This study used infrared thermography to monitor the immediate inflammatory reaction to an intratesticular injection of 20% (calcium chloride) CaCl with 0.5% dimethyl sulfoxide (DMSO) for chemical castration and to provide evidence of the treatment's clinical viability. Six animals received a 0.25 mL intratesticular injection of 20% CaCl with 0.5% DMSO in each testis. Thermographic imaging, testicular measurement, penile spine evaluation, electroejaculation, and sperm analysis were performed before the injection. Eighty days post-injection, the cats were evaluated again, then received an orchiectomy followed by histological analysis. Infrared thermography of the testicular area was performed before (M0) and after anesthesia (M0A); after electroejaculation (M0E); 10 min (M1), 1 h, (M2) and 6 h (M3) after the injection for seven consecutive days (M4-10); after 15 (M11) and 30 days (M12); and after 80 days, before and after anesthesia and after electroejaculation (M13, M13A and M13E). No cats had a significant increase in testicular temperature or behavioral changes. All animals were azoospermic and had a 50% reduction in total testicular volume after 80 days. Histologically, the testes showed different degrees of degeneration, necrosis, calcification and replaced connective tissue, as well as Leydig cell hyperplasia in 7/12 of the testes. Infrared thermography efficiently diagnosed and monitored scrotal inflammation caused by intratesticular injection of 20% CaCl solution with 0.5% DMSO, and it is suggested that a single injection of this compound can cause azoospermia after 80 days. We concluded by infrared thermography with a clinical exam, that the adverse reactions were minimal and did not interfere with animal welfare.