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Determination of viral production in aquatic sediments using the dilution-based approach

Nature Protocols volume 4, pages 1013–1022 (2009)Cite this article

Abstract

Viruses are the most abundant and dynamic biological entities in the world's ecosystems. Marine sediments, the largest biome in the world, have the potential to represent an optimal environment for viral development. To assess the viral effect on their hosts, and to understand the ecological role of the viruses in the benthic food webs and biogeochemical cycles, measurements of benthic viral production are needed. Different direct and indirect approaches have been proposed to estimate viral production in aquatic sediments, but a standardized protocol is not available yet. The method presented in this protocol relies on the short-time incubations of sediment samples with virus-free seawater, and the subsequent determination of the increase in viral abundance over time by epifluorescence microscopy. The protocol described here is highly reliable, inexpensive and easy to use. The entire procedure takes approximately 3 days to be completed, but the method allows the parallel processing of several sediment samples, which is recommended in ecological studies.

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Figure 1: Epifluorescence microscopy images of 0.02 μm Anodisc filters stained with SYBR Green I from sediment samples.
Figure 2: Changes in viral abundance (expressed as number of viruses per gram) during time-course incubation experiments carried out on different marine-sediment samples.

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Acknowledgements

This work was financially supported by the EU within the framework of the project HERMES (GOCE-CT-2005-511234-1).

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  1. All authors equally contributed to this work.

Authors and Affiliations

  1. Department of Marine Sciences, Polytechnic University of Marche, Via Brecce Bianche, Ancona, Italy

    Antonio Dell'Anno, Cinzia Corinaldesi, Mirko Magagnini & Roberto Danovaro

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  1. Antonio Dell'Anno
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  2. Cinzia Corinaldesi
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  4. Roberto Danovaro
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Correspondence to Roberto Danovaro.

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Dell'Anno, A., Corinaldesi, C., Magagnini, M. et al. Determination of viral production in aquatic sediments using the dilution-based approach. Nat Protoc 4, 1013–1022 (2009). https://doi.org/10.1038/nprot.2009.82

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