a1 Department of Microbiology, John Curtin School of Medical Research, Australian National University, Canberra
Strains of myxoma virus associated with a case-mortality rate in wild rabbits of 90—95% can be distinguished from those associated with a case-mortality rate of more than 99% by the use of a standardized virulence test carried out in groups of five laboratory rabbits, and tests carried out on laboratory rabbits are closely correlated with the results obtained with Australian wild rabbits. The test, which attempts to mimic natural mosquito transmission, consists of the intradermal inoculation in one site of about 5 rabbit-infectious doses of the virus, and the subsequent observation of the rate of progress of clinical symptoms and ultimate fate of the rabbit. The mean survival time provides a figure which allows satisfactory classification of all known myxoma virus strains into one of four grades of virulence, very attenuated strains being recognized by their low case-mortality rates.
Using this test, 92 strains of myxoma virus have been compared. These comprised 6 laboratory strains, 62 strains recovered from Australian wild rabbits or from wild caught mosquitoes, between February, 1951 and March, 1955, 19 strains recovered from naturally infected rabbits in Europe between October, 1953 and August, 1955, 3 strains from South America, and 2 from California. Eight types have been described and illustrated, differentiation being based on their virulence and the clinical picture of the disease they cause in laboratory rabbits. The differences are summarized in Text-fig. 2. Four of the virus types produce recognizably different pocks on the chorioallantois of developing chick embryos.
In Australia the repeatedly introduced standard laboratory strain of virus has been replaced by a moderately virulent strain, which appears to be better adapted for survival than either the highly virulent standard laboratory strain, or less virulent variants of which only a few had been recovered up to 1955. Strains of the type dominant in 1953–55 have appeared independently in many widely separated parts of Australia. Examination of these attenuated variants by pure clone techniques showed that the variability in symptomatology associated with them is not due to a mixture of virus strains, but to the expression of differences in innate resistance of the host animals which are obscured by the overwhelming virulence of the highly virulent strains.
Only one introduction of a different strain of virus was made in Europe. All field strains recovered during the first two years after the introduction were of high virulence, but a few attenuated strains appeared in 1955, and they have since become widespread. One of them appears to be a relatively stable mixture of a highly virulent and a greatly attenuated strain.
The collection of the large numbers of strains of virus described in this paper has been made possible only by the generous collaboration of many people. In Australia we would like to acknowledge the great help afforded us by Mr F. N. Ratcliffe, officer-in-charge of the Wildlife Survey Section, Commonwealth Scientific and Industrial Research Organization and his officers, especially Messrs J. H. Calaby, G. Douglas, A. L. Dyce, B. V. Fennessy, E. W. Lines, K. Myers, R. Mykytowycz, W. E. Poole and E. Waterhouse. Messrs B. D. Robinson and T. Pearce of the Lands Department of Victoria have also provided us with specimens, as have Mr G. Edgar, Director of the Veterinary Research Station, Glenfield, New South Wales, Dr D. Surrey Dane of the Institute of Medical and Veterinary Science, Adelaide, Mr D. Gooding, Department of Agriculture, Western Australia, and Messrs T. B. Alexander and A. F. Ryan, Department of Agriculture, Tasmania.
We are indebted to Dr H. de B. Aragão of the Instituto Oswaldo Cruz, Rio de Janeiro, and Dr B. Szyfres, of the Laboratorio de Biologia Animal, Uruguay, for specimens from South America; Dr D. G. McKercher of the University of California School of Veterinary Medicine, Davis, California, for the Californian strains; Drs P. Lépine and H. Jacotot, of the Institut Pasteur, Paris, for strains from France; and Mr J. R. Hudson, of the Veterinary Laboratory, Weybridge, and Dr C. H. Andrewes of the National Institute for Medical Research, Mill Hill, for material from England.
We are grateful to Dr S. Fazekas de St Groth and Dr G. S. Watson of this University for advice on the statistical treatment of the data.
Mr V. Paral took the photographs.
(Received July 30 1956)
* Aided by grants from the Rural Credits Development Fund of the Commonwealth Bank of Australia.