The host range of a mixture of Choristoneura fumiferana (Clemens) nucleopolyhedroviruses (CfMNPV and CfDefNPV) was investigated using a per os bioassay of larvae of 29 species of Lepidoptera and adult males of Megachile rotundata (F.) (Hymenoptera: Megachilidae). Using a whole-genomic DNA probe, positive results were obtained in 8 of 10 Tortricidae: Archips cerasivorana (Fitch), Choristoneura fractivittana (Clemens), C. fumiferana, Choristoneura occidentalis Freeman, Choristoneura pinus pinus Freeman, Choristoneura rosaceana (Harris), Clepsis persicana (Fitch), and Cydia pomonella (L.); one Crambidae: Ostrinia nubilalis (Hübner); one arctiine Erebidae: Estigmene acrea (Drury); and two Noctuidae: Oligia illocata (Walker) and Pyrrhia exprimens (Walker). Mortality rates were highest among C. fumiferana, C. occidentalis, C. pinus pinus, A. cerasivorana, and C. pomonella. Sequenced polymerase chain reaction (PCR) amplicons from infected individuals from several species confirmed that the primer sets amplified the target viruses. CfMNPV was consistently found in virus-fed C. fumiferana; whereas, CfDefNPV was present only occasionally. The presence of CfMNPV and CfDefNPV in A. cerasivorana was confirmed by PCR and DNA sequencing. Significant treatment-mortality rates were induced in the noctuids P. exprimens and Acronicta impleta Walker; PCR determined that both viruses were present in treated P. exprimens but only CfMNPV was present in A. impleta. No virus was detected in M. rotundata.