{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,26]],"date-time":"2026-09-26T00:16:25Z","timestamp":1790381785279,"version":"4.1.0"},"reference-count":45,"publisher":"MDPI AG","issue":"2","license":[{"start":{"date-parts":[[2024,2,5]],"date-time":"2024-02-05T00:00:00Z","timestamp":1707091200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"National Institute of Allergy and Infectious Diseases","award":["R21 AI149015"],"award-info":[{"award-number":["R21 AI149015"]}]},{"name":"National Institute of Allergy and Infectious Diseases","award":["R01 AI135005"],"award-info":[{"award-number":["R01 AI135005"]}]},{"name":"National Institute of Allergy and Infectious Diseases","award":["U19AI166761 357"],"award-info":[{"award-number":["U19AI166761 357"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["JoF"],"abstract":"<jats:p>Coccidioidomycosis (CM) can manifest as respiratory and disseminated diseases that are caused by dimorphic fungal pathogens, such as Coccidioides species. The inhaled arthroconidia generated during the saprobic growth phase convert into multinucleated spherules in the lungs to complete the parasitic lifecycle. Research on coccidioidal virulence and pathogenesis primarily employs murine models typically associated with low lethal doses (LD100 &lt; 100 spores). However, the Galleria model has recently garnered attention due to its immune system bearing both structural and functional similarities to the innate system of mammals. Our findings indicate that Coccidioides posadasii can convert and complete the parasitic cycle within the hemocoel of the Galleria larva. In Galleria, the LD100 is between 0.5 and 1.0 \u00d7 106 viable spores for the clinical isolate Coccidioides posadasii C735. Furthermore, we demonstrated the suitability of this model for in vivo antifungal susceptibility tests to validate the bioreactivity of newly discovered antifungals against Coccidioides. Additionally, we utilized this larva model to screen a Coccidioides posadasii mutant library showing attenuated virulence. Similarly, the identified attenuated coccidioidal mutants displayed a loss of virulence in a commonly used murine model of coccidioidomycosis. In this study, we demonstrated that Galleria larvae can be applied as a model for studying Coccidioides infection.<\/jats:p>","DOI":"10.3390\/jof10020131","type":"journal-article","created":{"date-parts":[[2024,2,5]],"date-time":"2024-02-05T05:22:44Z","timestamp":1707110564000},"page":"131","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":7,"title":["Galleria\u00a0mellonella Model of Coccidioidomycosis for Drug Susceptibility Tests and Virulence Factor Identification"],"prefix":"10.3390","volume":"10","author":[{"given":"Matthew","family":"Mendoza Barker","sequence":"first","affiliation":[{"name":"Department of Molecular Microbiology and Immunology, South Texas Center for Emerging Infectious Diseases, The University of Texas at San Antonio, San Antonio, TX 78249, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sarah","family":"Saeger","sequence":"additional","affiliation":[{"name":"Department of Molecular Microbiology and Immunology, South Texas Center for Emerging Infectious Diseases, The University of Texas at San Antonio, San Antonio, TX 78249, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Althea","family":"Campuzano","sequence":"additional","affiliation":[{"name":"Department of Molecular Microbiology and Immunology, South Texas Center for Emerging Infectious Diseases, The University of Texas at San Antonio, San Antonio, TX 78249, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8901-3366","authenticated-orcid":false,"given":"Jieh-Juen","family":"Yu","sequence":"additional","affiliation":[{"name":"Department of Molecular Microbiology and Immunology, South Texas Center for Emerging Infectious Diseases, The University of Texas at San Antonio, San Antonio, TX 78249, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1091-3420","authenticated-orcid":false,"given":"Chiung-Yu","family":"Hung","sequence":"additional","affiliation":[{"name":"Department of Molecular Microbiology and Immunology, South Texas Center for Emerging Infectious Diseases, The University of Texas at San Antonio, San Antonio, TX 78249, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2024,2,5]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"e00550-16","DOI":"10.1128\/mBio.00550-16","article-title":"Local Population Structure and Patterns of Western Hemisphere Dispersal for Coccidioides spp., the Fungal Cause of Valley Fever","volume":"7","author":"Engelthaler","year":"2016","journal-title":"mBio"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"e1","DOI":"10.1093\/cid\/ciu681","article-title":"Valley fever: Finding new places for an old disease: Coccidioides immitis found in Washington State soil associated with recent human infection","volume":"60","author":"Litvintseva","year":"2015","journal-title":"Clin. Infect. Dis."