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Two-spotted cricket as an animal infection model of human pathogenic fungi
Yuto Kochi1, Yasuhiko Matsumoto2, Kazuhisa Sekimizu2
1Laboratory of Microbiology, Graduate School of Pharmaceutical Sciences, The University of Tokyo.
Abstract:
Invertebrate infection models that can be evaluated at human body temperature are limited. In this study, we utilized the two-spotted cricket, a heat-tolerant insect, as an animal infection model of human pathogenic fungi. Injection of human pathogenic fungi, including Candida albicans, Candida glabrata, and Cryptococcus neoformans killed crickets within 48 h at both 27˚C and 37˚C. The median lethal dose values (LD50 values) of C. albicans and C. glabrata against crickets were decreased at 37˚C compared to that at 27˚C, whereas the LD50 value of C. neoformans was not different between 27˚C and 37˚C. Heat-killed cells of the three different fungi also killed crickets, but the LD50 value of the heat-killed cells was higher than 5-fold that of live fungal cells in the respective species. C. neoformans gene-knockout strains of cna1, gpa1, and pka1, which are required for virulence in mammals, had greater LD50 values than the parent strain in crickets. These findings suggest that the two-spotted cricket is a valuable infection model of human pathogenic fungi that can be used to evaluate fungal virulence at variable temperatures, including 37˚C, and that the killing abilities of C. albicans and C. glabrata against animals are increased at 37˚C.
Insights
The two-spotted cricket serves as a valuable infection model for human pathogenic fungi, enabling virulence studies at human body temperature (37°C). This model demonstrates increased fungal pathogenicity at higher temperatures, aiding in the development of new antifungal therapies.
Area of Science:
- Mycology
- Infectious Diseases
- Animal Models
Background:
- Limited invertebrate infection models exist for evaluating pathogens at human body temperature (37°C).
- The two-spotted cricket (Acheta domesticus) is a heat-tolerant insect suitable for infection studies.
Purpose of the Study:
- To establish and validate the two-spotted cricket as an infection model for human pathogenic fungi at varying temperatures.
- To assess the impact of temperature on fungal virulence and the efficacy of antifungal treatments.
Main Methods:
- Infection of crickets with human pathogenic fungi (Candida albicans, Candida glabrata, Cryptococcus neoformans) at 27°C and 37°C.
- Determination of median lethal dose (LD50) values for live and heat-killed fungal cells.
- Evaluation of Cryptococcus neoformans virulence gene knockouts in the cricket model.
Main Results:
- Human pathogenic fungi rapidly killed crickets at both 27°C and 37°C.
- Candida albicans and Candida glabrata showed increased virulence (lower LD50) at 37°C compared to 27°C.
- Cryptococcus neoformans virulence gene knockouts exhibited reduced pathogenicity in crickets.
Conclusions:
- The two-spotted cricket is a viable and versatile animal model for studying human fungal pathogens and their virulence at temperatures up to 37°C.
- Temperature significantly influences the pathogenicity of certain human fungal species, highlighting the importance of thermal factors in infection.
- This model facilitates the investigation of fungal virulence mechanisms and the screening of potential antifungal agents.
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