Epidemic Risks associated with high-temperature replicating infectious hematopoietic necrosis virus (IHNV) field isolates: from molecular characterization to host pathogen interactions

Abstract

Infectious hematopoietic necrosis virus (IHNV) is a WOAH-notifiable pathogen responsible for severe acute disease in salmonids at temperatures ranging from 4 to 14{degrees}. At higher water temperatures, the disease is generally associated with reduced mortality and lower viral replication. In the context of climate change, rising water temperatures could favor the emergence of thermotolerant strains while representing a major physiological and metabolic stress potentially altering fish's immune responses. This study aims to characterize the impact of increasing water temperatures on IHNV replication and host-pathogen interactions. Following pre-screening, the replication kinetics of the reference strain N61 and two field isolates RtNag0a and SA-22 were characterized in EPC (Epithelioma Papulosum Cyprini) and RTG-2 (Rainbow trout gonad) cell lines at 14, 20 and 22{degrees}. Virulence and host-pathogen interactions were subsequently investigated in vivo in zebrafish (Zf) larvae using complementary microinjection and bath immersion infection models at 21.5 and 23.5{degrees}. In vitro studies revealed distinct thermal replication profiles, distinguishing thermosensitive from thermotolerant isolates, although these phenotypes were influenced by the cellular environment. The zebrafish model enabled the characterization of virulence and antiviral host responses associated with thermotolerant IHNV isolates. Notably, waterborne infection proved effective, contrasting with previous reports describing this exposure route as unsuitable for IHNV studies. These findings expand the growing list of emerging European IHNV genotypes capable of replicating at elevated temperatures (>20{degrees}) and establish zebrafish larvae as a relevant model for virulence assessment of field isolates. Together, these results provide valuable insights into the potential impact of climate warming on IHNV epidemiology and support efforts to predict and anticipate future outbreaks under changing environmental conditions.