},{"key":"ref_3","unstructured":"Chiller, T. (2019). Vaccine Strategies for Endemic Fungal Pathogens, NIAID."},{"key":"ref_4","unstructured":"Einstein, H. (1996). Coccidioidomycosis, National Foundation for Infectious Disease."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"1217","DOI":"10.1086\/496991","article-title":"Coccidioidomycosis","volume":"41","author":"Galgiani","year":"2005","journal-title":"Clin. Infect. Dis."},{"key":"ref_6","first-page":"185","article-title":"Coccidioidomycosis: Epidemiology","volume":"5","author":"Brown","year":"2013","journal-title":"Clin. Epidemiol."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"372","DOI":"10.1093\/infdis\/147.3.372","article-title":"Coccidioidomycosis: Factors affecting the host-parasite interaction","volume":"147","author":"Drutz","year":"1983","journal-title":"J. Infect. Dis."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"6689","DOI":"10.1128\/IAI.73.10.6689-6703.2005","article-title":"A metalloproteinase of Coccidioides posadasii contributes to evasion of host detection","volume":"73","author":"Hung","year":"2005","journal-title":"Infect. Immun."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"3443","DOI":"10.1128\/IAI.70.7.3443-3456.2002","article-title":"A parasitic phase-specific adhesin of Coccidioides immitis contributes to the virulence of this respiratory Fungal pathogen","volume":"70","author":"Hung","year":"2002","journal-title":"Infect. Immun."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"4681","DOI":"10.1016\/j.vaccine.2012.04.084","article-title":"An agonist of human complement fragment C5a enhances vaccine immunity against Coccidioides infection","volume":"30","author":"Hung","year":"2012","journal-title":"Vaccine"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"3007","DOI":"10.1128\/IAI.00633-16","article-title":"A Coccidioides posadasii CPS1 Deletion Mutant Is Avirulent and Protects Mice from Lethal Infection","volume":"84","author":"Narra","year":"2016","journal-title":"Infect. Immun."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"5830","DOI":"10.1128\/IAI.68.10.5830-5838.2000","article-title":"Disruption of the gene which encodes a serodiagnostic antigen and chitinase of the human fungal pathogen Coccidioides immitis","volume":"68","author":"Reichard","year":"2000","journal-title":"Infect. Immun."},{"key":"ref_13","doi-asserted-by":"crossref","unstructured":"Mandel, M.A., Beyhan, S., Voorhies, M., Shubitz, L.F., Galgiani, J.N., Orbach, M.J., and Sil, A. (2022). The WOPR family protein Ryp1 is a key regulator of gene expression, development, and virulence in the thermally dimorphic fungal pathogen Coccidioides posadasii. PLoS Pathog., 18.","DOI":"10.1371\/journal.ppat.1009832"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"2188","DOI":"10.3389\/fimmu.2019.02188","article-title":"The Rise of Coccidioides: Forces Against the Dust Devil Unleashed","volume":"10","author":"Thompson","year":"2019","journal-title":"Front. Immunol."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"e02634-16","DOI":"10.1128\/AAC.02634-16","article-title":"Large-Scale Evaluation of In Vitro Amphotericin B, Triazole, and Echinocandin Activity against Coccidioides Species from U.S. Institutions","volume":"61","author":"Thompson","year":"2017","journal-title":"Antimicrob. Agents Chemother."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"912","DOI":"10.1128\/iai.40.3.912-916.1983","article-title":"Inbred mouse strains differ in resistance to lethal Coccidioides immitis infection","volume":"40","author":"Kirkland","year":"1983","journal-title":"Infect. Immun."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"S85","DOI":"10.1093\/mmy\/myy146","article-title":"A review of innate and adaptive immunity to coccidioidomycosis","volume":"57","author":"Hung","year":"2019","journal-title":"Med. Mycol."