smmococcnc LYMPHADENITIS
-IN THE GUINEA PIG

THESIS FOR THE DEGREE 0F 1!. S. _

» Claude H. Reading
1931

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STDSPTCCOCCIC LIKPHJDENITIS

IN THE GUINEA PIG

STRJPTOCOCCIC LIEPIADEIITIS

IN TWE GUINEA PIG

THESIS

Submitted to the faculty of the Kichigan
State College in partial fulfillment of the

requirements for the degree of faster of Science.

By

Claude Kifléeading

June 1951.

 

 

THESIS

 

 

CON ENTS

Introduction ............. 1
Review of literature --------- 3

Etiology and cultural characteristics- 7

Pathogenicity - ----------- 8
Acute lymphadenitis ————————— 9
‘Chronic lymphadenitis ........ 10
Technical methods ---------- 12
Experiment I ------------- l4
Experiment II ———————————— 21
Experiment III ------------ 28
Experiment IV — - - - - - - 4 - - - - 34
Experiment V ............. 40
Discussion -------------- 53
Summary - - ------------- 56
Tables ---------------- 58
Plates ---------------- 62
Acknowledgment - - - - - ; ------ 77
Bibliography ------------- 78

103400

INTRODUCTION

From time to time during the past eighteen months
several guinea pigs have been found in our colony with
abscesses in the lymph glands. In.the fall of 1930 this
condition became of considerable importance as it appeared
in our stock pigs with a tendency, in many cases to run an
acute course. Kany of the piss showing enlarged glands and
and many of those heretofore apparently healthy suddenly
became sick and died. Yew projects involving the use of
guinea pigs could not be started as the stock pigs were
badly affected and new cases were appearing rapidly. It was
due to this condition that an investigation of this problem
was undertaken.

There are no data available upon.uhich an accurate esti-
mate of the economic importance of this disease can be based.
Eouever, it occurs frequently in guinea pig colonies and,
once established, is a very stubborn condition to eradicate.

There appears thus far to be no effective prOphylactic
nor curative treatment. Furthermore, no diagnostic procedure
has been recommended that has a high degree of efficiency in
,severe epizootics. Megrail and Eoyt (14) found that the
serum of guinea pigs which had abscesses possessed no agglu-
tinins for the strains of streptococci isolated from.their
abscesses. Several have made frequent manual examinations
for swellings of the lymph glands and subsequent isolation
of all affected animals, together with the exercise of
hygienic methods of handling. Here again one meets with

difficulties. Due to the insidious onset and chronic course

usually manifested, the disease nay be present for a long
period of time in a latent form in some hidden part of the
body and the organism be eliminated from the body openings
such as the eyes, nasal passages, vagina, etc. from time to
time. It may even produce internal abscessation whidh es-
capes the most careful examination. As a chronic case may
at any time develop septicaemia std discharge numerous bac-
teria in the body excretions, an.animal thus affected may be
a severe menace to all guinea pigs tith which he comes in
contact.

Cne of the important requirements of the biological
sciences is the maintenance of good health in.laboratory
animals. Diseased animals are a severe handicap to any
scientist who has to use thenifor experimenta1.work. Their
use leads to incorrect interpretation of pathological lesions,
confuses clinical symptoms, and the presence of their
etiological organism.may make it impossible to obtain cul-
tures of the organism.used in the experiment. This is es-
pecially true with animals spontaneously infected with strap-
tococcus. For example, it is sometimes difficult, especially
on gross examination, to differentiate between streptococcus

lesions and those due to Brucella abortus.

 

REVIEW OF LITERATURE

Not a great deal of literature on Strepmococcus diseases
in the guinea pig is available. That covering its pathologic
aspects is especially meager. nest authors have dealt mainly
with the bacteriological phases, only mentioning the more
conspicuous gross pathological lesions.

Among the earlier reports Malassey and Vignal (2) in
1883 and 1884 described an infection.which they named
"Tuberculose Zoogleique". The lesions are large, mass-like
structures containing large numbers of an organimn which was
undoubtedly streptococcus. This was not, however, a spon-
taneous epidemic. The disease was produced by injection of
material from a subcutaneous abscess in a child dying of
tuberculous meningitis. ho tubercle bacilli were found in
the pus from this abscess, but after several animal passages
the coccoid organisms disappeared and the tubercle bacillus
was found. Also the animals survived longer after inocula—
tion. In 1885 Eberth (1) described gross and microscopic
lesions in the lungs, liver, abdominal lymph glands, and
spleen, a condition which he named pseudotuberculosis. He
also found chain formations of cocci. Chantemesse (5) in
1887 and Dor (4) in 1888 also reported infections Which were
probably the same as those of Malassey and Vignal. Dor's
infection.in gninea.pigs, however, was experimental. He
obtained the material from.an abscess in a rabbit.

