Nutrition Investigations. 327 



up to contain 2 per cent of the air dry extract, 1 per cent peptone, 

 \ per cent beef extract and \ per cent sodium chloride. This could be 

 filtered through paper only on a hot, water-jacketed funnel, from 

 which it dropped as a clear, amber-colored jelly. After standing 

 unsterilized over night in a warm room, this was found to be entirely 

 broken up by the formation of gas throughout the whole mass. The 

 reaction, which had been neutral, was now acid to litmus. This 

 material was placed in a flask and allowed to stand for two months, 

 at the end of which time, the greater portion was liquefied, the former 

 lumps of jelly being reduced to small particles distributed throughout 

 the liquefied portion. Alcoholic extracts did not reduce Fehling's 

 solution. A sterile preparation of the plain manauea extract in test 

 tubes was inoculated with some of this material, but without produc- 

 ing the same striking results. There were evidences of growth, but 

 none of liquefaction or gas formation, in the course of two weeks. 



TRIALS WITH ANAEROBES. 



The action upon Irish moss of pure cultures of the powerful putre- 

 factive organisms B. Putrificus, Bienstock, B. Maligni oedematis, 

 and B. Anthracis symptomatici, was tried in the following way. A 

 4 per cent solution of the moss was prepared, which would not become 

 liquefied at a temperature of 30°-35° C. From this material culture 

 media were prepared, neutral, alkaline, and acid in reaction, using 

 the solution plain, and with the addition of \ per cent beef extract 

 and \ per cent salt, or 1 per cent peptone and \ per cent salt. Test 

 tubes were inoculated from fresh, active cultures, and the organisms 

 allowed to grow for one to three weeks, being examined at first daily, 

 and later every three or four days, for liquefaction and gas formation. 

 The results were negative in all cases, save that in the peptone media 

 an occasional small bubble was seen, with cultures of the bacilli of 

 malignant oedema and symptomatic anthrax. However, the same 

 phenomena were observed in peptone-agar tubes used as controls. 



Mixtures of B. Anthracis symptomatici and B. Maligni oedematis 

 were tried upon solutions of dulse, Irish moss, salep and sinistrin, in 

 the following way: Small Erlenmeyer flasks containing 50 cc. of 

 1 per cent solutions of each of these carbohydrates, and 5 cc. of ordi- 

 nary nutrient bouillon, were inoculated with fresh cultures of these 

 organisms, rendered anaerobic, and incubated for four weeks at 37.5° 

 C. On inspection, no change was apparent. The carbohydrates 

 were removed, the alcoholic extracts examined for reducing sugar, and 