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"208","DOI":"10.1196\/annals.1406.029","article-title":"Experimental animal models of coccidioidomycosis","volume":"1111","author":"Clemons","year":"2007","journal-title":"Ann. N. Y. Acad. Sci."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"e00070-18","DOI":"10.1128\/IAI.00070-18","article-title":"Glucan-Chitin Particles Enhance Th17 Response and Improve Protective Efficacy of a Multivalent Antigen (rCpa1) against Pulmonary Coccidioides posadasii Infection","volume":"86","author":"Hung","year":"2018","journal-title":"Infect. Immun."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"ftaa056","DOI":"10.1093\/femspd\/ftaa056","article-title":"Galleria mellonella as an infection model: An in-depth look at why it works and practical considerations for successful application","volume":"78","author":"Pereira","year":"2020","journal-title":"Pathog. Dis."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"469","DOI":"10.1007\/978-1-61779-539-8_33","article-title":"Galleria mellonella as a model for fungal pathogenicity testing","volume":"845","author":"Fallon","year":"2012","journal-title":"Methods Mol. Biol."},{"key":"ref_22","doi-asserted-by":"crossref","unstructured":"Kavanagh, K., and Sheehan, G. (2018). The Use of Galleria mellonella Larvae to Identify Novel Antimicrobial Agents against Fungal Species of Medical Interest. J. Fungi, 4.","DOI":"10.3390\/jof4030113"},{"key":"ref_23","doi-asserted-by":"crossref","unstructured":"Wright, C.L., and Kavanagh, O. (2022). Galleria mellonella as a novel in vivo model to screen natural product-derived modulators of innate immunity. Appl. Sci., 12.","DOI":"10.3390\/app12136587"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"488","DOI":"10.1016\/j.micinf.2010.03.001","article-title":"Role of filamentation in Galleria mellonella killing by Candida albicans","volume":"12","author":"Fuchs","year":"2010","journal-title":"Microbes Infect."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"ftaa067","DOI":"10.1093\/femspd\/ftaa067","article-title":"The Galleria mellonella infection model as a system to investigate the virulence of Candida auris strains","volume":"78","author":"Romera","year":"2020","journal-title":"Pathog. Dis."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"818","DOI":"10.1080\/21505594.2021.1893945","article-title":"Galleria mellonella as a screening tool to study virulence factors of Aspergillus fumigatus","volume":"12","author":"Durieux","year":"2021","journal-title":"Virulence"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"351","DOI":"10.1093\/mmy\/myy042","article-title":"Galleria mellonella as a model system to study virulence potential of mucormycetes and evaluation of antifungal treatment","volume":"57","author":"Maurer","year":"2019","journal-title":"Med. Mycol."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"139","DOI":"10.4161\/viru.23047","article-title":"Galleria mellonella as a model host to study Paracoccidioides lutzii and Histoplasma capsulatum","volume":"4","author":"Thomaz","year":"2013","journal-title":"Virulence"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"3842","DOI":"10.1128\/IAI.73.7.3842-3850.2005","article-title":"Galleria mellonella as a model system to study Cryptococcus neoformans pathogenesis","volume":"73","author":"Mylonakis","year":"2005","journal-title":"Infect. Immun."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"4511","DOI":"10.1128\/IAI.05726-11","article-title":"Vaccine immunity to coccidioidomycosis occurs by early activation of three signal pathways of T helper cell response (Th1, Th2, and Th17)","volume":"79","author":"Hung","year":"2011","journal-title":"Infect. Immun."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"970","DOI":"10.1111\/j.1365-2958.2006.05436.x","article-title":"An alpha-(1,4)-amylase is essential for alpha-(1,3)-glucan production and virulence in Histoplasma capsulatum","volume":"62","author":"Marion","year":"2006","journal-title":"Mol. Microbiol."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"3206","DOI":"10.1002\/j.1460-2075.1995.tb07323.x","article-title":"Trans-kingdom T-DNA transfer from Agrobacterium tumefaciens to Saccharomyces cerevisiae","volume":"14","author":"Bundock","year":"1995","journal-title":"EMBO J."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"1575","DOI":"10.1242\/jeb.01589","article-title":"The origin of allometric scaling laws in biology from genomes to ecosystems: Towards a quantitative unifying theory of biological structure and organization","volume":"208","author":"West","year":"2005","journal-title":"J. Exp. Biol."