In vivo evaluation of phage therapy against Klebsiella pneumoniae using the Galleria mellonella model and molecular characterization of a novel Drulisvirus phage species
ABSTRACT Multidrug-resistant (MDR) Klebsiella pneumoniae is challenging to treat with conventional antibiotic regimens, posing a threat to healthcare systems. Phage therapy presents a promising alternative treatment strategy; however, characterization of its efficacy and safety is required. Here, we...
Saved in:
| Main Authors: | , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
American Society for Microbiology
2025-05-01
|
| Series: | Microbiology Spectrum |
| Subjects: | |
| Online Access: | https://journals.asm.org/doi/10.1128/spectrum.01145-24 |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1850203501353762816 |
|---|---|
| author | Gustavo Quispe-Villegas Gabriela I. Alcántara-Lozano Diego Cuicapuza Raúl Laureano Brenda Ayzanoa Pablo Tsukayama Jesús Tamariz |
| author_facet | Gustavo Quispe-Villegas Gabriela I. Alcántara-Lozano Diego Cuicapuza Raúl Laureano Brenda Ayzanoa Pablo Tsukayama Jesús Tamariz |
| author_sort | Gustavo Quispe-Villegas |
| collection | DOAJ |
| description | ABSTRACT Multidrug-resistant (MDR) Klebsiella pneumoniae is challenging to treat with conventional antibiotic regimens, posing a threat to healthcare systems. Phage therapy presents a promising alternative treatment strategy; however, characterization of its efficacy and safety is required. Here, we describe the microbiological and molecular characterization of a novel bacteriophage with activity against MDR K. pneumoniae using a greater wax moth (Galleria mellonella) model system. A bacteriophage was isolated from hospital wastewater. Viral kinetics and phage stability were evaluated under varied pH and temperature conditions. The therapeutic efficacy of the phage was evaluated using MDR Klebsiella-infected G. mellonella larvae as an in vivo model. Phage titers and larva survival were compared in phage-treated and control groups. Genomic sequencing (Nanopore and Illumina) was used to classify the bacteriophage and identify any resistance genes or virulence factors present in its genome. Functional characterization demonstrated effective lytic activity, favorable burst size (161 PFU/cell), and an optimal MOI of 0.1. The phage demonstrated stability across a wide range of temperatures (8°C–40°C) and pH levels (4–8). Experiments using the G. mellonella model showed improved larval survival with phage treatment. The novel bacteriophage was identified as a new species within the genus Drulisvirus with no lysogeny-associated, antimicrobial resistance, or virulence genes detected. The new Drulisvirus phage identified is a promising candidate for treatment of infections caused by MDR K. pneumoniae.IMPORTANCEThe study describes a bacteriophage with potential for use in phage therapy against Klebsiella pneumoniae, one of the most clinically significant bacterial pathogens today. Microbiological and genomic characterization of the phage revealed advantageous properties for therapeutic applications, while also identifying a novel species within the Drulisvirus genus. These findings significantly contribute to our understanding of bacteriophage diversity and their utility in combating antibiotic-resistant infections. Moreover, the authors developed an in vivo preclinical model of MDR infection using Galleria mellonella larvae and successfully applied it to study the bacteriophage’s therapeutic efficacy. This model offers a robust and efficient platform for preclinical testing. |
| format | Article |
| id | doaj-art-880d064f8faf4a46ae2e4db9ff738050 |
| institution | OA Journals |
| issn | 2165-0497 |
| language | English |
| publishDate | 2025-05-01 |
| publisher | American Society for Microbiology |
| record_format | Article |
| series | Microbiology Spectrum |
| spelling | doaj-art-880d064f8faf4a46ae2e4db9ff7380502025-08-20T02:11:30ZengAmerican Society for MicrobiologyMicrobiology Spectrum2165-04972025-05-0113510.1128/spectrum.01145-24In vivo evaluation of phage therapy against Klebsiella pneumoniae using the Galleria mellonella model and molecular characterization of a novel Drulisvirus phage speciesGustavo Quispe-Villegas0Gabriela I. Alcántara-Lozano1Diego Cuicapuza2Raúl Laureano3Brenda Ayzanoa4Pablo Tsukayama5Jesús Tamariz6Laboratorio de Resistencia Antibiótica y Fagoterapia, Laboratorios de Investigación y Desarrollo, Facultad de Ciencias e Ingeniería, Universidad Peruana Cayetano Heredia, Lima, PeruLaboratorio de Resistencia Antibiótica y Fagoterapia, Laboratorios de Investigación y Desarrollo, Facultad de Ciencias e Ingeniería, Universidad Peruana Cayetano Heredia, Lima, PeruLaboratorio de Resistencia Antibiótica y Fagoterapia, Laboratorios de Investigación y Desarrollo, Facultad de Ciencias e Ingeniería, Universidad Peruana Cayetano Heredia, Lima, PeruLaboratorio de Resistencia Antibiótica y Fagoterapia, Laboratorios de Investigación y Desarrollo, Facultad de Ciencias e Ingeniería, Universidad Peruana Cayetano Heredia, Lima, PeruLaboratorio de Genómica Microbiana, Laboratorios de Investigación y Desarrollo, Facultad de Ciencias e Ingeniería, Universidad Peruana Cayetano Heredia, Lima, PeruLaboratorio de Genómica Microbiana, Laboratorios de Investigación y Desarrollo, Facultad de Ciencias e Ingeniería, Universidad Peruana Cayetano Heredia, Lima, PeruLaboratorio de Resistencia Antibiótica y Fagoterapia, Laboratorios de Investigación y