Andrade, Mariana

Costa, SS

Couto, I

Marques, Joana

Rodrigues, L

Rodrigues, Telma

Silveira, H

Viveiros, M

English

Repositório da Universidade Nova de Lisboa

CONGRESS OF MICROBIOLOGY AND BIOTECHNOLOGY 2023BOOK OF7TH - 9THDECEMBERABSTRACTSUNIVERSIDADE DA BEIRA INTERIORCovilhãCongress of Microbiology and Biotechnology 2023  368             MicroBiotec 2023 - Covilhã, Portugal  P3.40 - IMPLEMENTATION OF A GALLERIA MELLONELLA INFECTION MODEL AT GHTM/IHMT-NOVA   Telma Rodrigues 1, Mariana Andrade 1, Joana Marques 1, Liliana Rodrigues 1, Miguel Viveiros 1, Henrique Silveira 1, Isabel Couto 1, Sofia Santos Costa 1(*) 1 Global Health and Tropical Medicine, GHTM, Associate Laboratory in Translation and Innovation Towards Global Health, LA-REAL, Instituto de Higiene e Medicina Tropical, IHMT, Universidade Nova de Lisboa, UNL, Lisbon, Portugal  (*) e-mail: scosta@ihmt.unl.pt   Keywords: Infection model; Galleria mellonella; pathogenicity assays; bacterial pathogens.    ABSTRACT Galleria mellonella represents a sustainable invertebrate infection model that is gaining increasing interest, particularly for pathogenicity and infection assays, host-pathogen interaction studies, and pharmaco-toxicological studies. This work describes the implementation of a G. mellonella colony at GHTM/IHMT-NOVA to provide standardized larvae for the research community. Infection protocols for relevant bacterial pathogens were optimized. G. mellonella at the last larval stage were acquired from a commercial house specialized in exotic animals and the species confirmed by PCR. The larvae were reared, in the dark, at 27-28 °C and 60-80% relative humidity with a high protein diet. Infection assays were optimized with reference strains Staphylococcus aureus ATCC25923, Pseudomonas aeruginosa ATCC27583, Neisseria gonorrhoeae ATCC49226 and Escherichia coli ATCC25922. A G. mellonella colony was established with parameters such as diet and routine maintenance tested. In-house reared G. mellonella presented a life cycle between 31 to 34 days (from egg to adult). Each female laid on average > 1,500 eggs with a rate of egg hatching of ≈34%. Compared to the commercially acquired, in-house reared larvae were healthier and more resistant to infection by S. aureus ATCC25923. The optimization of infection assays included pre-incubation conditions and bacterial inoculum size, among other parameters. By applying the optimized infection assays, at equivalent inoculums, the four bacterial pathogens tested could be differentiated and ranked according to their virulence potential, as follows: P. aeruginosa >> S. aureus ≈ E. coli >> N. gonorrhoeae. This work represents a build-up on the animal infection models available at GHTM/IHMT-NOVA for collaborators and the research community. Ongoing work focus on expanding this model to fungal and parasite pathogens, as well as additional functional assays, such as pharmaco-toxicological studies.  Acknowledgements:  Fundação para a Ciência e a Tecnologia (FCT, Portugal) through funds to Project DREBI Ref. 2022.07931.PTDC, to GHTM (UID/04413/2020), LA-REAL (LA/P/0117/2020) and grant UI/BD/154472/2022 awarded to MA.   

Implementation of a Galleria mellonella infection model at GHTM/IHMT-NOVA

https://run.unl.pt/bitstream/10362/164804/1/Rodrigues_MicroBiotec23_poster.pdf

Implementation of a Galleria mellonella infection model at GHTM/IHMT-NOVA

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