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Quantifying Ostreid Herpesvirus (OsHV-1) Genome Copies and Expression during Transmission

  • Host Microbe Interactions
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Abstract

Understanding the pathogenic potential of a new pathogen strain or a known pathogen in a new locale is crucial for management of disease in both wild and farmed animals. The Ostreid herpesvirus-1 (OsHV-1), a known pathogen of early-life-stage Pacific oysters, Crassostrea gigas, has been associated with mortalities of juvenile oysters in many locations around the world including Tomales Bay, California. In two trials, the California OsHV-1 strain was transmitted from infected juvenile C. gigas to naïve C. gigas larvae. Survival of control larvae was high throughout both trials (97–100%) and low among those exposed to OsHV-1. No OsHV-1-exposed larvae survived to day 9 in trial 1, while trial 2 was terminated at day 7 when survival was 36.90 ± 8.66%. To assess the amount of OsHV-1 DNA present, we employed quantitative polymerase chain reaction (qPCR) assays based on the A fragment and OsHV-1 catalytic subunit of a DNA polymerase δ (DNA pol) gene. Viral genome copy numbers based on qPCR assays peaked between 3 and 5 days. To measure the presence of viable and actively transcribing virus, the DNA pol gene qPCR assay was optimized for RNA analysis after being reverse transcribed (RT-qPCR). A decline in virus gene expression was measured using RT-qPCR: relative to earlier experimental time points copy numbers were significantly lower on day 9, trial 1 (p < 0.05) and day 7, trial 2 (p < 0.05). Peaks in copies of active virus per genome occurred during two periods in trial 1 (days 1 and 5/7, p < 0.05) and one period in trial 2 (day 1, p < 0.05). Transmission electron microscopy confirmed OsHV-1 infection; herpesvirus-like nucleocapsids, capsids, and extracellular particles were visualized. We demonstrated the ability to transmit OsHV-1 from infected juvenile oysters to naïve larvae, which indicates the spread of OsHV-1 between infected hosts in the field and between commercial farms is possible. We also developed an important tool (OsHV-1-specific RT-qPCR for an active virus gene) for use in monitoring for active virus in the field and in laboratory based transmission experiments.

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Acknowledgments

The authors wish to Washington state and Tomales Bay shellfish growers for continued support and supplying oysters, S. White and S. Roberts for technical assistance and design of 18S C. gigas primers, R. Strenge for technical assistance and editorial comments, and K. Reece for editorial comments. Transmission electron microscopy was performed by P. Mason at the Virginia Institute of Marine Sciences; J. Baines and T. Renault provided assistance with interpretation. Funding was provided by National Sea Grant as administrated by Washington Sea Grant NA04OAR4170032 AMD 12 and NA04OAR4170032 MOD05.

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  1. Colleen A. Burge

    Present address: Department of Ecology and Evolutionary Biology, Cornell University, Ithaca, NY, 14853, USA

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  1. School of Aquatic and Fishery Sciences, University of Washington, Box 355020, Seattle, WA, 98195, USA

    Colleen A. Burge & Carolyn S. Friedman

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Correspondence to Carolyn S. Friedman.

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Burge, C.A., Friedman, C.S. Quantifying Ostreid Herpesvirus (OsHV-1) Genome Copies and Expression during Transmission. Microb Ecol 63, 596–604 (2012). https://doi.org/10.1007/s00248-011-9937-1

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  • DOI: https://doi.org/10.1007/s00248-011-9937-1

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