Epizootics have apparently taken different forms as
reported by various twentieth century writers. In 1901 Weber

(5) reported an epizootic affecting chiefly the lungs.

Eagner (6) in 1904 found the uterus involved, mainly causing
abortion of fetuses and puerperal endometritis. Teacher and
Burton (8), in 1914, reported an epizootic Which caused
nearly all of their breeding sows to abort or give birth to
immature young.- The placentas showed degeneration and exten—
sive necrosis with large numbers of streptococci in the
uterus and on the placenta. Boxmeyer (7) in 1907 reported
an extensive epizootic of lymphadenitis due to a gram posi—
tive staining hemolytic streptococcus in which over 3000
guinea pigs were affected. Studies were made on over 100 of
these in which he found 90 % to have affected lymph glands,
and occasionally the liver, uterus, and lungs were involved.
A few cases developed septicaemia and pyemia. In pigs three
to four weeks of age the infected cervical glands appeared
as palpable, shot-like nodules about the size of small peas
beneath the skin. These, on sectioning, were found to be
edematous and congested. As the pigs grew older, the glands
increased in size and developed areas of softening which
fused to form abscesses. The abscesses enlarged rapidly,
occasionally reaching the size of a hen'ssegg. Endothelial
cells and fibroblasts formed large groups of mononuclear
cells between the lymph follicles which involved them as
they increased in size. In more advanced cases these foci
softened in their centers, increased in size, and fused
together to produce abscesses. Infection.was found to take
place early in the life of the animal largely through
abrasions in the throat from.eating contaminated rough foods.
Experimental infection was produced in this manner. The or-

ganism was found to be pathogenic for-guinea pigs, rabbits,

and mice. Adult guinea pigs were much less susceptible than
immature animals.

In 1916 an article was published by Holman (10) on
"Spontaneous infection in guinea pigs", in which streptococcus
infections were found in 94 guinea pigs. Streptococcus pyo-
genes was isolated from 49 cases of pneumonia, ten cases of
acute endometritis,and occasionally from.various other parts
of the body. Only one cervical abscess was found.

Zardinbergh (11) in 1926 reported an epizootic in.whiCh
65 guinea pigs out of a.colony of 750 were affected. In 58
guinea pigs a total of 89 abscesses was found. The sub-
maxillary lymph nodes in 45 cases were involved, the cervical
in 26, the precrural in fourteen, and the parotid and ingui-
nal each in two. Pure cultures of hemolytic streptococcus
were found in 51 of these cases. Subcutaneous inoculation
killed two out of three guinea pigs and produced a local
abscess in the third. Three guinea pigs were fed cultures
of hemolytic streptococcus. Two of these remained well, and
the third developed pneumonia,and streptococcus was obtained
from.the lungs. Inoculation.via the conjunctiva did not
develop any disease. Spontaneous infection ran a chronic
course (only two cases died) while the artificial infection
usually ran an acute fatal course. (The organism.was patho-
genic for white mice, white rats, and rabbits.)

In 1928 Parsons and Hyde (12) examined 189 cases of
streptococcio infection in a colony of complement deficient
guinea pigs. A combination of lymphadenitis, pneumonia, and
puerperal types was found. From.these, 77 strains of strep-

tococcus were studied. According to growth on blood agar,

twelve were A, 49 were B, and fifteen were Y types. All of
these strains produced capsules.

Experimental intraperitoneal inoculation produced sub-
cutaneous abscesses at the point of injection in one half of
the cases and chronic lymphadenitis in a few cases. Acute
types were produced experimentally only in new born guinea
pigs.

Another epizootic was reported in 1928 by Megrail and
Hoyt (14) in which 35 animals with abscesses mostly in.the
cervical region were examined. Every animal but one showed
the presence of both streptococcus and staphylococcus or—
ganisms. This one gave a pure culture of streptococcus.

