Reference values for buffaloes especially those at late pregnancy are not yet established. The aim of this study was to establish serum biochemical and hematological reference intervals for water buffaloes (Bubalus bubalis) during the late pregnancy. A total number of 125 clinically healthy late pregnant (7.0-10.5 months) buffaloes (3-8 years old) were included in the study. The inclusion of animals in the study was based on numbers of basic selection criteria. Animals were examined at buffaloes' farms that belong to Assiut Governorate at the mid of Egypt. Three types of samples were collected; serum samples for biochemical analysis, whole blood samples for hematological analysis and fecal samples for parasitological examination. A total of 55 blood analytes were measured during this study. The 95% reference intervals for each serum biochemical and hematological constituents were calculated by removing the upper and lower 2.5% of the interval to give the 2.5 and 97.5 percentiles. The results revealed that serum biochemical and hematological constituents in late pregnant buffaloes showed some differences when compared with previously published data. In conclusion, the established reference values will be a useful guide for interpreting serum biochemical and hematologic data in late pregnant buffaloes, especially those live under the Egyptian environment