Praziquantel (PZQ), long-used in veterinary and human medicine for the treatment of helminth parasites, is known to enhance humoral and cellular immune responsiveness in mammals but has unknown direct immunomodulatory capabilities in fish. In the present study, we examined the ability of PZQ to induce gene transcriptional changes in immune-competent primary tissue/organ cultures of two highly important yet evolutionarily discrete fish species – Southern bluefin tuna Thunnus maccoyii and Atlantic salmon Salmo salar. These cultures consisted of mixed blood cell population for both species, as well as intestinal explants from bluefin. Although expression profiles varied between species and tissue/organ type, PZQ induced both T-cell receptor (more than twofold) and IL-8 transcriptional expression (more than fourfold). Additionally, increased expression of other inflammatory cytokines including IL-1β was detected in blood cell cultures from both species, and a general pattern of heightened antiviral signaling was observed. Specifically, elevated transcription of Type I (IFNα) and Type II (IFNγ) interferon in Atlantic salmon blood cultures along with elevated expression of MHC class I in blood cultures of both species. These findings provide preliminary evidence for direct immunomodulation by PZQ in fish and insight into its potential capacity as an immune stimulant/adjuvant in the rapidly expanding aquaculture industry.