Abstract
Bluetongue virus (BTV) is an arbovirus that has been associated with dramatic epizootics in both wild and domestic ruminants in recent decades. As a segmented, double-stranded RNA virus, BTV can evolve via several mechanisms due to its genomic structure. However, the effect of BTV’s alternating-host transmission cycle on the virus’s genetic diversification remains poorly understood. Whole genome sequencing approaches offer a platform for investigating the effect of host-alternation across all ten segments of BTV’s genome. To understand the role of alternating hosts in BTV’s genetic diversification, a field isolate was passaged under three different conditions: (i) serial passages in Culicoides sonorensis cells, (ii) serial passages in bovine pulmonary artery endothelial cells, or (iii) alternating passages between insect and bovine cells. Aliquots of virus were sequenced, and single nucleotide variants were identified. Measures of viral population genetics were used to quantify the genetic diversification that occurred. Two consensus variants in segments 5 and 10 occurred in virus from all three conditions. While variants arose across all passages, measures of genetic diversity remained largely similar across cell culture conditions. Despite passage in a relaxed in vitro system, we found that this BTV isolate exhibited genetic stability across passages and conditions. Our findings underscore the valuable role that whole genome sequencing may play in improving understanding of viral evolution and highlight the genetic stability of BTV.
Highlights
Bluetongue virus (BTV; family Reoviridae, genus Orbivirus) is a globally distributed, arthropod-borne virus that can cause profound disease in both domestic and wild ruminants
We suspect that samples may have been compromised during storage, during storage, or that BTV passaged serially on Bovine pulmonary artery endothelial cells (BPAEC) or CuVaW3 cells may have diminished or that BTV passaged serially on BPAEC or CuVaW3 cells may have diminished growth kinetics in growth kinetics in BHK 21 cells, thereby compromising our ability to accurately titer this virus using
By using a relaxed system of propagation, we sought to remove the impact of varying host-derived features on BTV’s genetic diversification, instead capturing the virus’s inherent capacity to diversify in vertebrate and invertebrate cells
Summary
Bluetongue virus (BTV; family Reoviridae, genus Orbivirus) is a globally distributed, arthropod-borne virus that can cause profound disease in both domestic and wild ruminants. BTV is the etiologic agent of bluetongue disease and is transmitted by biting midges in the genus Culicoides. Bluetongue is listed as a notifiable disease by the World Organisation for Animal Health (OIE). Sheep and white-tailed deer are considered to be relatively more susceptible to severe or even fatal disease compared to other ruminants, cattle and other wildlife species are vulnerable to infection and illness. The global distribution of bluetongue virus is defined by the presence of a competent insect vector, which is—with few exceptions—necessary for virus transmission between ruminants [3,4].
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