To validate radiation dose of cattle affected by the Fukushima Daiichi Nuclear Power Plant (FNPP) accident, we applied electron spin resonance (ESR) tooth dosimetry. Teeth were collected from cattle that had stayed continuously after the accident in Okuma Town within the ex-evacuation zone of the FNPP accident. Radiation exposure to cattle attributed to the FNPP accident was confirmed retrospectively by X-band ESR tooth dosimetry, which was almost consistent with the estimated radiation dose from airborne and individual cattle, whereas positive radiation-induced signals (RIS) were not detectable in any sample by nondestructive measurement using L-band ESR tooth dosimetry. Although ESR tooth dosimetry reflects total radiation doses of affected animals, the uncertainty of measurement was relatively large. Therefore, in order to accurately measure the additional radiation dose from the nuclear accident, it is necessary to clarify possible causes of the uncertainty. Making continuous improvements, X-band ESR tooth dosimetry for animals in the ex-evacuation zone is ongoing.