Low-titer group O whole blood (LTOWB) is increasingly being used in the civilian trauma setting, although there is a risk of hemolysis. This study evaluated the impact on hemolytic markers following the transfusion of 4 or more units of uncrossmatched LTOWB. Civilian adult trauma patients who received four or more units of leukoreduced group O+, low-titer (<50 anti-A and anti-B), platelet-replete uncrossmatched whole blood during their initial resuscitation and who survived for more than 24 hours after the transfusion were included in this retrospective study. Lactate dehydrogenase (LDH), total bilirubin, haptoglobin, potassium, and creatinine were evaluated on the day of LTOWB transfusion (Day 0) and the next 3 days. Blood product administration over the first 24 hours of admission was recorded. There were 54 non-group O and 23 group O recipients of four or more LTOWB units. The median (interquartile range [IQR]) number of transfused LTOWB units was 4 (4-5) and 4 (4-4), respectively, the maximum number in both groups was eight. The non-group O patients received a median (IQR) volume of 1470 mL (1368-2052) of ABO-incompatible plasma. Comparing the non-group O to the group O recipients, there were no significant differences in the haptoglobin, LDH, total bilirubin, potassium, or creatinine concentrations at any of the time points. There were no reported transfusion reactions. Receiving at least four LTOWB units was not associated with biochemical or clinical evidence of hemolysis. © 2019 AABB.