Method for serial passage of infectious hematopoietic necrosis virus (IHNV) in rainbow trout

Transmission is a fundamental component of pathogen fitness. A better understanding of pathogen transmission can greatly improve disease management. In particular, controlled studies of multiple rounds of natural transmission (i.e. serial passage) can provide powerful epidemiological and evolutionary inferences. However, such studies are possible in only a few systems because of the challenges in successfully initiating and maintaining transmission in the laboratory. Here we developed an efficient and reproducible cohabitation method for conducting controlled experiments investigating the effects of serial passage on infectious hematopoietic necrosis virus (IHNV) in rainbow trout. This method was used to investigate the transmission efficiency and kinetics of viral shedding of IHNV over 3 serial passages. Transmission efficiency decreased from 100 to 62.5% over the passage steps and was associated with a decrease in virus shedding into water. A shift in the peak of viral shedding was also observed, from Day 2 post immersion for passage 0 to at least 24 h later for all subsequent passages. Finally, the characterization of viruses after 1 round of transmission and propagation on cells showed no change in glycoprotein (G gene) sequences or viral virulence compared to the ancestral virus stock. The methods developed provide valuable tools for reproducible population-level studies of IHNV epidemiology and evolution.