2l6 MEDICAL BACTERIOLOGY 



peptone is shown in small amount. The phosphorous content is 0.5 per cent., the reaction 

 acid. 



" Protein No. 4 passes the filter after an addition of an 0.4 per cent, solution of sodic hydrate 

 and gives all reactions for nucleo protein. The remaining bacillary residue amounting to about 

 5 per cent, weight of fat from tubercle bacilli, is free from proteins (nitrogen determination), 

 may contain traces of fats, and is probably cellulose. The secondary proteoses give reaction 

 for sugar. 



"The fatty extractives of tubercle bacilli may be obtained separately as neutral fats and 

 fatty acids by their saponification and subsequent extraction of the acid precipitate with alco- 

 hol and with ether, or in order to avoid undesirable chemical changes by saponification, the 

 alcohol fats and the ether fats may be obtained respectively by first complete extraction with 

 ether, followed by alcohol and vice versa, which to me seems preferable. When in solution they 

 can be shaken out with distilled water, the later holding about 0.5 per cent, in free suspension, 

 which is opalescent in appearance in that concentration, the opalescence disappears in the con- 

 centration employed by me. The several proteins and fatty extractives having been obtained 

 separately and having each been standardized, any free alkali in protein No. 4 having been 

 neutralized by addition of weak solution of HC1, just short of causing precipitation, the several 

 constituents are assembled to represent the formula (given below). 



"The precautions employed in preventing injury or undesirable modifications of the several 

 bacillary products have not all been demonstrated as absolutely requisite for their highest 

 efficiency, my purpose in their adoption having been to take no chances of an inferior prepara- 

 tion by their omission. 



"Formula. This vaccine is absolutely free from real or supposed danger, powerful enough 

 to act by one or two applications, uniform in action by producing the desired immunity in all 

 cases, simple enough to make it available for use of the general practitioner, without the 

 necessity of elaborate examinations or investigations in selecting suitable cases. 



"The vaccine ready for use contains per cubic centimeter 10 milligrams of proteins of 

 tubercle bacilli and a small amount of their fatty extractives, proportioned quantitatively 

 as follows: 



Protein No. i, o. 25 mg. 

 Protein No. 2, 2.75 mg. 

 Protein No. 3, i . oo mg. 

 Protein No. 4, 6 . oo mg. 

 Fatty extractives, o.oi mg. 



In the adoption of the amounts of the several constituents present in the preparation, I was 

 governed by the study of their action when employed separately in animal experiments as well 

 as in some children. Finding that with the same doses but little effect was noted in the produc- 

 tion of specific antibodies in the use of protein No. i, this is present in the smallest amount, 

 whereas protein No. 2, and especially the nucleoprotein designated as No. 4, appearing most 

 effective, they are quantitatively in excess. The addition of fatty extractives may be even- 

 tually found unnecessary, it appearing in my studies of complement fixation with the several 

 constituents of the tubercle bacillus as antigens, that such fixation occurs with all of them 

 inclusive of the fatty extractives with the sera of non-tuberculous children after their vaccina- 

 tion with protein Nos. 2, 3 and 4, irrespective of the presence of one or the other of the fatty 

 extractives in the preparation. 

 "The dose of vaccine is: 



0.05 to o. 10 to nurslings. 



0.02 to 0.60 cc. to children. The higher dosage is given to those 18 to 20 years old. 



"Usually at point of injection in arm there isn't much swelling or tenderness unless the 

 patient is suffering from an active tuberculous lesion." 



