Ten 250-day-old broiler breeders, seven 16-day-old broiler chicks, and three 25-day-old broiler chicks suffering from hydropericardium syndrome (HPS) in Japan were examined histologically, immunohistochemically, and ultrastructurally. Clinically, the chickens died suddenly without apparent signs. The mortality rates were 6.4%, 20.2%, and 26.1%, respectively. The common characteristic histologic lesion was necrosis of hepatocytes, accompanied by intranuclear inclusions of hepatocytes and hemorrhages. In the spleen, there were activation of macrophages in splenic sinus and ellipsoids and erythrophagocytosis in the splenic sinus. The interlobular interstitium of the lung showed marked edema. The air and blood capillary areas of parabronchi included many macrophages with yellow pigments. With immunoperoxidase staining, intranuclear inclusion bodies within degenerating hepatocytes stained positively for group I adenovirus antigen. Ultrastructurally, numerous viral particles (65-70 nm in diameter) were demonstrated in the intranuclear inclusions of hepatocytes. Group I adenovirus (serotype 4) was isolated from liver samples of adult broiler breeders and broiler chicks with HPS. This study suggests that HPS may be caused by group I adenovirus.