Home | Accessory Kit | Marsh CD Collection | Library | Contact Us |
Dr. O. Cameron. Gruner |
On the Marsh Rife Audio CD's Dr. Rife talks at great length about his work with Dr. Guner on the Cryptomyces Pleomorpha Fungus. Below is Dr. Gurners written paper on this organism. CRYPTOMYCES PLEOMORPHA: A NEW ORGANISM ISOLATED FROM THE BLOOD OF A THIS report is presented for the following reasons: Isolation.-From the blood of a case of carcinoma of the breast (excised five years ago) with intrathoracic metastasis; previously, in other cases, from the tumour tissue. Cultural characteristics.-The initial growth appeared only after seven days' incubation at room temperature. After developing a raised formation like that of sporotrichum, but extremely hard in consistence, it remained stationary for two weeks, despite repeated attempts at subculture on a great variety of media of all gradations of pH. Finally, the use of the patient's serum, and asparagus extract to enrich the glycerine-glucose-peptone water, proved to induce cultural success, good growths now appearing in all subsequent cultures in 48 hours. The organism was strictly aerobic. Room temperature was best. The optimum pH was 6.8 to 7.0. Bacterial contamination effectually antagonized the cultures. Fig 1.-Petri dish culture, 12 days old. The central raised growth consists of the torula-like spore forms, and is of coral colour in the original. The delicate hairy outgrowth is the mycelial phase. On asparagus glucose glycerine peptone water, a pellicle forms in 48 hours; this is in the form of a ring at the surface, adhering to the glass, and made up almost entirely of mycelia, the spores falling to the bottom of the tube in large numbers. A pink colour appears in 7 to 10 days, and the ring can be detached from the glass, when it will remain intact and hanging in the medium. Gelatine.-Not liquefied; very slow growth. Meat-extract media.-Very slow growth; colonies very small; whitish; no colour. Plain glucose glycerine agar.-Extremelyslow growth; small glistening colonies; no colour; no mycelium. Sabouraud.-Very slow colourless growth; very small and few hyphre; spores much smaller than those on asparagus media. Dextrose-tartaric acid.-No growth (in distinction from usual fungi). Potato.-Very scanty dull yellow growth, slowly turning pinkish. Tomato.-Very luxuriant growth in 48 hours; pink colour slightly paler than the flesh of the fruit. The condensation water rapidly fills with torula forms and turns pink. Milk.-Budding torula forms develop in 72 hours; no coagulation; no acid. Peptone water.-Noindol. Nitrate media.-No nitrite formation. Sugar-reactions.-No fermentation of glucose; gas, but no acid in lactose; saccharose, nil. Stainability.-Dilutecarbol-fuchsin gives the best results. Methylene blue stains relatively feebly. It is Gram-positive. It is not acid-fast. Rossophilia is almost absent. Morphology.-This organism is very pleomorphic. Thus at different times, in the same culture tube, one may see transient micrococci like Doyen's (1) M. neoformans of 1904 (Fig. 8), spherical forms like Sanfelice's blastomyces (1896) (Fig. 7), tubular forms apparently answering v. Brehmer's (2) description of his Siphonospora (Fig. 8) ; oval resting spores like those found in a specimen of Schmidt’s (3) "Blastolysin" (Fig. 9); combined with mycelial formations (Fig. 11). The following forms of reproduction are observable, as met with in other fungi: (a) fission forms or oidia, or diplospores (Fig. 10) suggestive of schiwsaccharomyces; (b) bud-formation as usual in torula or saccharomyces (Fig. 11); (c) ascospores, or "megaspores" (Fig. 12) in keeping with zygosaccharomyces; (d) very minute microspores (Fig. 14), which are highly refractile and arise by detachment from (b); these are metachromatic with Leishman and with methylene blue; (e) chlamydospore formation (Fig. 15); (f) lateral buds (Fig. 28), as in typical sporotrichum; (g) sessile sporangia in the aerial mat (Fig. 16), suggestive of some mucors; (h) still smaller particles occur, possibly filter-passing. There is a fairly regular sequence. Thus from b to d, from b to mycelium (first week), from mycelium to f, and to a, and to e (second week); from a to c and to g. The whole series ends in about three weeks, when a new subculture becomes essential. DISCUSSION Of some interest is the experience of the animal lesions produced, in that the organism is found closely mingled with the reactive infiltration of predominantly monocytic type, and shows in the sections appearances which strongly recall those seen in sections of carcinomas and sarcomas, though such particles are usually regarded as unessential bacterial or degenerative components. Applied to human tumour histology, the presence of such microscopic objects would 'seem to require an explanation. Of further interest is the evidence of sexuality in this organism (in common with other fungi), though the question of haploid and diploid phases has not been taken up. The delay in obtaining good subcultures would be explained by the difficulty of finding a medium favourable for the conjugation of the + and - elements. This provides a useful suggestion that such occupants of tumour tissue (whatever their presence means) present a sexuality factor which determines ease or difficulty of culture outside the body. The repeated observation of the megaspore forms inside the phagocytic cells in the infected animals with their mimicry of the Plimmer-body and other inclusions in human cancers supports this suggestion quite definitely. Very grateful thanks are due to Dr. Archibald and to Dr. Mark Kaufmann for enabling this case to be worked out, and also to Dr. Pauline Beregoff, for recently undertaking further tests on animals. REFERENCES 2. V. BREHMER, W.: Krebs eine Erregerkrankheit, Fortschrit. der Medizin, 1932, 50: 697; edit. in Brit. M. J., 1934, 2: 520; annotation in The Lancet, 1934, 2: 609. 3. SCHMIDT, O.: Ewing's Neoplastic Diseases, W. B. Saunders, Phila., 3rd ed., 1928, p. 125. |