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THE JOHNS HOPKINS HOSPITAL REPORTS

MONOGRAPHS. NEW SERIES No. VIII.

LEUKA:MIA OF THE FOWL:
SPONTANEOUS AND
EXPERIMENTAL

BY
HARRY C. SCHMEISSER, M. D.
+ (From the Department of Pathology of The Johns Hopkins University)

Illustrated: 4 Plates

BALTIMORE
THE JOHNS HOPKINS PRESS
1915

[Copyright, 1915, by The Johns Hopkins Press]

Monograph

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CONTENTS.

PAGE
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1. Preparation of Material, Methods of Injection, etc............ 20
eI RIS RTD oo odo. cle Fd ie ats wo nk Pe GER lo eee ecg 21
PERC FPR Ne oi. Cree Eis ee ego 8d 22
4. Autopsy Findings (gross and microscopical)................. 24

5. Table: Weights of Body, Organs and Relation of Organs to
ase Ne eR Seo Sk nee ance-e we Ae Se Wisse vleidie 0.4 0 apCetaens 30
ULE ea Rai aie BIE RS eG A pie RR SRM = SED 30
mR OMER ME SNENENE NS Og cng sake Sock SE ok We Oa Eo whi 8 co aon ae 31

V. Conclusion

LEUKAMIA OF THE FOWL: SPONTANEOUS AND
EXPERIMENTAL. * +

By HARRY C. SCHMEISSER, M. D.
(From the Department of Pathology of The Johns Hopkins University.)

I. REVIEW OF THE LITERATURE.

The first publication was made by Moore’ in 1895-1896, in which
he reported observations upon an epizootic disease of fowls which he
designated “infectious leukemia.” His studies were based upon cases
obtained from five different outbreaks, and upon those artificially
produced by means of the specific organism (Bacterium sanguinarium )
isolated from the organs of these fowls. The descriptions of the
spontaneous and experimental cases clearly show that he was not
_ dealing with a true leukemia of the fowl, but rather with an acute
infectious disease, characterized by fever, diminution in the number
of red blood cells, and increase in number of white blood cells, this
increase being “ restricted” to the polymorphonuclear leucocyte with
the eosin staining spindles, i. e., the phagocyte of the normal fowl’s
blood.

Butterfield,’ in 1905, reported three cases of aleukemic lymph-
adenoid tumors of the hen. His studies were limited to the livers
of these animals. They correspond in every detail, both grossly and
microscopically; were uniformly and strikingly enlarged (340, 420,
270 gm. respectively) and of a diffuse, light, cream, yellow color.
Histologically, the liver tissue was extensively replaced by large oval
or round, sometimes irregular, collections of cells. The intralobular
capillaries were filled with the usual nucleated red blood cells. Most
of the nodules of tumor cells were thought to be in connection
with the portal spaces. The cells resembled in general the “large

* Submitted for publication, October, 1914.

+ Dissertation submitted to the Board of University Studies of The Johns
Hopkins University, in conformity with the requirements for the Degree of
Doctor of Philosophy, 1914.

2 Harry C. Schmeisser.

lymphocyte” of the hen’s blood.* Allowing for flattening due to
pressure, these cells were round or oval, with round or oval nuclei,
mostly eccentric. The nuclear chromatin appeared granular or
reticular. The nucleus was not infrequently pale and vesicular with
one or more bright chromatin bodies. The cytoplasm was present as
a narrow rim, non-granular, and basophilic. Mitoses were common.
Between these cells was seen a delicate reticulum. In association with
these tumor masses were quite large collections of eosinophilic cells,
the majority showing a single, pale, round or oval, eccentric nucleus
and a relatively abundant cytoplasm, filled with large, round, elongated,
or spindle-shaped oxyphilic structures. They varied greatly in size
and shape. The lymphocytes and eosinophiles were occasionally
seen in the capillaries, but there was “certainly no definite increase
within the blood-vessels.” From the character of the cells, their rela-
tion to the portal spaces and absence of the lymphzmia, the condi-
tion was regarded by Butterfield as a proliferation of periportal
lymphadenoid tissue, and thought by him to merit the designation
“ aleukeemic lymphadenoma.”

Mohler had under observation five similar cases showing lymphemia
and the same lymphadenoid condition. Mohler’s description of two
of his cases, as presented by Butterfield, is as follows: The process
showed a tendency to involve several organs. In one case the liver
and spleen were the principal seats, the lungs and intestine were
also involved. The liver was enlarged to twice the normal size, of
very light color and mottled. Throughout its surface and within
its parenchyma were whitish lymphomatous deposits of varying
diameters. A section shows the liver tissue only partly obliterated
as a result of a filling up of the interacinous capillaries with leucocytes.
The spleen was about the size of an ordinary blue plum. Each lung
contained a white spherical area, the size of a pea and composed
of hyperplastic lymphoid tissue, infiltrating white cells. In the wall
of the large intestine, involving the lymphadenoid tissue, were round,
slightly raised nodules. In the second case the changes were most
marked in the kidney and spleen. The liver, intestine and heart
were also involved. The kidneys were three or four times the normal
size and had macroscopically very much the same appearance as the
liver in the first case. The myocardium contained some small, whitish

* Obviously the large mononuclear cell of the classification presented in
this paper.

.

f

DEC 28 1915

©cla418249

Leukemia of the Fowl. 3

areas. Butterfield, in comparing the histological findings in Mohler’s
cases with his own, finds a convincing similarity between, or identity
of, the infiltrating cells. In Mohler’s cases, the infiltration was more
diffuse and not so regularly arranged about the portal tissue. Above
all, the white blood cells were markedly increased in all the blood-
vessels, from the intraacinar capillaries to the largest veins. The
predominating white cell was indistinguishable from the cells in the
tumor masses of both these and Butterfield’s cases. From the fore-
going, Butterfield felt justified in granting “the existence of a
leukemic and aleukemic lymphadenoid hyperplasia in hens.”

Butterfield and Mohler deserve the credit of showing that leukemia
occurs in the fowl. Their studies were limited to post-mortem
findings.

To Warthin,’ in 1907, we are indebted for the first complete study
of this disease in fowls. He had the good fortune to secure a living
animal, which he kept under observation for two and a half months
before it died. During this time he had an opportunity of care-
fully studying the clinical course. The blood showed a great reduction
in the total number of red blood cells. These varied greatly in
size and shape, and contained less hemoglobin than normal. The
total number of white cells was greatly increased. Differential counts
showed a striking increase of the “large lymphocyte” at the ex-
pense of the other cells of the normal blood. Numerous mononuclear
forms of the eosinophiles, cells abnormal to the blood, were seen.
From the blood changes Warthin felt justified in making a diagnosis
of “lymphatic leukemia of the large cell type.” A careful autopsy
revealed the following findings: The liver was greatly increased
in size and was pale yellow in color. The spleen and kidneys were
also enlarged; both showed numerous yellowish spots from pin-
point to pin-head in size. In the hemolymph nodes small yellowish
nodules were found. The bone-marrow appeared as solid plugs, red-
dish-yellow and friable. The tissue changes were summarized as
follows: (1) Tumor-like nodules and infiltrations of lymphoid cells
in all the organs, particularly in the liver, spleen, kidneys, bone-
marrow and hemolymph nodes; (2) replacement of the normal white
cells of the blood by cells of the large lymphocyte type, more or less
atypical in character, but identical with cells in the tumor masses.
From the findings, both in the blood and in the tissue, Warthin made
the diagnosis of leukemic lymphocytoma.

+ Harry C. Schmetsser.

He likewise examined the material from two additional cases of
lymphadenoid tumors of the fowl. One proved to be of the aleukeemic
type, as described by Butterfield; the other was of the leukemic form,
as described above. The histological picture, apart from the increase
of white cells in the blood-vessels in the one case, was identical in
both cases and also with that in the cases given above (Butterfield’s,
Mohler’s and Warthin’s). From these cases Warthin concludes, there-
fore, that there occurs in the common fowl a condition of disseminated
lymphocytoma, sometimes aleukemic, at other times associated with
an increase of the white cells of the blood, this increase consisting in
the appearance of great numbers of cells of the large lymphocyte type
and apparently identical with the lymphoid cells of the tumor nodules.
He also regards the aleukemic and leukemic conditions as genetically
related, if not actually one and the same process in different stages.
Both conditions, he thinks, are in all respects analogous to the similar
ones existing in man.

The preceding cases appear to be very similar to those reported by
later investigators and designated by them as splenomedullary in
type.

Kon * in 1907, after the appearance of Warthin’s article, reported
what he considers a true case of leukemic disease of the chicken.
The gross and microscopical changes, which were studied and reported
in great detail, are in general identical with those of the cases above.
In addition, he found giant cells, though only a few, in the bone-
marrow, and mitoses in the basophiles as well as in the eosinophiles
in the kidneys. Because of the lack of smears, he determined the
ratio W/R and made differential counts from sectioned blood-vessels
of the liver. He found that the white cells, as compared with the
reds, were greatly increased and that the large mononuclear was the
predominating cell of the whites. Mitoses of this cell were observed.
The evidence given, he thinks, is sufficient to consider this a case of true
leukemia. Because of the enormous swelling of the spleen and the
plentiful occurrence of the characteristic cell in the pulp of the normal
spleen and to a lesser degree in the normal bone-marrow, and in the
absence of lymphatic swelling and the occurrence of this cell in the nor-
mal lymph tissue, it seems to him most likely that this is a case of true
“ splenic leukeemia.”

Soshestrenski, in May, 1908, reported a case of spontaneous
leukemia of the fowl, very similar to the case described by Kon.

