R. E. O. Williams

Standardized methods are essential if phage typing of staphylococci is to be reliable and if the results obtained in different laboratories are to be compared. This paper, prepared on behalf of the Subcommittee on Phage Typing of Staphylococcus of the Nomenclature Committee of the International Association of Microbiological Societies, gives a detailed account of methods that have been found satisfactory for propagating the phages and defines a standard testing routine by which the stability of the phage preparations can be verified. Technical details are given of methods recommended for determining the phage type of staphylococci and the interpretation of differences between the types is discussed. The need for close collaboration between the epidemiologist and the laboratory worker in the use of phage typing is emphasized.

Cancer of the large bowel shows marked variations in geographical distribution (Doll, 1967; Doll, Payne, and Waterhouse, 1966; Davis, Knowelden, and Wilson, 1965) and, with the exception of Japan, the disease is more prevalent in developed than in underdeveloped countries. The reason for this variation is not known but epidemiological evidence suggests that environmental factors may be involved. It is claimed that immigrants from areas with a low incidence of cancer of the large bowel tend to show the same high incidence of this cancer as the local population (Haenszel and Dawson, 1965; Buell and Dunn, 1965). Changes in dietary habit may be especially important (Wynder and Shigematsu, 1967;

The routine methods for propagation of staphylococcal typing bacteriophages, and for their use in identifying strains of staphylococci, are described. Most of the phages can be propagated in fluid media as well as on agar, and for some glucose-peptone-water is a better medium than nutrient or Todd-Hewitt broth. Many phage filtrates derived from broth or agar propagation contain, in addition to the phage, an agent that inhibits the growth of staphylococci. Investigations of variations in the routine typing technique showed that considerable latitude is permissible in the age of cultures used for typing and in the inoculation procedures. It is, however, important to test all phage filtrates after propagation for purity and freedom from the inhibitory agent; and to repeat the tests frequently during use to detect alterations in titre. About 40% of staphylococci are not lysed by the phages used at their test dilution; about half of these untypable strains are lysed by undiluted phage filtrates. An analysis was made of the results of typing 567 independent strains of staphylococci; 229 of these showed strong lysis by one or more phages, but there were no fewer than 82 distinct phage patterns represented, and only 118 strains were lysed strongly by a single phage. Certain phages tend to appear together in patterns and on the basis of such associations three main phage groups can be distinguished; they are known respectively as the ‘3A’, ‘6/47’ and ‘52’ groups. No method was discovered for segregating patterns into ‘types’, but conventions have been devised on the basis of the variation observed in sets of strains from various sources, for distinguishing between different patterns. We are deeply indebted to Dr V. D. Allison for teaching us the bacteriophage typing methods and for giving us the benefit of his wide experience. We are grateful to the following for sending us phages, and in some cases discussing their results with us: Dr Phyllis Rountree, Sydney; Dr R. Wahl, Paris; Dr G. Wallmark, Stockholm; Dr H. Williams Smith, Poultry Research Station, and Mr A. M. Hood, Birmingham. Our thanks are also due to Miss S. Mayo, Mrs E. Lyons and Mr D. Woodroof for technical assistance. The photographs for PI. 14 were taken by Mr W. Clifford.