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The phagosome compartment is a nutrient-limited environment, requiring<jats:italic>Histoplasma<\/jats:italic>yeasts to be able to assimilate available carbon sources within the phagosome to meet their nutritional needs. In this study, we showed that<jats:italic>Histoplasma<\/jats:italic>yeasts do not utilize fatty acids or hexoses for growth within macrophages. Instead,<jats:italic>Histoplasma<\/jats:italic>yeasts consume gluconeogenic substrates to proliferate in macrophages. 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