Fluctuating Bacteriophage-induced galU Deficiency Region is Involved in Trade-off Effects on the Phage and Fluoroquinolone Sensitivity in Pseudomonas aeruginosa

Nakamura, Keisuke; Fujiki, Jumpei; Nakamura, Tomohiro; Furusawa, Takaaki; Gondaira, Satoshi; Usui, Masaru; Higuchi, Hidetoshi; Tsutsumi, Yutaka; Iwano, Hidetomo

doi:10.1016/j.virusres.2021.198596

Cited by 13 publications

(24 citation statements)

References 28 publications

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“…In order to address this issue, new strategies, which reverse antibiotic resistance, have been focused 50 . Actually, we have been previously reported that a type of P. aeruginosa variant acquires phage resistance and loses multi-drug efflux system MexXY-encoding gene at the same time, leading to restoration of fluoroquinolone sensitivity instead of a loss of phage sensitivity 51 . Similar with this strategy, restoration of antibiotics sensitivity of MRSA by ΦSA012 is expected through further analysis.…”

Section: Discussionmentioning

confidence: 99%

“…A phylogenetic tree of the phage whole genome was constructed by VICTOR 53 using this phage sequence. In addition, Staphylococcus viruses ΦSA039 18 , 21 , 48 and ΦMR003 26 and Pseudomonas viruses ΦS12-3 51 , 54 and ΦR18 54 , 55 , which were isolated and characterized previously, were used in this study. Escherichia viruses T1-T7 were obtained from the National Institute of Technology and Evaluation Biological Resource Center (NBRC, Chiba, Japan).…”

Section: Methodsmentioning

confidence: 99%

“…The lytic activity of ΦSA012 against SA003 under several conditions was evaluated in turbidity assays by monitoring the OD 590 for 24 h using a plate reader (Sunrise Rainbow Thermos RC, TECAN, Austria) as previously reported 51 . In brief, ΦSA012 was inoculated into exponentially growing SA003 cultures in a 96-well plate at a multiplicity of infection of 0.01.…”

Section: Methodsmentioning

confidence: 99%

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Biological properties of Staphylococcus virus ΦSA012 for phage therapy

Fujiki

Nakamura

et al. 2022

Sci Rep

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Staphylococcus virus ΦSA012 has a wide host range and efficient lytic activity. Here, we assessed the biological stability of ΦSA012 against temperature, freeze-thawing, and pH to clinically apply the phage. In addition, inoculation of ΦSA012 through i.p. and i.v. injections into mice revealed that phages were reached the limit of detection in serum and accumulated notably spleens without inflammation at 48 h post-inoculation. Furthermore, inoculation of ΦSA012 through s.c. injections in mice significantly induced IgG, which possesses neutralizing activity against ΦSA012 and other Staphylococcus viruses, ΦSA039 and ΦMR003, but not Pseudomonas viruses ΦS12-3 and ΦR18 or Escherichia viruses T1, T4, and T7 in vitro. Immunoelectron microscopic analysis showed that purified anti-phage IgG recognizes the long-tail fiber of staphylococcus viruses. Although S. aureus inoculation resulted in a 25% survival rate in a mouse i.p. model, ΦSA012 inoculation (i.p.) improved the survival rate to 75%; however, the survival rate of ΦSA012-immunized mice decreased to less than non-immunized mice with phage i.v. injection at a MOI of 100. These results indicated that ΦSA012 possesses promise for use against staphylococcal infections but we should carefully address the appropriate dose and periods of phage administration. Our findings facilitate understandings of staphylococcus viruses for phage therapy.

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Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

See 1 more Smart Citation

Biological properties of Staphylococcus virus ΦSA012 for phage therapy

Fujiki

Nakamura

et al. 2022

Sci Rep

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“…SNPs analysis of Kp21 and Kp21r suggests that Kp21r possesses deficient capsular polysaccharide, and thus, phage tail protein encoded in Myoviridae may more efficiently degrade Kp21r peptidoglycan and result in rapid lysis. Some studies have reported that phage-resistant bacteria become susceptible to antibiotic due to mutation in the genes involved in antibiotics resistance and showed that phages and antibiotics combination effectively kills the target bacteria (35–38). Our results demonstrate that the combinations of phage and phage-encoded tail lysozyme efficiently eliminate and/or inhibit the phage-resistant bacterial growth (39).…”

