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The subject of the isolation ofanaerobes is one which the worker is inclined to approach with apologies. Every month or so a paper appears in some journal in which a new and expeditious procedure for the separation of anaerobes is. described. There are many successful ways of isolating anaerobes and it is unwise torecommend any one method above all others. I have succeeded with various arrangements; and wish in this paper to analyze some of the principles governing the isolation of these organisms and to explain a few of the pitfalls which have caused many workers to believe that the securing of "absolutely pure" anaerobic cultures is a difficult matter. With a little practice and with the exercise of much discrimination, anaerobes may be isolated as quickly, or nearly as quickly, as aerobes. Contamination occurs somewhat more frequently in anaerobic cultures than in those of aerobes. Contamination of originally pure cultures may be attributed to the following causes: (1) Insufficiently sterilized media; anaerobe media are usually pasty and require more careful sterilization than others. (2) Inoculation transfer involving the exposure of the cotton plug and of the inoculum to the air. I have noted in working in London and near the sea in San Francisco, that the more dusty the air, the more frequent are contaminations, and the contamination flora may vary according to location. (3) During incubation in closed jars the cotton plugs may become sufficiently moist for

PRINCIPLES CONCERNING THE ISOLATION OF ANAEROBES STUDIES IN PATHOGENIC ANAEROBES. II