Development of a bacterial infection model of Galleria mellonella larvae (greater wax moth)

Original article
УДК 615.076.7: 615.076.9: 59.083: 59.084
DOI: 10.57034/2618723X-2022-03-05

J.V. Salmova*, microbiologist, https://orcid.org/0000-0001-9891-8634
L.R. Nikiforova, microbiologist, https://orcid.org/0000-0001-8710-2023
K.Е. Borovkova, head of microbiological laboratory, https://orcid.org/0000-0003-1571-6549

Research and manufacturing company «Home оf Pharmacy»,
188663, Russia, Leningrad oblast, Vsevolozhskiy district, Kuzmolovskiy t.s., Zavodskaya st. 3-245

* E-mail: [email protected]

Keywords: antimicrobial activity alternative models virulence antibiotic sensitivity
Acknowledgements

The study was performed without external funding.

For citation:

Salmova J.V. , Nikiforova L.R., Borovkova К.E. Development of a bacterial infection model of Galleria mellonella larvae (greater wax moth). Laboratory Animals for Science. 2022; 3. https://doi.org/10.29296/2618723X-2022-03-05

Abstract

In recent years,_ Galleria mellonella_, the larvae of a large wax moth, has been widely used as an alternative model for studying bacterial infections, as well as for evaluating the effectiveness of antibacterial drugs. The advantages of this model system are the absence of bioethical restrictions, the possibility of using a large number of animals in the experiment, as well as a short life cycle. The larvae can grow in a wide temperature range from 18 to 37 °C, allowing the study of human pathogens in this model, which in most cases require incubation at 37 °C. Despite the absence of acquired immunity in insects, the innate immunity of wax moths has significant similarities with the immune response of vertebrates, including humans.

The aim of the work was to develop a model of bacterial infection of larvae of large wax moths including comparative evaluation of the efficiency of larval infection with reference strains of microorganisms and their clinical isolates as well as evaluation of the effectiveness of Rifampicin (PJSC Krasfarma, Russia) and Ciprofloxacin (Elfa Laboratories, India) drugs on infected larvae.

This study included three consecutive stages; at the first stage, comparative assessment of the virulence of reference strains of Staphylococcus aureus, Streptococcus pneumoniae, Escherichia coli and their clinical isolates was performed, and an infectious dose of bacterial suspension for injection in larvae was determined. Thus, it was found that the studied clinical isolates of St. aureus, Str. pneumoniae and E. coli were more virulent relative to the reference strains of microorganisms.

At the second stage of the study, in vitro minimum suppressive concentration (MSC) was determined for each microorganism strain, which allowed to determine the sensitivity of the strains to the studied antibiotics, namely Ciprofloxacin and Rifampicin.

At the third stage, after infecting the larvae with an infectious dose, antibiotic therapy was administered. The results obtained in the third stage confirmed the results of the second stage in vitro. The death of larvae infected with resistant strains of St. aureus and E. coli to Ciprofloxacin and Rifampicin, respectively.

This model was characterized by changes in the general state of larvae (melanization, motility, cocoon formation) and animal death depending on the concentration of microorganisms in the inoculum. The applicability of this model to assess the sensitivity of bacteria to antibiotics was demonstrated using Ciprofloxacin and Rifampicin as examples.

Conflict of interest

The authors declare no conflict of interest.

Authors contribution

J.V. Salmova — research design, experimental part, collection and systematization of material, analysis and statistical processing of data, writing the text of the article.
L.R. Nikiforova — performing the experimental part, collecting and systematizing the material.
K.E. Borovkova — research concept, editing the text of the article.

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Received: 2022-05-23
Reviewed: 2022-06-21
Accepted for publication: 2022-09-01

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