SOME EXPERIMENTS ON THE CULTIVATION OF BACILLUS 



LEPR^.^ 



By Moses T. Clegq. 

 {From the Biological Laboratory, Bureau of Science, Manila, P. I. 



The following experiments on the cultivation of the leprosy bacillus 

 were confined entirely to an attempt to grow the organism in symbiosis. 

 Believing that the leprosy bacillus derives its nutrition from the products 

 of the tissue cells in the lesions in which it is encountered, and from 

 the fact that it occurs so frequently within the protoplasm of these cells, 

 an organism was selected which could be cultivated and which corres- 

 ponded in a degree to the living tissue cells; that is, an organism con- 

 taining a nucleus and protoplasm, the amoeba. It was first necessary to 

 procure a suitable growth of amoebae and in this step the technique em- 

 ployed was the same as that recommended previously by Dr. Musgrave 

 and myself in the cultivation of these protozoa. The culture media was 

 composed of agar 20 grams, sodium chloride 0.3, extract of beef 0.3 ; it 

 was otherwise prepared in the same manner as ordinary agar media for 

 bacteria, the finished product giving a reaction of 1 per cent alkaline to 

 phenolpthalein. The media was then placed in tubes and sterilized. 

 The culture media when it is to be used is melted and poured into sterile 

 Petri dishes and allowed to harden before being inoculated. The material 

 containing the amoebae is then spread in a thin layer over the surface of 

 the media. If the proper symbiotic bacteria are present, the amcebas will 

 develop within two to ten days, depending upon the source of the amoebae 

 and the number and character of the bacteria present. 



As intimated this media, an amoeba, and its symbiotic bacteria were 

 used in an attempt to cultivate the leprosy bacillus. A culture of 

 amoebae was obtained from a dysentery stool and after a sufficient growth 

 of the amoebae had occurred to overbalance the original symbiotic bacteria 

 growing with the amoebae, that is to say, when it would have been neces- 

 sary to add a fresh supply of the symbiotic bacteria to the culture in order 

 to obtain a further development of the amoebae, leprosy bacilli were added 

 instead. The inoculation of the plates with leprosy bacilli was accom- 

 plished by smearing the surface of the media containing the amoebae with 

 a portion of the pulp of a leper-'s spleen. The first series of plates was 



' Read at the Sixth Annual Meeting of the Philippine Islands Medical As- 

 sociation, February 11, 1909. 



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