Abstract
<jats:p> Pathogenic <jats:italic>Leptospira</jats:italic> transitions between environmental reservoirs and mammalian hosts, expose the bacterium to distinct temperature conditions. Although <jats:italic>Leptospira</jats:italic> is generally considered to rely its growth primarily on long-chain fatty acids and to have limited capacity to use glucose, whether host-like temperature alters glucose-associated metabolic capacity remains unclear. Here, we used transcriptome-integrated genome-scale metabolic modeling to examine temperature-dependent metabolic states in pathogenic <jats:italic>Leptospira</jats:italic> . Models contextualized with transcriptomic data from cultures at 37°C predicted increased flux through glucose transport, glucose phosphorylation, and downstream glucose-associated reactions than it did at 29–30°C. Similar temperature-dependent predictions were obtained for another pathogenic <jats:italic>Leptospira</jats:italic> species under <jats:italic>in vitro</jats:italic> conditions. Consistent with these findings, analysis of leptospires cultivated intraperitoneally in dialysis membrane chambers also predicted enhanced glucose uptake and metabolism under host-temperature conditions. <jats:italic>In vitro</jats:italic> validation experiments showed increased bacterial uptake or accumulation of a fluorescent glucose analogue, elevated expression of candidate glucose transporter genes, and glucose-dependent enhancement of bacterial growth at 37°C. Treatment with a glucose degradation pathway inhibitor, 2-deoxy-D-glucose further supported a contribution of glucose-associated processes to proliferation at 37°C. Together, these findings indicate that pathogenic <jats:italic>Leptospira</jats:italic> displays condition-dependent glucose uptake and glucose-associated metabolic activity that become apparent at host-like temperature, opposed to glucose-incompetent in an environmental-like temperature. This prediction-driven framework refines the conventional view of <jats:italic>Leptospira</jats:italic> carbon metabolism and provides a basis for future studies of glucose-associated metabolic capacity during mammalian infection. </jats:p>