Section: Discussionmentioning

confidence: 99%

Isolation and characterization of bacteriophages against IMP-6-producingKlebsiella pneumoniaeisolated from clinical settings in Japan

Kondo

Nakano

Hisatsune

et al. 2022

Preprint

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Carbapenemase-producing Enterobacteriaceae (CPE) are one of the most detrimental species of antibiotic-resistant bacteria worldwide. Phage therapy has emerged as an effective strategy for the treatment of infections caused by CPE pathogens. In west Japan, the increasing occurrence of Klebsiella pneumoniae harboring the pKPI-6 plasmid, which encodes blaIMP-6, is a growing concern. To manage such major antimicrobial-resistant pathogens, we isolated 29 novel phages from sewage in Japan, targeting 31 strains of K. pneumoniae and one strain of Escherichia coli harboring the pKPI-6 plasmid. Electron microscopy analysis indicated that of the 29 isolated phages, 21 (72.4%), 5 (17.2%), and 3 (10.3%) belonged to Myoviridae, Siphoviridae, and Podoviridae, respectively. Host range analysis revealed that 20 Myoviridae members in isolated phages infected 25-26 strains of K. pneumoniae, indicating that most of the isolated phages have a broad host range. The K. pneumoniae Kp21 can only be infected by phage øKp_21, while Kp22 can be infected by more than 20 phages. We applied a phage cocktail, which consists of 10 phages, against Kp21 and Kp22 and found that the phage cocktail delayed the emergence of phage-resistant bacteria for Kp21 strain but not for the Kp22 strain. Furthermore, phage-resistant Kp21 (Kp21r) became prone to be infected from other bacteriophages as a "trade-off" of resistance to phage øKp_21. Our proposed phage set has an adequate number of phages to combat the K. pneumoniae strain isolated in Japan. Notably, our work demonstrates how a suitable phage cocktail diminishes the occurrence of phage-resistant bacteria.

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“…The lytic activity of ΦSA012 against SA003 under several conditions was evaluated in turbidity assays by monitoring the OD 590 for 24 h using a plate reader (Sunrise Rainbow Thermos RC, TECAN, Austria) as previously reported 53,58 . In brief, ΦSA012 was inoculated into exponentially growing SA003 cultures in a 96-well plate at a multiplicity of infection of 0.01.…”

Section: Monitoring Of Phage Lytic Activity By a Plate Readermentioning

confidence: 99%

Biological properties of Staphylococcus virus ΦSA012 for phage therapy

Fujiki

Nakamura

et al. 2022

Preprint

Self Cite

View full text Add to dashboard Cite

Staphylococcus virus ΦSA012 has a wide host range and efficient lytic activity. Here, we assessed the biological stability of ΦSA012 against temperature, freeze-thawing, and pH to clinically apply the phage. In addition, inoculation of ΦSA012 through i.p. and i.v. injections into mice revealed that phages were reached the limit of detection in serum and accumulated notably spleens without inflammation at 48 h post-inoculation. Furthermore, inoculation of ΦSA012 through s.c. injections in mice significantly induced IgG, which possesses neutralizing activity against ΦSA012 and other Staphylococcusviruses, ΦSA039 and ΦMR003, but not Pseudomonasviruses ΦS12-3 and ΦR18 or Escherichiaviruses T1, T4, and T7 in vitro. Immunoelectron microscopic analysis showed that purified anti-phage IgG recognizes the long-tail fiber of staphylococcus viruses. Although S. aureus inoculation resulted in a 25% survival rate in a mouse i.p. model, ΦSA012 inoculation (i.p.) improved the survival rate to 70%; however, the survival rate of ΦSA012-immunized mice decreased to less than non-immunized mice with phage i.v. injection at a MOI of 100. These results indicated that ΦSA012 possesses promise for use against staphylococcal infections but we should carefully address the appropriate dose and periods of phage administration. Our findings facilitate understandings of staphylococcus viruses for phage therapy.

show abstract

Fluctuating Bacteriophage-induced galU Deficiency Region is Involved in Trade-off Effects on the Phage and Fluoroquinolone Sensitivity in Pseudomonas aeruginosa

Cited by 13 publications

References 28 publications

Biological properties of Staphylococcus virus ΦSA012 for phage therapy

Biological properties of Staphylococcus virus ΦSA012 for phage therapy

Isolation and characterization of bacteriophages against IMP-6-producingKlebsiella pneumoniaeisolated from clinical settings in Japan

Biological properties of Staphylococcus virus ΦSA012 for phage therapy

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