Reference values for 18 plasma chemical variables in blue neck ostriches (Struthio camelus australis, n = 60, age 24–36 mo) were established for use in veterinary clinical practice using nonparametric statistics. The following values were established for the percentiles P_2.5 and P_97.5: sodium 147–157 mmol/L, calcium 2.4–4.8 mmol/L, inorganic phosphate 1.3–2.3 mmol/L, chloride 94–105 mmol/L, glucose 10.3–13.7 mmol/L, urea 0.5–0.8 mmol/L, uric acid 351–649 μmol/L, bile acids 8–33 μmol/L, total protein 39–56 g/L, albumin-globulin ratio 0.45–0.59, osmolality 304–330 mOsm/kg, alkaline phosphate 69–217 IU/L, aspartate aminotransferase 243–418 IU/L, gamma-glutamyltransferase 0–1 IU/L, creatine kinase 1648–4894 IU/L, glutamate dehydrogenase 8–17 IU/L, and lactate dehydrogenase 860–2236 IU/L. The plasma calcium concentration was significantly (P < 0.001; r = 0.74) related to the total protein concentration and an adjustment-formula for calcium was derived: adjusted Ca (mmol/L) = Ca (mmol/L) – 0.09 TP (g/L) 4.4. The influence of blood sample treatment on the plasma potassium concentration as seen in other avian species was demonstrated in a separate experiment, emphasizing the need to separate plasma and cells immediately after collection in avian blood samples.