A permanent immunity must depend on a stimulation or education of the system to the production of these protective agents _de novo_ or in increased quantity. This must be done by exposure of the tissues to the toxins of the bacillus anthracis, and is accomplished slowly. Precisely what tissues are stimulated to the production of the defensive agents is not fully known, though certain indications may be drawn from observant facts. The eosinophile cells of the blood are presumably important factors as the natural sources of the leucomaines. The spleen as the seat of important blood changes and as preëminently the seat of election of internal anthrax is probably involved. The dogs from which Bardach had removed the spleen were found to be three times as susceptible to anthrax as were the dogs that had not been operated on. Leo’s rats, in which he had produced mellituria by the administration of phloridzin were found to be much more susceptible to anthrax. The liver is also a favorite seat of election of internal anthrax. The products of the healthy liver, are probably in some measure protective. No matter where the defensive products are formed, the practical problem is to secure their production without imperilling life. _By Minimum Dose._ Chauveau and Colin secured this in the larger animals by intravenous injection of a minimum dose,—one or two bacilli. This is more lasting in effect if a second and stronger dose is injected some days later. _By Weakened Virus._ This has been secured by heating the defibrinated blood to 55° C. for ten minutes (Toussaint); Pasteur, Chamberland and Roux accomplished the same end by making anthrax cultures at 42° to 43° C. in presence of air; Chauveau by subjecting the virulent culture for eight days to oxygen under a pressure of 8 atmospheres at a temperature of 38° C.; Chamberland, Roux and others have cultivated the bacillus in weak antiseptic bouillons as phenic acid (1:600 or 1200), bichromate of potash (1:2000 or 5000), sulphuric acid (2:100). Other methods have been followed, as growing the bacillus in the blood or serum of immune animals (dog, chicken, pigeon, white rat, frog).