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A Simple Way of Purifying Several Insect Viruses

Rothamsted Experimental Station, Harpenden, Hertfordshire

Various methods have been reported for purifying ‘non-occluded’ viruses of insects but they have seldom been compared. We find that one we use to purify acute beeparalysis virus (crytogram R/1:2/(25):8/3:I/O) and sacbrood virus (R/*:*/(35):S/S:I/O) (Bailey, Gibbs & Woods, 1963, 1964) is suitable for many other viruses, and is as effective but simpler than other methods. For example, we have used it to purify Galleria dense nucleus virus (GDNV) (D/*:*/37:S/S:I/*) (Meynadier et al. 1964). Larvae of Galleria mellonella that had been injected with the virus and kept at 30° for 7 to 14 days were ground in a 4/1 mixture of water and carbon tetrachloride (1 larva/ml. of water) and centrifuged at 8000 g for 10 min. The supernatant fluid contained about 10¹³ virus particles/ml., which separated in the analytical centrifuge into two components with S_20,w values of 120 and 60 (Fig. 1b); the median lethal doses, by injection, of these two components, separated by centrifuging in sucrose density gradients, were about 10² and 10⁵ particles respectively.

As proposed by Gibbs et al. (1965).

^* Present address: Department of Microbiology, John Curtin School of Medical Research, Australian National University, Canberra.

Received 19 August 1969; accepted 8 September 1969.