Desarrollo, Facultad de Ciencias e Ingeniería, Universidad Peruana Cayetano Heredia, Lima, PeruABSTRACT Multidrug-resistant (MDR) Klebsiella pneumoniae is challenging to treat with conventional antibiotic regimens, posing a threat to healthcare systems. Phage therapy presents a promising alternative treatment strategy; however, characterization of its efficacy and safety is required. Here, we describe the microbiological and molecular characterization of a novel bacteriophage with activity against MDR K. pneumoniae using a greater wax moth (Galleria mellonella) model system. A bacteriophage was isolated from hospital wastewater. Viral kinetics and phage stability were evaluated under varied pH and temperature conditions. The therapeutic efficacy of the phage was evaluated using MDR Klebsiella-infected G. mellonella larvae as an in vivo model. Phage titers and larva survival were compared in phage-treated and control groups. Genomic sequencing (Nanopore and Illumina) was used to classify the bacteriophage and identify any resistance genes or virulence factors present in its genome. Functional characterization demonstrated effective lytic activity, favorable burst size (161 PFU/cell), and an optimal MOI of 0.1. The phage demonstrated stability across a wide range of temperatures (8°C–40°C) and pH levels (4–8). Experiments using the G. mellonella model showed improved larval survival with phage treatment. The novel bacteriophage was identified as a new species within the genus Drulisvirus with no lysogeny-associated, antimicrobial resistance, or virulence genes detected. The new Drulisvirus phage identified is a promising candidate for treatment of infections caused by MDR K. pneumoniae.IMPORTANCEThe study describes a bacteriophage with potential for use in phage therapy against Klebsiella pneumoniae, one of the most clinically significant bacterial pathogens today. Microbiological and genomic characterization of the phage revealed advantageous properties for therapeutic applications, while also identifying a novel species within the Drulisvirus genus. These findings significantly contribute to our understanding of bacteriophage diversity and their utility in combating antibiotic-resistant infections. Moreover, the authors developed an in vivo preclinical model of MDR infection using Galleria mellonella larvae and successfully applied it to study the bacteriophage’s therapeutic efficacy. This model offers a robust and efficient platform for preclinical testing.https://journals.asm.org/doi/10.1128/spectrum.01145-24bacteriophagesphage therapyKlebsiella pneumoniaeGalleria mellonella |
| spellingShingle | Gustavo Quispe-Villegas Gabriela I. Alcántara-Lozano Diego Cuicapuza Raúl Laureano Brenda Ayzanoa Pablo Tsukayama Jesús Tamariz In vivo evaluation of phage therapy against Klebsiella pneumoniae using the Galleria mellonella model and molecular characterization of a novel Drulisvirus phage species Microbiology Spectrum bacteriophages phage therapy Klebsiella pneumoniae Galleria mellonella |
| title | In vivo evaluation of phage therapy against Klebsiella pneumoniae using the Galleria mellonella model and molecular characterization of a novel Drulisvirus phage species |
| title_full | In vivo evaluation of phage therapy against Klebsiella pneumoniae using the Galleria mellonella model and molecular characterization of a novel Drulisvirus phage species |
| title_fullStr | In vivo evaluation of phage therapy against Klebsiella pneumoniae using the Galleria mellonella model and molecular characterization of a novel Drulisvirus phage species |
| title_full_unstemmed | In vivo evaluation of phage therapy against Klebsiella pneumoniae using the Galleria mellonella model and molecular characterization of a novel Drulisvirus phage species |
| title_short | In vivo evaluation of phage therapy against Klebsiella pneumoniae using the Galleria mellonella model and molecular characterization of a novel Drulisvirus phage species |
| title_sort | in vivo evaluation of phage therapy against klebsiella pneumoniae using the galleria mellonella model and molecular characterization of a novel drulisvirus phage species |
| topic | bacteriophages phage therapy Klebsiella pneumoniae Galleria mellonella |
| url | https://journals.asm.org/doi/10.1128/spectrum.01145-24 |
| work_keys_str_mv | AT gustavoquispevillegas invivoevaluationofphagetherapyagainstklebsiellapneumoniaeusingthegalleriamellonellamodelandmolecularcharacterizationofanoveldrulisvirusphagespecies AT gabrielaialcantaralozano invivoevaluationofphagetherapyagainstklebsiellapneumoniaeusingthegalleriamellonellamodelandmolecularcharacterizationofanoveldrulisvirusphagespecies AT diegocuicapuza invivoevaluationofphagetherapyagainstklebsiellapneumoniaeusingthegalleriamellonellamodelandmolecularcharacterizationofanoveldrulisvirusphagespecies AT raullaureano invivoevaluationofphagetherapyagainstklebsiellapneumoniaeusingthegalleriamellonellamodelandmolecularcharacterizationofanoveldrulisvirusphagespecies AT brendaayzanoa invivoevaluationofphagetherapyagainstklebsiellapneumoniaeusingthegalleriamellonellamodelandmolecularcharacterizationofanoveldrulisvirusphagespecies AT pablotsukayama invivoevaluationofphagetherapyagainstklebsiellapneumoniaeusingthegalleriamellonellamodelandmolecularcharacterizationofanoveldrulisvirusphagespecies AT jesustamariz invivoevaluationofphagetherapyagainstklebsiellapneumoniaeusingthegalleriamellonellamodelandmolecularcharacterizationofanoveldrulisvirusphagespecies |