EXPERIMENTAL TERATOMA TESTIS I N FOWL PRODUCED BY
INJECTIONS OF ZINC SULPHATE SOLUTION
PRELIMINARY
COMMUNICATION
L. I. FALIN
AND
K. E. GROMZEWA
(From the Histological Section of the Medical Institute in Smolensk, U. S. S. R.,
Director, L . I . Falin)
In 1925 Michalowsky produced a teratoid tumor of the genital glands of
the fowl by the injection of a small amount of 5 per cent zinc chloride solution. During the following five years he obtained 9 more experimental zinc
teratomas in fowl, and his observations were subsequently confirmed by his
associates Anissirnova and Falin, as well as by a number of foreign authors,
including Bagg and Ljvraga.
Since in all these experiments a zinc chloride solution was used the question naturally arises as to its specific nature in the production of these complex tumors arising in the fowl from the pluripotential cells of the testes. I t
may be asked: (1) Is it possible to obtain teratoid tumors of the testes by
applying other salts of zinc, particularly zinc sulphate, or is this effect peculiar to zinc chloride? ( 2 ) Is the zinc itself a specific agent or can teratoid
tumors of the testes in fowls be produced by the injection of other substances
exerting a similar chemical action on the tissues. The present communication
is concerned only with the first of these questions, describing experiments with
a 10 per cent zinc sulphate solution.
Taking into account the seasonal factor in the formation of experimental
teratomas, because of the development, in the spring, of active spermatogenesis in the fowl, as noted by Michalowsky, we carried out our experiments
in March and April, 1938. Twenty-nine roosters were given injections: 2 2
011 March 17, March 21, and March 28, and 7 on April 17. In the course of
the first two weeks after treatment 3 died.
The April series ( 7 fowls) gave negative results. Three of the fowls
(Nos. 2, 3, 4) treated in March, however, had tumors originating from the
testis, In all cases the tumors were unilateral. The histories of these birds
follow :
BIRDNo. 2 ( 5 7 9 7 ) : M a r c h 77, 7938: Each testis was injected with 0.15 C.C. of 10 per
cent zinc sulphate solution.
M a y 28, 7938: An exploratory laparotomy was done: The left testis was large, but
showed no particular changes. I n the lower part of the right testis was a large tuberous
tumor of a deep pink color.
M a y 29, 1938: The fowl was sacrificed, seventy-four days after injection of the testes.
I n the right half of the peritoneal cavity was a large, soft tumor arising from the middle of
the right testis and covering the whole anterior surface of its lower pole (Fig. 1). The
tumor with the remnant of testis weighed 66.15 gm. The left testis weighed 7.8 gm.
Microscopic examination showed a large amount of fatty tissue distributed in islands
and richly supplied with blood vessels and capillaries. The tumor also contained great
233
234
L. I. FALIN AND K. E. GROMZEWA
numbers of glandular ducts lined with columnar or sometimes with ciliated or goblet epithelium. I n all preparations many islands of hyaline cartilage of various size and form
were found. There was no evidence of osteogenesis. I n some places in the connective
tissue there were accumulations of pigment cells with long outgrowths and dark-brown
granules of pigment in the protoplasm. There were also present irregular areas of clear
FIG. 1.
BIRDNO. 2. TERATOID
TUMOR
OF
THE
RIGHTTESTIS
tissue rich in nuclei and resembling the embryonic anlagen of nerve tissue. I n these areas
there were often ii rosettes ” formed by columnar cells located around a small cavity (glial
elements?).
BIRD No. 4 (1823): March 28, 1938: Each testis was injected with 0.2 C.C.of 10 per
cent zinc sulphate solution (pH 6.4).
May 26, 1938: The fowl died, fifty-nine days after injection. It was much emaciated,
weighing 1176 gm. I n the peritoneal cavity was a tumor of enormous size, of blood-red
color (Fig. 2 ) . Examination showed that this originated from the left testis, a remnant of
which was found in the upper pole of the tumor on its dorsal side. The substance of the
testis was densely adherent to the tumor substance and gradually merged with it. The tumor measured 11.5 X 7.0 X 5.0 cm. and weighed 238 gm. The right testis showed no significant change. Its weight was 4.1 gm. The left lung showed inflammatory changes.
Other organs exhibited no alterations.
The stroma of the tumor consisted chiefly of embryonic connective tissue and great
numbers of blood vessels which, particularly in deep portions of the tumor, showed stasis
or even thrombosis. I n tissue taken from the more superficial portions of the tumor there
were great numbers of peculiar dark cells previously described by Michalowsky, Bagg, and
Anissimova (in our laboratory) in the experimental teratoma testis of the fowl produced
by injections of zinc chloride. These dark cells were distributed in the connective tissue
of the tumor in the form of voluminous nests and foci. I n some cases they showed a
special orientation suggesting glandular ducts. Among the accumulations of dark cells were
numerous epithelial strands and tubes lined with clearer cells. Many of them showed intensive cornification which resulted in the formation of typical “ horny pearls.” These
areas composed of nests of dark cells and accumulations of “ horny pearls ’’ gave a cancerous aspect to the tumor. Numerous islands of hyaline cartilage were present, however,
and in many of them a process of endochondral ossification developed. Beside the foci of
osteogenesis, areas consisting of smooth muscle tissue were found. The muscle fibers appeared as bundles running in various directions, at times piercing the foci of dark cells
described above. I n places were groups of typical pigment cells located in the embryonic
connective tissue or embedded in the basal layers of the epithelial lining of the glandular
ducts.
