Chlorotetracycline hydrochloride is widely used in veterinary medicine as an antibiotic to treat bacterial infections in animals. It is particularly effective against a range of Gram-positive and Gram-negative bacteria, making it a valuable tool in managing diseases in livestock, poultry, and aquaculture. The compound is often administered through feed or water for therapeutic, prophylactic, and growth promotion purposes to ensure proper dosage and absorption.

In addition to its veterinary applications, it has been utilized in research settings as a selective agent in microbiology to study bacterial resistance and antibiotic mechanisms. Its ability to inhibit protein synthesis in bacteria makes it a useful compound for experimental purposes.

However, its use is carefully regulated to prevent the development of antibiotic resistance and to ensure food safety, as residues can potentially affect human health. Proper withdrawal periods are observed before treated animals are processed for food.