Melarsomine is used intramuscularly to destroy adult heartworms when treating canine heartworm disease (HWD). This drug is highly irritative and can elicit local complications. Therefore, melarsomine should be injected into the paralumbar muscles by strictly adhering to the manufacturers' prescriptions. However, it is not known how to determine the optimal location of the needle during the injection process. Ultrasonography (US) of the epaxial (paralumbar) musculature was used as a new method to measure the cross-sectional diameter of the paralumbar musculature, to determine the required location of the injection needle, and to study the local side effects in two dogs with HWD. The macroscopic appearance of the melarsomine solution during injection was demonstrated by video imaging. Melarsomine was not fully gravitating, but its majority was spreading along the thickest fascia of the musculature. Three minutes thereafter, no ultrasound signs of the melarsomine solution were seen, suggesting a full absorption at least ultrasonographically. This procedure was simulated in vitro with methylene blue solution having the same appearance. Removing the injection needle only after 5 min post-injection could prevent undesirable leakage of the drug through the injection channel into the subcutaneous tissue. Ultrasonography can be a useful aid during the treatment of HWD with melarsomine according to this preliminary study.