BIRD No. 3 (492): March 28, 1938: Each testis was injected with 0.2 C.C. of 10 per
cent zinc sulphate solution (pH 6.4).
EXPERIMENTAL TERATOMA TESTIS IN FOWL
235
May 28, 1938: Exploratory laparotomy showed a large tumor of the right testis, of a
brown-red color.
Jane 7, 1938: The fowl died, seventy-one days after injection. I t weighed 1560 gm.
I n the peritoneal cavity was a fairly large amount of liquid and coagulated blood. I n the
right half was a round tumor about the size of a child’s fist, of a dark red color.
FIG.2. BIRDNo. 4. TERATOID
TUMOROF
THE
LEFTTESTIS,POSTERIOR
SURFACE
TUMOROF
FIG.3. BIRDNo. 3. TERATOID
THE
RIGHTTESTIS
The tumor arose from the right testis, the remnant of which, in the form of a prominent white body, was located on the medial surface of the tumor, gradually merging with
it. The tumor had a free surface throughout, except at the upper pole, where it was adherent to the testis. I t was found to arise from the middle portion of the lateral border of
the testis, widely separating its upper and lower poles (Fig. 3). The bulk of the tumor
consisted of a very soft, spongy tissue of a red color. The remaining substance, particularly close to the testis, showed a clearer tint and contained a number of cysts. The weight
of the tumor was 91.5 gm. and it measured 7.0 X 5.5 X 3.7 cm.
236
L. I. FALIN AND K. E. GROMZEWA
Microscopic examination showed a stroma consisting of an embryonic connective tissue
rich in cells and containing many hematogenic elements (eosinophil leukocytes). Great
numbers of blood vessels and capillaries piercing the tumor substance showed stasis and
thrombosis, as in the previous case. It is of interest that the walls even of the largest vessels dilated by blood consisted of but one layer of flat endothelial cells.
In various portions of the tumor were many cavities of diverse size and form lined
with cuboidal or columnar epithelium. There occurred, also, epithelial strands, consisting
of stratified epithelium, in the center of which a process of cornification took place, with
the formation of “ horny pearls ” of various sizes and of the same type as i i Bird No. 4.
In the central portions of the tumor, which appeared macroscopically as a red spongy
tissue, there were many large cavities filled with blood. Their walls often consisted of a
single epithelial layer. I n larger cavities the wall was composed of several layers of smooth
muscle cells. Between these cavities lay great numbers of rather thick bundles of smooth
muscle fibers running in different directions and in their totality having some resemblance
to the corpora cavernosa.
Our observations may be summarized as follows. The injection of small
amounts of 10 per cent zinc sulphate solution (0.15-0.2 c.c.) into the genital
glands of the fowl produces in a considerabli percentage of cases (15 per cent
of all birds treated in March) rapidly growing tumors of these glands. Being
and glandular elements,
composed of a great variety of tissues-epithelial
cartilage, developing bone, smooth muscle tissue, pigment cells, embryonic
connective tissue, anlagen of nerve elements-these tumors must be classified
as teratoid neoplasms and in this respect are quite similar to the teratomas
produced by Michalowsky and others with zinc chloride.
The production of teratoid tumors of the testes by injections of both zinc
chloride and zinc sulphate solution shows that the ions C1, and SO, do not
play any particular r61e in the mechanism of the tumor-producing effect by
the zinc salts in the genital glands of the fowl. Further experiments must
show whether other substances having a similar influence on the tissues, but
having nothing in common with the zinc salts in their chemical structure, may
produce a similar effect.
The teratomas obtained by us are characterized by rather rapid but unequal growth. One of-the tumors (Bird No. 4 ) attained a weight of 238 gm.
in fifty-nine days. The other two weighed 66.5 and 91.5 gm., seventy-one and
seventy-four days after injection. The tumor distinguished by its rapid
growth (Bird No. 4) contained an enormous quantity of dark cells and horny
pearls,” which gave to it some similitude to cancer.
As in the experiments performed by Professor Michalowsky, the seasonal
factor, associated with the beginning of spermatogenesis in the fowl, plays an
important r61e in the origin of the zinc sulphate teratomas. All three tumors
of the testes described by us were obtained in birds treated in March. This
emphasizes once more the significance of the hormonal factor in the pathogenesis of the experimental zinc teratomas.
‘(
REFERENCES
BAGG,H. J.: Am. J. Cancer 26: 69, 1936.
LJVRAGA,
P.: Pathologica 26: 726, 1934.
MICHALOWSKY,
I.: Centralbl. f. allg. Path. u. path. Anat. 38: 585, 1926; Virchows Arch. f
path. Anat. 267: 27, 1928; 274: 319, 1929.