Beef carcasses were examined to explore the effects of stunning methods on central nervous system tissue (CNST) dissemination on the surface during the slaughtering process. The frequency of occurrence of CNST contamination on four defined parts, each on the interior and exterior surfaces of the split carcass, and their level of the contamination were measured by an ELISA test. The effect of hot carcass weight was also examined. The results showed that the frequency of contamination occurrence was not affected by the stunning method. However, the penetrating captive bolt stunning method resulted in a higher level of CNST contamination than non-penetrating sledge-hammer stunning (P<0.001). The higher level of contamination occurred on the interior surface of carcasses along the vertebral area. Therefore, the dissemination of CNST on carcasses seemed to be affected by the stunning method, carcass splitting, and carcass washing. The carcass weight significantly affected the level of CNST contamination on the carcass (P<0.01) but there was no interaction with the stunning method.