Isolation of Bacteriophages from Environmental Sources, and Creation and Functional Screening of Phage DNA Libraries

Pelzek, Adam J.; Schuch, Raymond; Schmitz, Jonathan E.; Fischetti, Vincent A.

doi:10.1002/9780470089941.et1303s07

Cited by 7 publications

(8 citation statements)

References 39 publications

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“…Temperate phages, integrated within their bacterial host genomes as prophages, also pose a challenge for isolation, but can be induced from lysogenised isolates using UV light (Furuse et al, 1983), mitomycin C, and heat treatment (Pelzek et al, 2013). Induced prophages can then be separated from their hosts using the same methods as for virulent phages.…”

Section: Culture-dependent Methodsmentioning

confidence: 99%

The Human Gut Phageome: Origins and Roles in the Human Gut Microbiome

Townsend

Kelly

Muscatt

et al. 2021

Front. Cell. Infect. Microbiol.

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The investigation of the microbial populations of the human body, known as the microbiome, has led to a revolutionary field of science, and understanding of its impacts on human development and health. The majority of microbiome research to date has focussed on bacteria and other kingdoms of life, such as fungi. Trailing behind these is the interrogation of the gut viruses, specifically the phageome. Bacteriophages, viruses that infect bacterial hosts, are known to dictate the dynamics and diversity of bacterial populations in a number of ecosystems. However, the phageome of the human gut, while of apparent importance, remains an area of many unknowns. In this paper we discuss the role of bacteriophages within the human gut microbiome. We examine the methods used to study bacteriophage populations, how this evolved over time and what we now understand about the phageome. We review the phageome development in infancy, and factors that may influence phage populations in adult life. The role and action of the phageome is then discussed at both a biological-level, and in the broader context of human health and disease.

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Section: Culture-dependent Methodsmentioning

confidence: 99%

The Human Gut Phageome: Origins and Roles in the Human Gut Microbiome

Townsend

Kelly

Muscatt

et al. 2021

Front. Cell. Infect. Microbiol.

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“…The fact that most phages are discovered in locations such as bodies of water, sewage, and infected animals is a testament to the number of new phages that can be found in these niches, which can potentially be isolated and applied in a useful context. The self-replicating nature of phages makes their isolation simple and straightforward [22,23], with propagation being fairly trivial as well [24]. Their remarkable host specificity is an additional benefit, although this may also pose a drawback for broad-spectrum therapies [25,26].…”

Section: Broader Applications Of the Viruses Of Bacteriamentioning

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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“…Phages were isolated from the Athlone wastewater treatment plant (Cape Town, South Africa), using the enrichment protocol previously described [31]. Wastewater was centrifuged (8000× g, 15 min, 4 • C), the supernatant filtered through 0.2 µm GVS filters (Lasec, Cape Town, South Africa), and 1.0 mL inoculated into a 100 mL culture of P. mirabilis B16 in exponential phase, supplemented with 1 mM CaCl 2 .…”

Section: Phage Isolation and Purificationmentioning

confidence: 99%

Isolation and Characterization of Lytic Proteus Virus 309

Aaron

Zyl

Dicks

2022

Viruses

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Proteus mirabilis is frequently associated with complicated urinary tract infections (UTIs) and is the main cause of catheter-associated urinary tract infections (CAUTIs). Treatment of such infections is complicated and challenging due to the biofilm forming abilities of P. mirabilis. If neglected or mistreated, infections may lead to life-threating conditions such as cystitis, pyelonephritis, kidney failure, and bacteremia that may progress to urosepsis. Treatment with antibiotics, especially in cases of recurring and persistent infections, leads to the development of resistant strains. Recent insights into phage therapy and using phages to coat catheters have been evaluated with many studies showing promising results. Here, we describe a highly lytic bacteriophage, Proteus_virus_309 (41,740 bp), isolated from a wastewater treatment facility in Cape Town, South Africa. According to guidelines of the International Committee on Taxonomy of Viruses (ICTV), bacteriophage 309 is a species within the genus Novosibovirus. Similar to most members of the genus, bacteriophage 309 is strain-specific and lyse P. mirabilis in less than 20 min.

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Isolation of Bacteriophages from Environmental Sources, and Creation and Functional Screening of Phage DNA Libraries

Cited by 7 publications

References 39 publications

The Human Gut Phageome: Origins and Roles in the Human Gut Microbiome

The Human Gut Phageome: Origins and Roles in the Human Gut Microbiome

Flagellotropic Bacteriophages: Opportunities and Challenges for Antimicrobial Applications

Isolation and Characterization of Lytic Proteus Virus 309

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