Danovaro R, Dell'Anno A, Trucco A, Serresi M, Vanucci S
Institute of Marine Science, University of Ancona, 60131 Ancona, Italy.
Appl Environ Microbiol. 2001 Mar;67(3):1384-7. doi: 10.1128/AEM.67.3.1384-1387.2001.
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.
在本研究中，我们优化了通过落射荧光显微镜使用SYBR Green I作为染色剂从海洋沉积物中计数病毒的程序。通过利用焦磷酸盐和3分钟的超声处理，从大部分沉积物中获得了最高的病毒产量。焦磷酸盐-超声处理提取底栖病毒的效率约为可提取病毒颗粒的60%。用核酸酶处理的样品病毒计数增加，表明细胞外DNA具有掩盖作用。落射荧光显微镜和透射电子显微镜获得的病毒计数之间未观察到显著差异。甲醛和戊二醛在沉积物储存仅24小时后均使病毒计数显著降低，但7天后未发生进一步损失。
