550 



NA TURE 



[April 6, 1899 



March 9. —"A rrelimiiiary Note upon certain Organisms 

 isolated from Cancer, and their Pathogenic EfTecIs upon 

 Animals." By li. G. Plimmer, Pathologist, and Lecturer on 

 Pathology and Bacteriology, St. Mary's Hospital, London. 

 Communicated by Prof. J. Rose Bradford, K. R.S. 



During the past six years I have been studying the cell- 

 inclusions found in cancer, and for this work I have had to 

 examine 1278 cancers taken from various organs and parts. 

 Out of this large number of cases there have been a few — nine 

 in all — in which the cell-inclusions have been extremely 

 numerous ; so that at the growing edge, and even far into the 

 tumour, scarcely a cell could be found without an inclusion, 

 sometimes with as many as thirty-six, even, of these inclusions 

 in one cell ; and these bodies have been similar to those which 

 Metschnikoff, Ruffer, and others, as well as myself, have re- 

 garded and described as parasites, standing in causal relationship 

 to the disease. 



It will be seen that I mean by " cell inclusions" those bodies 

 found in cancer cells which are not recognisable as any known 

 degeneration, and which do not form any part of the cell. 



I have succeeded in isolating from the last of these cases, an 

 organism, which is pathogenic, in a peculiar manner, to certain 

 animals, and whose virulence I have been able to keep un- 

 impaired for some months. 



Previous Work on the Experimental Production of Tumours 

 itt Animals, 



The only work, I think, that needs mention here is that of 

 Sanfelice, in Cagliari, and of Roncali, in Rome. Sanfelice has 

 produced tumours in animals with organisms which he isolated 

 from infusions of various fruits ; and .they both have isolated 

 organisms from cancers. But Sanfelice's organism appears to 

 have been very difficult to isolate in a virulent form from human 

 cancer, and to keep virulent ; so that in his last paper,' he 

 treats only of the organisms derived from fruit infiisions, and of 

 their effects upon animals. Most of their statements are doubted 

 by the German pathologists, including such a good observer as 

 Baumgarten. But I do not find any reason to doubt any of 

 Sanfelice's statements ; and I think that he deserves the greatest 

 credit for removing the study of the ajtiology of cancer from 

 the histological to the experimental region of work. 



On the Method of Isolation adopted. 



The cancer, from which the organisms described were 

 isolated, and with which my experiments have been made, was 

 taken from the breast of a woman aged thirty-five years ; it had 

 a history of only two months' duration, and it was growing 

 rapidly at the lime of the operation. Immediately after re- 

 moval, I examined a fresh scraping, and, finding such an 

 extraordinary number of the bodies I have mentioned in the 

 cells, I cut thin slices from the growth, which I placed with a 

 little of the juice scraped from the cut surface in a flask con- 

 taining the following liquid, which was of course carefully 

 sterilised. This medium consisted of an infusion made from 

 cancer, just as the ordinary beef infusion is made, to which was 

 added, after careful neutralisation, 2 per cent, of glucose and I 

 per cent, of tartaric acid. Upon this medium scarcely any 

 bacteria, however hardy, will grow ; so that contaminations are 

 not common. 



Then, remembering that in the body these organisms were 

 under anaerobic conditions, I exhausted the air from my flasks, 

 and passed hydrogen into them, finally sealing them up. This 

 I have found is of great importance as regards the maintenance 

 of the virulence ; and I find that my cultures are as active now as 

 they were four months ago. liy these means, I got, after from 

 three to five days, a pure culture of the organism described, 

 which has been kept growing in this, and various other media, 

 ever since. 



Morphology and Relation to Media. 



The organism is apparently a saccharomyces ; but, according 

 to some authorities (such as De Bary, Cubrmi, and Duclaux), 

 the saccharomyceles are nothing but the developmental stages of 

 fungi which really belong to cither the I'hyco-, Asco-, or 

 Basidio-mycetes. Moreover, they state that in some species of 

 mycelium-forming fungi, single parts, especially conidia, can 



' "Ziilschiifl for Hygiene," 1899. 



