Galleria mellonella as an experimental model to study human oral pathogens

Rossoni, Rodnei Dennis; Ribeiro, Felipe de Camargo; Santos, Hanna Flávia Santana dos; Santos, Jéssica Diane dos; Oliveira, Nicássia de Sousa; Dutra, Marignês Theotonio dos Santos; Lapena, Simone Aparecida Biazzi de; Junqueira, Juliana Campos

doi:10.1016/j.archoralbio.2019.03.002

Cited by 27 publications

(18 citation statements)

References 128 publications

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“…The use of the invertebrate Galleria mellonella has become an alternative experimental in vivo infection model to traditional mammals. The larvae are easy to maintain and manipulate, and have similarities with the mammalian innate immune system, including the cellular immune response performed by hemocytes present in the insect's hemolymph …”

Section: Introductionmentioning

confidence: 99%

“…The larvae are easy to maintain and manipulate, and have similarities with the mammalian innate immune system, including the cellular immune response performed by hemocytes present in the insect's hemolymph. 23,24 Treating oral diseases with a natural and non-invasive method has great appeal and can prevent problems related to pharmacological treatments, such as antibiotic resistance. Considering that conventional periodontal treatment fails to control periodontal disease progression in some patients and that L reuteri has been successfully used as an adjunct alternative in some clinical studies, we have hypothesized that heat-killed L reuteri (paraprobiotic) and L reuteri cell-free supernatant may be effective in cases of P gingivalis infection.…”

Section: Introductionmentioning

confidence: 99%

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Heat‐killed Lactobacillus reuteri and cell‐free culture supernatant have similar effects to viable probiotics during interaction with Porphyromonas gingivalis

Geraldo

Batalha

Milhan

et al. 2019

J of Periodontal Research

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Background and objective In the last decade, numerous studies have been published to clarify the role of probiotics, especially Lactobacillus reuteri, as an adjunct to conventional periodontal treatment. Although the health benefits of probiotics are numerous, they are live bacteria, and the administration of live organisms is not risk‐free. We evaluated the antimicrobial effect of L reuteri and its cell‐free culture supernatant on Porphyromonas gingivalis, a keystone periodontal pathogen, in vitro. We also evaluated the influence of this probiotic in its live, heat‐killed (HKL, paraprobiotic) form and its supernatant on the Galleria mellonella invertebrate model after infection by P gingivalis. Methods The interaction assay was conducted with P gingivalis and L reuteri preparations (live cells and supernatant preparation). For this, P gingivalis and L reuteri preparations were added to tubes containing Brain Heart Infusion broth and incubated for 3 days. The suspensions were then seeded onto appropriate culture media for the calculation of colony‐forming units per mL (CFU/mL). An in vivo assay with the G mellonella model was also performed. Live L reuteri, HKL, or supernatant was inoculated 2 hours prior to infection with P gingivalis. Survival was evaluated over 7 days, and the number of hemocytes in the hemolymph was estimated 3 hours after P gingivalis infection. Data were then subjected to statistical testing (α = 5%). Results Both live L reuteri and its supernatant had antimicrobial activity against P gingivalis (CFU reduction up to 86%, P < .05). Moreover, treatment with live and HKL had similar effects on G mellonella survival (increased survival up to 46%, P < .05). However, only live L reuteri was able to significantly increase the hemocyte density in this invertebrate model. Conclusion Lactobacillus reuteri antimicrobial activity against P gingivalis and its effects on G mellonella survival after infection with a periodontopathogen do not depend on cell viability. This allows the development of products without live bacterium while maintaining similar effects.

