).…”

Section: Phage Efficacy Assaymentioning

confidence: 99%

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In vitroandin vivoevaluation of the biofilm-degradingPseudomonasphage Motto, as a candidate for phage therapy

Manohar

Loh

Turner

et al. 2022

Preprint

Self Cite

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Infections caused byPseudomonas aeruginosaare becoming increasingly difficult to treat due to the emergence of strains that have acquired multidrug resistance. Therefore, phage therapy has gained attention as an alternative to the treatment of pseudomonal infections. Phages are not only bactericidal but occasionally show activity against biofilm as well. Here, we describe thePseudomonasphage Motto which has the ability to clearP. aeruginosainfections in an animal model and also exhibited biofilm-degrading properties. The phage has substantial antibiofilm activity against strong biofilm-producing isolates (n = 10), with at least a 2-fold reduction within 24 hours. To demonstrate the safety of using phage Motto, we performed cytotoxicity studies with human cell lines (HEK 293 and RAW 264.7 macrophages). Using a previously establishedin vivomodel, we demonstrated the efficacy of Motto inC. elegans, with a 90% survival rate when treated with the phage at an MOI of 10.

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Tracking Chlamydia – Host interactions and antichlamydial activity in Caenorhabditis elegans

Reigada,

Kapp,

Kaudela

et al. 2024

Biomedicine & Pharmacotherapy

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A Multiwell-Plate Caenorhabditis elegans Assay for Assessing the Therapeutic Potential of Bacteriophages against Clinical Pathogens

Abstract: Validating the efficacy and safety of phages prior to clinical application is crucial to see phage therapy in practice. Current animal models include mice and insect larvae, which pose ethical or technical challenges.

Cited by 10 publications

References 47 publications

Flagellotropic Bacteriophages: Opportunities and Challenges for Antimicrobial Applications

Flagellotropic Bacteriophages: Opportunities and Challenges for Antimicrobial Applications

In vitroandin vivoevaluation of the biofilm-degradingPseudomonasphage Motto, as a candidate for phage therapy

Tracking Chlamydia – Host interactions and antichlamydial activity in Caenorhabditis elegans

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