The organism showed a definite tissue localization, only one
case showing any involvement of internal organs. After a
series of experiments, they concluded that the disease was
due to both B and Y types of streptococcus and that the
staphylococcus was merely a secondary invader. Experimental
intradermic inoculation of animals gave the highest percen—
tage of infection. Most cases deve10ped local abscesses at
the point of inoculation.and in the regional lymph glands.

a few cases ran an acute course, resulting in septicaemia
and pyemia. The serum of pigs with abscesses did not agglu-
tinate streptococci isolated from their abscesses. The
organism.was found to be pathogenic for white mice, white
rats, and rabbits. An epizootic was described by Cunningham
(15) in 1929 which involved the cervical lymph glands prin—
cipally. The organism was an incapsulated streptococcus

similar to that found by Parsons and Hyde.

ETIOLOGY AND CULTURAL CHARACTERISTICS

The material used in.this investigation, except for the
experimental animals, had its origin in a lot of 65 guinea
pigs which were intended to be used for another project.
From.time to tire infected pigs from.the stock colony were
added, bringing the total number of animals kept under obser-
vation up to 80. Many other guinea pigs in the stock colony
developed abscesses in the lymph glands, but due to the amount
of routine necessary for a detailed study it was not possible
to utilize all of the available material.

Out of the 65 original guinea pigs only nine remained
well, gave negative cultures, and Showed no pathological
lesions attributable to streptococcus infection.

Because of limited housing conditions, it was necessary
to keep these animals for three months in.a basement room of
one of the campus buildings. During the early fall it was
difficult to maintain an even temperature in this room, which
resulted in the room.becoming overheated a few times. How—
ever, this condition was promptly corrected, and no further
trouble of that kind developed. The high morbidity of the
infection in this group of animals may possibly be due to
the animals' becoming overheated a few times.

2, streptococcus was isolated, in most cases in pure
cultures, from.a11 cases examined. All strains were gram
positive, aerobic, and non capsulated and produced acid on
all culture media used. No hydrogen sulphide or other gases
could be detected. Nitrates were reduced to ammonia. Three

strains reduced nitrates to nitrites. The size of the cocci

varied considerably with different strains and on different
culture media, averaging about .7 microns. The length of
the chains varied from 2 to 50 or more elements, usually
growing singly and in chains of of two to six. Acid was
produced in litmus milk, but there was no coagulation of the
casein nor reduction of litmus. On nutrient agar, the
growth was visible after 18 to 24 hours as minute, round,
smooth, finely granular, semi-translucent, discrete, raised
colonies. They showed a marked tendency to remain discrete
for several days even when sown thickly. Nutrient broth
remained clear with a fine granular sediment at the bottom
and small flakes adhering to the sides of the tube. On
blood agar, all types produced a wide zone of hemolysis
which was noticeable after six to eighteen hours. Twenty
five strains were tested on the sugars according to Holman's
(59) classification. Mannite, raffinose, and inulin were
not fermented, saccharose Only was fermented by five, lactose

and saccharose by two, and salicin and saccharose by one.

PATHOGENICITY

Pus containing hemolytic streptococcus from.the lymph
glands of an infected guinea pig was found to be pathogenic
for mice and rabbits. Two mice injected subcutaneously
developed large, local, subcutaneous, inflammatory swellings
and died of septicaemia in 72 hours. One rabbit, inoculated
subcutaneously, develOped a local inflammatory swelling and

died of septicaemia in twelve days.

AC 33 LY PIA SHITIS

The period of incubation in acute cases varies from one
to ten days. The course is rapid, terminating in death in
from eight to 48 hours after the first noticeable symptoms.
Indeed, in the majority of cases, the animal nay appear well
and be found dead on the following day'without siving one a
chance to observe clinical symptoms.

TAG clinical symptoms of acute lymphadenitis occurring
as a primary condition or developing secondarily in a case
of chronic lymphadenitis are those of septicaemia. The onset
is sudden. In casesfinhich we have had an opportunity to
observe the animal, the general symptoms of an acute febrile
affection such as debility, weakness, and anorexia are pre—
sented. Rather marked loss of weight is noticeable even in
animals which have been sick but a short time. In case of
respiratory complications there may be a purulent nasal dis—
charge which adheres to the nostrils, usually respiratory
sounds are heard, coughing and sneezing are common, and in
case of pneumonia or pleuritis a marked dyspnea is noticed.
There may be conjunctivitis involving either one or both
eyes with reddened mucous membranes and a mucous or muco-
purulent discharge which adheres to the hair below the eyes.
Upon handling the animal does not make the usual vi:orous
effort to escape and will often lie limp in the hand. The
temperature as taken per rectum bears no relation to the
condition of he animal or stage of the disease. It fluc-
tuates considerably from.day to day, anywhere from.57° to

