Bacteriophage-mediated decolonization of Klebsiella pneumoniae in a novel Galleria mellonella gut colonization model with Enterobacteriaceae.

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Kamran A Mirza, Sandor Nietzsche, Oliwia Makarewicz, Mathias W Pletz, Lara Thieme

Author Information

Kamran A Mirza: Jena University Hospital, Institute of Infectious Diseases and Infection Control, Friedrich-Schiller-University Jena, Am Klinikum 1, 07747, Jena, Germany. kamran.mirza@med.uni-jena.de.
Sandor Nietzsche: Center for Electron Microscope, Jena University Hospital, Ziegelmühlenweg 1, 07743, Jena, Germany.
Oliwia Makarewicz: Jena University Hospital, Institute of Infectious Diseases and Infection Control, Friedrich-Schiller-University Jena, Am Klinikum 1, 07747, Jena, Germany.
Mathias W Pletz: Jena University Hospital, Institute of Infectious Diseases and Infection Control, Friedrich-Schiller-University Jena, Am Klinikum 1, 07747, Jena, Germany.
Lara Thieme: Jena University Hospital, Institute of Infectious Diseases and Infection Control, Friedrich-Schiller-University Jena, Am Klinikum 1, 07747, Jena, Germany.

PMID: 38172281 DOI: 10.1038/s41598-023-50823-9

Galleria mellonella larvae have emerged as an invertebrate model for investigating bacterial pathogenesis and potential therapies, addressing ethical concerns related to mammalian models. This model has the advantage of having a simple gut microbiome, which is suitable for gut colonization studies. Intestinal colonization by Enterobacteriaceae significantly contributes to the spread of antibiotic resistance. This study aimed to establish a novel Enterobacteriaceae gut colonization larval model and assess its suitability for evaluating distinct antimicrobial efficacies. Larvae were force-fed sequentially with bacterial doses of K. pneumoniae and E. coli at 0, 24, and 48 h, with survival monitoring at 24 h intervals. Bacterial counts were assessed after 48 h and 120 h of force-feeding. Successfully colonized larvae were subjected to one-time force feeding of a bacteriophage cocktail (10 PFU/larvae) or MIC-based meropenem and ciprofloxacin. The colonized bacterial load was quantified by CFU count. Three doses of 10 CFU/larvae resulted in stable gut colonization, independent of the K. pneumoniae or E. coli strain. Compared with the control, force-feeding of the bacteriophage reduced the colonization of the strain Kp 419614 by 5 log CFU/larvae, while antibiotic treatment led to a 3 log CFU/larval reduction. This novel G. mellonella model provides a valuable alternative for gut colonization studies, facilitating proof-of-concept investigations and potentially reducing or replacing follow-up experiments in vertebrate models.

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Animals

Anti-Bacterial Agents

Bacteria

Bacteriophages

Enterobacteriaceae

Escherichia coli

Klebsiella pneumoniae

Larva

Mammals

Moths

Anti-Bacterial Agents

Journal Article

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