192 Investigations in Plant Pathology 



anilin by Koch when isolating the tubercle bacillus, to the fact 

 that fuchsin might be substituted for methyl violet, to projection 

 eye pieces and their most effective use, how to make holes through 

 glass, and such, appeared in his " Record Book of Culture Media." 

 By far the most important references were those to a formula for 

 preparing an "Agar Medium with Beef Broth," supplied to him 

 on February 26, 1890, by Moore of the Bureau of Animal Industry, 

 and another agar formula supplied by Dr. W. M. Gray, micro- 

 scopist of the Army Medical Museum at Washington, on March 

 25 of the same year. Smith chose his magnum opus, his monu- 

 mental three-volume work, Bacteria in Relation to Plant Diseases 

 (1905), in which to acknowledge his debt to Dr. Theobald Smith 

 and Dr. Moore. In its preface, he thanked each of these distin- 

 guished men for " friendly advice and helpful suggestions [given] 

 at a time when the writer was beginning his bacteriological studies 

 and was perplexed in many ways." ^" 



Dr. Moore's formula for nutrient media read: 



To each gram of ground or minced meat add 2 cc of distilled water. Set 

 away 18 hours, i.e. overnight, in ice box. Strain through a towel. If 

 necessary add enough water to bring the fluid up to the original amount 

 of water added. Add |% sodium chloride, and 1% of peptone. Add of 

 normal solLution] of sodium carbonate (53 grams flame dried salt per litre 

 of distilled water) . l^nough to render the acid liquid slightly alkaline. Boil 

 30-35 minutes on water bath. Filter. 



The stock flasks not needed for immediate use may now be boiled 1^ 

 hours on each of three consecutive days, and then set away. 



To the flasks wanted for immediate use add 1% of agar, dry and cut 

 very fine. Boil briskly 2 hours on the covered water bath in Erlenmeyer 

 flask, keeping the water up to level of the fluid in the flask. Cool down 

 to 40° or 45 °C and add white of Qgg at rate of one white to each 200 cc 

 of the liquid. Mix thoroughly by beating with a spoon, or by pouring 

 from one dish to another. Again boil 2 hours briskly on the waLer bath. 

 Filter on hot water filter through 2 layers of Japanese filter paper (it will 

 not pass in any quantity through ordinary filter paper). Then fill tubes 

 or plates. 



If rolls are made, the tubes must lie jiat and must be examined in that 

 position, as where the agar cools some water always exudes. 



On July 8, 1885, Dr. William M. Gray had been appointed by 

 Dr. John S. Billings to perform some bacteriological analyses 

 incident to the work of the Army Medical Museum. A skillful 



^^ Page V. 