40° with an average of 59°. This temperature range is found

10

to be the same in normal animals as in sick animals.
_Anatomigal changes - The acute forms assure the role
of a septicaemia. Hyperemia and inflammatory edema tith
numerous streptococci in the superficial lymph glands are
constant symptoms with a slight increase in.their size,
which is due mainly to edematous swelling as there is
usually no hyperplasia of the cells. There may or may not
be exudation of leucocytes. Parenchymatous organs manifest
chanfes usually associated uith septicaemia and pyemia.
Streptococcus bronchopneuuonia is the most frequent general
manifestation. hext in importance are fibrinous, sero-
fibrinous, or fibrinopurulent pleuritis and peritonitis.
Other organs occasionally affected are in order of their
frequency: liver, heart, uterus, spleen, small intestine,
bone marrow, thyroid, and adrenal glands. The lesions in
these organs are usually focal in.character, suggestive of
pyemic origin. Often in acute cases there is little or no
exudation of blood cells into the affected part. In cases
of septicaemia developing in an animal with chronic lymph-
adenitis we find a very active exudation of leucocytes.
Degenerative changes including cloudy swelling, hydropic
and fatty degeneration are frequently seen in the liver,

kidneys, and heart muscle.

CHRONIC LY? PELDITITITIS

The period of incubation has not been established.
Apparently it varies considerably.

Abscesses may develop in a few weeks and rupture or may

11

remain visible to the eye for several months.

The only clinical symptom that can be observed in chronic
cases is swelling of the superficial lymph glands. In the
early stages this can be detected by palpation only as firm,
shot-like nodules in the submaxillary, cervical, sublingual,
prescapular, postscapular, and prefanoral regions. These
occur single or multiple and vary in size from two to five
mm. in the early cases up to as much as 40 mm. in those
showing advanced abscessation. As the nodules enlarge in
size, an area of softening and fluctuation appears over which
the skin becomes adherent. Eventually the abscess ruptures
at this point and discharges a large quantity of thick,
creamy white, homogeneous, finely granular pus which on
stained smear shows large numbers of streptococci and necro-
tic debris. The abscess continues to drain for several days
after Which the wound closes and complete healing takes place.
Another abscess may develop in the same region or in other
glands. No cases have been observed in this epizootic in
which an animal has recovered completely from.the infection
even after one or several abscesses have ruptured and healed.

Out of 82 cases observed, 65 involved the cervical
glands, 42 the submaxillary, 14 the prefemoral, 20 the sub-
lingual, 6 the prescapular, 2 the postscapular, and 3 the
internal illiac lymph gland.

Anatomical changes — Scattered throughout the cortex

 

of the gland and arrising independently of the germinal cen—
ters are foci in which the normal lymphoid cells are entirely
replaced by large mononucleated cells with pale staining

cytoplasm.and large reticular nuclei. These cells have a

12

very marked phagocytic activity for streptococci which are
often found scattered throughout the central portion of the
lesions. For convenience these till be called macrOphages.
As the lesions increase in size, they become confluent and
deve10p areas of necrosis in their centers. At this time
polymorphonuclear leucocytes invade the lesions, the bacteria
multiply, and the suppurative process develOps rapidly. At
the same time there is great fibroblastic and vascular endo—
thelial cell hyperplasia at the margin of the lesion showing
a tendency to incapsulate the developing abscess. Soon the
primary lesion is entirely obliterated and reduced to pus.
The necrotic process attacks the inner layers of the fibrous
tissue capsule, and at the same time more layers of fibrous
tissue are laid down at the periphery. Thus the abscess
increases in size until the overlying skin becomes involved.
Due to inadequate circulation no further incapsulation can
take place at this point, and the suppurative process erodes

its way to the surface with subsequent rupture of the abscess.

TECENICAL KETKODS

All living animals were killed by a blow on the head.
Sections from the heart, liver, spleen, kidneys, lungs, lymph
glands, and any other organs or tissues Showing evidence of
disease were removed with sterile instruments and smeared
over beef liver agar plates containing 5 % defibrinated
cow's blood. These were incubated for 48 hours at 37° and
then 48 hours at room.temperature. In every case a markedly

hemolytic streptococcus was obtained often in pure culture

13

and in a few cases a non-hemolytic dyplococcus was obtained

from the lung.

Subsequent inoculations were made on nutrient agar
slants containing 5 5 defibrinated cow's blood.

