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Hundreds of thousands of new infections are reported annually around the world. The etiological agent of the disease,\n              <jats:italic>Histoplasma,<\/jats:italic>\n              is a dimorphic fungus commonly found in the soil where it grows as mycelia. Humans can become infected by\n              <jats:italic>Histoplasma<\/jats:italic>\n              through inhalation of its spores (conidia) or mycelial particles. The fungi transition into the yeast phase in the lungs at 37\u00b0C. Once in the lungs, yeast cells reside and proliferate inside alveolar macrophages. Genomic work has revealed that\n              <jats:italic>Histoplasma<\/jats:italic>\n              is composed of at least five cryptic phylogenetic species that differ genetically. Three of those lineages have received new names. Here, we evaluated multiple phenotypic characteristics (colony morphology, secreted proteolytic activity, yeast size, and growth rate) of strains from five of the phylogenetic species of\n              <jats:italic>Histoplasma<\/jats:italic>\n              to identify phenotypic traits that differentiate between these species:\n              <jats:italic>Histoplasma capsulatum sensu stricto<\/jats:italic>\n              ,\n              <jats:italic>Histoplasma ohiense<\/jats:italic>\n              ,\n              <jats:italic>Histoplasma mississippiense<\/jats:italic>\n              ,\n              <jats:italic>Histoplasma suramericanum<\/jats:italic>\n              , and an African lineage. We report diagnostic traits for three species. The other two species can be identified by a combination of traits. Our results suggest that (i) there are significant phenotypic differences among the cryptic species of\n              <jats:italic>Histoplasma<\/jats:italic>\n              and (ii) those differences can be used to positively distinguish those species in a clinical setting and for further study of the evolution of this fungal pathogen.\n            <\/jats:p>\n            <jats:sec>\n              <jats:title>IMPORTANCE<\/jats:title>\n              <jats:p>\n                Identifying species boundaries is a critical component of evolutionary biology. Genome sequencing and the use of molecular markers have advanced our understanding of the evolutionary history of fungal pathogens, including\n                <jats:italic>Histoplasma<\/jats:italic>\n                , and have allowed for the identification of new species. This is especially important in organisms where morphological characteristics have not been detected. In this study, we revised the taxonomic status of the four named species of the genus\n                <jats:italic>Histoplasma, H. capsulatum sensu stricto<\/jats:italic>\n                (\n                <jats:italic>ss<\/jats:italic>\n                ),\n                <jats:italic>H. ohiense<\/jats:italic>\n                ,\n                <jats:italic>H. mississippiense<\/jats:italic>\n                , and\n                <jats:italic>H. suramericanum,<\/jats:italic>\n                and propose the use of species-specific phenotypic traits to aid their identification when genome sequencing is not available. These results have implications not only for evolutionary study of\n                <jats:italic>Histoplasma<\/jats:italic>\n                but also for clinicians, as the\n                <jats:italic>Histoplasma<\/jats:italic>\n                species could determine the outcome of disease and treatment needed.\n              <\/jats:p>\n            <\/jats:sec>\n          <\/jats:sec>","DOI":"10.1128\/msphere.00009-24","type":"journal-article","created":{"date-parts":[[2024,5,21]],"date-time":"2024-05-21T13:01:48Z","timestamp":1716296508000},"update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":13,"title":["Phenotypic characterization of cryptic species in the fungal pathogen\n            <i>Histoplasma<\/i>"],"prefix":"10.1128","volume":"9","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-2085-9667","authenticated-orcid":true,"given":"Victoria E.","family":"Sep\u00falveda","sequence":"first","affiliation":[{"name":"Department of Biology, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2419-3399","authenticated-orcid":false,"given":"Jonathan A.","family":"Rader","sequence":"additional","affiliation":[{"name":"Department of Biology, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jingbaoyi (Janet)","family":"Li","sequence":"additional","affiliation":[{"name":"Department of Biology, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"William E.","family":"Goldman","sequence":"additional","affiliation":[{"name":"Department of Microbiology and Immunology, University of North Carolina at Chapel Hill, Chapel Hill, North 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