The objective of this study was to evaluate the effect of castration on dimensions of the prostate by B‐mode ultrasound for 90 days, as well as to evaluate changes in serum concentrations of testosterone and oestradiol in clinically healthy canines and those with benign prostatic hyperplasia (BPH). Three groups of animals were used: healthy canines castrated on day 0 (HCC, n = 8), canines with a diagnosis of benign prostatic hyperplasia (BPHC, n = 9) castrated on day 0 and healthy uncastrated canines (IHCC, n = 10). Prostate size was obtained by B‐mode ultrasound, while blood samples were collected (days 0, 7, 14, 28, 60 and 90 post‐castration) to determine the serum testosterone and oestradiol concentration. Castration did not affect serum oestradiol concentrations. Castration led to a decrease in serum testosterone (reaching values <20 ng/dl in the first week, p < .05), causing a gradual decrease in dimensions of prostate parameters relative to body weight (p < .05). Canines with BPHC had similar prostate dimensions as the IHCC group 2 weeks post‐castration. In relation to the prostate's relative parameters, the decrease with time was greater in canines with BPHC than those in the HCC group. When evaluating parameters as a percentage of relative change, the decrease with time was similar in both castrated groups, except for relative length percentage, which presented a more pronounced decline in BPHC canines than HCC canines. In conclusion, castration is an effective, fast and long‐lasting treatment for canine patients with BPH, as it causes a rapid decrease in blood testosterone concentrations and a decrease in glandular volume, but does not affect serum oestradiol concentrations. This study did not determine the mechanisms of action, but allowed us to observe (based on morphometric variables) that characteristics of prostate involution after castration differed between canines with BPH and healthy canines.