Gallibacterium, a gram-negative coccobacillus, is a genus of the Pasteurellaceae family with capability of infecting a wide range of avian host species. In 2003, Gallibacterium was selected instead of different previous names. Four different species has been recognized in Gallibacterium genus so far. Members of the genus can cause a wide range of pathological lesions, from upper respiratory tract lesions, follicle degeneration, enteritis, pericarditis, hepatitis, oophoritis, septicemia to more important problems such as salpingitis and peritonitis. Salpingitis and peritonitis of layer farms caused by Gallibacterium result in decreased egg-production and increased mortality. Gallibacterium is the most common single bacterial infection in egg-laying farms with reproductive disorders. During a microbiological survey on bacterial infectious agents of reproductive tract of layers, some isolates from salpingitis cases were suspicious of Gallibacterium. According to culture and biochemical properties, the primary identification was in support of genus Gallibacterium. A polymerase chain reaction (PCR) test using primers specific for the genus identification was carried out. The suspected isolate showed bands of 970 and 1080 bp, which are specific for Gallibcterium. To confirm the identification, a positive sample was sent to University of Copenhagen for repeating the PCR test. To our knowledge, this is the first report of isolation and identification of Gallibacterium in Iran. This study shows the possibility of a role for Gallibacterium in poultry industry of Iran. This study also shows the need for further investigations on epidemiological situation of the infection, as well as the isolation and identification of different species of the genus Gallibacterium.