The objective of this study was to isolate and identify suspected pathogens from peacocks and peacock farmers with severe pneumonia and to investigate its potential association with peacocks' pneumonia, caused by Chlamydophila psittaci infection. A clinical examination of infected peacocks identified birds with symptoms of anorexia, weight loss, yellowish droppings, airsacculitis, sinusitis, and conjunctivitis, whereas the infected farmers showed high fever and respiratory distress. Immunofluorescence tests detected chlamydial antigens in pharyngeal swabs (12 of 20) and lung tissue samples (four of five) from peacocks. One of four swabs taken from farmers was also positive by the same test. Specific anti-chlamydia immunoglobulin G was detected in 16 of 20 peacocks and four of four peacock farmers. The isolated pathogen was able to grow in specific-pathogen-free (SPF) chicken embryos and McCoy cell lines and was identified as Chlamydiae by immunofluorescence assay and PCR. Avian influenza virus, Newcastle disease virus, and infectious bronchitis virus were eliminated as potential causative agents after pharyngeal swabs inoculated onto the chorioallantoic membrane of embryonate eggs failed to recover viable virus. PCR and restriction fragment length polymorphism indicated the ompA gene from the isolate was similar to that of avian C. psittaci type B. Three-week-old SPF chickens challenged with the peacock isolate via intraperitoneal injection showed a typical pneumonia, airsacculitis, and splenitis. Subsequently, the inoculating strain was recovered from the lungs of challenged birds. This is the first report of C. psittaci infection in peacocks and peacock farmers.