Leukemia of the Fowl. 5

His study was limited to gross and microscopical findings at autopsy.
The bone-marrow was omitted. The contents of the blood-vessels
of the organs were found to be leukemic. He designates the case
also as “ splenic leukemia,” but in so doing he calls attention to the
fact that the process involves many of the other organs and appears
to be only primary and most marked in the spleen. There was
no enlargement of the lymph-glands.

With Ellermann and Bang,’" in 1908, the subject takes on a
different phase. Up to this time, the communications dealt entirely
with the recording of spontaneous cases of leukemia in the fowl.
Ellermann and Bang were the first to successfully transmit the
disease by experimental inoculations from a spontaneous case to
other, healthy, fowls. In fact, they claim to have been the first to
successfully produce the disease experimentally in any animal. They
report the findings in two typical spontaneous cases giving identical
pictures. The blood study during hfe was found to be practically
the same as that of Warthin’s case, with the exception that the large
mononuclears frequently contained in their cytoplasm many granules,
and thus represented cells which are not found in the normal blood—
~ myelocytes. Small granules alternated with very large ones. The
autopsy showed an enlarged spleen and liver, the latter with white
dots and streaks. The bone-marrow was gray-red. The other organs
presented nothing of interest. The microscopical changes were very
similar to those of Warthin’s case. The experimental leukemia they
transmitted to the third generation, producing a blood picture and
organic findings identical with those of the spontaneous cases.

Pseudoleukeemia, which is characterized by the same organic lesions
as are found in true leukemia, but in which the blood picture is
normal, they found to occur spontaneously among chickens. They
received from the same flock, at the same time, a leukemic and
pseudoleukemic animal. From the pseudoleukemic animal, by inocu-
lation, they produced a leukemic animal, and conclude, therefore,
that the leukemia and pseudoleukemia of chickens are etiologically
identical.

Multiple sarcomatosis of the peritoneum occurs epidemically in the
fowl. They consider this a manifestation of leukemia, because by
inoculation from such a case they produced an atypical leukemia and
carried this to the second generation.

They were successful in two out of five inoculations with a filtrate
from an emulsion passed through a “ Kerze aus Infusorierde” and

6 Harry C. Schmeisser.

conclude, therefore, that a cell-free filtrate is effective and that the
cause must be an organized virus.

In the same year, Ellermann and Bang * reported that the blood of
leukemic fowls contained the virus. They also showed that among
transmitted cases the disease may appear as a typical leukemia or as
a pseudoleukemia, or only as an anemia with changes in the bone-
marrow. The injection of five animals with a Berkefeld filtrate re-
sulted in one early leukemia. The injected fowls were isolated and
great care was exercised during the experiment.

The following year, 1909, Ellermann and Bang,” reported the
transmission of the disease into the sixth generation, and also an addi-
tional positive Berkefeld filtrate series. Because the disease can be
produced by a cell-free filtrate, they conclude: (a) Leukemia must
be an infectious disease; (b) it is to be placed among the diseases due
to a filterable virus. They call attention to the fact that mitoses in
the blood are pathological and always present in leukemia.

Schriddle,* in 1909, was the first to question whether, in leukemia,
we are really dealing with an infectious etiology. Basing his con-
clusions upon experimental work, he claims that chickens, injected
with extracts of entirely normal organs, present the same changes
as Ellermann and Bang have reported for leukemia. He thinks that
their findings are not leukemic and that, therefore, there is no proof
in favor of the infectious etiology of this disease.

Hirschfeld and Jacoby” in 1909, report a spontaneous case of
leukemia showing changes in the blood and organs which, they claim,
agree entirely with the description of Ellermann and Bang. In a
second case, the blood picture, although not typical, appeared to
them leukemic. At autopsy this fowl showed a typical chicken tuber-
culosis. They succeeded in transmitting this tuberculosis, in asso-
ciation with the apparently leukemic blood picture, into the fourth
generation. They did not know whether they were dealing with
a combination of tuberculosis and leukemia in the same animal, or
with pure tuberculosis. An animal injected with a pure culture of
the chicken tubercle bacillus developed a blood picture identical with
that of the second spontaneous case. Therefore it seems highly prob-
able that this case may have been one of pure tuberculosis. At any
rate, it cannot be accepted as a definite and pure case of leukemia.

The following year (1910) the same authors“ report the injection
of fowls subcutaneously from their first spontaneous case. They

0

Leukemia of the Fowl. vi

obtained no takes and their strain died out. From a leukemic animal
given to them by Ellermann and Bang, they transmitted the disease
into the fifth generation, obtaining both leukemic and pseudoleukemic
eases. In the blood, they found normoblasts and megaloblasts, 1. e.
small and large, round, nucleated, mostly polychromatophilic red blood
cells. Although they state that the disease begins in the bone-
marrow, they leave it an open question whether in the chicken one
is dealing with a myeloid or a lymphoid leukemia.

Burckhardt,* in 1910, advanced the theory, based upon investiga-
tions of his own on transmissible chicken leukemia, that a particular
leukemic virus does not exist, and that in the so-called chicken
leukeemia one is very probably dealing with a very chronic chicken
tuberculosis. As proof, he claims that with a pure culture from such
an animal (leukemic) and also with older pure cultures of chicken
tubercle bacilli, one can produce the same blood picture described by
Ellermann and Bang for chicken leukemia. The anatomical findings,
he states, likewise correspond. Gratz” suggested that possibly in
the culture of chicken tubercle bacilli from the leukemic animal, there
may have been cultivated, in addition to the tubercle virus, also the
- virus of leukemia. Friedberger™ considered this objection invalid,
because, as he claimed, one could produce the leukemic blood picture
also by using old laboratory tubercle cultures. ;

Burckhardt,* in 1912, still adheres to his theory, but admits that
he is unable ever to bring the blood into a leukemic condition by
inoculating with tubercle bacilli.

Hirschfeld and Jacoby” in the same year inform us that they
presented to Burckhardt and Friedberger the chicken with which
these investigators started their transmissions, and that on the day
of delivery it showed a pronounced leukemic blood picture. The
animal, in addition, surely had tuberculosis, because all the transmitted
leukemic animals of the stock from which this animal originated
had tuberculosis. They feel confident that in Burckhardt and Fried-
berger’s stock the tuberculosis had crowded out the leukemia, until
finally only tuberculous and not leukeemic animals were obtained. They
had a similar experience with the stock from which Burckhardt and
Friedberger’s animal originated.

Against the existence of an etiological relationship between tuber-
culosis and leukeemia, they present the following arguments: (1) The
blood picture in tuberculosis is never to be confused with that of

8 Harry C. Schmeisser.

leukemia. It is characterized by a hyperleucocytosis due to the
polymorphs. (Hllermann and Bang” had also stated this fact at an
earlier date.) (2) The organs of the tuberculous chickens were entirely
free from the changes which Ellermann and Bang, and they themselves,
found to be characteristic of leukemia and which correspond entirely
with leukemic lesionsin man. (3) By injections of organic emulsions
obtained from spontaneous tuberculous chickens, or of pure cultures
of chicken tubercle bacilli, they uniformly: produced a typical tuber-
culous blood picture and organic changes, always without any signs
of leukemia.

Ellermann “” in 1913 and 1914, in answer to the question whether
this chicken disease is really leukemia, shows that it has all the
symptoms of human leukemia. He meets Schridde’s objection by the
argument that this investigator’s claim was limited to the blood picture,
and in the absence of the characteristic organic changes, of which he
makes no mention, his experiments are of no importance. The in-
jection of an organic emulsion Ellermann never found to cause any
change in the blood, provided the material was not virulent. Ellermann
and Bang had previously produced the disease with Berkefeld filtrates
in three different experiments. In this paper Ellermann reports two
more successful series. All other investigators have had only negative
results. Thus, Hirschfeld and Jacoby “ were unsuccessful in two experi-
ments and Burckhardt ™ in one.

Ellermann is convinced that the filtrate experiments prove the
theory of infection, for all the cells were surely removed. He feels
that the fact that the virus passed through a rather thick-walled Berke-
feld filter demonstrates that it is an invisible filtrate virus. He clearly
shows that the leukemic virus can be separated from the virus
of tuberculosis by filtration and that, therefore, the two diseases
are distinct. Starting with an emulsion of spleen taken from an
animal, both leukemic and tuberculous, in which tubercle bacilli had
been demonstrated, he passed this through a porcelain (Reichel) filter
and with the filtrate he produced leukemia with complete absence of
tuberculosis in all the inoculated animals.

Ellermann finally states that both the spontaneous and the trans-
mitted leukemia occur in two types; (a) myeloid, (b) lymphatic. In
the first the blood is characterized by the presence of numerous
myelocytes and transitional cells, occurring in association with a pro-
nounced myelosis (large deposits of myelocytes) in the organs. In the

18 19

Leukemia of the Fowl. 9

second, the blood is characterized by a predominance of small and
large lymphocytes (90 per cent of the total number) occurring in
association with a pronounced lymphomatosis (numerous infiltrations
with lymphoid cells) of the organs. Of interest is the fact that a
myeloid type may occur in one generation and a lymphatic in the
next, or that both types may occur in the same generation. The
occurrence of both types in the same stock suggests very strongly
that both forms of leukemia in man are due to one and the same
infection.

Il. SrupiEs oN THE NorMAL Fowt.

With the hope of making the findings in the leukemic animals more
clear, the following brief data of the normal fowl are presented. These
are limited to those portions of the body which are involved in
leukemia. They are compiled from a large number of chickens.

EXTERNAL APPEARANCE,

Healthy, young, adult Plymouth Rock hen. Comb, featherless
area about eyes, and wattles bright red. Conjunctive and buccal
mucous membrane pink. Skin of body white with a faint yellow
tint. Well nourished, sides of sternum covered with a thick pad of
tissue, ribs not palpable. Average rectal temperature, 42 ° C.

