Refined methodology for quantifying Pseudomonas aeruginosa virulence using Galleria mellonella
ABSTRACT Larvae of Galleria mellonella (the greater wax moth) are being increasingly used as a model to study microbial pathogenesis. In this model, bacterial virulence is typically measured by determining the 50% lethal dose (LD50) of a bacterial strain or mutant. The use of G. mellonella to study...
Saved in:
| Main Authors: | , , , , , , , , , , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
American Society for Microbiology
2025-02-01
|
| Series: | Microbiology Spectrum |
| Subjects: | |
| Online Access: | https://journals.asm.org/doi/10.1128/spectrum.01666-24 |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1832540832916307968 |
|---|---|
| author | Christopher M. R. Axline Travis J. Kochan Sophie Nozick Timothy Ward Tania Afzal Issay Niki Sumitra D. Mitra Ethan VanGosen Julia Nelson Aliki Valdes David Hynes William Cheng Joanne Lee Prarthana Prashanth Timothy L. Turner Nathan B. Pincus Marc H. Scheetz Kelly E. R. Bachta Alan R. Hauser |
| author_facet | Christopher M. R. Axline Travis J. Kochan Sophie Nozick Timothy Ward Tania Afzal Issay Niki Sumitra D. Mitra Ethan VanGosen Julia Nelson Aliki Valdes David Hynes William Cheng Joanne Lee Prarthana Prashanth Timothy L. Turner Nathan B. Pincus Marc H. Scheetz Kelly E. R. Bachta Alan R. Hauser |
| author_sort | Christopher M. R. Axline |
| collection | DOAJ |
| description | ABSTRACT Larvae of Galleria mellonella (the greater wax moth) are being increasingly used as a model to study microbial pathogenesis. In this model, bacterial virulence is typically measured by determining the 50% lethal dose (LD50) of a bacterial strain or mutant. The use of G. mellonella to study Pseudomonas aeruginosa pathogenesis, however, is challenging because of the extreme sensitivity of larvae to this bacterium. For some P. aeruginosa strains, as few as 1–5 colony-forming units are sufficient to kill G. mellonella, which poses challenges for determining LD50 values. For this reason, some groups have used time-to-death as a measure of P. aeruginosa virulence, but methodologies have not been standardized. We provide a detailed protocol for using the time at which 50% of larvae have died (LT50) at a particular inoculum as a measure of P. aeruginosa virulence. We also describe a quality control metric for enhancing the reproducibility of LT50 values. This approach provides an accurate and reproducible methodology for using G. mellonella larvae to measure and compare the virulence of P. aeruginosa strains.IMPORTANCEPseudomonas aeruginosa is a significant cause of morbidity and mortality. The invertebrate Galleria mellonella is used as a model to determine the virulence of P. aeruginosa strains. We provide a protocol and analytical approach for using a time-to-death metric to accurately quantify the virulence of P. aeruginosa strains in G. mellonella larvae. This methodology, which has several advantages over 50% lethal dose approaches, is a useful resource for the study of P. aeruginosa pathogenicity. |
| format | Article |
| id | doaj-art-2ff80e4d9a85452f95ec8aa657bcabe1 |
| institution | Kabale University |
| issn | 2165-0497 |
| language | English |
| publishDate | 2025-02-01 |
| publisher | American Society for Microbiology |
| record_format | Article |
| series | Microbiology Spectrum |
