This serum had no neutralizing effect on cultures of non-toxip:enie strains. A considerable proportion of the bacterin-containing liquid could then be squeezed out with a press, and the organisms could then be washed and spun in a centrifuge. Intradermal titration of a toxin transferred to rabbit inner hind leg skin.
Unless otherwise stated, it must be assumed that the pH of the medium was corrected and the medium boiled just prior to inoculation. Accordingly, the supernatant of the alum precipitate was dialyzed for 48 h and then dried down. To the supernatant of the first precipitate, a further 20 g per 100 c.c. aclclecl, aud t:the precipitation dialyzerl nn1l the n•snltant fluid 1hierl. 'l'he M:.R.J).
The reaction produced by the toxin and arenalin will be labeled "etheric ions to:s.:in". In these tests, the hemolytic power of the pulp culture and this pulp filtrate after preliminary dialysis. In addition, large quantities of antitoxin are required for neutralization, more so to neutralize that part of the toxin which causes .,;\;iH l'Padion,;.
The ability of serum from cattle over t"·0 years of age and rabbits to neutralize 1 to (j ~LR.J).
PART II
For the Lr. titrations, 0 2 c.c. of the toxin was titrated against the antitoxin, the levels of which differed by 20-30 percent. The literature hears that pigs, sheep and pigs can be immunized by solitary means or against the JJ:dmal Although the sheep were immune to intramuscular inoculation of li1·ing culture, no anh-to.--;in \Yas present in the bloo1l mtd L The sheep 1lid uot the intradertuil' o1· intn11·eJtOlb injel' resisted. tion of tosin. H OII'E'YPl', hun(hPrls ot l·xuntple;; coulrl he giYen of ihP ahilit.~· of one injection of formol- fil!ratP to itttlll1llti:tP aguin:<. Lteclainche and Vallee (19002) fondd tl1<1t Cl.,)uwroei antiserum aclmiuistererl to guinea pigs 1lid not iJJJillllllize ihem against (:f. 1·haunJei hut nol against Cl. se;Jt/c/1111 nncl that chauvoei agglutinating serum does not contain 011 ;ppticum hacilli. septil'llill SPnlm nP In the second test, adrenaline was omitted because it was found that toxin was a suitable activator in itself and did not introduce a non-specific factor. After three injections, the chauvoei antitoxin was detectable in the sera of all five sheep tested. No indications were found for a relationship between the amount of septic and chauYoei antitoxins in the different sheep. However, there is a clear indication that the amount of circulating chauvoei antitoxin was closely related to the amount of toxin (contained in the activated spore suspension) carried by the sheep. Cl. chauvoei with injections of Cl. septicum anaccultu1·e. A septic toxin user! for antitoxin detection was noted in the text; 37 L was used for the detection of chauvoei antitoxin). Experiment in sheep (a). - The sheep received 12 dry injections of septicum anaculture (strain R 27), at two-year intervals. Before starting immunization, uo septi<:um or cbanvoPi :mtito:xin \HIS demonstrable in 0 ·1 e. Primary stimulus and ,;secondary- dried d1alized ammonium sulfate precipitates of bncl an If only the good and the bad vaccines had been used, the result of the test would have been considered highly satisfactory; 0 ·1 e.r.;. However, the introduet1011 of the fair and fairly good vaccines show that no correlation exists between the amount of circulating antitoxin and the amount of culture resisted. Knowing the toxin's Lr, a simple calculation showed how much antitoxin the toxin had jumped. Furthermore, by knowing the volume of the initially precipitated toxoid and the weight of the dry powder, one could calculate the amount of antitoxin, which 1·0 c.c. of liquid toxoid would hixul.. roxoid 86 C bound more antitoxin than toxoicl 87 and stimulated the formation of more antitoxin 1:n vivo. Also in experiment 6, it was shown that the toxoid from the virulent strain precipitated 45 times more antitoxin than the avirulent strain and stimulated the formation, in sheep, of more antitoxin. Furthermore, since one injection elicited fixed immunity, testing of the immunity after 2 i-injections was not performed. An injection of the boiled supernatant did not so immunize 2 sheep as resisted 2} and 1 M.L.D. There is a possibility that heating the antigen will destroy some of the heat-stable material. Power of the Unwanted will lloiled :iupe1·natant of Frozen and 1'lw7cecl Cl. Some support for this is provided by the fact that after one injection there is no trace of antitoxin, as is not detectable in any of the four sheep. The effect of this substance appears to be the localization of the toxin in the tissues. long enough to cause visible damage, but it is admitted that the study of the lysine was not given the same attention as the toxin. The immunity produced by toxoids from a virulent and a relatively avirulent culture of the same strain "·as of about the same order of magnitude. The presence of the heat-stable antigen is demonstrated if toxin-free (heated) bacilli are injected into sheep; after a suitable interval is those immune to live culture inoculation.PART III
