552 A MANUAL OF BACTERIOLOGY 



Clinical Examination 



1. Pour out the pus or discharge into a large capsule or Petri 

 dish so that it forms a thin layer, look for any yellowish or other 

 granules, pick them out with a needle, and place on a clean slide 

 in a drop of 50 per cent, glycerin. If no granules can be found, a 

 little of the discharge may be spread on a slide with a drop of 

 50 per cent, glycerin. Cover with a cover-glass, and apply a 

 little pressure. Examine with a f -in. objective. If any actino- 

 mycotic tufts are present they will be seen as yellowish or pale 

 brownish, spheroidal, ovoid, or reniform masses, and with a higher 

 power will be found to consist of tufts of filaments with, perhaps, 

 stunted clubs (in the bovine form rosettes of clubs will be seen). 



2. Stain films of the discharge, by Gram's method, with eosin. 

 The actinomycotic tufts will generally be found to consist of little 

 masses of tangled filaments stained violet, and surrounded by a 

 pink zone which has an indistinct radiating structure. 



N.B. In most instances the clubs in Actinomycosis hominis 

 do not stain by Gram's method. 



3. If the pus be otherwise sterile, cultures may sometimes be 

 obtained by inoculation into blood- or serum -broth and incubating 

 anaerobically. 



4. Sections of actinomycotic tissue are best prepared by the 

 paraffin method. If frozen, the actinomycotic nodules are very 

 apt to fall out. Sections may be stained by any of the following 

 ways: 



(a) By Gram's method, with eosin or orange-rubin. 



(6) With the Ehrlich-Biondi triple stain. Stain for from half 

 an hour to two hours. Place in methylated spirit until the 

 sections appear greenish, then pass through absolute alcohol and 

 xylol. The clubs are stained yellowish-brown, and are sometimes 

 shown in human cases when unstained by Gram's method. 



(c) By Plant's method. Stain in warm carbol-fuchsin for ten 

 minutes, rinse well in water, stain in a saturated solution of picric 

 acid in methylated spirit for five to ten minutes, rinse well in 

 water, place in 50 per cent, alcohol for ten minutes, pass through 

 absolute alcohol and xylol. 



(d) Good preparations may be obtained by staining in Ehrlich's 

 haematoxylin and counter-staining with orange-rubin. This may 

 also show the clubs when they are unstained by Gram's method. 