Histological technique — The tissues were fixed in
Zenker's fluid, imbedded in paraffin, and stained by the
following method:

1. Stain to a deep blue color in Harris's hematoxylin and
differentiate until the section shows a pale pink color
in 95 fl alcohol containing about ten drops of HCl to each
50 cc.

2. Neutralize acidity in weak aqueous NHQOH solution.

3. flash in water.

4. Stain lightly in 1 % eosin.

5. Kix 3 cc. of 5 fl aqueous solution of KahOOa with 6 cc. of
l % aqueous solution of Gentian violet. Pour onto slides
immediately and allow to stain for two minutes.

0. Pour off excess stain and cover with Lugol's solution,
(one third U. S. P. strength) one minute.

7. Rinse off excess iodine and stains under tap.

8. Decolorize slightly by dropping a mixture of one part
ether and three parts acetone on the slide until the
excess of stain begins to come out of the section. This
should be watched very closely so as not to decolorize
too much, thereby removing the hematoxylin stain. Two or
three seconds time is usually long enough.

9. Wash in water immediately.

10. Dehydrate by dipping the section four or five times in

95 fl and than absolute alcdhol.

14

11. Clear in xylol.

l2. Iount in xylol Canada balsam.

From 8 to 10 inclusive the slides should he handled
individually since this is the most critical part of the
staining technique, and these steps must be accompliShed
rapidly or a part of the nuclear stain will be lost. One
should not attempt to remove all of the excess stain by
washing in acetone—ether. This is not necessary. The
acetone-ether quickly dissolves all of the precipitated gen-
tian violet except that which is contained in Gram positive
bacteria. It is then readily removed by the water, alcohol,
and xylol baths. This method is very simple, and, if direc-
tions are followed, one should be able to obtain very good
histological sections with the bacteria well stained in

$1131.15.

ER IELEI'IT I

Feeding Experiment

 

Guinea pigs 101, 102, and 103, weighing 750, 620, and
740 grams respectively, were prepared by feeding each 1 cc
of bile to render the mucous membranes more permeable. Two
hours later each animal received orally with a medicine
dropper 1 cc. of a 48 hour liver infusion broth culture of
hemolytic streptococcus isolated from a case of lymphadenitis.
Guinea pig 101 was found dead with streptococcus septi-
caemia on the 117th day after exposure. The principal ana-

tomical lesions found were unilateral bronchOpneumonia with

serofibrinous pleuritis. A pure culture of hemolytic strep-

15

tococcus was isolated from the lung, pleural fluid, and cer-
vical lymph glands. All other organs cultured were negative.
The long incubation period and the acuteness of the patholo—
gical conditions found, together with the total absence of
any long standing lesions, makes it doubtful that infection
could have been due to inoculation and suggests them it

might be due to natural exposure.

Guinea pig 102 died seventeen days after exposure with
acute bronchopneumonia, acute lymphadenitis, acute myocar—
ditis, and nephrosis. Pure cultures of hemolytic strepto-
coccus were isolated from.the lungs, heart, and cervical
lymph glands.

Guinea pig 105 remained well 208 days and showed no

clinical evidence of infection at any tine.

Protocols
Guinea pig 101.
Sex - female.
Weight - 750 grams.
History - On the 117th day after exposure the animal was
found dead.
Autopsy findings:
The general condition is fair. The animal has lost 60 grams
in weight and shows a slight serous nasal died arge.

Thoracic cavity - Contains about 10 cc. of serosanguinous
fluid. The dorsal aspect of the right lung with the adjacent
parietal pleura and the apical portion of the pericardium with
the adjacent parietal pleura are covered With a heavy layer
of fibrinous exudate. All of the right apical, cardiac, and

about one half of the right diaphragmatic lobes are consoli-

16

dated and show a mottling of red and grey hepatization. The
entire left lung shows variable amounts of edema and conges-
tion. The bronchial mucous membrane is somewhat hyperemic.
The lumina of the bronchi contain a considerable amount of
frothy sanguinous watery fluid. The bronchial lymph glands
are somewhat enlarged and congested.

Peritoneal cavity - Contains no excess of fluid. The
surfaces are smooth.

.EEEEE - Apparently normal.

Liyg£,— Marked cloudy swelling and slight fatty changes.

Kidneys - Marked cloudy swelling.

Uterus - Slightly hyperemic.

Cervical lymph glands - Slightly enlarged.

All other organs - Apparently normal.