| spelling | doaj-art-2ff80e4d9a85452f95ec8aa657bcabe12025-02-04T14:03:41ZengAmerican Society for MicrobiologyMicrobiology Spectrum2165-04972025-02-0113210.1128/spectrum.01666-24Refined methodology for quantifying Pseudomonas aeruginosa virulence using Galleria mellonellaChristopher M. R. Axline0Travis J. Kochan1Sophie Nozick2Timothy Ward3Tania Afzal4Issay Niki5Sumitra D. Mitra6Ethan VanGosen7Julia Nelson8Aliki Valdes9David Hynes10William Cheng11Joanne Lee12Prarthana Prashanth13Timothy L. Turner14Nathan B. Pincus15Marc H. Scheetz16Kelly E. R. Bachta17Alan R. Hauser18Department of Microbiology-Immunology, Feinberg School of Medicine, Northwestern University, Chicago, Illinois, USADepartment of Microbiology-Immunology, Feinberg School of Medicine, Northwestern University, Chicago, Illinois, USADepartment of Microbiology-Immunology, Feinberg School of Medicine, Northwestern University, Chicago, Illinois, USADepartment of Microbiology-Immunology, Feinberg School of Medicine, Northwestern University, Chicago, Illinois, USADepartment of Microbiology-Immunology, Feinberg School of Medicine, Northwestern University, Chicago, Illinois, USADepartment of Microbiology-Immunology, Feinberg School of Medicine, Northwestern University, Chicago, Illinois, USADepartment of Microbiology-Immunology, Feinberg School of Medicine, Northwestern University, Chicago, Illinois, USADepartment of Microbiology-Immunology, Feinberg School of Medicine, Northwestern University, Chicago, Illinois, USADepartment of Microbiology-Immunology, Feinberg School of Medicine, Northwestern University, Chicago, Illinois, USADepartment of Microbiology-Immunology, Feinberg School of Medicine, Northwestern University, Chicago, Illinois, USADepartment of Microbiology-Immunology, Feinberg School of Medicine, Northwestern University, Chicago, Illinois, USADepartment of Microbiology-Immunology, Feinberg School of Medicine, Northwestern University, Chicago, Illinois, USADepartment of Microbiology-Immunology, Feinberg School of Medicine, Northwestern University, Chicago, Illinois, USADepartment of Microbiology-Immunology, Feinberg School of Medicine, Northwestern University, Chicago, Illinois, USADepartment of Microbiology-Immunology, Feinberg School of Medicine, Northwestern University, Chicago, Illinois, USADepartment of Microbiology-Immunology, Feinberg School of Medicine, Northwestern University, Chicago, Illinois, USADepartment of Pharmacy Practice, Midwestern University Colleges of Pharmacy and Pharmacology, Downers Grove, Illinois, USADepartment of Microbiology-Immunology, Feinberg School of Medicine, Northwestern University, Chicago, Illinois, USADepartment of Microbiology-Immunology, Feinberg School of Medicine, Northwestern University, Chicago, Illinois, USAABSTRACT Larvae of Galleria mellonella (the greater wax moth) are being increasingly used as a model to study microbial pathogenesis. In this model, bacterial virulence is typically measured by determining the 50% lethal dose (LD50) of a bacterial strain or mutant. The use of G. mellonella to study Pseudomonas aeruginosa pathogenesis, however, is challenging because of the extreme sensitivity of larvae to this bacterium. For some P. aeruginosa strains, as few as 1–5 colony-forming units are sufficient to kill G. mellonella, which poses challenges for determining LD50 values. For this reason, some groups have used time-to-death as a measure of P. aeruginosa virulence, but methodologies have not been standardized. We provide a detailed protocol for using the time at which 50% of larvae have died (LT50) at a particular inoculum as a measure of P. aeruginosa virulence. We also describe a quality control metric for enhancing the reproducibility of LT50 values. This approach provides an accurate and reproducible methodology for using G. mellonella larvae to measure and compare the virulence of P. aeruginosa strains.IMPORTANCEPseudomonas aeruginosa is a significant cause of morbidity and mortality. The invertebrate Galleria mellonella is used as a model to determine the virulence of P. aeruginosa strains. We provide a protocol and analytical approach for using a time-to-death metric to accurately quantify the virulence of P. aeruginosa strains in G. mellonella larvae. This methodology, which has several advantages over 50% lethal dose approaches, is a useful resource for the study of P. aeruginosa pathogenicity.https://journals.asm.org/doi/10.1128/spectrum.01666-24Pseudomonas aeruginosaGalleria mellonellavirulence |
