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Background: Ki67 index, tumor associated macrophages (TAMs) and mast cells (MCs) are associated with malignancies in animal and human neoplasms including colorectal carcinomas (CRC). This has not been assessed in canine CRC. Given similar genetic abnormalities between human and canine CRC, we assessed Ki-67 and mitotic indices, TAMs and MC count (MCC) in canine CRC (n = 17). TAMs and MCC were compared with those in adenomas (n = 13) and control (n = 9). Results: Ki-67 index in CRC (17.13 ± 11.50) was strongly correlated (r = 0.98, p < 0.05) with mitotic index (3.52 ± 1.80). MCC was higher (p < 0.05) in CRC (6.30 ± 3.98) than in adenomas (0.78 ± 0.77) and control (0.35 ± 0.33). The results suggest that Ki-67 index and MCC are associated with malignancy in canine CRC. Higher average TAMs were counted in adenomas (21.30 ± 20.70) and in CRC (11.00 ± 9.82) than in the control (7.69 ± 7.26), although the differences were not significant (p > 0.05). Conclusion: Ki-67 index, TAMs and MCC in canine CRC were recorded for the first time in this study. Ki-67 index and MCC are associated with malignancy in canine CRC. Quantitative assessment of MCs and Ki-67 coupled with mitotic index and other clinical parameters may help in evaluating malignancy in canine CRC. TAMs likely play a role in the development of canine colorectal tumors. Further studies to determine the clinical significance of these parameters for prognostic, chemo-preventive and chemotherapeutic purposes in canine colorectal tumors are recommended. [ABSTRACT FROM AUTHOR]