 

Histological examination:

 

Lppg§.- There is great thickening of the pleura with
fibroblastic proliferation involving the subpleural lung
tissue to a small extent. The pleural surface is covered
with a dense layer of fibrinous exudate heavily infiltrated
with polymorphonuclear leucocytes. The subpleural zones are
greatly congested and consolidated with fibrinous exudate in
the alveoli. The bronchial and vascular walls are someWhat
thickened and edematous and contain a small number of leuco-
cytes. The mucous membrane of the bronchi is in most cases
almost completely eroded and contains a few leucocytes and
streptococci. The alveoli in the lung tissue surrounding
the bronchi contain an exudate mainly leucocytic but with
some fibrin. This region also contains dense chains of

streptococci. Few foci show a.hemorrhagic exudate in the

17

alVeoli and in a few places mainly laked'blood., Throughout
the lung variable amounts of edema are found.

Bronchial lymph glands - Swollen, congested, and edema-

 

tous.

E3233 — Apparently normal.

Lgygg - Extensive passive congestion.and edema.with con-
siderable atrOphy of the liver cords especially noticed in
the central part of the lobe. Occasionally a.focus of necro-
sis is found.

K dney - The cortex and medulla are congested and con—
tain a few scattered small intertubular hemorrhages. The
tubular epithelium shows marked cloudy swelling. Hany glo-
meruli present degenerative changes as evidenced by hydropic
appearance of some of the endothelial cells of the capillary
tuft with the presence of erythrocytes in the subcapsular
space of Bowman's capsule. A few glomeruli show slight evi-
dence of proliferative changes but not enough to classify as
acute diffuse proliferative glomerulonephritis. There is
also a considerable amount of finely granular pink staining
homogeneous material in the glomerular subcapsular spaces
and tubular lumina.

Cervical lymph glands - Moderately swollen, moderately
congested, and show diffuse inflammatory edema with some
fibrin, numerous macrophages, and polymorphonuclear leuco-
cytes in the exudate. There are a few streptococci in the
gland almost completely phagocytosed by the macrophages.
Occasionally a polymorphonuclear leucocyte is found with
bacteria in its cytoplasm. The capsule and marginal sinuses

are very edematous.

18

All other organs - Apparently normal.

 

Anatomical diagnosis: Streptococcus septicaemia with bron.

 

chopneumonia, serofibrinous pleuritis, acute lymphadenitis,

and acute diffuse degenerative glomerulonephritis.

Guinea pig 102.

§§§ - female.

Weight at time of exposure - 620 grams.

History - On the twelfth day after exposure the aninal showed
a general lassitude and a greatly reduced reflex excitability.

Respiration lOO. Died on the seventeenth day.

AutOpsygfindings:

 

The animal is slightly emaciated, loss in.weight 135 grams.
Pleura - Somewhat dry and dull in appearance.
Lu :3 — Both apical, one half of right cardiac, and one
third of right diaphragmatic ldbes are consolidated, shoring
a mottling of red and grey hepatization. The left diaphrag-
matic lobe is fluctuating, edematous, and congested. The cut
surface presents numerous grayish-white translucent glistening
foci around the bronchi. A considerable quantity of sero—
sanguinous, frothy fluid oozes from the cut surface then the
organ is compressed.

Bronchial glands - Enlarged, edematous, and congested.

 

Bronchi and trachea - Contain a cepious amount of turbid

 

fluid.
Heart - A few small greyish spots on the myocardium,
Larypx — Shows a marked congestion.

Peritoneal cavity — Contains no excess of fluid. The

 

surfaces are smooth.

19

Liver - Normal in size, grayish brown, showing some con-
gestion and a few petechial hemorrhages on the surface of the
liver. The interlobular spaces appear swollen, translucent,
glistening. The cut surface is flat or slightly raised and
shows no hemorrhages.

Spleen — Apparently normal.

kidney — Apparently normal.

Stomach - Slightly hyperemic.

Small intestine — Slightly hyperemic.

 

Uterus - Pregnant, contains two fetuses each 1 1/3 cm.
in length. The placenta and the adjacent uterine wall are
speckled with pin point hemorrhages.

Cervical glands — Enlarged about 100 fi. Bdematous and

congested.

Histological examination:

 

Pleura - Infiltrated with leucocytes. It contains a
thin deposit of fibrin on the surface.

Lang — Right apical, right cardiac, and right diaphrag—
matic lobes show considerable red hepatization with fibrinous
exudate, edema, a few erythrocytes, and large tangled chains
of streptococci in the alveoli, and marked congestion of the
capillaries. In areas surrounding the small bronchioles the
exudate is made up of leucocytes and macrOphages with a
slight thickening of the alveolar wall. Other areas more
remote fronxthe bronchi shOW'considerable congestion and
edema, but no increased cellular material of the alveolar
wall.

