Wommack K E, Hill R T, Muller T A, Colwell R R
Center of Marine Biotechnology, University of Maryland Biotechnology Institute, Baltimore 21202, USA.
Appl Environ Microbiol. 1996 Apr;62(4):1336-41. doi: 10.1128/aem.62.4.1336-1341.1996.
Current estimates of viral abundance in natural waters rely on direct counts of virus-like particles (VLPs), using either transmission or epifluorescence microscopy. Direct counts of VLPs, while useful in studies of viral ecology, do not indicate whether the observed VLPs are capable of infection and/or replication. Rapid decay in bacteriophage viability under environmental conditions has been observed. However, it has not been firmly established whether there is a corresponding degradation of the virus particles. To address this question, viable and direct counts were carried out employing two Chesapeake Bay bacteriophages in experimental microcosms incubated for 56 h at two depths in the York River estuary. Viruses incubated in situ in microcosms at the surface yielded decay rates in full sunlight of 0.11 and 0.06 h-1 for CB 38 phi and CB 7 phi, respectively. The number of infective particles in microcosms in the dark and at a depth of 1 m was not significantly different from laboratory controls, with decay rates averaging 0.052 h-1 for CB 38 phi and 0.037 h-1 for CB 7 phi. Direct counts of bacteriophages decreased in teh estuarine microcosms, albeit only at a rate of 0.028 h-1, and were independent of treatment. Destruction of virus particles is concluded to be a process separate from loss of infectivity. It is also concluded that strong sunlight affects the viability of bacteriophages in surface waters, with the result that direct counts of VLPs overestimate the number of bacteriophage capable of both infection and replication. However, in deeper waters, where solar radiation is not a significant factor, direct counts should more accurately estimate numbers of viable bacteriophage.
目前对天然水体中病毒丰度的估计依赖于使用透射或落射荧光显微镜对病毒样颗粒(VLP)进行直接计数。VLP的直接计数虽然在病毒生态学研究中很有用,但并不能表明观察到的VLP是否能够感染和/或复制。已经观察到噬菌体在环境条件下活力的快速下降。然而,病毒颗粒是否存在相应的降解尚未得到确凿证实。为了解决这个问题,在约克河河口的两个深度对实验微宇宙中的两种切萨皮克湾噬菌体进行了56小时的培养,并进行了活菌计数和直接计数。在微宇宙中于水面原位培养的病毒,在充分阳光下,CB 38 phi和CB 7 phi的衰减率分别为0.11和0.06 h-1。在黑暗中以及1米深度的微宇宙中,感染性颗粒的数量与实验室对照没有显著差异,CB 38 phi的平均衰减率为0.052 h-1,CB 7 phi的平均衰减率为0.037 h-1。河口微宇宙中噬菌体的直接计数有所下降,尽管速率仅为0.028 h-1,且与处理无关。得出结论,病毒颗粒的破坏是一个与感染力丧失分开的过程。还得出结论,强烈阳光会影响地表水噬菌体的活力,结果是VLP的直接计数高估了能够感染和复制的噬菌体数量。然而,在更深的水域,太阳辐射不是一个重要因素,直接计数应该能更准确地估计活菌噬菌体的数量。