| spellingShingle | Christopher M. R. Axline Travis J. Kochan Sophie Nozick Timothy Ward Tania Afzal Issay Niki Sumitra D. Mitra Ethan VanGosen Julia Nelson Aliki Valdes David Hynes William Cheng Joanne Lee Prarthana Prashanth Timothy L. Turner Nathan B. Pincus Marc H. Scheetz Kelly E. R. Bachta Alan R. Hauser Refined methodology for quantifying Pseudomonas aeruginosa virulence using Galleria mellonella Microbiology Spectrum Pseudomonas aeruginosa Galleria mellonella virulence |
| title | Refined methodology for quantifying Pseudomonas aeruginosa virulence using Galleria mellonella |
| title_full | Refined methodology for quantifying Pseudomonas aeruginosa virulence using Galleria mellonella |
| title_fullStr | Refined methodology for quantifying Pseudomonas aeruginosa virulence using Galleria mellonella |
| title_full_unstemmed | Refined methodology for quantifying Pseudomonas aeruginosa virulence using Galleria mellonella |
| title_short | Refined methodology for quantifying Pseudomonas aeruginosa virulence using Galleria mellonella |
| title_sort | refined methodology for quantifying pseudomonas aeruginosa virulence using galleria mellonella |
| topic | Pseudomonas aeruginosa Galleria mellonella virulence |
| url | https://journals.asm.org/doi/10.1128/spectrum.01666-24 |
| work_keys_str_mv | AT christophermraxline refinedmethodologyforquantifyingpseudomonasaeruginosavirulenceusinggalleriamellonella AT travisjkochan refinedmethodologyforquantifyingpseudomonasaeruginosavirulenceusinggalleriamellonella AT sophienozick refinedmethodologyforquantifyingpseudomonasaeruginosavirulenceusinggalleriamellonella AT timothyward refinedmethodologyforquantifyingpseudomonasaeruginosavirulenceusinggalleriamellonella AT taniaafzal refinedmethodologyforquantifyingpseudomonasaeruginosavirulenceusinggalleriamellonella AT issayniki refinedmethodologyforquantifyingpseudomonasaeruginosavirulenceusinggalleriamellonella AT sumitradmitra refinedmethodologyforquantifyingpseudomonasaeruginosavirulenceusinggalleriamellonella AT ethanvangosen refinedmethodologyforquantifyingpseudomonasaeruginosavirulenceusinggalleriamellonella AT julianelson refinedmethodologyforquantifyingpseudomonasaeruginosavirulenceusinggalleriamellonella AT alikivaldes refinedmethodologyforquantifyingpseudomonasaeruginosavirulenceusinggalleriamellonella AT davidhynes refinedmethodologyforquantifyingpseudomonasaeruginosavirulenceusinggalleriamellonella AT williamcheng refinedmethodologyforquantifyingpseudomonasaeruginosavirulenceusinggalleriamellonella AT joannelee refinedmethodologyforquantifyingpseudomonasaeruginosavirulenceusinggalleriamellonella AT prarthanaprashanth refinedmethodologyforquantifyingpseudomonasaeruginosavirulenceusinggalleriamellonella AT timothylturner refinedmethodologyforquantifyingpseudomonasaeruginosavirulenceusinggalleriamellonella AT nathanbpincus refinedmethodologyforquantifyingpseudomonasaeruginosavirulenceusinggalleriamellonella AT marchscheetz refinedmethodologyforquantifyingpseudomonasaeruginosavirulenceusinggalleriamellonella AT kellyerbachta refinedmethodologyforquantifyingpseudomonasaeruginosavirulenceusinggalleriamellonella AT alanrhauser refinedmethodologyforquantifyingpseudomonasaeruginosavirulenceusinggalleriamellonella |