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Applied and Environmental Microbiology, March 2001, p. 1384-1387, Vol. 67, No. 3
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.
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
*
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.
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