BLOOD.

The blood for counting and smears was obtained from the vein under
the wing, which always appears to be well filled. From a small needle
puncture the blood flows under pressure, is thick, dark red and clots
quickly.

Blood-count.—The nuclei-containing red blood cells of the fowl
make it practically impossible to count the white blood cells by the
usual direct method. After laking the former, one cannot differ-
entiate between the nuclei of the two types of cells. It was there-
fore necessary to resort to the indirect method, 1. e., to determine the
proportion of white to red cells from a stained smear, count the total
number of cells, both whites and reds, by the direct method, and then
calculate the total number of each present.

The red blood cells vary somewhat: lowest limit, 2,500,000 ; highest,
4,500,000; average, 3,000,000-4,000,000 per c.mm. White blood cells:

10 Harry C. Schmeisser.

the extreme limits are between 20,000 and 80,000 per ec. mm. Propor-
tion of whites to reds, lowest limit, 1/40; average, 1/50-1/150.
Hemoglobin (Sahlv).—This shows considerable variation, lowest
limit, 45 per cent; highest, 75 per cent; average, 60-70 per cent.
Blood Smears.—Wilson’s stain was found to give the best results,
especially when applied immediately after the smears were made.
Classification of blood cells (Fig. 1).

Erythrocyte.

1. Normocyte (a): Elliptical disk. Nucleus, same shape as cell,
deep blue, slightly picnotic. Cytoplasm yellow and glassy. (Both cell
and nucleus uniform in size, shape and staining).

Blood Platelet (b).

Length of normocyte, width less than that of normocyte. Nucleus
round, purple, chromatin diffuse, diameter equal to width of its
cell. Cytoplasm pale gray with vacuoles about nucleus, frequently con-
taining small circumscribed red structures. May vary in size and
shape.

Leucocytes.

1. Polymorphonuclear leucocyte with eosinophilic rods (c) : Round,
diameter about length of normocyte. Nucleus has two or more lobes,
pale blue, chromatin diffuse. Cytoplasm colorless with bright red,
spindle-shaped rods.

2. Polymorphonuclear leucocyte with eosinophilic granules (d) :
About the same in shape and size. Nucleus has two or more lobes,
purple, slightly picnotic. Cytoplasm faintly blue with dull red
granules.

3. Lymphocyte (e, e’): Round, diameter about width of normocyte.
Nucleus round, purple, chromatin diffuse. Cytoplasm small in amount,
to one side of nucleus, pale blue. Same cell may be slightly larger.
Thus a division into the small (e) and large (e’) lymphocyte may be
made.

4. Large mononuclear cell (f): Round or oval, diameter about
length of normocyte (at times, more or less). Nucleus round, oval or
slightly irregular, and larger, otherwise similar to nucleus of
lymphocyte. Cytoplasm abundant, completely surrounds nucleus, pale
blue. (A suggestion of fine granules.)

Leukemia of the Foul. sta)
5. Mast cell (g): About same size and shape. Nucleus round or

oval, very pale blue. Cytoplasm abundant, colorless, mostly to one
side of nucleus with purple granules, some scattered over nucleus.

Differential Count, 300 Cells.

Per cent.
Polymorphonuclear with eosinophilic rods......... 29.6
Polymorphonuclear with eosinophilic granules..... 4.3
Lymphocyte .........csecccecnecccceccncecssccces 42.3
Large mononuclear Cell.......-.-+eseeeeeeeeeeeees 19.4
WES ae OSB ORE SD bn ODOG cle Linnea acm icranda tcno.o 2.2
Mnelassified sCClIS. oe ac os <1 ccc ce ele cc sie se soe cieielsims 2.2

100.0

AUTOPSY PROTOCOL.

The following findings would correspond to those in a fowl of the
average size and weight (1760 gm.) of the animals injected.

Subcutaneous Fat Abundant (Fig. 4). Thick muscle pad covering
' the skeleton (Fig. 4). Inner surface of skin slightly yellow.

Cervical Lymph-glands * (Figs. 3,a and b). Sometimes difficult to
find. Present as two chains, each consisting of six to ten glands,
one on each side of neck, lying upon the internal jugular vein, buried
in fat, and extending from the middle of neck to base of heart. They
are delicate, elliptical, flat, lobulated bodies, averaging 1 x .5 x .2 cm.
The parenchymatous lobules are pink and separated from each other
by narrow septa of fat. No other lymph-glands were found in the
entire body.

Thyroids and Parathyroids—Two of each gland. The thyroid, a
dark red, translucent, spindle-shaped body, 1 x .4 x .4 cm., with
just visible colloid-filled acini, separated by delicate gray septa. The
parathyroid is attached to its lower pole. This is a small bluish-
white sphere, 2 mm. in diameter. Thus, in pairs, they are found, one
pair on each side of the body, just above the heart.

Peritoneal Cavity (Fig. 4) —An omentum composed entirely of fat,
1 cm. in thickness, takes its origin from the anterior surface and lower
margin of the gizzard, extending over coils of intestine, ete., holding

* Kon‘ considers these structures the thymus. He‘ and Ellermann Bue
state that the fowl has no lymph-glands. Soshestrenski® asserts their
existence by stating that in his case they were not enlarged.

12 Harry C. Schmeisser.

them against the posterior peritoneal wall. It is fastened to the
parietal peritoneum, laterally, and at the pubis. The lower margins of
the right and left lobes of the liver extend down between the sternum
and omentum to within 3 cm. of the tip of the xiphoid.

Inver (Fig. 4) —The first organ which presents itself on removing
the sternum. Right lobe measures 7.5 x 4.5 x 2 cm.; left lobe, 6.5 x 5 x 2
cm. Total weight 50 gm. or 2.8 per cent of body weight. Uni-
formly reddish brown with smooth and glistening surface, soft and
friable. Lobule about pin-head in size, center darker than periphery ;
usually distinct, at times difficult to see. The cut surface shows small
blood-vessels on cross and longitudinal section. The lobulation is less
readily seen.

Spleen (Fig. 6).—Lies just behind the liver. Measures 2 x 1.5 x 1
cm., weighs 1 gm., or .05 per cent of body weight. It is small, soft,
reddish brown. Beneath its smooth and glistening capsule, may be
seen bluish white Malpighian bodies, slightly larger than pin-points, at
times indistinct. On section, the capsule appears very delicate;
trabeculz are few, but usually definite, containing gaping blood-vessels.
Malpighian bodies at times are prominent. Pulp does not rise above
edge of capsule.

Heart.—Lies within a delicate, transparent pericardium, containing
a small amount of clear serum. Myocardium is uniformly reddish
brown.

Lungs.—Bright red and of a characteristic construction, consisting
of a mass of entwined air tubules with large lumina and spongy walls,
separated from each other by a small amount of vascular connective
tissue. On section, this construction becomes more pronounced. The
wall of each air tubule projects as a pale ridge, separated from its
neighbor by a red linear depression, in which blood-vessels are common.

Gastro-intestinal Tract, Pancreas and Adrenals.—These are of little
interest except that beneath the serosa of the lower half of the intestine
are frequently seen pearly nodules, less than a pin-head in size.

Mesentery.—A delicate, cobweb-like membrane with some fat, but
no demonstrable lymph-glands.

Kidneys.—Measure 6.5 x 1.7 x 1 cm., and weigh together 12 gm., or
7 per cent of body weight. They are uniformly reddish brown with a
slightly nodular surface.

Bone-marrow.—Taken from long bones of upper and middle third
of leg. It is very soft, semifluid, bright red, mottled with yellow, i. e.,

Leukemia of the Fowl. 13

rich in red blood cells and fat. Lower part of middle third frequently
consists of a solid, yellow column of fat. The marrow cavity may be
almost occluded by bony septa.

MICROSCOPICAL EXAMINATION.

Cervical Lymph-gland.—Section shows lobules of parenchyma,
separated by fatty tissue. Each lobule is surrounded by a delicate
fibrous capsule and contains a very fine reticulum, in which the
lymphocyte is diffusely scattered. This is a small, round cell with a
round, deeply staining, picnotic nucleus, usually incompletely sur-
rounded by a narrow rim of pink cytoplasm. Sometimes red blood
cells are associated with the lymphocyte, but mostly they are confined
to and form the sole occupants of the numerous capillaries. They
appear as elongated, yellowish-pink cells, with a solid black rod or dot
for nucleus, according to whether this is seen in longitudinal or cross
section: Scattered through the lobule are small, sharply outlined,
hyaline structures, which Kon * considered as corresponding to Hassall’s
corpuscles of the thymus.

Thyroid.—Consists of polyhedral acini, which vary in size, are lined
- with cubical epithelium and filled with granular colloid. They are
separated from each other by a small amount of fibrous tissue, rich
in blood capillaries. Sometimes accumulations of lymphocytes may be
seen near the larger vessels.

Parathyroid.—Within its capsule is seen a prominent reticulum filled
with closely packed cells, whose nucleus is round or oval, semivascular
and larger than that of the lymphocyte.

Liver.—The lobule is difficult: to limit. Periportal spaces are not
easy to find. They consist of one or several arteries, a vein, one or
several gall-ducts, and very little or practically no fibrous tissue, usually
free from infiltrated cells, although lymphocytes may be present dif-
fusely or in small follicles. The liver cells are arranged in trabecule,
separated by capillaries. All blood-vessels and capillaries are filled
exclusively with red blood cells.

Spleen—The Malpighian bodies are indistinct, numerous, closely
packed, composed of masses of lymphocytes surrounding very small
arteries. Red blood cells are limited to the pulp, where they occur
more or less in clusters, although one is unable to demonstrate sinuses
or inclosures of any kind. The lymphocytes are also diffusely scattered
throughout the pulp. Veins tend to be rich in lymphocytes and
poor in red blood cells; the artery shows the opposite picture.

