Benign prostatic hyperplasia (BPH) is an androgen-related non-neoplastic enlargement of the prostate gland that commonly affects both reproductive capabilities and the general health of intact dogs. The subclinical form of BPH can be challenging to diagnose due to a lack of clinical signs, even if rectal palpation is performed. Left untreated, this condition poses risks to the dogs’ health and breeding status. This study, involving 65 male dogs, aimed to investigate subclinical BPH through rectal palpation, ultrasonography, and analysis of canine prostatic-specific esterase (CPSE). Of the participants, 35 had subclinical BPH, and 30 served as a healthy control group. Dogs suspected of subclinical BPH, as determined by examination results from ultrasonography and CPSE analysis, underwent fine needle aspiration (FNA) guided by ultrasound to enhance diagnostic precision. Findings revealed distinct differences in rectal palpation and ultrasonography between subclinical BPH and healthy dogs. This study established diagnostic thresholds based on prostatic volume and CPSE values and proposed new thresholds for subclinical BPH. Additionally, results showed that prostate gland volume depended on the weight and the age of the dog. In conclusion, early detection of this condition is possible through various examinations, such as changes in ultrasound features, CPSE levels, and rectal palpation. All together, these methods can aid practitioners in early detection of BPH and assist with scheduling screening programs for dogs, ultimately promoting their overall health and reproductive well-being. In conclusion, we advocate for routine, non-invasive prostate screenings in breeding males, underlining the effectiveness of a combination of various multiple techniques for early subclinical BPH detection.