Objective: To report and compare the hematological and serum biochemical profile of goat and sheep from Rahim Yar Khan district in Southern Punjab. Methods: One hundred and twenty blood samples (98 goat and 22 sheep) were collected and their blood glucose, hemoglobin and serum biochemical parameters, cholesterol, aspartate transaminase (AST), alanine aminotransferase (ALT) and lactate dehydrogenase (LDH) were determined by using spectrophotometer. Parameters were compared between goat and sheep. Results: It was observed that glucose (P=0.001), LDH (P=0.001) and ALT (P=0.001) concentrations differed significantly between sheep and goats. The studied parameters were compared between buck and goats and also between young goats (less than 12 months old) and adults (more than 12 months old). It was found that age and gender did not affect these hematological and serum biochemical parameters.Conclusions: It can be concluded that sheep have overall higher values than goats for studied hematological and serum biochemical parameters.