Blood samples collected from captive Southern lechwes (Kobus leche) were used to develop reference intervals for complete blood count and serum biochemistry values. The study population consisted in 108 healthy individuals, including 62 females and 46 males and 38 neonates (<2 days old), 27 juveniles (2 mo to 2 yr old), and 43 adults (>2 yr old) from the Réserve Africaine de Sigean, Sigean, France, between 2014 and 2016. No significant effects of sex and season of sampling were found. No significant differences were found for all hematology and biochemistry parameters between juveniles and adults, and their results were then combined. Significant differences were found between animals >2 mo old and neonates for most tested parameters; in particular, neonates had lower hemoglobin, eosinophil count, alanine transaminase, and total protein values, but higher alkaline phosphatase values. The reference intervals obtained were similar to values previously reported for other Bovidae species.