14 Harry C. Schmeisser.

Lung.—Presents a characteristic structure, entirely different from
that in the human lung. The tubules mentioned in the macroscopical
description are seen in cross, oblique, or longitudinal section, forming
large polyhedral units with wide walls composed of capillaries. The
intercapillary air spaces communicate with each other and with a large
central lumen. The latter is lined with a narrow band of smooth
muscles surmounted by endothelium, frequently cuboidal. The inter-
tubular fibrous tissue is small in amount, and contains large blood-
vessels, whose branches extend into the capillary bed. These structures
are closely packed with red blood cells. Only occasionally does one see a
lymphocyte. The bronchi resemble those of the human lung. Follicles
of lymphoid cells are frequently found in the submucosa or outside of
the bronchi near arteries.

Bone-marrow.—A low-power picture is very similar to human resting
marrow, 1. é€., one sees a framework of fatty tissue inclosing nests of
marrow cells. Arteries are filled solely with red blood cells. Venous
spaces are difficult to determine..

Classification of Marrow Cells——Very thin paraffin sections were
stained with hematoxylin and eosin and studied under the oil-
immersion lens. The same cells appear slightly different under these
conditions than when seen in the Wilson smears of normal and leukemic
blood.

Erythrocytes.

1. Normocyte (a): Elongated, varying in shape, due to pressure.
Nucleus a solid, black rod or dot (longitudinal or cross section).
Cytoplasm yellowish-pink and glassy.

%. Normoblast (b): Round, diameter about width of normocyte.
Nucleus round, uniformly black. Cytoplasm pink or faintly blue, and
glassy. Frequently a narrow clear zone is seen about the nucleus.

3. Megaloblast (b): Same, larger, about length of a normocyte.
Nucleus slightly picnotic.

Leucocytes.

1. Polymorphonuclear myelocyte with eosinophilic rods* (c):
Round, Nucleus, two or more solid black lobes. Cytoplasm colorless
with bright red, spindle-shaped rods.

* These are obviously the polymorphonuclear with eosinophilic rods, the
polymorphonuclear with eosinophilic granules and the large mononuclear
cell of the normal blood.

Leukemia of the Fowl. 15

2. Polymorphonuclear myelocyte with eosinophilic granules * (0):
Round, about same size. Nucleus has two or more slightly vesicular
lobes. Cytoplasm colorless with bright red granules.

3. Mononuclear myelocyte with eosinophilic granules (a): Round,
or oval, varying in size, mostly larger than 1 and 2. Nucleus round,
oval, or horseshoe-shaped, eccentric, slightly or very vesicular. Cyto-
plasm colorless with bright red granules.

4, Large mononuclear myelocyte* (d): Round, slightly larger.
Nucleus round, very vesicular, one or more nucleoli. Cytoplasm baso-
philic, moderate in amount.

Reticular Cell—Branched. Nucleus elongated, vesicular, several
nucleoli. Cytoplasm pink, giving off delicate fibers to form reticulum.

Lymphocytes, mast cells, platelets, mitoses of red and white cells
were not seen. Letters in ( ) =order of predominance.

Heart, Pancreas and Kidney: For all practical purposes similar to
the human. ~

Ill. A Case or SPontTANEOUS LEUKAZMIA.

On October 31, 1912, a typical case of leukemia of the fowl was
brought to the pathological laboratory. The animal, a Plymouth Rock
hen, had just been killed, and its body was still warm. Nothing is
known of its clinical history. An abstract of the findings is here
given.

BLOOD.

The study of the blood was limited to smears taken from the heart
just after the animal had been received at the laboratory. They were
stained by Wilson’s method. A detailed description of the cells of
leukeemic blood will be reserved for the experimental leukemia where
far better opportunities for careful study of the same were offered.
Suffice it here simply to enumerate and briefly comment upon those
cells which were present.

Blood Smears. There was an enormous increase in the white blood
cells, the proportion of whites to reds being 1: 1.3. The predomina-
ting cells were the large mononuclear and the mononuclear myelocyte
with eosinophilic granules. The latter is abnormal to the blood,

* These are obviously the polymorphonuclear with eosinophilic rods, the
polymorphonuclear with eosinophilic granules and the large mononuclear
cell of the normal blood.

2

16 Harry C. Schmeisser.

normal to the bone-marrow, and when extramedullar is typical of
leukemia. Lymphocytes and polymorphs were strikingly decreased.
Of the latter those with red granules and the mast cell were rare.
Mitoses of the large mononuclear were common. The red blood cells
appeared to be poor in hemoglobin and showed anisocytosis,
poikilocytosis and polychromatophilia. These cytoplasmatic changes
were usually associated with an increase in the size of the nucleus.
Premature red blood cells, normoblasts and megaloblasts were present in
large numbers.

Differential Count, 300 Cells.

Per cent.

Polymorphonuclear with eosinophilic rods..........
Polymorphonuclear with eosinophilic granules...... 0
TGV MPN OCY CC osha oe oh pes oe lee aerey aiisliacst a se auwaleyer el eledeileyeuene 2
Large mononuclear Celli. .ccsistosnie choke vi cssvets sim eichome alee 30
Mats GOLD is sre rats oie w ete teres Leteca anh aL outtake tszehe bayections one 0
Monuclear myelocyte with eosinophilic granules..... 52
Unelassified), (Cells seis sctercysieccysscte sleet teteveretore caste ove Retametalee 8

100

AUTOPSY PROTOCOL.

The animal was of medium size ; the weight was not taken.

External Appearance——Comb, featherless area about eyes, and
wattles of an ash color. Buccal mucous membrane and conjunctive,
very pale. Anterior chamber of left eye filled with an old blood-
clot. Right eye normal. A moderately firm slightly nodular tumor,
3.7 x 2 x 1.5 cm., occupied the triangular space on the left side of
head between the angle of the mouth, ear and angle of the lower jaw,
extending below the ramus of the latter. Subcutaneous fat was
practically absent. Emaciation was extreme and there was marked
muscular atrophy. A second, slightly smaller tumor, 1.7 x 1 x 1 cm.,
was found just inside the ramus and below the orbit, apparently
communicating over the ramus with the first growth and continuous
below with a slightly larger, third mass.*

Cervical Lymph-glands—Somewhat enlarged. Lobulation indis-
tinct.

* A more detailed description and study of these tumors will be reserved
for a future communication.

Leukemia of the Fowl. 17

Peritoneal Cavity Omental fat entirely absent. Lower margins of
right and left lobes of liver extended almost to the pubes, 1. e., far
below xiphoid.

Liver—Enormous. Dimensions and weight not taken. Its surface
was extremely mottled. It was reddish brown and specked with
innumerable gray or slightly yellow spots from a pin-point to a few
millimeters in diameter, frequently closely packed to form irregular
areas, the largest 1 cm. in diameter. In addition there were scat-
tered gray or slightly yellow nodules, averaging about 2 mm. in
diameter. Sectioned surface similar in appearance. The blood-vessels
were surrounded by a gray zone.

Spleen.—Enormous. Dimensions and weight not taken. It was
about the size of a small hen’s egg. Diffusely gray.

Heart—At the apex there were several gray spots, pin-point in size.

Kidneys.—Greatly enlarged. Both showed nodules, similar to those
in liver.

Bone-marrow.—Abundant, gray, with absence of fat.

Thyroids, Parathyroids, Lungs, Gastro-intestinal Tract, Pancreas
_and Adrenals.—Apparently uninvolved.

MICROSCOPICAL. EXAMINATION.

Cervical Lymph-glands—The normal stricture was somewhat
changed. There was complete disappearance of the interlobular fat
with approximation of the greatly swollen parenchymatous lobules.
Here and there were intra- and inter-lobular foci of myeloid tissue.
These frequently enclosed an artery or vein. There was also a more
diffuse infiltration of the parenchyma by myeloid cells, filling capillaries,
veins, arteries and in places breaking through the lobular capsule into
the interlobular tissue. Of the infiltrating cells two predominated: (1)
A large mononuclear, usually round, at times slightly polygonal due to
pressure, with a single, round, oval, or indented, vesicular, at times
multiple, nucleus with one or more nucleoli. Its cytoplasm was non-
granular and slightly basophilic. (2) The mononuclear myelocyte
with eosinophilic granules, described under normal bone-marrow.
Mitoses of these two cells were quite common. Normoblasts and
megaloblasts appeared to be present.

Itwer.—Here the process was most extreme. Very little liver tissue
remained. Everywhere were closely packed masses of myeloid cells
from which the liver trabecule had completely disappeared. These

18 Harry C. Schmeisser.

masses of cells surrounded blood-vessels, both arteries and veins, infil-
trating the walls of the latter and filling the lumina of both. Within
the vessels the white blood cells were present in about equal proportion
with the red blood cells. Between these more focal accumulations the
myeloid cells had infiltrated in a more diffuse manner between the liver
columns, spreading them apart. It was usually difficult to demonstrate
the capillary wall. Hepatic cells in these locations did not suffer so
much. Some of the above-mentioned, circumscribed infiltrations
showed a coarse sclerosis. The same infiltrating cells were present as
in the cervical lymph-glands, with a prevalence of the large mononu-
clear, which was very rich in mitoses, and the mononuclear myelocyte
with eosinophilic granules. The two types of polymorphonuclear cells
and the premature red blood cells were present in fewer numbers.

Spleen.—Pulp diffusely infiltrated with closely packed leukemic
cells, which crowded the reticulum, distended the blood-vessels and
separated and compressed the Malpighian bodies. The mononuclear
myelocyte with eosinophilic granules, although present, was scarce.
The large mononuclear, frequently showing mitoses, was the common
cell.

