Abstract: Hemoglobin (Hb) and Packed Cell Volume (PCV) values were compared in blood collected from the following four sites of the Japanese quail: jugular, brachial, medial metatarsal veins and the heart. Ten 25-week-old hatch-mate layers of similar body weight having a hard shelled egg in the uterus were used in this study. Hb and PCV values were found to be the highest in the medial metatarsal vein followed by, in descending order, the brachial vein, the jugular vein and the heart. The heart had significantly lower values (p<0.05) compared with the jugular, brachial and medial metatarsal veins with the latter two having almost similar values. Blood samples from the jugular vein exhibited the least variability compared to other sites. The brachial vein followed by the jugular vein, were found to be suitable sites for blood sampling as well as for making injections without any significant complication. The heart puncture technique was very risky, traumatic and resulted in many side effects and should not be used, particularly in on-going experiments. Scientists should be aware of possible differences in blood constituents as a result of sampling sites. For obtaining accurate and reliable results, these points should be taken into consideration while designing an experiment or assessing comparative hematological data collected using different collection techniques.