Isolation, characterization and molecular identification of a novel aquareovirus that infects endangered fountain darter Etheostoma fonticola

• Published in: Diseases of aquatic organisms Vol. 130; no. 2; pp. 95 - 108
• Main Authors: Iwanowicz, L R, Iwanowicz, D D, Adams, C R, Lewis, T D, Brandt, T M, Sanders, L R, Cornman, R S
• Format: Journal Article
• Language: English
• Published: Germany Inter-Research Science Center 10.09.2018
• Subjects: Animals; Biosecurity; Diagnostic systems; Embryos; Endangered & extinct species; Endangered Species; Etheostoma fonticola; Fish; Gene sequencing; Genomes; Next-generation sequencing; Oncorhynchus mykiss; Oncorhynchus tshawytscha; Perches - virology; Phylogeny; Refugia; Reoviridae - genetics; Reoviridae - isolation & purification; Ribonucleic acid; Rivers; RNA; RNA viruses; rRNA; Salmon; Trout; Viruses; Virus; Etheostoma fonticola · Fountain darter; Endangered; Aquareovirus
• ISSN: 0177-5103; 1616-1580
• DOI: 10.3354/dao03261

The fountain darter Etheostoma fonticola (FOD) is a federally endangered fish listed under the US Endangered Species Act. Here, we identified and characterized a novel aquareovirus isolated from wild fountain darters inhabiting the San Marcos River. This virus was propagated in Chinook salmon embryo (CHSE)-214, rainbow trout gonad-2 and fathead minnow cells at 15°C. The epithelioma papulosum cyprini cell line was refractory at all temperatures evaluated. High throughput sequencing technologies facilitated the complete genome sequencing of this virus utilizing ribosomal RNA-depleted RNA extracted from infected CHSE-214 cells. Conventional PCR primer sets were developed for the detection and confirmation of this virus to assist diagnostic screening methods. Phylogenetic analysis suggests this virus belongs to the Aquareovirus A genus. This research provides requisite initial data critical to support hatchery and refugia biosecurity measures for this endangered species.