Heart—The intramuscular capillaries, were moderately distended
with the large mononuclears and red blood cells, present in about equal
proportion. The former had also infiltrated between the muscle fibers.
The mononuclear myelocyte with eosinophilic granules was only occa-
sionally seen, both within and without the blood-vessels.

Iungs.—The intertubular blood-vessels, their branches which extend
into the capillary bed, and also the capillaries, were distended with
leukemic blood, in which the large mononuclear seemed to exceed the
red blood cell. Mitoses were common. Infiltration of the tissue was
limited to two foci of myeloid cells. The one was composed principally
of the mononuclear myelocyte with eosinophilic granules and the large
mononuclear. Mitoses, especially of the first cell, were seen. Both
types of polymorphs were also present. The other focus was composed
exclusively of premature red blood cells, normoblasts and megaloblasts
showing mitoses.

Kidneys.—All the blood-vessels, from the largest to the capillaries of
the glomerular tuft, were distended with the characteristic leukemic
blood. Throughout the section there was a diffuse infiltration of
myeloid cells, especially between the convoluted tubules. Here and
there were foci, in some of which the mononuclear myelocyte with

Leukemia of the Foul. 19

eosinophilic granules predominated, frequently showing mitoses ; others
seemed to be composed exclusively of the large mononuclear. Here
mitoses were also common. In these foci the convoluted tubules had
either been pushed aside or had atrophied.

Bone-marrow.—This was greatly changed. It consisted of a closely
packed mass of white marrow cells, with complete atrophy of the fat
and great rarity of normocytes. One or two small focal areas of
sclerosis were present. The arteries were mostly empty. Some showed
a little blood containing a few large mononuclears. Those marrow cells,
which were present, both red and white, answered to their descriptions
given under normal bone-marrow, with the exception of the large
mononuclear myelocyte, which showed considerable variation in its
nucleus. This was either single or multiple, round, oval, or horseshoe-
shaped. The order of predominance had changed greatly. The large
mononuclear myelocyte was present in far greater numbers than any
other cell, and showed extensive mitoses. The normoblast and megalo-
blast with mitoses followed next in frequency; then the normocyte.
Polymorphonuclear myelocytes, those with eosinophilic rods and those
with eosinophilic granules, had disappeared entirely. Curious is the
fact that the mononuclear myelocyte with eosinophilic granules could
not be demonstrated with certainty. As in the normal marrow lymph-
ocytes, mast cells and platelets were not seen.

Thyroids, Parathyroids, Pancreas and Adrenals——These were not
studied.

SUMMARY.

In summarizing the changes in the blood and organs, there was
found :

1. In the Blood—(a) A great increase in the total number of white
blood cells. Although a total count was not made, this increase was
evident from the appearance of the blood smears, and of the blood-
vessels in sections. The actual proportion of white to red cells sub-
stantiated this fact. The differential count showed a marked increase
of the large mononuclear cell at the expense of the other white blood
cells of the normal blood. In addition, a true myelocyte appeared in the
circulation. Mitoses of the large mononuclear were common. (b) A
corresponding decrease in the total number of red blood cells. These
showed a drop in hemoglobin, variation in size and shape, basophilic
staining of the cytoplasm, and swelling of their nuclei. Premature
red cells, normoblasts and megaloblasts made their appearance.

20 Harry C. Schmeisser.

2. In the Organs.—(a) A diffuse and circumscribed, at times
nodular, infiltration of myeloid cells, in which the large mononuclear,
and mononuclear myelocyte with eosinophilic granules predominated,
although the other cells of the normal bone-marrow were at times
present. This myelosis involved especially the liver, spleen, kidneys and
bone-marrow, resulting in an extreme enlargement of the first three
organs. Almost all of the remaining organs showed infiltrations, but to
a less degree. (b) A change in the contents of the blood-vessels. The
proportion of white to red cells was greatly increased. The predomina-
ting cells were the same as those of the infiltration.

CONCLUSION.

In conclusion, if we consider that the cells, characteristic of both
leukeemic infiltrate and blood, were the same, and that under normal
conditions the mononuclear myelocyte with eosinophilic granules
occurs only in the bone-marrow and the large mononuclear only in the
marrow and blood, it is evident from the summarized facts, that the
above case must be considered a typical case of myeloid leukemia.

TV. EXPERIMENTAL LEUK@MIA.

With material from the above case, the disease was successfully trans-
mitted into the fifth generation. A total of 105 animals were used in
conducting many different kinds of experiments. Of this number, 22
in all have developed leukemia. In four additional animals, a definite
diagnosis could not be established, although the appearances were highly
suggestive. The strain has now been running for more than 13 months.

This paper will be confined to a report of those series which deal with
simple transmission of the disease by the injection of an organic
emulsion.

Five of these series were conducted, each consisting of 5, 10 or 15
fowls. The percentage of positives per series varied from 20-40 per
cent. Of the total number of 40 chickens injected, 13 became leukemic,
i.e. 32.5 per cent. In addition, a definite diagnosis could not be made
in three, although the findings were very suggestive.

PREPARATION OF MATERIAL, METHODS OF INJECTION, ETC.

The material used for transmission was an emulsion of liver, and
sometimes of spleen also. It was prepared by thoroughly macerating

Leukemia of the Fowl. 21

pieces of the organ in a mortar, after the addition of .9 per cent normal
salt solution. This was filtered through a single layer of fine linen or
a small amount of raw cotton, thus removing the fibrous tissue suf-
ficiently for intraperitoneal injection, although, when it was used in-
travenously, instant death frequently resulted. Hence it was found best
to filter also through two layers of filter paper with the aid of a suction
pump. This filtrate was found to be absolutely fiber-free, but still con-
tained blood and parenchymatous cells. A 15 per cent emulsion gave
the best results. The entire preceding process, as well as the following
steps, were carried out aseptically.

The fowls were injected either intravenously or intraperitoneally
with 10 cc. Both methods were sometimes combined, when a total of
20 ce. was administered. The vein selected for injection was the large
one under the wing.

Animals used for transmission were young, adult hens of the same
breed as the spontaneous case, and exemplified by the normal control.

CLINICAL HISTORY.

Incubation Pertod.—This is usually from five to six weeks. In rare
cases, it may reach a maximum of 16 weeks.

Signs and Symptoms.—The onset is usually rather abrupt, preceded
by no definite premonitory symptoms. The animal, which has pre-
viously appeared entirely normal, begins to show a slight but progres-
sive, at times a sudden and intense, pallor of its comb, the featherless
area about the eyes and the wattles. This is almost invariably associated
with jaundice, which is likewise progressive and usually reaches an
extreme grade. The combination of intense pallor and jaundice results
in a characteristic, yellowish-pink color, which gives the fowl a ghastly
appearance. The comb may be surprisingly red or extremely pale with-
out jaundice. The animal emaciates rapidly and to an extreme degree.
The actual loss in weight during the disease is striking and best illus-
trated by the following figures. No. 28, on the day of the onset of the
disease, weighed 1484.5 gm. The duration of the disease was three
weeks, four days. At death the weight was 897 gm. Therefore, there
was a loss of 587.5 gm. in 25 days, or at rate of 23.5 gm. per day.

At first the bird acts entirely normally, or it may appear slightly ill.
In a few days, however, it acts as if it were very sick, stops eating and
drinking, and stands about with head retracted, eyes closed and tail
drooping. It prefers to assume a squatting position. If made to move

22 Harry C. Schmeisser.

it does so very slowly and carefully. Weakness becomes more and more
marked, until on the last day it lies prostrated, with its head on the floor,
wings drooping, eyes closed and at times is dyspneeic.

Fever has never been observed.

Duration of the Disease—This is most frequently from one to two
weeks. One case exceptionally acute lasted only 33 hours. Two cases,
three days. The longest course was four weeks, four days.

‘Prognosts.—Having once begun, the disease progresses steadily to a
fatal issue. Only in one case out of 23 leukemic animals did a spon-
taneous cure result.

BLOOD.

In Gross.—With the onset of anzemia, the vein under the wing col-
lapses and the blood undergoes a change. It soon flows with the great-
est ease and rapidity, resembles pale yellow water and shows no tendency
to clot. The animal apparently would bleed to death from a pin-point
wound, if the hemorrhage were not artificially arrested.

Blood-count.—In most cases there is a progressive and extreme de-
crease in the total number of red blood cells. At onset the count may
be but slightly below normal, 2,224,000, while just before death it, at
times, reaches 630,000. More commonly, the lowest point ranges
between 1,112,000 and 1,408,000 per c. mm. In one case, the count
remained normal.

The count of white cells is invariably high, between 131,200 and
210,000 per c. mm.

Ratio of W/R=1/3—1/9.

Hemoglobin.—This usually falls steadily and reaches a very low
point. From slightly below normal, 40-50 per cent at onset, it fre-
quently drops to 10-15 per cent just before death. In several cases it
fell only to 23 per cent. In one case there was no change at all.

Blood Smears.—The morphology of the leukemic blood is excep-
tionally interesting. In addition to the enormous increase in the num-
ber of white cells, all of the cellular elements show marked changes and
many new forms make their appearance. Smears were stained by Wil-
son’s method.

Classification of blood cells (Fig. 2).

Erythrocytes.

1. Normocyte: (a) Normal (a), except for variation in the
amount of hemoglobin. (b) Showing anisocytosis, poikilocytosis and

Leukemia of the Fowl. 23

polychromatophilia, mostly associated with a swelling of the nucleus
and separation of its chromatin (0).

2. Normoblast (c): Round, diameter less than length of normal
normocyte. Nucleus round, of same color or slightly purple, but larger
and with more scattered chromatin than nucleus of normal normocyte.
Cytoplasm greenish-blue and glassy. Frequently with clear zone about
nucleus.

