Infectious diseases have been responsible for large-scale declines in many endangered animals. Disease outbreaks in small populations have probably led to the eventual extinction of such endangered animals in the wild. The endangered Asiatic lion (Panthera leo persica) population may also face such threats. This was evident from this study on captive Asiatic lions from western India, which were sampled from December 1998 to March 1999. Fifty-six Asiatic lions, including 17 hybrid lions (Afro–Asian crosses) from six captive centers in western India, were tested for antibodies against canine distemper virus (CDV), feline parvovirus (FPV), feline immunodeficiency virus (FIV), and feline leukemia viral (FeLV) antigen. Agar gel immunodiffusion test and dot immunobinding assay were employed for CDV and FPV antibody detection. Commercial enzyme-linked immunosorbent assay kits were used for FIV antibody and FeLV antigen detection. Forty-nine of 56 lions (87.5%) were positive for CDV. All 56 (100%) lions were positive for FPV antibodies. There were no detectable levels of FIV antibodies and FeLV antigens. It was observed that CDV and FPV, two viruses known to cause high mortality in captive carnivores, were widely prevalent in these captive Asiatic lions. It is suggested that these seropositive animals will have the potential to pose a risk of infection to other seronegative animals. Hence, it is imperative to carefully consider any movement, translocation, or reintroduction of these animals to new regions. It is also recommended that these animals be required to undergo standard quarantine and disease screening protocols. The lack of exposure to pathogens, such as FIV and FeLV, would also be a risk, and, hence, identification of reservoirs and screening of in-contact animals is highly recommended. Vaccinations must be considered, using killed or other suitable viral vaccines, which have been proved to be safe, effective, and efficacious in endangered felids.