Acute necrotic proventriculitis is a naturally occurring disease of broiler chickens that causes proventricular rupture during routine evisceration. Although infectious bursal disease virus (IBDV) has been implicated, it has not been proven to be a direct cause of this disease. To further study the role of IBDV in proventriculitis, proventriculi and bursas were collected during both acute and chronic phases of naturally occurring proventriculitis and from chickens experimentally infected with seven different IBDV strains. All tissues were examined for IBDV by light microscopy, immunohistochemistry (IHC), and real time reverse transcriptase (RT)–polymerase chain reaction (PCR) and for apoptosis by the terminal deoxynucleotidyl transferase–mediated deoxyuridine triphosphate nick end labeling method (TUNEL). Tissues from naturally occurring proventriculitis had bursal and proventricular lesions. Two out of four bursas had no IHC-stainable IBDV antigen or RT-PCR detectable IBDV sequences. No proventriculus had IBDV detectable by any of these methods. Bursas from chickens experimentally infected with IBDV had microscopically evident lesions, IBDV was detectable by IHC and RT-PCR, and strong IHC staining for apoptosis was present. Proventriculi from these experimentally exposed chickens had no lesions, low levels of IBDV detectable by IHC or RT-PCR, and very little IHC-stainable apoptosis. We conclude that naturally occurring proventriculitis can occur in the absence of IBDV and that the IBDV strains tested do not directly produce proventriculitis or induce increased proventricular apoptosis.