Afzal, Tania

Hauser, Alan

Kochan, Travis J.

Lee, Joanne J. H.

Mitra, Sumitra D.

Turner, Timothy L.

Voisine, Cindy

English

NEIU Digital Commons (Northeastern Illinois University)

Northeastern Illinois University NEIU Digital Commons NEIU Student Research and Creative Activities Symposium 2021 NEIU Student Research and Creative Activities Symposium Apr 22nd, 9:45 AM Use of Galleria mellonella Larvae for Assessing Pseudomonas aeruginosa Virulence Tania Afzal Northeastern Illinois University Timothy L. Turner Northwestern University Sumitra D. Mitra Northwestern University Travis J. Kochan Northwestern University Joanne J. H. Lee Northwestern University See next page for additional authors Follow this and additional works at: https://neiudc.neiu.edu/srcas Afzal, Tania; Turner, Timothy L.; Mitra, Sumitra D.; Kochan, Travis J.; Lee, Joanne J. H.; Voisine, Cindy; and Hauser, Alan, "Use of Galleria mellonella Larvae for Assessing Pseudomonas aeruginosa Virulence" (2021). NEIU Student Research and Creative Activities Symposium. 1. https://neiudc.neiu.edu/srcas/2021/s38/1 This Event is brought to you for free and open access by the Conferences and Symposia at NEIU Digital Commons. It has been accepted for inclusion in NEIU Student Research and Creative Activities Symposium by an authorized administrator of NEIU Digital Commons. For more information, please contact h-owen3@neiu.edu,l-wallis@neiu.edu. Presenter Information Tania Afzal, Timothy L. Turner, Sumitra D. Mitra, Travis J. Kochan, Joanne J. H. Lee, Cindy Voisine, and Alan Hauser This event is available at NEIU Digital Commons: https://neiudc.neiu.edu/srcas/2021/s38/1 USE OF GALLERIA MELLONELLA LARVAE FOR ASSESSING PSEUDOMONAS AERUGINOSA VIRULENCE  Tania Afzal1, Timothy L. Turner2, Sumitra D. Mitra2, Travis J. Kochan2, Joanne J. H. Lee4, Cindy Voisine1, and Alan R. Hauser2,3  1Department of Biology, Northeastern Illinois University, Chicago, IL 60625 2Department of Microbiology-Immunology and 3Department of Medicine, Division of Infectious Diseases, Northwestern University, Feinberg School of Medicine, Chicago, IL 60611 4Department of Biology and Department of Psychology, Northwestern University, Weinberg College of Arts and Sciences, Evanston, IL 60201  Pseudomonas aeruginosa is a gram-negative opportunistic pathogen that is ubiquitous in the environment. Due to its multi-drug resistance mechanisms and virulence factors, P. aeruginosa has become a growing concern in the health care community and can be life-threatening in compromised individuals. High-throughput in vivo models are needed to assess the virulence of the wide variety of P. aeruginosa strains. Galleria mellonella has several advantages as a model organism in bacterial virulence studies, such as its affordability, minimal ethical restrictions, and ease of use. Our preliminary data indicated a low correlation (R2: 0.33) between the lethal dose for 50% mortality in mice (mLD50) and 50% mortality at 18 hours post-infection in G. mellonella (gLD50) infected with P. aeruginosa bloodstream isolates. Also, poor correlation (R2 = 0.34) was noted between the preliminary data's two biological replicates, indicating a problem with reproducibility. In this study, the protocol for virulence assays using G. mellonella as a model for P. aeruginosa infections was optimized and the sources of variability and unintended deaths in negative controls of preliminary studies were investigated. Injection site and depth, microbial contamination, and the impact of G. mellonella weight were investigated to determine if these factors created variation in preliminary experimentation. Mortality checks were changed to a 24-hour time course starting at 8 hours post-infection and continuing every hour until 24 hours post-infection or until 100% mortality was achieved. Five bacterial doses were checked, and the lethal time to achieve 50% mortality in G. mellonella (gLT50) was determined for each dose. Exponential decay curves were fit to colony-forming units (CFUs) vs. gLT50s plots, allowing for an accurate strain to strain comparisons at ~2,000 CFUs. The new methodology was then used to assess the relative gLT50 of  ~49 strains of P. aeruginosa. The virulence assay retest demonstrated an increase in correlation between gLT50 and mLD50 (R2: 0.61). These findings suggest that G. mellonella is a promising model organism for studying bacterial infections when utilizing our time course and dose-response curve protocol.   

Use of Galleria mellonella Larvae for Assessing Pseudomonas aeruginosa Virulence

https://neiudc.neiu.edu/cgi/viewcontent.cgi?article=1392&context=srcas

Use of Galleria mellonella Larvae for Assessing Pseudomonas aeruginosa Virulence

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