Abstract
New diseases in marine animals are emerging at an increasing rate, yet methodological limitations hinder characterization of viral infections. Viral metagenomics is an effective method for identifying novel viruses in diseased animals; however, determining virus pathogenesis remains a challenge. A novel anellovirus (Zalophus californianus anellovirus, ZcAV) was recently reported in the lungs of captive California sea lions involved in a mortality event. ZcAV was not detected by PCR in the blood of these animals, creating the inability to assess the prevalence of ZcAV in live sea lions. This study developed an enzyme-linked immunosorbent assay (ELISA) to detect antibodies to ZcAV in sea lion serum. To assess ZcAV prevalence, paired serum and lung samples (n = 96) from wild sea lions that stranded along the California coast were tested through ELISA and PCR, respectively. Over 50% of the samples tested positive for ZcAV by ELISA (34%), PCR (29%), or both (11%) assays. ZcAV is prevalent in stranded wild sea lion populations and results suggest that PCR assays alone may grossly underestimate ZcAV exposure. This ELISA provides a tool for testing live sea lions for ZcAV exposure and is valuable for subsequent studies evaluating the potential pathogenicity of this anellovirus.
Highlights
New diseases in marine animals are emerging at an increasing rate, yet methodological limitations hinder characterization of viral infections
Viral metagenomics is an effective method for identifying viruses involved in mortality events in marine animals[6,7,8,9], yet it remains difficult to establish a connection between a novel virus and disease due to limitations of culturing the viruses as well as difficulties in obtaining fresh diagnostic tissues from wild marine mammals
Viral metagenomics performed on lung tissue of several necropsied captive California sea lions (Zalophus californianus) with signs of respiratory disease that were involved in a mortality event of unknown cause revealed the presence of a novel anellovirus, Zalophus californianus anellovirus (ZcAV)[7]
Summary
New diseases in marine animals are emerging at an increasing rate, yet methodological limitations hinder characterization of viral infections. There is a critical need for an assay to detect ZcAV exposure in blood samples to investigate the epidemiology of this virus, understand its association with disease, and preemptively develop management strategies that can prevent the spread of this virus in captive and rehabilitation animals To overcome these technical limitations of studying the role of ZcAV in disease, here we describe the development of an enzyme-linked immunosorbent assay (ELISA) for ZcAV, and demonstrate that sea lions mount an immune response against ZcAV. We developed an ELISA based on hydrophilic regions of the ORF 1 gene product of the ZcAV genome and demonstrated that this assay is capable of detecting anti-ZcAV antibodies in sea lion serum This ELISA provides a tool for studying ZcAV epidemiology and identifying seroconversion upon symptom development in captive sea lions, enabling future research investigating the pathogenesis of ZcAV. The creation of this assay lays the groundwork for bridging the gap between genome discovery via viral metagenomics and assessing the epidemiology of novel viruses and their significance for wild populations
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