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Applied and Environmental Microbiology, March 2001, p. 1384-1387, Vol. 67, No. 3
0099-2240/01/$04.00+0   DOI: 10.1128/AEM.67.3.1384-1387.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Determination of Virus Abundance in Marine Sediments

R. Danovaro,1,2,* A. Dell'Anno,1 A. Trucco,1 M. Serresi,3 and S. Vanucci4

Institute of Marine Science, University of Ancona, 60131 Ancona,1 Department of Zoology, University of Bari, 70125 Bari,2 Faculty of Medicine and Surgery, University of Ancona, 60020 Ancona,3 and Department of Animal Biology and Marine Ecology, University of Messina, Salita Sperone, 90100 Messina,4 Italy

Received 28 August 2000/Accepted 13 December 2000

In this study, we optimized procedures to enumerate viruses from marine sediments by epifluorescence microscopy using SYBR Green I as a stain. The highest virus yields from the bulk of the sediments were obtained by utilizing pyrophosphate and 3 min of sonication. The efficiency of extraction benthic viruses by pyrophosphate-ultrasound treatment was about 60% of the extractable virus particles. Samples treated with nucleases had increased virus counts, suggesting a masking effect of extracellular DNA. No significant differences were observed between virus counts obtained by epifluorescence microscopy and transmission electron microscopy. Both formaldehyde and glutaraldehyde gave significant reductions of virus counts after only 24 h of sediment storage, but no further loss occurred after 7 days.

* Corresponding author. Mailing address: Institute of Marine Science, University of Ancona, Via Brecce Bianche, 60131 Ancona, Italy. Phone: 39 71 220 4654. Fax: 39 71 220 4650. E-mail: danovaro{at}popcsi.unian.it.

Applied and Environmental Microbiology, March 2001, p. 1384-1387, Vol. 67, No. 3
0099-2240/01/$04.00+0   DOI: 10.1128/AEM.67.3.1384-1387.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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