NO. 1536, VOL. 59J 



grow in the saccharomyces form on certain nutrient media ; so 

 I will not attempt to locate this organism at present. Sanfelice 

 and Roncali, however, definitely state that the organisms they 

 have isolated are Blastomycetes. 



When grown in the medium described, these organisms 

 produce a cloudiness which becomes visible in about forty-eight 

 hours, and increases till about the sixth day, when the growth 

 sinks to the bottom, the medium then becoming clear ; no scum 

 or pellicle is formed. 



When grown on this medium solidified with agar, the 

 organisms form small round colonies which remain separate ; 

 after some weeks the colour, which was originally w hite, becomes 

 yellow ; the colonies do not attain a large size at any time. 



Gelatin is not liquefied, but the growth on this medium is 

 never luxuriant. On potato a thick white layer is formed, 

 which in about two weeks will cover the entire surface, changing 

 then to a yellowish brown colour. 



They will grow aerobically, but not well, at any rate at first ; 

 and they lose their virulence in a short time, when grown in 

 this way. 



Microscopically they are round bodies, frequently growing in 

 clumps, with a central portion which stains deeply, and, in 

 most cases, with a thin, strongly refractile capsule, which some- 

 times shows a double contour ; but young forms can be seen 

 which are without a capsule. The size varies from 0"004 mm. 

 to o'04 mm. 



Their reproduction appears to be by budding ; but I have 

 fancied that I have also seen, in a few instances, endogenous 

 budding. 



These bodies correspond morphologically with those found in 

 the original tumour, and also with those described by Ruflfer 

 and myself, and by some others of those who have worked at 

 the microscopical appearances of cancer. 



Experimental Results. 



These can be summarised under the four following divisions. 

 Up to the present, I have not been able to make any such 

 experiments upon animals as would allow of the easy bringing 

 of the organisms into contact with a likely epithelial surface, 

 with the exception of the cornea ; but, through the kindness of 

 Dr. Bradford, I have been enabled now, at the Brow n Institution, 

 to inoculate a hitch in the mamma;, but the time is as yet too 

 short to enable me to make any statement as to the result. 



The cultures used in the experiments were made in the 

 medium previously described. 



(1) Negative results. Rabbits inoculated intravenously and 

 intraperitoneally ; and rabbits and guinea-pigs inoculated sub- 

 cutaneously. The animals were killed in from fifteen days to 

 fourteen weeks. I have left none longer than this. 



(2) Those animals in which death was produced without any 

 obvious lesion ; but from the organs of which pure cultures 'of 

 the organism vvere made. These consisted of rabbits which 

 were trephined, the organisitis being then placed under the 

 dura mater. The organisms were present in the brain, cord, and 

 viscera. 



(3) Corneal inoculation, in rabbits, in which true neoplasms 

 were produced. There was considerable proliferation of the 

 corneal epithelium, which had forced its way in all directions 

 from the point of inoculation, deep down into, and between, 

 the fibrous layers of the corneal tissue. The organisms were 

 found in the epithelial cells. 



(4) Positive results : the animal dying with the production of 

 new growths. These results are found in guinea-pigs inoculated 

 intraperitoneally. Death ensues in from thirteen to twenty 

 days : and the liver, lungs, and peritoneum are found studded 

 with new growths of a w hite colour, which are of an endothelial 

 nature. Pure cultures could be made from the growths. 



The important point of this work is : the experimental pro- 

 duction of malignant tumours in animals by an organism isolated 

 from a malignant tumour in man. That these experimental 

 tumours are, so far w ith one exception, of endothelial origjn is 

 due to the fact that until I was enabled to inoculate a dog, I 

 found it very difticull to get the organism in cont.act with likely 

 epithelium ; all the above methods of inoculation, save one, 

 could only bring them into contact with endothelial surfaces. 

 The corneal experiment is the only one in which an epithelial 

 surface was tried ; and in this case the great proliferation of the 

 epithelium, the appearances of the organisms in the cells, and 

 the irritation produced, are very striking. 



