Definitive radiotherapy refers to delivery of large doses, typically 48–62 Gray, of ionizing radiation over several weeks using a daily or alternate-day fractionation schedule. The impact of definitive radiotherapy alone on haematopoiesis in tumour-bearing dogs is unknown. Medical records from 103 dogs receiving definitive 60Cobalt teletherapy for cancer over a 5-year period were reviewed for signalment, tumour type and location, total radiotherapy dose and fractionation scheme. Complete blood count data were collected before, halfway through, and at the end of radiation treatment, and analysed for changes associated with patient variables. The results demonstrate significant reductions in haematocrit, total white blood cell count, neutrophils, eosinophils, monocytes, lymphocytes and platelets occurred during definitive radiotherapy but remained within laboratory reference intervals. These data are important for anticipation of toxicity associated with combinations of radiotherapy and chemotherapy in dogs but do not support the routine monitoring of haematology parameters during definitive radiotherapy.