Bronchi - The epithelium is greatly desquamated and

shows some evidence of albuminous degeneration. The lumina

20

are partly filled with desquamated cells and a few leucocytes.

Bronchial lymph glands - ire increased about lCO'f in

 

size, very congested, and edematous. i few small foci in
which numerous macrOph gee are found are seen in the marginal
sinus. The capsule is congested and shows slight evidence

of fibroblastic hyperplasia.

heart - There is cloudy swelling of the muscle cells of
the myocardium.with some edaua between the muscle bundles.

Liver - There is well marked cloudy snelling throughout
with moderate fatty changes in the central portion of the
lobules. Two small focal hemorrhages l mm. in diameter are
found near Glisson's capsule.

Spleen - Apparently normal.

Kidneys — There is marked cloudy swelling of the con-
voluted tubules with necrosis of occasional tubular cells.
Hany of the tubules are dilated and filled with a fine granu-
lar material taking a pale eosin stain suggestive of albumin.
The glomeruli are swollen and congested but show no evidence
of severe degenerative ckaqges.

Stomach - Apparently normal.

Small intestine - Apparently normal.

Colon - A mild mucoii degeneration of the mucosa.

Uterus - The fetus is apparently normal. There is con-
siderable hemorrhage involving the fetal and neternal placenta.

Cervical lymph glands - Kiddle cervical glands are
swollen, someWhat edematous, aid congested. A severe in-
flammatory reaction is found involving a portion of the mar-
ginal sinus and following the septa down into the cortical

portions of the gland. Some necrosis and suppuration.with

21

an exudate made up chiefly of polymorphonuclear leucocytes
involves the marginal sinus and adjacent lymph tissue. A
considerable number of macrOphages and a slight fibroblastic
hyperplasia is found in the zone surrounding the necrosis.
The lymph sinuses are impacted by thrombi composed of lympho-
cytes and fibrin. A

Anatomical diagnosis: Acute progressive bronchopneumonia

 

with acute lymphadenitis and nephrosis.

E: BEECH-IT II

Subcutaneous inoculation

Two guinea pigs, 201 and 202, weighing 265 and 400
Grams respectively were injected subcutaneously over the
ventral.wall of the abdomen with 1 cc. of a physiological
NaCl solution containing approximately 30 fl of pus taken
from a case of chronic lymphadenitis.

Guinea pig 201 develOped a local swelling which dis—
appeared in a short time. At the end of 40 days an abscess
developed in front of the Shoulders from.thich a pure culture
of hemolytic streptococcus was isolated. All other organs
cultured were negative.

Guinea pig 202 died of septicaemia in fourteen days.
Pure cultures of hemolytic streptococcus were isnlated from
the peritoneal cavity, liver, spleen, kidney, adrenal gland,

lung, heart blood, and precrural lymph glands.

Protocols

 

Guinea pig 201.

Sex _ male.

22

grams a

x.)

height at time of exposure — 265
History - Animal was examined daily, and a local swelling
was noticed in 24 hours, which continued for seven days. No
further clinical symptoms were noticed for 40 days when an
abscess was observed in front of the left shoulder. The
animal was killed on the 53rd day.

Auto pay findin 19:3

 

An abscess 1 cm. in diameter is found in the subscapular
region and one .5 cm. in diameter involving the prescapular
lmmhgflmm.

Pleura — Apparently normal.

Lppgg - The right cephalic and dorsal one half of the
right cardiac lobes are moderately congested.

Trachea and larynx — Apparently nornml.

M - Apparently normal.

Peritoneal cavity _ Contains no excess of fluid. The
serous membranes are smooth.

2&122.' Light greyish brown color, some mottling with
congestion. There is slight cloudy swelling.

Spleen ~ Apparently normal.

Kidneyp - mhe capsule strips easily, and the cortex has
a dull grayish brown, slightly parboiled appearance sugges—
tive of cloudy swelling. The cut surface of the cortex:$hows
some mottling of hyperemia.

Stomach - Apparently normal.

Small intestine — Apparently normal.

291-29. — Apparently normal.

Testes - Apparently normal.

Cervical glands - Apparently normal.

 

23

All other organs - Apparently normal.

 

histological examination:

Pleura - Apparently normal.

Lungs - There are a few areas of congestiOh. One focus
4 mm. in diameter of atalectasis and fibrous thickening of
the alveolar walls.

