EXAMINATION OF SPUTUM 391 



If more antiformin is used the specific gravity is too much increased and the bacilli 

 are damaged. The fluidification is hastened at incubator temperature. 



To 5 parts of sputum add i part of antiformin, shake well and place in incu- 

 bator for one hour. To 10 c.c. of the homogeneous mixture add 1.5 c.c. of a 

 solution made up of i part chloroform and 9 parts alcohol. Shake violently and 

 centrifuge for fifteen minutes. Mix the sediment with egg albumin, smear out and 

 stain. 



When it is desired to culture the tubercle bacilli mix 20 c.c. of sputum with 65 c.c. 

 sterile water and add 15 c.c. antiformin. Stir the mixture with a glass rod. After 

 thirty minutes to two hours we should have a homogeneous mixture. Centrifuge 

 for fifteen minutes or longer, wash the sediment twice with sterile salt solution and 

 smear out the well- washed sediment over serum or glycerine egg slants. The tubes 

 should be covered with black paper and the plugs paraffined. It must be remem- 

 bered that for culturing tubercle bacilli we must protect the growth from sunlight 

 as this will kill the organism. If fluid culture media are inoculated the transferred 

 material should be deposited on the surface. Should the particle sink growth will 

 not occur. 



Sputum smears stained by some Romanowsky method or by the 

 haematoxylin-eosin stain are best adapted for the study of various 

 cells, and in particular of the eosinophile cells so characteristic of 

 bronchial asthma. In sputum from cancer of the lungs the large 

 vacuolated cells may be found. 



When examining the sputum of the bronchopneumonia of influenza the formol 

 fuchsin gives the best results. The influenza bacilli are found in little masses, fre- 

 quently grouped about small collections of M. tetragenus. The cocci stain a rich 

 purplish-red, while the small influenza bacilli take on a light pink color. 



A greenish yellow, nummular sputum, often profuse, is frequently noted in in- 

 fluenza. 



T. B. sputum showing a mixed infection with streptococci or pneumococci or with 

 the influenza bacillus makes for a bad prognosis. M. tetragenus, which often is 

 present when cavities exist, does not seem to be so unfavorable prognostically. 



Red cells show up well in specimens stained by the Romanowsky method; if 

 rouleaux formation is marked, it may indicate pulmonary infarction. 



In culturing sputum a mucopurulent mass should be washed in 

 sterile water and should then be dropped into a tube of sterile bouillon. 

 With a sterile swab it should be emulsified and successive streaks made 

 along the surface of an agar, blood agar or glycerine agar plate. In 

 obtaining cultures from influenza sputum, first smear the material 

 thoroughly over a blood-serum slant; then inoculate, by thorough 

 smearing over the surface of successive blood-streaked agar slants, the 

 material on the surface of the blood-serum slant. The platinum loop 

 should be transferred from one slant to another without recharging. 

 The influenza bacillus seems to grow better if the blood-streaked agar 



