MATERIAL FROM THE LIVING SUBJECT 233 



cavity, and of the pathogenic organisms, pneumococci, streptococci, 

 and occasionally diphtheria bacilli are found. Usually clinical signs 

 or an abnormal appearance of the sputum, mouth, or throat lead 

 to a microscopic examination of the material from this region and, as 

 a rule, the nature of the symptomatology is a reliable guide to the 

 stain to be used. Among the organisms which stain by Gram's method, 

 pneumococci, streptococci, staphylococci, Micrococcus tetragenus, 

 and occasionally Diplococcus crassus are the more common spherical 

 organisms. Micrococcus catarrhalis, the meningococcus and para- 

 meningococcus are the only Gram-negative cocci, so far as is known. 



Of the Gram-staining bacilli, the diphtheria and pseudodiphtheria 

 bacilli together with Bacillus subtilis and rarely Bacillus anthracis 

 may be found. The bacillus of Friedlander, typhoid, influenza, pertus- 

 sis, plague and glanders bacilli are Gram-negative. Bacillus fusiformis 

 and Vincent's spirillum are Gram-negative as well. They color some- 

 what indistinctly with Loffler's methylene blue and very distinctly 

 with Wright's or Giemsa's stain. Mouth spirals and Treponema 

 pallidum are best stained with the latter method. Tubercle, leprosy 

 and nasal secretion bacilli (Karlinski) stain with the acid-fast stain. 



Higher bacteria and moulds are occasionally identified in material 

 from the buccal cavity. Actinomyces, Oidium albicans, aspergillus, 

 mucor, streptothrix, and yeasts have been detected. The virus of 

 poliomyelitis has also been demonstrated in material from the naso- 

 pharynx which has been freed from bacteria by passage through a 

 Berkefeld filter and injected into a monkey. 



For the routine examination of sputum, three stains are ordinarily 

 employed Ziehl-Neelsen for tubercle bacilli, Loffler's alkaline methyl- 

 ene blue for diphtheria, pseudodiphtheria, and fusiform bacilli (and 

 Vincent's spirillum), and the Gram stain, using dilute carbol fuchsin 

 as a counterstain for pneumococci, streptococci, influenza, and per- 

 tussis bacilli principally. Smith's stain for sputum (see page 186) 

 is advantageous for pneumonic sputum. 



The organisms mentioned previously but not detailed in the routine 

 examination of sputum are of comparatively rare occurrence. They 

 must be studied by purely cultural methods. 



Cultural Methods. Antiseptic gargles should not be used before 

 collecting sputum or material from the mouth or pharynx for cultural 

 examination. Sputum or exudate, obtained in a suitable manner, is 

 first washed through six or seven portions of sterile salt solution, if its 

 cohesiveness permits, to remove or diminish surface contamination. 



