Tissue Distribution of the Piscine Novirhabdovirus Genotype IVb in Muskellunge (Esox masquinongy)

SIMPLE SUMMARY: A novel strain of viral hemorrhagic septicemia virus was discovered in the Great Lakes. This Great Lakes strain of the virus infects a broad range of fish species with outcomes ranging from clinical and subclinical disease, persistent viral shedding, and/or death. Among the most susceptible species to the Great Lakes strain of the virus are juvenile muskellunge. Increased susceptibility to the virus in this regionally and economic important species generated a multitude of research questions to include but not limited to host range, pathogenesis, and diagnostic tools to efficiently detect and minimize viral spread into captive fish stocks. The overarching aim of the current study focuses on assessing the early and latter stages of disease progression in a battery of traditional and non-traditional diagnostically relevant tissues in juvenile muskellunge. Tissue damage from the virus and amount of live virus in each tissue were evaluated in conjunction with advanced diagnostic methods to identify cells targeted by the virus when possible. ABSTRACT: A novel sublineage of the piscine novirhabdovirus (synonym: viral hemorrhagic septicemia virus), genotype IVb, emerged in the Laurentian Great Lakes, causing serious losses in resident fish species as early as 2003. Experimentally infected juvenile muskellunge (Esox masquinongy) were challenged with VHSV-IVb at high (1 × 10(5) PFU mL(−1)), medium (4 × 10(3) PFU mL(−1)), and low (100 PFU mL(−1)) doses. Samples from spleen, kidneys, heart, liver, gills, pectoral fin, large intestine, and skin/muscle were collected simultaneously from four fish at each predetermined time point and processed for VHSV-IVb reisolaton on Epitheliosum papulosum cyprini cell lines and quantification by plaque assay. The earliest reisolation of VHSV-IVb occurred in one fish from pectoral fin samples at 24 h post-infection. By 6 days post-infection (dpi), all tissue types were positive for VHSV-IVb. Statistical analysis suggested that virus levels were highest in liver, heart, and skin/muscle samples. In contrast, the kidneys and spleen exhibited reduced probability of virus recovery. Virus distribution was further confirmed by an in situ hybridization assay using a VHSV-IVb specific riboprobe. Heart muscle fibers, hepatocytes, endothelia, smooth muscle cells, and fibroblast-like cells of the pectoral fin demonstrated riboprobe labeling, thus highlighting the broad cellular tropism of VHSV-IVb. Histopathologic lesions were observed in areas where the virus was visualized.