3. Megaloblast (d): Same, except diameter equal to or greater than
length of normal normocyte.

4, Mitotic Cells (e): All stages, from monaster to complete division
of nucleus. (a) Round or elliptical, diameter about length of normal
normocyte. Dense, deep blue chromosomes, massed in center of cell.
Cytoplasm greenish-blue and glassy. (b) Same, with two masses of
chromosomes in opposite extremes of cell. (c) Same (at times with a
slight constriction in the middle of the cell) with irregular, dense, deep
blue or slightly purple chromatin masses in place of individual chromo-
somes. (d) Same, with two nuclei, similar in appearance to those of
normoblast. (e) Same as normal normocyte with two nuclei, similar in
appearance to those of normoblast, but only slightly larger than nucleus
of normal normocyte. (a), (b), (c) and (d) may show polychroma-
tophilic cytoplasm.

Blood Platelets (f).

1. Same as normal, except that cell and nucleus are both larger.
2. Larger than normal with two nuclei.

Leucocytes.

1. Polymorphonuclear leucocyte with eosinophilic rods (g): Same
as in normal blood, at times possibly a little smaller.

2. Polymorphonuclear leucocyte with eosinophilic granules (h):
Same as in normal blood, at times possibly a little smaller.

3. Lymphocyte (7) : Same as in normal blood.

4, Large mononuclear cell: (a) As described in normal blood, with
a little less cytoplasm. (b) Same size to one half times larger than (a),
with both nucleus and cytoplasm paler (7). (a) and (b) cannot be
separated absolutely. Every gradation. (c) Mitoses (4): All stages,
from monaster to complete separation of nucleus. (a’) Elliptical,
diameter about one and a half length of normal normocyte. Dense,
purple chromosomes centrally massed. Cytoplasm pale bluish-gray,

24 Harry C. Schmeisser.

granular. (b’) Same, with two masses of chromosomes in opposite ex-
tremes of the cell. (c’) Same, with two purple nuclei. (d’) Same, with
three purple nuclei.

5. Mast Cell (2): As described in normal blood.

6. Mononuclear myelocyte with eosinophilic granules (m): Round
or slightly elliptical, diameter about length of normal normocyte or
longer. Nucleus, oval, pale blue with dense chromatin, eccentric.
Cytoplasm colorless with small and large, bright red granules, some
scattered over granules.

Cytoplasmatic Masses without Nuclei (nm): Round or slightly oval;
diameter varies, usually about width of normal normocyte. No nucleus.
Cytoplasm grayish-blue, sometimes with vacuoles.

This, then, is the blood picture common to all the leukemic cases.
The order of predominance of the white cells is typical of leukaemia,
differing from the normal, but agreeing with that of the spontaneous
case. The large mononuclear is present, by far, in greatest numbers.
The other cells of the normal blood are decreased. The polymorphonu-
clear with eosinophilic granules and the mast cell are very scarce. The
mononuclear myelocyte with eosinophilic granules, although not as
common as in the spontaneous case, can usually be demonstrated in
every leukemic animal.

The platelets are greatly increased in number.

Differential Count, 300 Cells.

Per cent.

Polymorphonuclear with eosinophilic rods.......... 6
Polymorphonuclear with eosinophilic granules...... 0
ADA g 06] NOTA TE eecee leone SIO GOL 6 Uslaidy Gee Gen OD OIacra a 4
Arsen MONOnUGlEAraCel lade ce eteeriers ices cieleretaiel etn 86
IESE COL G ieak este scuey een oom che vom elioteere lalate a ioneseachs ots lelieiteveveoecrenenele 1
Mononuclear myelocyte with eosinophilic granules... il
Wneclassitied eCell sir varclescic chars occa ey ale doioonaveneronet helena 2

100

AUTOPSY FINDINGS.

The weight of the animal in every case was far below its weight before
injection.* Two lowest weights, at autopsy, were 675 and 897 gm.
External Appearance.—The jaundice was found at times to involve
the skin of the entire body.+ The conjunctive and buccal mucous mem-
* Animals which remained negative invariably gained in weight.

+ Probably this was more frequent but not recognized, except when very
grave, because of the normal yellow tint of the skin.

Leukemia of the Foul. 25

branes were always very pale. The eyes, nose and mouth were never
involved. There was extreme emaciation, and great scarcity or entire
absence of subcutaneous fat. Muscular atrophy was usually marked, at
times practically only the skeleton remained (Fig. 5).

Cervical Lymph-glands.—Macroscopically, these were usually unin-
volved. In two cases they were definitely enlarged (Fig. 3, a and b), in
the one very much so, measuring 1.5x8x4 cm. and 2x1x.5 cm.,
respectively. They appeared uniformly gray. The lobules of the par-
enchyma were enlarged and the interlobular fat had disappeared. Ina
third case, they were of normal size and appearance, but showed several
gray nodules, 1 mm. in diameter. Microscopically, those glands which
appeared normal in gross were usually free from myeloid infiltration,
although their blood-vessels contained leukemic blood. The diffusely
enlarged glands showed a marked swelling of their parenchyma, with
complete atrophy of the interlobular fat. Myeloid cells, both the large
mononuclear and the mononuclear myelocyte with eosinophilic granules,
were scattered through the lobules. Foci, some composed entirely of
the first, and others exclusively of the second, were localized principally
in the interlobular connective tissue. They were rich in mitotic figures.
The gray nodules seen in gross proved to be masses of proliferating
myeloid cells. All the blood-vessels were filled with the characteristic
blood.

Thyroids and Parathyroids.—These were grossly and microscopically
uninvolved, with the exception of the blood-vessels, which were filled
with leukemic blood.

Peritoneal Cavity.—In every case the omental fat was greatly de-
creased. At best, it was present only in moderate amount. As a rule
it was replaced by a thin yellow membrane. Ascites occurred in six out
of 13 cases, at times in sufficient quantity to distend the abdomen. It
was always associated with a sero-fibrinous mass, which covered not only
the liver and most of the other abdominal viscera, but also spread over
the outer surface of the pericardium.

With enlargement of the liver, the lower margins of its two lobes
change their position in relation to the tip of the xiphoid. They
descend to this landmark and frequently extend far beyond. With the
exception of Nos. 31 and 70, every animal of the 13 had an enlarged liver
(Table 1). The margins of the right and left lobes of the smallest of the
enlarged livers were, respectively, 1 and 3 cm. above the xiphoid, while
the margins of the largest liver extended 3.5 and 3 cm., respectively,

26 Harry C. Schmeisser.

below it. In fact, the latter organ im situ (Fig. 5) was so enormous that
it filled the entire peritoneal cavity and was the only viscus visible on
entering the abdomen.

Liver—As mentioned, the liver was greatly enlarged in 11 cases
(Table 1), frequently enormous (Fig. 5). In the smallest of these
the right lobe measured 9 x 4.5 x 3.8 cm.; the left lobe, 7 x 5 x 3.8 cm.;
in the largest, right lobe 13.5 x 4.5 x 4 cm.; left lobe 12.5 x 4.x 3.5 cm,
On comparing them with the normal, these dimensions are not very con-
vincing, with the exception of the length of the lobes. As the organ
enlarges, it becomes distorted, because the limited cross-diameter of the
peritoneal cavity prevents its expansion in this direction and forces the
lobes to follow the costal framework, posteriorly. The long diameters
of the lobes are more reliable, because the organ can enlarge freely in
this direction.

The weight of the liver is a more accurate indication of its size,
especially when expressed in per cent of body weight, 7. e., the number of
grams of weight of organ per 100 gm. of weight of the animal at death.
Thus, the weights were found to range from 80-265 gm, 1. e., they were
over five times the normal or 6.2-10.3 per cent of body-weight, an
increase of 3.6+ times the normal (Table 1).

The external appearance of the liver may not differ much from the
normal, but usually it is very characteristic (Fig. 5). It may appear
diffusely gray, due to very closely packed subcapsular spots, pin-point to
2mm. in diameter. These are usually more scattered and translucent,
at times slightly yellow and opaque. They may fuse to form larger
areas with a diameter of 0.5-1 cm., or they may be arranged in a delicate
gray network. In addition, numerous gray or slightly yellow nodules,
ranging from 2-0.5 cm. in diameter are often present. The whole gives
the surface an extremely mottled appearance. Lobulation at times is
visible, usually indistinct. The liver may be slightly firm or friable.
The sectioned surface appears very similar to the external surface. Dots
and lines often form gray borders along the blood-vessels. The latter
also occupy the centers of some of the larger, circumscribed, gray areas.
The perivascular connective tissue in rare instances is bile-stained.
Microscopically, there is usually a diffuse infiltration of myeloid cells.
They are closely packed within and without the intralobular capillaries.
The liver trabecule show fatty degeneration and atrophy. The large
mononuclear, rich in mitotic figures, is by far the predominating cell.

Leukemia of the Fowl. 27

The mononuclear myelocyte with eosinophilic granules is very scarce.
Scattered through the section are also foci of large mononuclears, or of
mononuclear myelocytes with eosinophilic granules. Some contain both
types of cells. They abound in mitotic figures. The liver cells in these
foci have entirely disappeared. The small, gray nodules consist of cir-
cumscribed masses of myeloid tissue, usually occupying the periportal
spaces and composed of a central portion of large mononuclears sur-
rounded by the mononuclear myelocyte with eosinophilic granules. In
the blood-vessels and capillaries, the large mononuclear, at times, seems
to greatly exceed the red blood cell in numbers.

Spleen.—Except in Nos. 31 and 70, the spleen in every case was en-
larged (Table 1); usually very much (Fig. 6). The dimensions, in
contrast to those of the liver, are of definite comparative value. The
smallest of the enlarged organs measured 3x2x1.7 cm., the largest
4 x 3 x 2.3 cm., the latter being just twice the normal in every
dimension.

