The effect on canine erythrocyte osmotic fragility under different temperature variations and storage times

The erythrocyte osmotic fragility (EOF) test has been used to determine the integrity of human and animal red blood cell (RBC) membranes, and to observe the mechanical quality of blood cells meant for transfusion. Existing studies proposed 1-6˚C as an optimal range of blood bag storage temperature to minimize risk of hemolysis. However, temperature fluctuations during blood bag delivery may induce denaturation of RBC membrane, resulting to in vitro hemolysis, and subsequent compromise of RBC function and viability. The purpose of this study was to determine the effect of time and temperature on the quality of canine whole blood (WB) used for transfusion. The specific aim is to utilize EOF analysis to determine alterations of osmotic fragility under different temperatures at selected storage time points. Fifteen healthy dogs were recruited to donate WB (250 mL) collected in CPDA-1 (citrate, phosphate, dextrose, and adenine). Fifteen segments of blood samples from each blood bag were stored at 4˚C. Over a period of 35 days, samples were analyzed at 3 different time points: days 0, 28 and 35. On each testing day, 5 blood segments of each blood bags were separately stored at 1, 4, 10, 25 and 35˚C for 24 h before EOF analysis.

Findings of our study showed slightly increased osmotic resistance amongst storage temperatures, 1, 4 and 10˚C, but increased osmotic fragility were observed at 25 and 35˚C through all testing days. When storage time points were compared, the mean corpuscular fragility (MCF) of samples did not significantly differ regardless of temperature; however, when considering both temperature and time, samples stored over 28 days and incubated at 35˚C showed the most significant differences.

Canine whole blood with CPDA-1 stored over 28 days and exposed to temperature over 25˚C may have resulted in the assumed loss of membrane integrity, causing hemolysis. The observed outcome of the study implies that EOF underwent statistically significant influence by increasing temperatures and was only affected in lesser degrees by storage time.