Serum biochemical and hematological reference values are used to establish normality and to diagnose disease and physiological alterations. Up till now there are no reference values for different blood biochemical and hematological variables in lactating pregnant buffaloes, consequently the current study considered the first one that established those values in lactating pregnant water buffaloes (Bubalus bubalis). Animals were examined at buffalo farms that belong to Assiut Governorate, Egypt. A total of 148 pregnant lactating buffaloes (5-10 years old) were examined, of these, 20 buffaloes did not meet the selection criteria and excluded from study. The remained animals (N.=128) were clinically healthy and included in the study. A total of 55 hematological and serum biochemical variables were measured in blood of lactating buffaloes. Mean reference values, reference intervals (2.5 and 97.5 percentiles) and their 90% confidence intervals, for the measured hematological and serum biochemical constituents were statistically calculated. Data for the serum biochemical and hematological variables were presented in tables and compared with previously published data. The established reference values will be a useful guide for interpreting serum biochemical and hematologic data in lactating pregnant water buffaloes
