768 PARASITES IN TUMOURS 



witch the parasite penetrates into the interior of the cell, and in growing displaces 

 and compresses against the cell wall first the nucleus and afterwards the protoplasm : 

 the parasite is set free by the destruction of the infected cell, after which the capsule 

 of the parasite itself bursts and the spores thus set free infect neighbouring cells. 

 The most important method of propagation is said to be that which takes place in 

 the infected cell : the cancer cell divides by karyokinesis, giving rise to two daughter 

 cells, both of which are infected. 



Ruffer's method. Examination of fresh preparations. With a scalpel remove a 

 little of the cancer juice, transfer it to a slide, cover with a cover-glass, and examine 

 with an high power dry objective. Using an Abbe condenser rounded spaces can be 

 seen within some of the epithelial cells resembling, at first sight, vacuoles, but 

 surrounded by a membrane with a double contour and containing a body the 

 structure of which is difficult to make out with the condenser. If however the 

 condenser be removed it will be seen that this body consists of a nucleus surrounded 

 by a ring of homogeneous protoplasm. 



Stained preparations can be obtained by mixing a drop of cancer juice with a 

 drop of the stain, covering with a cover-glass and luting with paraffin. The best 

 stain for the purpose is an aqueous solution of methylene blue containing a little 

 aqueous solution of methyl green and very slightly acidified with acetic acid. This 

 mixture stains the cell nucleus green and the cell protoplasm very pale blue, while 

 the nucleus of the parasite is stained pink and its protoplasm pale blue. 



Sections. Fixing. Place small pieces (4-5 mm.) of the tumour for 1224 hours in 

 a saturated solution of perchloride of mercury. Wash in running water. Harden 

 in different strengths of alcohol. Mount in xylol-paraffin. 



Staining. Fix the section on a slide, remove the paraffin, pass through alcohol, 

 water, Gram's solution, alcohol and water successively. Stain by the Ehrlich- 

 Biondi method 1 which stains the nuclei of the epithelial cells green, the nucleolus 

 intense red, and the protoplasm red ; while the nucleus of the parasite is stained 

 red and its protoplasm is practically unstained. 



The following is a better method : 



(a) Stain for 1 or 2 minutes in a 5 per cent, aqueous solution of hsematoxylin. 

 Wash in water. 



(&) Wash the section in a concentrated solution of copper sulphate until it becomes 

 black. 



(c) Transfer to a 1 per cent, solution of hydrochloric acid until the section is pale 

 yellow. 



(d) Wash again in the copper sulphate solution for a few seconds until it assumes 

 a blue colour. Wash in a large quantity of water. 



(e) Stain with an acid dye, e.g. a concentrated tincture of cochineal. 

 Preparations stained in this way show the cell-nucleus blue and the protoplasm 



reddish blue. The parasite is red. 



At the present time very few pathologists regard the formations which have just 

 been described as Coccidia. Borrel, Fabre-Domergue, Duplay and Cazin, A. Brault, 

 Sikorsky, De Quervain have all expressed themselves as opposed to the theory that 

 a coccidium is the cause of cancer. Without going into details of the objections 

 which they have raised against the parasitic origin of malignant disease it may be 

 said that the different descriptions which have been given of the supposed parasites 

 in tumours are mutually conflicting and in no way recall the forms characteristic 

 of the development of Coccidia ; and that on the other hand there is a complete 

 morphological similarity between the pseudo-parasites and the various appearances 

 presented by cells undergoing degeneration. According to A. Brault the figures 

 described as Coccidia in tumours are cellular modifications of one or other of the 

 following types : 1. encapsulation of certain cells : 2. hyaline degeneration : 3. pro- 

 duction of endogenous cells : 4. excessive budding of the nuclei : 5. multiple 

 degenerations of the nuclei and nucleoli. 



1 The method is as follows : Prepare saturated aqueous solutions of methyl- violet, acid- 

 fuchsin (Rubin S.) and aniline-orange. Leave for 3 or 4 days. Then mix 100 c.c. of 

 the orange solution, 50 c.c. of methyl-green and 20 c.c. of acid-fuchsin. Filter. Dilute 

 1 volume of the mixture with 60-100 volumes of water. Leave the preparations in the 

 diluted mixture for 24 hours, wash in strong alcohol, absolute alcohol and xylol and 

 mount in balsam. 



