Development and evaluation of the Galleria mellonella (greater wax moth) infection model to study Brucella host-pathogen interaction

Elizalde-Bielsa, Aitor; Aragón-Aranda, Beatriz; Loperena-Barber, Maite; Salvador-Bescós, Miriam; Moriyón, Ignacio; Zúñiga-Ripa, Amaia; Conde-Álvarez, Raquel

doi:10.1016/j.micpath.2022.105930

Cited by 8 publications

(6 citation statements)

References 48 publications

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“…Therefore, the larval model might be a good substitute for the early evaluation of potential virulence genes when using a mammalian model, which might not be morally or practically appropriate. While preparing this article, a study was published by Aitor et al, which focused on utilizing the larval model to screen for potential Brucella factors modulating innate immunity and yielded similar findings to our own [ 46 ].…”

Section: Discussionsupporting

confidence: 67%

A novel Galleria mellonella experimental model for zoonotic pathogen Brucella

Wang,

Yin,

Zai

et al. 2023

Virulence

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Brucellosis is a major threat to public health and animal husbandry. Several in vivo vertebrate models, such as mice, guinea pigs, and nonhuman primates, have been used to study Brucella pathogenesis, bacteria-host interactions, and vaccine efficacy. However, these models have limitations whereas the invertebrate Galleria mellonella model is a cost-effective and ethical alternative. The aim of the present study was to examine the invertebrate G. mellonella as an in vivo infection model for Brucella . Infection assays were employed to validate the fitness of the larval model for Brucella infection and virulence evaluation. The protective efficacy of immune sera was evaluated by pre-incubated with a lethal dose of bacteria before infection. The consistency between the mouse model and the larval model was confirmed by assessing the protective efficacy of two Brucella vaccine strains. The results show that G. mellonella could be infected by Brucella strains, in a dose- and temperature-dependent way. Moreover, this larval model can effectively evaluate the virulence of Brucella strains in a manner consistent with that of mammalian infection models. Importantly, this model can assess the protective efficacy of vaccine immune sera within a day. Further investigation implied that haemolymph played a crucial role in the protective efficacy of immune sera. In conclusion, G. mellonella could serve as a quick, efficient, and reliable model for evaluating the virulence of Brucella strains and efficacy of immune sera in an ethical manner.

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Section: Discussionsupporting

confidence: 67%

A novel Galleria mellonella experimental model for zoonotic pathogen Brucella

Wang,

Yin,

Zai

et al. 2023

Virulence

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“…Apx toxins (ApxI and ApxII are produced by MIDG2331) are unlikely mediators, as culture supernatants did not induce the melanization or killing of G. mellonella larvae (Pereira et al, 2015), and Apx toxins have a tropism for porcine cells (Kuhnert et al, 2003). Culture supernatants would also have contained EVs and their constituent LPS, which in other Gram-negatives has proved to be a potent inducer of the G. mellonella immune response, while triggering an early-melanization response (Wu et al, 2015;Elizalde-Bielsa et al, 2023). Further work is Gene organization and relative abundance of novel sRNA candidates from EVs and cognate whole cells of A. pleuropneumoniae.…”

Section: Discussionmentioning

confidence: 99%

Identification of novel small RNAs in extracellular vesicles produced by Actinobacillus pleuropneumoniae

da Silva,

Rosa,

Fontes

et al. 2023

Front. Microbiol.

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Extracellular vesicle (EV) production by bacteria is an important mechanism for microbial communication and host-pathogen interaction. EVs of some bacterial species have been reported to contain nucleic acids. However, the role of small RNAs (sRNAs) packaged in EVs is poorly understood. Here, we report on the RNA cargo of EVs produced by the pig pathogen Actinobacillus pleuropneumoniae, the causal agent of porcine pleuropneumonia, a disease which causes substantial economic losses to the swine industry worldwide. The EVs produced by aerobically and anaerobically grown bacteria were only slightly different in size and distribution. Total cell and outer membrane protein profiles and lipid composition of A. pleuropneumoniae whole cell extracts and EVs were similar, although EVs contained rough lipopolysaccharide compared to the smooth form in whole cells. Approximately 50% of Galleria mellonella larvae died after the injection of EVs. RNAseq, RT-PCR, protection from nuclease degradation, and database searching identified previously described and 13 novel A. pleuropneumoniae sRNAs in EVs, some of which were enriched compared to whole cell content. We conclude that A. pleuropneumoniae EVs contain sRNAs, including those known to be involved in virulence, and some with homologs in other Pasteurellaceae and/or non-Pasteurellaceae. Further work will establish whether the novel sRNAs in A. pleuropneumoniae EVs play any role in pathogenesis.

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“…All of these invertebrate models share common innate immune system features with vertebrates, including tolllike receptors, microbial killing pathways, C-lectins, and apoptotic pathways [67,[71][72][73][74][75]. Additionally, invertebrate models are highly economical, commercially available, and allow for a robust sample size, allowing researchers to conduct high throughput screening of mutant libraries, strains, and clinical isolates [71,73,[76][77][78][79][80][81][82]. In a recent review [83], 72% of the bacterial species studied were Gram-negatives, including species in the genera Pseudomonas, Acinetobacter, Klebsiella, Escherichia, Burkholderia, and Campylobacter; Gram-positive bacteria comprised 26% of the studied genera, including Staphylococcus, Enterococcus, Bacillus, Listeria, and Streptococcus.…”

Section: Introductionmentioning

confidence: 99%

Galleria mellonella Invertebrate Model Mirrors the Pathogenic Potential of Mycoplasma alligatoris within the Natural Host

Burne,

Richey,

Schoeb

et al. 2024

Transboundary and Emerging Diseases

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Most mycoplasmal infections result in chronic, clinically silent disease. In direct contrast, Mycoplasma alligatoris elicits a fulminant, multisystem disease in the natural host, Alligator mississippiensis (American alligator). The goals of the study were to better understand the disease in the natural host and to determine if the invertebrate model G. mellonella could serve as a surrogate alternate host. The survival of alligators infected intratracheally was dose dependent (p=0.0003), ranging from no mortality (102 CFU) to 100% mortality (108 CFU), with 60% mortality at the 104 and 105 CFU infectious dose. Microbial load in blood, joints, and brain was dose dependent, regardless of whether alligators were infected intratracheally or intravenously (p <0.002). Weight loss was similarly impacted (p <0.001). Experimental infection of the invertebrate Galleria mellonella mirrored the result in the natural host. In a dose response infection study, both larval survival curves and successful pupation curves were significantly different (p≤0.0001) and dose dependent. Infected insects did not emerge as moths (p <0.0001). Here, we describe the first study investigating G. mellonella as a surrogate model to assess the pathogenic potential of M. alligatoris. G. mellonella survival was dose dependent and impacted life stage outcome.

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Development and evaluation of the Galleria mellonella (greater wax moth) infection model to study Brucella host-pathogen interaction

Cited by 8 publications

References 48 publications

A novel Galleria mellonella experimental model for zoonotic pathogen Brucella

A novel Galleria mellonella experimental model for zoonotic pathogen Brucella

Identification of novel small RNAs in extracellular vesicles produced by Actinobacillus pleuropneumoniae

Galleria mellonella Invertebrate Model Mirrors the Pathogenic Potential of Mycoplasma alligatoris within the Natural Host

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