Stingrays are prominent marine animals; however, there are few published reference values for their blood chemistry and hematology. Twenty-eight southern stingrays (Dasyatis americana) were caught using the bottom trawl nets of fishery-independent boats operated by the South Carolina Department of Natural Resources during June and July 2002 from Winyah Bay, South Carolina, to St. Augustine, Florida. Median values of blood and plasma obtained from live animals promptly after capture are as follows: packed cell volume = 0.22 L/L (22%), total solids (TS) = 56.5 g/L (5.65 g/dl), total protein (TP) = 26 g/L (2.6 g/dl), sodium = 315 mmol/L, potassium = 4.95 mmol/L, chloride = 342 mmol/L, calcium = 4.12 mmol/L (16.5 mg/dl), phosphorus = 1.5 mmol/L (4.7 mg/dl), urea nitrogen = 444 mmol/L (1,243 mg/dl), glucose = 1.69 mmol/L (30 mg/dl), aspartate aminotransferase = 14.5 U/L, creatine phosphokinase = 80.5 U/L, osmolality = 1065 mOsm/kg, and lactate = 3.1 mmol/L. Bicarbonate was less than the low end of the instrument range (5 mmol/L) in all but three samples. Anion gap was negative in all samples. Albumin was less than the low end of the instrument range (1 g/dl) in all except one sample. Osmolality was significantly higher in the rays caught in the southern region. TS and TP values were linearly related to each other, and the equation for the fitted line is TS = (11.61 × TP) 25.4 (in g/L) [or TS = (1.161 × TP) 2.54 (in g/dl)]. The reference ranges reported in this study can be used to aid in the management of aquarium stingrays and to create a baseline for health monitoring of the wild Dasyatis spp.