Viral hemorrhagic septicemia virus (VHSV) is the etiologic agent of a disease of great concern to aquaculture. We performed experimental infections with intraperitoneal VHSV IVb injections at 106 pfu/fish in four species: tiger muskellunge, ♂Esox lucius×♀Esox masquinongy; Atlantic salmon, Salmo salar; channel catfish, Ictalurus punctatus; and walleye, Sander vitreus. The fish were thermal shocked concurrent with infection to cause immunosuppression and increase the probability of eliciting disease. Each species was observed for morbidity and mortality and periodic samples taken. Virus was detected using virus isolation and real-time quantitative reverse-transcription polymerase chain reaction (qRT-PCR). Histopathology confirmed lesions in tiger muskellunge and Atlantic salmon, and immunohistochemistry showed virus associated with some lesions. Tiger muskellunge were highly susceptible to VHSV IVb, and high viral levels were detected in exposed fish. A single mortality was observed in exposed Atlantic salmon with a short viral replication period. Channel catfish and walleye were resistant to developing clinical signs of VHS. The results show that these species vary in the development of VHS under these conditions, and qRT-PCR testing enabled virus detection in subclinically affected fish. Basic concordance statistics between the qRT-PCR test results and the virus isolation results were evaluated.