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Enumeration of Marine Viruses in Culture and Natural Samples by Flow Cytometry
Open accessAbstract
Flow cytometry (FCM) was successfully used to enumerate viruses in seawater after staining with the nucleic acid-specific dye SYBR Green-I. The technique was first optimized by using the Phaeocystis lytic virus PpV-01. Then it was used to analyze natural samples from different oceanic locations. Virus samples were fixed with 0.5% glutaraldehyde and deep frozen for delayed analysis. The samples were then diluted in Tris-EDTA buffer and analyzed in the presence of SYBR Green-I. A duplicate sample was heated at 80°C in the presence of detergent before analysis. Virus counts obtained by FCM were highly correlated to, although slightly higher than, those obtained by epifluorescence microscopy or by transmission electron microscopy ( r = 0.937, n = 14, and r = 0.96, n = 8, respectively). Analysis of a depth profile from the Mediterranean Sea revealed that the abundance of viruses displayed the same vertical trend as that of planktonic cells. FCM permits us to distinguish between at least two and sometimes three virus populations in natural samples. Because of its speed and accuracy, FCM should prove very useful for studies of virus infection in cultures and should allow us to better understand the structure and dynamics of virus populations in natural waters.
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Cited by
Interactions between microplastics and phytoplankton aggregates: Impact on their respective fates
Long, M., Moriceau, B., Gallinari, M., Lambert, C., Huvet, A., Raffray, J., & Soudant, P. (2015) · Marine Chemistry · doi:10.1016/j.marchem.2015.04.003 · Cited by 711
Viral Control of Phytoplankton Populations—a Review1
BRUSSAARD, C. P. D. (2004) · Journal of Eukaryotic Microbiology · doi:10.1111/j.1550-7408.2004.tb00537.x · Cited by 501
Self-citationYoch, D. C. (2002) · Applied and Environmental Microbiology · doi:10.1128/aem.68.12.5804-5815.2002 · Cited by 452
Past, present and future applications of flow cytometry in aquatic microbiology
Wang, Y., Hammes, F., De Roy, K., Verstraete, W., & Boon, N. (2010) · Trends in Biotechnology · doi:10.1016/j.tibtech.2010.04.006 · Cited by 247
Flow cytometry and the single cell in phycology
Collier, J. L. (2000) · Journal of Phycology · doi:10.1046/j.1529-8817.2000.99215.x · Cited by 136
Metadata
- Authors
- Marie, D., Brussaard, C. P. D., Thyrhaug, R., Bratbak, G., & Vaulot, D.
- Venue
- Applied and Environmental Microbiology
- Publisher
- American Society for Microbiology
- Published
- 1999-01
- Volume
- 65
- Issue
- 1
- Pages
- 45-52
- ISSN
- 0099-2240 (ISSN-L)
- 1098-5336
- OpenAlex
- W2102820270
- Cited by
- 646
- References
- 0
- FWCI
- 20.30
- Type
- journal article
- Language
- en
- Open access
- green
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- Marie, D., Brussaard, C. P. D., Thyrhaug, R., Bratbak, G., & Vaulot, D. (1999). Enumeration of Marine Viruses in Culture and Natural Samples by Flow Cytometry. Applied and Environmental Microbiology. https://doi.org/10.1128/aem.65.1.45-52.1999
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