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Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Heat‐killed Lactobacillus reuteri and cell‐free culture supernatant have similar effects to viable probiotics during interaction with Porphyromonas gingivalis

Geraldo

Batalha

Milhan

et al. 2019

J of Periodontal Research

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“…mutans extracts using two different in vivo models. The G. mellonella model is a prompt and reliable method to evaluate toxicity and efficacy of antifungal agents ( Junqueira and Mylonakis, 2019 ; Lin et al, 2019 ; Rossoni et al, 2019 ). The S. mutans extract tested concentrations were not toxic to the host.…”

Section: Discussionmentioning

confidence: 99%

Streptococcus mutans Secreted Products Inhibit Candida albicans Induced Oral Candidiasis

et al. 2020

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In the oral cavity, Candida species form mixed biofilms with Streptococcus mutans, a pathogenic bacterium that can secrete quorum sensing molecules with antifungal activity. In this study, we extracted and fractioned culture filtrate of S. mutans, seeking antifungal agents capable of inhibiting the biofilms, filamentation, and candidiasis by Candida albicans. Active S. mutans UA159 supernatant filtrate components were extracted via liquid-liquid partition and fractionated on a C-18 silica column to resolve S. mutans fraction 1 (SM-F1) and fraction 2 (SM-F2). We found anti-biofilm activity for both SM-F1 and SM-F2 in a dose dependent manner and fungal growth was reduced by 2.59 and 5.98 log for SM-F1 and SM-F2, respectively. The SM-F1 and SM-F2 fractions were also capable of reducing C. albicans filamentation, however statistically significant differences were only observed for the SM-F2 (p = 0.004). SM-F2 efficacy to inhibit C. albicans was confirmed by its capacity to downregulate filamentation genes CPH1, EFG1, HWP1, and UME6. Using Galleria mellonella as an invertebrate infection model, therapeutic treatment with SM-F2 prolonged larvae survival. Examination of the antifungal capacity was extended to a murine model of oral candidiasis that exhibited a reduction in C. albicans colonization (CFU/mL) in the oral cavity when treated with SM-F1 (2.46 log) and SM-F2 (2.34 log) compared to the control (3.25 log). Although both SM-F1 and SM-F2 fractions decreased candidiasis in mice, only SM-F2 exhibited significant quantitative differences compared to the non-treated group for macroscopic lesions, hyphae invasion, tissue lesions, and inflammatory infiltrate. Taken together, these results indicate that the SM-F2 fraction contains antifungal components, providing a promising resource in the discovery of new inhibitors for oral candidiasis.

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“…Waxworms are captive-bred worldwide as food for amphibians, reptiles, and fish, and are easy to purchase and maintain (Table 1). For injection of waxworms, the worms are anesthetized on ice and injected using a syringe with a 30-gauge needle, 18,19 or held in a special apparatus and injected. 20 Because waxworms can grow at 37°C, 21 infection experiments can be performed at 37°C.…”

Section: Waxworm (Galleria Mellonella)mentioning

confidence: 99%

Animal infection models using non‐mammals

Kaito

Murakami

Imai

et al. 2020

Microbiology and Immunology

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The use of non‐human animal models for infection experiments is important for investigating the infectious processes of human pathogenic bacteria at the molecular level. Mammals, such as mice and rabbits, are also utilized as animal infection models, but large numbers of animals are needed for these experiments, which is costly, and fraught with ethical issues. Various non‐mammalian animal infection models have been used to investigate the molecular mechanisms of various human pathogenic bacteria, including Staphylococcus aureus, Streptococcus pyogenes, and Pseudomonas aeruginosa. This review discusses the desirable characteristics of non‐mammalian infection models and describes recent non‐mammalian infection models that utilize Caenorhabditis elegans, silkworm, fruit fly, zebrafish, two‐spotted cricket, hornworm, and waxworm.

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Galleria mellonella as an experimental model to study human oral pathogens

Cited by 27 publications

References 128 publications

Heat‐killed Lactobacillus reuteri and cell‐free culture supernatant have similar effects to viable probiotics during interaction with Porphyromonas gingivalis

Heat‐killed Lactobacillus reuteri and cell‐free culture supernatant have similar effects to viable probiotics during interaction with Porphyromonas gingivalis

Streptococcus mutans Secreted Products Inhibit Candida albicans Induced Oral Candidiasis

Animal infection models using non‐mammals

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