Galleria mellonella as an infection model: an in-depth look at why it works and practical considerations for successful application

Pereira, Monalessa Fábia; Rossi, Ciro César; Silva, Giarlã Cunha da; Rosa, Jéssica Nogueira; Bazzolli, Denise Mara Soares

doi:10.1093/femspd/ftaa056

Cited by 68 publications

(69 citation statements)

References 114 publications

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“…There are no ethical constraints and their short life span makes them ideal for large-scale studies. While insects do not possess an adaptive immune response, their innate immune response is quite similar to mammals 67 . Insects possess Toll-like receptors, CAMPs, and neutrophil-like cells called hemocytes 68 .…”

Section: Discussionmentioning

confidence: 99%

A Klebsiella pneumoniae DedA family membrane protein is required for colistin resistance and for virulence in wax moth larvae

Tiwari

Panta

Billiot

et al. 2021

Sci Rep

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Ineffectiveness of carbapenems against multidrug resistant pathogens led to the increased use of colistin (polymyxin E) as a last resort antibiotic. A gene belonging to the DedA family encoding conserved membrane proteins was previously identified by screening a transposon library of K. pneumoniae ST258 for sensitivity to colistin. We have renamed this gene dkcA (dedA of Klebsiella required for colistin resistance). DedA family proteins are likely membrane transporters required for viability of Escherichia coli and Burkholderia spp. at alkaline pH and for resistance to colistin in a number of bacterial species. Colistin resistance is often conferred via modification of the lipid A component of bacterial lipopolysaccharide with aminoarabinose (Ara4N) and/or phosphoethanolamine. Mass spectrometry analysis of lipid A of the ∆dkcA mutant shows a near absence of Ara4N in the lipid A, suggesting a requirement for DkcA for lipid A modification with Ara4N. Mutation of K. pneumoniae dkcA resulted in a reduction of the colistin minimal inhibitory concentration to approximately what is found with a ΔarnT strain. We also identify a requirement of DkcA for colistin resistance that is independent of lipid A modification, instead requiring maintenance of optimal membrane potential. K. pneumoniae ΔdkcA displays reduced virulence in Galleria mellonella suggesting colistin sensitivity can cause loss of virulence.

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Section: Discussionmentioning

confidence: 99%

A Klebsiella pneumoniae DedA family membrane protein is required for colistin resistance and for virulence in wax moth larvae

Tiwari

Panta

Billiot

et al. 2021

Sci Rep

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“…Although Galleria is a pest of honeybee ( Apis mellifera ) hives worldwide [14], in recent years it has gained popularity as a model host for a range of human pathogens. It has the advantages of being inexpensive, easy to use, and able to grow at 37 °C, while not being subject to the same regulations and ethical concerns as mammalian models such as mice [15–17]. It has also been of interest due to the ability of the larvae to metabolize polyethylene [18].…”

Section: Introductionmentioning

confidence: 99%

Enterococcus innesii sp. nov., isolated from the wax moth Galleria mellonella

Gooch

Kiu

Rudder

et al. 2021

International Journal of Systematic and Evolutionary Microbiology

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Four bacterial strains were isolated from two different colony sources of the wax moth Galleria mellonella. They were characterized by a polyphasic approach including 16S rRNA gene sequence analysis, core-genome analysis, average nucleotide identity (ANI) analysis, digital DNA–DNA hybridization (dDDH), determination of G+C content, screening of antibiotic resistance genes, and various phenotypic analyses. Initial analysis of 16S rRNA gene sequence identities indicated that strain GAL7T was potentially very closely related to Enterococcus casseliflavus and Enterococcus gallinarum , having 99.5–99.9 % sequence similarity. However, further analysis of whole genome sequences revealed a genome size of 3.69 Mb, DNA G+C content of 42.35 mol%, and low dDDH and ANI values between the genomes of strain GAL7T and closest phylogenetic relative E. casseliflavus NBRC 100478T of 59.0 and 94.5 %, respectively, indicating identification of a putative new Enterococcus species. In addition, all novel strains encoded the atypical vancomycin-resistance gene vanC-4. Results of phylogenomic, physiological and phenotypic characterization confirmed that strain GAL7T represented a novel species within the genus Enterococcus , for which the name Enterococcus innesii sp. nov. is proposed. The type strain is GAL7T (=DSM 112306T=NCTC 14608T).

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“…Despite the increasing popularity of G. mellonella for general screening of microbes, toxins, and food additives (Champion et al 2016;Maguire et al 2016;Pereira et al 2020;Emery et al 2021), most experimental approaches inoculate directly into the body cavity (intrahaemocoelic injection)-circumventing natural routes of exposure for the GI tract-and relatively few studies monitor the microbiome. There is consensus, however, that enterococci are the most abundant and prevalent bacterial resident in the gut (Fig.…”

Section: Discussionmentioning

confidence: 99%

The diarrhetic shellfish-poisoning toxin, okadaic acid, provokes gastropathy, dysbiosis and susceptibility to bacterial infection in a non-rodent bioassay, Galleria mellonella

et al. 2021

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Diarrhetic shellfish-poisoning (DSP) toxins such as okadaic acid and dinophysistoxins harm the human gastrointestinal tract, and therefore, their levels are regulated to an upper limit of 160 μg per kg tissue to protect consumers. Rodents are used routinely for risk assessment and studies concerning mechanisms of toxicity, but there is a general move toward reducing and replacing vertebrates for these bioassays. We have adopted insect larvae of the wax moth Galleria mellonella as a surrogate toxicology model. We treated larvae with environmentally relevant doses of okadaic acid (80–400 μg/kg) via intrahaemocoelic injection or gavage to determine marine toxin-related health decline: (1) whether pre-exposure to a sub-lethal dose of toxin (80 μg/kg) enhances susceptibility to bacterial infection, or (2) alters tissue pathology and bacterial community (microbiome) composition of the midgut. A sub-lethal dose of okadaic acid (80 μg/kg) followed 24 h later by bacterial inoculation (2 × 105Escherichia coli) reduced larval survival levels to 47%, when compared to toxin (90%) or microbial challenge (73%) alone. Histological analysis of the midgut depicted varying levels of tissue disruption, including nuclear aberrations associated with cell death (karyorrhexis, pyknosis), loss of organ architecture, and gross epithelial displacement into the lumen. Moreover, okadaic acid presence in the midgut coincided with a shift in the resident bacterial population over time in that substantial reductions in diversity (Shannon) and richness (Chao-1) indices were observed at 240 μg toxin per kg. Okadaic acid-induced deterioration of the insect alimentary canal resembles those changes reported for rodent bioassays.

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Galleria mellonella as an infection model: an in-depth look at why it works and practical considerations for successful application

Cited by 68 publications

References 114 publications

A Klebsiella pneumoniae DedA family membrane protein is required for colistin resistance and for virulence in wax moth larvae

A Klebsiella pneumoniae DedA family membrane protein is required for colistin resistance and for virulence in wax moth larvae

Enterococcus innesii sp. nov., isolated from the wax moth Galleria mellonella

The diarrhetic shellfish-poisoning toxin, okadaic acid, provokes gastropathy, dysbiosis and susceptibility to bacterial infection in a non-rodent bioassay, Galleria mellonella

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