The efficacy of intraosseous catheterization has not been described previously in the desert tortoise (Gopherus agassizii). The goal of this study was to describe and compare the efficacy of four intraosseous catheter sites (humerus, femur, plastocarapacial junction [bridge], and gular region of the plastron) to jugular catheterization. Five adult tortoises were catheterized in each of the sites at least once. The distribution of a bolus injection of radiopharmaceutical (technetium-99m–diethylenetriaminepentaacidic acid [^99mTc -DTPA]) was monitored via gamma camera over 2-min periods at five time intervals over 24 min. Compared to jugular catheterization, the humerus and femur sites provided the next best vascular access, with 84.4 and 61.8% of activity reaching the systemic circulation by 7 min, respectively. The bridge and gular catheter sites were less effective with only 41.9 and 40.8% systemic activity, respectively. Intraosseous catheters were no more technically difficult to place than jugular catheters and were less commonly dislodged, making them a viable option for vascular access in tortoises.