Molecular and microscopic evidence of viruses in marine copepods
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Edited by Edward F. DeLong, Massachusetts Institute of Technology, Cambridge, MA, and approved December 12, 2012 (received for review September 23, 2012)

Abstract
As dominant members of marine mesozooplankton communities, copepods play critical roles in oceanic food webs and biogeochemical cycling. Despite the ecological significance of copepods, little is known regarding the causes of copepod mortality, and up to 35% of total copepod mortality cannot be accounted for by predation alone. Viruses have been established as ecologically important infectious agents in the oceans; however, viral infection has not been investigated in mesozooplankton communities. Here we used molecular and microscopic techniques to document viral infection in natural populations of the calanoid copepods Acartia tonsa (Dana) and Labidocera aestiva (Wheeler) in Tampa Bay, FL. Viral metagenomics revealed previously undocumented viruses in each species, named Acartia tonsa copepod circo-like virus (AtCopCV) and Labidocera aestiva copepod circo-like virus (LaCopCV). LaCopCV was found to be extremely prevalent and abundant in L. aestiva populations, with up to 100% prevalence in some samples and average viral loads of 1.13 × 105 copies per individual. LaCopCV transcription was also detected in the majority of L. aestiva individuals, indicating viral activity. AtCopCV was sporadically detected in A. tonsa populations year-round, suggesting temporal variability in viral infection dynamics. Finally, virus-like particles of unknown identity were observed in the connective tissues of A. tonsa and L. aestiva by transmission electron microscopy, demonstrating that viruses were actively proliferating in copepod connective tissue as opposed to infecting gut contents, parasites, or symbionts. Taken together, these results provide strong independent lines of evidence for active viral infection in dominant copepod species, indicating that viruses may significantly influence mesozooplankton ecology.
Footnotes
↵1Present address: Blood Systems Research Institute, San Francisco, CA 94118.
- ↵2To whom correspondence should be addressed. E-mail: hewson{at}cornell.edu.
Author contributions: M.B. and I.H. designed research; D.S.D., T.F.F.N., J.G.B., A.M.G., M.B., and I.H. performed research; D.S.D., T.F.F.N., K.R., M.B., and I.H. analyzed data; and D.S.D., K.R., M.B., and I.H. wrote the paper.
The authors declare no conflict of interest.
This article is a PNAS Direct Submission.
Data deposition: The sequences reported in this paper have been deposited in the GenBank database (accession nos. JF912805 and JQ837277; metagenomic sequences have been deposited under GenBank accession nos. JY253391–JY253440).
This article contains supporting information online at www.pnas.org/lookup/suppl/doi:10.1073/pnas.1216595110/-/DCSupplemental.
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