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"235","DOI":"10.3109\/03602538708998307","article-title":"The metabolism and fate of closantel (Flukiver) in sheep and cattle","volume":"18","author":"Michiels","year":"1987","journal-title":"Drug Metab. Rev."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"208","DOI":"10.1111\/jvp.12835","article-title":"Sanguinarine metabolism and pharmacokinetics study in vitro and in vivo","volume":"43","author":"Wu","year":"2020","journal-title":"J. Vet. Pharmacol. Ther."},{"key":"ref_36","doi-asserted-by":"crossref","unstructured":"McMahon, C.L., Esqueda, M., Yu, J.J., Wall, G., Romo, J.A., Vila, T., Chaturvedi, A., Lopez-Ribot, J.L., Wormley, F., and Hung, C.Y. (2023). Development of an Imaging Flow Cytometry Method for Fungal Cytological Profiling and Its Potential Application in Antifungal Drug Development. J. Fungi, 9.","DOI":"10.3390\/jof9070722"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"1166","DOI":"10.1128\/IAI.01066-15","article-title":"Card9- and MyD88-Mediated Gamma Interferon and Nitric Oxide Production Is Essential for Resistance to Subcutaneous Coccidioides posadasii Infection","volume":"84","author":"Hung","year":"2016","journal-title":"Infect. Immun."},{"key":"ref_38","doi-asserted-by":"crossref","unstructured":"Munoz-Hernandez, B., Palma-Cortes, G., Cabello-Gutierrez, C., and Martinez-Rivera, M.A. (2014). Parasitic polymorphism of Coccidioides spp. BMC Infect. Dis., 14.","DOI":"10.1186\/1471-2334-14-213"},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"342","DOI":"10.1111\/1744-7917.12325","article-title":"Immunity of the greater wax moth Galleria mellonella","volume":"24","author":"Wojda","year":"2017","journal-title":"Insect Sci."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"ftab013","DOI":"10.1093\/femspd\/ftab013","article-title":"Fungal immunity and pathogenesis in mammals versus the invertebrate model organism Galleria mellonella","volume":"79","author":"Smith","year":"2021","journal-title":"Pathog. Dis."},{"key":"ref_41","doi-asserted-by":"crossref","unstructured":"Campuzano, A., Pentakota, K.D., Liao, Y.R., Zhang, H., Wiederhold, N.P., Ostroff, G.R., and Hung, C.Y. (2024). A Recombinant Multivalent Vaccine (rCpa1) Induces Protection for C57BL\/6 and HLA Transgenic Mice against Pulmonary Infection with Both Species of Coccidioides. Vaccines, 12.","DOI":"10.3390\/vaccines12010067"},{"key":"ref_42","doi-asserted-by":"crossref","unstructured":"Garcia, J.A., Vu, K., Thompson, G.R., and Gelli, A. (2022). Characterization of the Growth and Morphology of a BSL-2 Coccidioides posadasii Strain That Persists in the Parasitic Life Cycle at Ambient CO2. J. Fungi, 8.","DOI":"10.1101\/2022.03.29.486294"},{"key":"ref_43","doi-asserted-by":"crossref","unstructured":"Jemel, S., Guillot, J., Kallel, K., Botterel, F., and Dannaoui, E. (2020). Galleria mellonella for the Evaluation of Antifungal Efficacy against Medically Important Fungi, a Narrative Review. Microorganisms, 8.","DOI":"10.3390\/microorganisms8030390"},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"107327","DOI":"10.1016\/j.jip.2020.107327","article-title":"Immune response of Galleria mellonella after injection with non-lethal and lethal dosages of Candida albicans","volume":"170","author":"Vertyporokh","year":"2020","journal-title":"J. Invertebr. Pathol."},{"key":"ref_45","doi-asserted-by":"crossref","unstructured":"Cruz, L.I.B., Lopes, L.F.F., de Camargo Ribeiro, F., de Sa, N.P., Lino, C.I., Tharmalingam, N., de Oliveira, R.B., Rosa, C.A., Mylonakis, E., and Fuchs, B.B. (2018). Anti-Candida albicans Activity of Thiazolylhydrazone Derivatives in Invertebrate and Murine Models. J. Fungi, 4.","DOI":"10.3390\/jof4040134"}],"container-title":["Journal of Fungi"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2309-608X\/10\/2\/131\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T13:55:09Z","timestamp":1760104509000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2309-608X\/10\/2\/131"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2024,2,5]]},"references-count":45,"journal-issue":{"issue":"2","published-online":{"date-parts":[[2024,2]]}},"alternative-id":["jof10020131"],"URL":"https:\/\/doi.org\/10.3390\/jof10020131","relation":{},"ISSN":["2309-608X"],"issn-type":[{"value":"2309-608X","type":"electronic"}],"subject":[],"published":{"date-parts":[[2024,2,5]]}}}