The weights were strikingly increased. The lowest was 6 gm., the
highest, 18 gm., 7. e. 18 times the normal, or .6-1.6 per cent of the body .
weight, 1. e., 32 times the normal.

The surface of the spleen may look normal, be reddish brown and
show beneath its capsule pin-point, gray Malpighian bodies. More fre-
quently one sees scattered, gray or slightly yellow spots and nodules
(Fig. 6), the first from 1-3 mm., and the largest of the second from
0.5-1 cm. in diameter. Again, the organ appears just diffusely gray. A
section (Fig. 6) appears usually the same as the surface. When dif-
fusely gray, the pulp is somewhat granular and bulges. Microscopically,
the entire pulp may be diffusely infiltrated with the large mononuclear
cell, separating and compressing the Malpighian bodies. In addition,
circumscribed, closely packed masses of pure large mononuclears or
mononuclear myelocytes with eosinophilic granules are scattered
through the section. Again, the myelosis may be present only in
foci, with practically no disturbance of the normal splenic structure.
Mitoses are abundant wherever the large mononuclear occurs. The con-
tents of the blood-vessels are the same as in the liver.

Heart.—As mentioned above, the external surface of the pericardium
is frequently covered with a serofibrinous mass. The heart itself may be
somewhat enlarged, otherwise it appears normal (Fig. 5). Micro-
scopically also there is little to be seen besides a great excess of the large

28 Harry C. Schmeisser.

mononuclear in the intramuscular capillaries, with an occasional small
extravascular focus of infiltration, consisting of the same cell.

Lungs.—These are seldom involved. In two cases they were cedemat-
ous. In one, the tubular walls were wide and gray, occluding the
lumina. On section, the latter case presented a similar but more marked
picture. Microscopically the intertubular blood-vessels, their branches
which extend into the capillary bed, and the capillaries themselves, are
always filled with leukemic blood. At times they are greatly distended.
The white cells are always increased in number, and sometimes the
blood seems to consist solely of closely packed, large mononuclears.
Mitoses are everywhere abundant. Very rarely, a few mononuclear
myelocytes with eosinophilic granules or large mononuclears are gath-
ered in small masses in the intertubular connective tissue.

Gastro-intestinal Tract, Pancreas and Adrenals.—These in gross
showed nothing of interest. No mesenteric lymph-glands were demon-
strated. The pancreas, microscopically in no case showed any infiltra-
tion of its parenchyma proper, but in several there were numerous nests
of myeloid cells in the interlobar fat tissue, usually surrounding one or
more capillaries, and composed of the large mononuclear, eosinophilic
myelocytes, both mononuclear and polymorphonuclear. Mitoses of both
the mononuclears were frequent. The blood-vessels in the parenchyma
and fatty tissue contained, as usual, the predominating large mononu-
clear.

The adrenals also reveal interesting findings histologically. It is
quite common for the intertrabecular capillaries to be enormously dis-
tended with the characteristic leakeemic blood and for the myeloid cells
to have infiltrated between the capillary wall and parenchymatous cells.
These nests contain mostly the large mononuclear, but now and again
they seem to be very rich in lymphocytes. The parenchyma has suffered
greatly; its nuclei stain poorly and frequently its cells show disinte-
gration.

Kidneys.—Of the 13 cases, the kidneys of five were enlarged. The
smallest measured 7 x 1.5 x 1.2 cm., the largest 7.2 x2.2x1.4cm. The
weight of both kidneys ranged from 12-26 gm., or 0.8-1.4 per cent of
the body-weight. In one case the kidneys were extremely oedematous
and weighed 32 gm. or 2 per cent of the body-weight.

The surface and section of the kidney may appear entirely normal,
but usually one can see, below its capsule, scattered, gray pin-point dots,

Leukemia of the Fowl. 29

delicate lines, or even one or two nodules, the largest 0.5 cm. in diameter.
Again, they may appear diffusely gray. In the one case, they were large,
succulent and translucent, due to edema. Microscopically, the kidneys
always show more or less distension of their blood-vessels and inter-
tubular capillaries with the characteristic leukemic blood, in which
white blood cells, at times, seem even to exceed the normocytes, and in
which the large mononuclear predominates. When capillary distension
is moderate, the tubules appear normal; but at times they are so enor-
mously over-filled as to cause extreme atrophy and degeneration of the
tubular epithelium, resulting frequently in circumscribed areas devoid
of any parenchyma. These areas tend to form a kind of zone about the
larger blood-vessels. The glomeruli are surprisingly free. The walls
and perivascular tissue of some of the vessels are infiltrated with
actively generating myeloid cells, especially the large mononuclear.
Bone-marrow.—This is always involved, usually more or less char-
acteristically, being increased in amount, moderately soft and red, mot-
tled with countless gray dots to slightly larger areas, or it may be dif-
fusely gray. In either case the fat is absent entirely and red blood
cells are very scarce. The marrow may be involved as a whole or only
in part. Now and again it has been converted into a uniformly gray,
dry and slightly firm column, which bulges from the marrow cavity.
In such cases it is obviously very rich in white cells, poor in red cells and
fat. Microscopically, the normal structure of the marrow is greatly
changed. The fatty tissue has completely disappeared and the marrow
cells are closely packed in a solid mass, in which the normocyte is
rather inconspicuous. The arteries in the center of the marrow are
poor in blood. One or two contain a few normocytes and large mononu-
clears. The description of the red and white cells, which are present
agrees with that given under normal marrow. Possibly the large
mononuclear is more commonly polygonal, due to pressure. Mitoses of
both the large mononuclear and the erythrocytes are common especially
the first. The order of predominance has changed from the normal.
The large mononuclear myelocyte is present by far in yreatest numbers ;
the normoblast, megaloblast and mononuclear myelocyte with eosino-
philic granules follow in about equal numbers; the normocyte comes
last. Both types of polymorphonuclear cells, as well as the lymphocyte,
mast cells and platelets were not seen at all. Mitoses of the mononu-
clear myelocyte with eosinophilic granules were not demonstrated.

30 Harry C. Schmeisser.

TABLE 1.—EXPERIMENTAL LEUKAMIA.

WEIGHTS, IN GRAMS OF BoDy, ORGANS AND RELATION OF ORGANS TO Bopy
AT AUTOPSY.

ae Body. Liver. ree Spleen. arta Kidneys. pace
Normal 1760 50 2.8 1 0.05 2: 0.7
1 1515 117 ites ll 0.7 20 its)
6 1200 124 10.3 15 12 10 0.8
12 MOpen 95 MOEN 15 sore ss a2
14 1573 125 7.9 18 aE By A 220
15 1660 116 6.9 10 0.6 20 12
248 x. Sets eee Seeks me Ace ne ote
28 897 85 9.6 6 ORF * ae
30 1410 107 163 17 0.6 ee As
31 675 20 2.9 3 4 5 0.7
53 alas 265 en 12 a arene
57 1070 91 8.5 18 1.6 15 1.4
704 AA 65 ane 4 ee 26 sents
103 1282 80 6.2 8 0.6 12 0.9

1 Not weighed.

2Very edematous.

3 Whole animal kept as a museum specimen. Illustrated, Fig. 5.
4 Spontaneously cured. Not leukemic at death.

SUMMARY.

In summarizing the essential points deduced by the simple trans-
mission of leukemia by the injection of an organic emulsion, we shall
consider :

1. Clinical History.—After an incubation period, usually from five to
six weeks, the fowl suddenly becomes pale, jaundiced, emaciates rapidly,
loses weight and shows signs of extreme weakness, followed almost with-
out exception by death in one to two weeks.

2. Blood.—(a) The total number of white blood cells is greatly in-
creased, resulting in a proportion of one white to three red blood cells.
The differential count shows a marked increase and predominance of
the large mononuclear over the other white cells of the normal blood,
which have decreased. The mononuclear myelocyte with eosinophilic
granules is present in more or less numbers in practically every case.
Besides the normal-appearing large mononuclear, one sees many larger,
pale forms. In these cells, mitoses, in all stages, typical and atypical,
are common. (b) The total number of red blood cells is correspond-
ingly decreased, with a marked fall in the hemoglobin. They present

Leukemia of the Fowl. 31

variations in size, shape and staining, associated with swelling of the
nucleus. Normoblasts, megaloblasts, with mitoses in all stages make
their appearance. (c) There is an increase in the number of the blood
platelets, associated with an increase both in the size of the cell and the
nucleus. The cells frequently contain more than one nucleus. (d) The
blood loses its power of clotting.

3. Organic Findings.—(a) There is a diffuse, focal, less frequently
a nodular infiltration of marrow cells, in which the large mononuclear
and mononuclear myelocyte with eosinophilic granules predominate.
The foci may be limited to the large mononuclear or granular eosino-
philic myelocytes, commonly the mononuclear, or may be composed of
both types. Extensive mitoses testify to active proliferation. This
myelosis affects most particularly the liver, spleen, kidneys and bone-
marrow, causing a great increase in the size of the first three organs.
Rarely, the cervical lymph-glands are also very much enlarged. Most
of the remaining organs and neighboring tissues may contain infiltra-
_ tions but not so extensive. (b) The relative proportion of white to red
- blood cells is greatly increased in the blood-vessels. The same cells pre-
dominate as in the infiltrations. (c) There are atrophic and degenera-
tive changes of the parenchyma, general atrophy of the adipose tissue,
ascites etc.

CONCLUSION.

The injection of an organic emulsion causes a picture of myeloid
leukemia in every respect similar to that of spontaneous leukemia as it
occurs in the fowl.

