Design of a Bacteriophage Cocktail Active against Shigella Species and Testing of Its Therapeutic Potential in Galleria mellonella

Abstract

Shigellosis is a leading global cause of diarrheal disease and travelers’ diarrhea now being complicated by the dissemination of antibiotic resistance, necessitating the development of alternative antibacterials such as therapeutic bacteriophages (phages). Phages with lytic activity against Shigella strains were isolated from sewage. The genomes of 32 phages were sequenced, and based on genomic comparisons belong to seven taxonomic genera: Teetrevirus, Teseptimavirus, Kayfunavirus, Tequatrovirus, Mooglevirus, Mosigvirus and Hanrivervirus. Phage host ranges were determined with a diverse panel of 95 clinical isolates of Shigella from Southeast Asia and other geographic regions, representing different species and serotypes. Three-phage mixtures were designed, with one possessing lytic activity against 89% of the strain panel. This cocktail exhibited lytic activity against 100% of S. sonnei isolates, 97.2% of S. flexneri (multiple serotypes) and 100% of S. dysenteriae serotypes 1 and 2. Another 3-phage cocktail composed of two myophages and one podophage showed both a broad host range and the ability to completely sterilize liquid culture of a model virulent strain S. flexneri 2457T. In a Galleria mellonella model of lethal infection with S. flexneri 2457T, this 3-phage cocktail provided a significant increase in survival.

Appelmans Protocol forin vitro Klebsiella pneumoniaephage host range expansion leads to induction of a novel temperate linear plasmid prophagevB_KpnS-KpLi5

“…Our results support linear use of phages in therapy, as recently advocated by the Turner lab [12], instead of simultaneous application in cocktails to limit the likelihood of stress inductions of possibly pathogenic temperate phages. Simultaneous use of phages in cocktails is still considered state of the art [13,14]. Our findings may be useful to inform therapeutic approaches of our clinical partners [12][13][14], and the Phage4_1Health program [15].…”

“…Simultaneous use of phages in cocktails is still considered state of the art [13,14]. Our findings may be useful to inform therapeutic approaches of our clinical partners [12][13][14], and the Phage4_1Health program [15]. ( 903)…”

Isolation and characterization of bacteriophages against Burkholderia species

“…For instance, Rai et al ( 2022) [39] demonstrated the isolation of phages targeting methicillin-resistant S. aureus from sewage, while Sjahriani et al (2021) [40] reported a high abundance of phages against E. coli in environmental sewage. Additionally, Klai (2020) [41] noted that phages are prevalent in hospital waste, and research has shown their presence in wastewater treatment plants [42].…”

UsingGalleria mellonellato study pathogen dissemination and anti-infective surface coatings

“…G. mellonella is a robust model organism that has gained popularity in recent decades. This model organism has been used for a wide range of assays including virulence assays (bacterial and fungal), immunotoxicity testing (concerning the adaptive immune system), drug toxicity assays, testing the effects of ultraviolet radiation, probiotics, and bacteriophage treatment efficacy (Cunha et al ., 2023, Stempinski, Smith and Casadevall 2022, Erbas et al ., 2023, Demirtürk, Uçkan, and Mert, 2023, Sabockyte et al ., 2023, Filippov et al ., 2022, Maslova et al ., 2023). G. mellonella are affordable, have low ethical considerations associated with their use and have low maintenance and handling requirements which could facilitate faster early-stage pathogen dissemination and prevention strategy testing.…”