This study assessed the effects of five different transport periods on physiometabolic responses and gas exchange in ostriches. It included 138 ostriches that were assigned to five experimental groups. G1 included 78 birds that were set aside as a reference group (RG). Each one of the four remaining experimental groups included 20 ostriches, which were transported in the following manner: G1 on one occasion for a period of 1 h; G2 on one occasion for 2 h; and G3 on one occasion for 3 h; G4a was made up of the ostriches from G1 but they were shipped on a second occasion, for 2 h (making a total of 3 h); finally, G4b was that same group, but after a third transport period, on this occasion during 3 h (for a total of 6 h). Groups G2 and G3 presented the most marked blood alterations (P < 0.05), including an increase in pH, hypocapnia (27.8 ± 0.80 mmHg), hypernatremia (171.75 ± 1.84 mmol/L), hypocalcemia (0.95 ± 0.03 mmol/L), and hyperglycemia (224.05 ± 3.94 mg/dL). Also, group G3 presented the lowest hematocrit values (26.5 ± 0.47 HTC%). Therefore, transporting ostriches with no prior experience for 3 h caused the most pronounced physiometabolic changes.