The chemical picture and changes produced in the blood and organs
are analogous to those which occur in human leukemia.

V. CoNCLUSION.

1. The spontaneous occurrence of myeloid leukemia of the fowl is
confirmed.

2. Myeloid leukemia in the fowl is transmissible by the intravenous
or intraperitoneal injection of an organic emulsion. The latter is in
confirmation of the work by Ellermann and Bang who, as I have stated,
were the first to successfully transmit the disease. They were followed
by Hirschfeld and Jacoby, whose successful transmissions, however,
seem to be limited to a strain, which had its origin in a fowl presented
to them by Ellermann and Bang. Burckhardt likewise transmitted the

3

32

Harry C. Schmeisser.

disease, but here again the stock animal came from Hirschfeld and
Jacoby, and therefore indirectly from Ellermann and Bang.

The above reported transmission is of special interest, because it

originated from an animal entirely independent and far remote from
those of the previous investigators.

i

je)

£0:

Til,

LITERATURE OF LEUK@MIA IN THE FOWL.

Moors, V. A.: Infectious Leukaemia in Fowls—A Bacterial Dis-
ease Frequently Mistaken for Fowl Cholera. Twelfth and
Thirteenth Annual Reports, Bureau of Animal Industry,
U. S., 1895-1896, 185-205.

. Burrerrietp, E. E.: Aleukemic Lymphadenoid Tumors of the

Hen. Folia hemat., 1905, 11, 649-657.

. Wartutin, A. 8.: Leukemia of the Common Fowl. Jour. Infect.

Dis., 1907, IV, 369-381.

. Kon, J.: Ueber Leukimie beim Huhn. Virchows Arch. f. path.

Anat., 1907, CXC, 338-349.

. SOSHESTRENSKI, N. A.: (Leukemia of the Chicken.) Uchen.

zapiski Kazan. Vet. Inst., 1908, X XV, 215-222.

. ELLERMANN, V., and Bane, O.: Experimentelle Leukimie bei

Hiihnern. Vorliufige Mitteilung. Centralbl. f. Bakteriol.,

1908, Theil 1, Orig., XLVI, 4-5.

Experimentelle Leukamie bei Hiihnern. Centralbl. f.

Bakteriol., 1908, Theil 1, Orig., XLVI, 595-609.

Experimentelle Leukaémie bei Hiihnern. Verhandl. d. d.

path. Gesellsch., 1908, XII, 224-225.

(Experimental Leukemia in Fowls.) Overs. v. d. k.

Danske Vidensk. Selsk. Torh., Kbenk., 1908, 65-94; ibid.,

1909, p. 261. é

Experimentelle Leukimie bei Hiihnern. Ztschr. f. Hyg.
u. Infektionskrankh., 1909, LXITI, 231-272.

SCHRIDDE: Gibt es eine infektiose Aetiologie der Leukamie?
Deutsche med. Wchnschr., 1909, XX XV, 280.

. HirrscHrexp, H., and Jacopy, M.: Zur Kenntnis der tibertrag-

baren Hiihnerleukaémie. Berl. klin. Wchnschr., 1909, XLVI,
159-160.

Uebertragbare Hiihnerleukimie. Berl. klin. Wehnschr.,
1909, XLVI, 314.

Leukemia of the Fowl. 33

14. Hirscuretp, H., and Jacopy, M.: Uebertragungsversuche mit
Hihnerleukimie. Ztschr. f. klin. Med., 1910, LXIX, 107-
120.

15. BurckHarpt, J. L.: (Beider 82. Versammlung Deutscher Natur-
forscher und Aerzte zu Konigsberg.) Ztschr. f. Immunitits-
forsch. u. exper. Therap., 1910, Theil 2, 810-811.

Ueber das Blutbild bei Hiihnertuberkulose und dessen
Beziehungen zur sogenannten Hiihnerleukaémie, nebst Bemer-
kungen tiber das normale Hiihnerblut. Ztschr. f. Immunitits-
forsch. u. exper. Therap. 1912, XIV, 544-604.

17. HrrscHFre.p, H., and Jacosy, M.: Uebertragbare Hiihnerleukaémie
und ihre Unabhangigkeit von der Hiihnertuberculose. Ztschr.
f. klin. Med., 1912, LX XV, 501-505.

18. ELLERMANN, V: Underségeler over Hgnseleukaemiens Virus.
Urgesk. f. Laeger, 1913, LX XV, 1685-1691.

19. ——— Untersuchungen iiber das Virus der Hiihnerleukimie.
Ztschr. f. klin. Med., 1914, LX XIX, 43-48.

16. —

EXPLANATION OF FIGURES.

Fic. 1—Normal blood of the fowl. Wilson’s stain. Zeiss objective 1/12,
eye-piece No. 3. Composite picture. (a) Normocytes. (b) Blood plate-
lets. (c) Polymorphonuclear leucocyte with eosinophilic rods. (d) Poly-
morphonuclear leucocyte with eosinophilic granules. (e, e’) Lymphocytes
(small and large). (f) Large mononuclear cell. (g) Mast cell.

Fie. 2.—Leukemic blood of the fowl. Wilson’s stain. Zeiss objective
1/12, eye-piece No. 3. Composite picture. (a) Normal normocytes. (b)
Normocytes showing anisocytosis, poikilocytosis, polychromatophilia,
with swelling of nucleus and separation of its chromatin. (c) Normoblast.
(d@) Megaloblast. (e) Mitoses of erythrocytes; all stages from monaster
to complete division of nucleus. (f) Blood platelets; both cell and nucleus
increased in size; increase in size with two nuclei. (g) Polymorphonu-
clear leucocyte with eosinophilic rods. (h) Polymorphonuclear leucocyte
with eosinophilic granules. (i) Lymphocyte. (j) Large mononuclear
cell; same size and larger than normal with both nucleus and cytoplasm
paler. (k) Mitoses of large mononuclear cell; all stages, from monaster
to complete separation of nucleus. (1) Mast cell. (m) Mononuclear
myelocyte with eosinophilic granules. (n) Cytoplasmatic masses without
nuclei.

Fie. 3.—Cervical lymph-glands of the fowl; normal and leukemic. Actual
size and X 2. Note the great increase in size; replacement of fat by the
greatly swollen parenchymatous lobules.

Fie. 4—Normal fowl with organs in situ. Weight, 1750 gm. Note the
well-nourished condition of the body; the abundance of fat, subcutaneously,

34 Harry C. Schmeisser.

in the neck, and in the omentum; the large muscles; size of heart and
liver.

Fic. 5—Leukemic fowl with organs in situ. Weight, 1350 gm. Note
great emaciation of body; almost entire absence of fat; atrophy of muscles;
large heart; enormous liver with the characteristic myeloid infiltrations.

Fie. 6.—Spleen of the fowl; normal and leukemic, actual size; surface
and section. Note the enormous increase in size; in upper half of surface,
circumscribed areas of myeloid infiltration; the swollen pulp and infiltra
tions on section.

DESCRIPTION OF PLATE I.

Fic. 1—Normal blood of the fowl. Wilson’s stain. Zeiss objective 1/12,
eye-piece No. 3. Composite picture. (a) Normocytes. (b) Blood platelets.
(c) Polymorphonuclear leucocyte with eosinophilic rods. (d) Polymor-
phonuclear leucocyte with eosinophilic granules. (e. é) Lymphocytes
(small and large). (f) Large mononuclear cell. (g) Mast cell.

Fig. 2.—Leukemic blood of the fowl. Wilson’s stain. Zeiss objective
1/12, eye-piece No. 3. Composite picture. (qa) Normal normocytes. (0b)
Normocytes showing anisocytosis, poikilocytosis, polychromatophilia, with
swelling of nucleus and separation of its chromatin. (c) Normoblast.
(d) Megaloblast. (e) Mitoses of erythrocytes, all stages from monaster
to complete division of nucleus. (f) Blood platelets; both cell and nucleus
increased in size; increase in size with two nuclei. (g) Polymorphonu-
clear leucocyte with eosinophilic rods. (nh) Polymorphonuclear leucocyte
with eosinophilic granules. (i) Lymphocyte. (j) Large mononuclear
cell; same size and larger than normal with both nucleus and cytoplasm
paler. (k) Mitoses of large mononuclear cell; all stages, from monaster
to complete separation of nucleus. (1) Mast cell. (m) Mononuclear
myelocyte with eosinophilic granules. (n) Cytoplasmatic masses without
nuclei.

THE JOHNS HOPKINS HOSPITAL REPORTS. PLATE 1,

Fig. 1.

Fic. 2. Dorothea Pennington fec.

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THE JOHNS HOPKINS HOSPITAL REPORTS. PLATE Il.

ene ere rey

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Actual size.

glands of the fowl; nor-
mal and leukemic. Actual size and xX 2. Note
the great increase in size; replacement of fat by
the greatly swollen parenchymatous lobules.

Was rods 14

pal

Normal spleen
actual size

Surface Section

Leukzemic spleen
actual size

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Surface Section

kes ad uy

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Fic. 6.—Spleen of the fowl; normal and leukemic, actual
size; surface and section. Note the enormous increase in
size; in upper half of surface circumscribed areas of myeloid
infiltration; the swollen pulp and infiltrations on section.

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Fig. 4.—Normal fowl with organs in situ. Weight, 1750 gm. Note the
well-nourished condition of the body; the abundance of fat, subcutaneously,
in the neck, and in the omentum; the large muscles; size of heart and liver.

THE JOHNS HOPKINS HOSPITAL REPORTS. PLATE IV.

Fie. 5.—Leukemic fowl with organs in situ. Weight, 1350 gm. Note great
emaciation of body; almost entire absence of fat; atrophy of muscles; large
heart; and enormous liver with the characteristic myeloid infiltrations.

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