Bronchial glaids - Apparently normal.

 

.EEEEE — Very extensive hydropic degeneration of the
parenchyma.

Spleen - Moderately congested.

K dney — where is well marked tubular cloudy swelling
with well marked congestion in.the cortex and medulla.

Bladder - Apparently normal.

Stomach - Apparently normal.

Small intestine - Apparently normal.

'0-

Colon - Apparently normal.

 

Testicles - Apparently normal.

 

Cervical lymph glands - Apparently normal.

The abscesses were collapsed after culturing and were not
in good condition for histological study.
Anatomical diagnosis: Chronic streptococcus lymphadenitis

with abscessation.

Guinea pig 202.

§p_x_ — female.

Weight at time of exposure - 400 grams.

History — The animal developed a firm, hot, painful swelling
at the point of inoculation.three days after exposing, which
rapidly became necrotic and caused a complete perforation of

the epidermis on the seventh day. The left precrural lymph

24

gland was found, by palpation, to be somewhat enlarged and
fimm on the fifth day. This condition persisted until death.
After thirteen days the aninml refused to move when disturbed
and would lie completely relaxed in the hand. It was found
dead the following morning, fourteen days after inoculation.
Autopsy findings:

A scab covering a perforating area of necrosis 5 mm. in
diameter is found on the skin of the abdomen about 5 mm. to
the left of the median line and about one cm. in.front of

the flank.

Pleura - Apparently normal.

[Lppgp - There are a few scattered foci of congestion
from 1 to 10 mm. in diameter and a few petechial hemorrhages
on the surface of the lung just beneath the pleura.

Trachea - Contains some mucus.

Eggpp - Apparently normal.

Peritoneum - Peritoneal cavity contains about 5 cc. of

 

slightly turbid, flocculent serous fluid. Most of the loops
of the small intestine are adherent to each other, and some
parts are adherent to the liver and spleen by a thick layer
of fibrin.

piggy - Shows some hypostatic congestion. A few
slightly raised translucent, glistening white areas are found
on the surface. The cut surface is convex, glistening, and
appears somewhat translucent.

Spleen - Slightly enlarged.

Kidney - Capsule strips easily, light chocolate color.
The cut surface of the cortex is slightly raised and has a

someWhat parboiled appearance suggestive of cloudy swelling.

25

Adrenal gland - Tie left adrenal shows a well marked
congestion and is enlarged about 50 3 in size.

Stomach - Peritoneal surface is somewhat roughened.
Other than this the organ appears normal.

Small intestine — The peritoneal surface is dull, some—

 

what hyperemic, and covered in patches with fibrin. The
mucosa appears normal.

Colon - Peritoneal inflammation same as above but much
less fibrin. The mucosa is somewhat swollen, hyperemic, and
covered with a layer of tenacious mucus.

Uterus - Shows a very marked congestion and edema of the
mucosa.

Cervical lymph glands — Apparently normal.

 

Precrural lymph glands — The right gland is apparently

 

normal. The left is slightly'enlarged but normal in con—
sistency.
Histological examination:

§§in_- In the region.of inoculation the Skin presents
an ulcer with necrosis extending completely through the
epidermis and derma. The necrotic and inflammatory process
appears to involve the derma more extensively than.the
epidermis so as to undermine the latter. Long tangled
chains of gram positive staining streptococci and numerous
macrophages are found invading the necrotic tissue debris
and involving the margin of healthy tissue. The macrOphages
show a very active phagocytic activity usually containing
countless numbers of bacteria and often so many as to com—
press the nucleus against the cell wall.

Pleura - Apparently normal.

26

Lungs — There are a few foci of mild congestion.

Bronchi and trachea - The lumina of the trachea and of

 

the large bronchi contain a large quantity of mucus in which
are numerous chains of streptococci and many desquamated
epithelial cells. The mucous membrane of the trachea appears
normal but that of the bronchi is partly desquanated and some-
what thickened by lymphoid infiltration.

Heart - There is well marked cloudy swelling and moder-
ate fatty changes in the myocardium.

Peritoneum — Described with the organs involved.

 

Liver - Toderate fatty changes in the tubular cells.
There are numerous subcapsular foci one half to one mm. in
diameter, of metastatic infection, with degeneration and
necrosis of the liver cells but no hematogenous reaction.
Some phagocytic activity is shown by Kupffer's cells. The
peritoneum is covered with a heavy layer of fibrin containing
many tangled chains of gram.staining streptococci and
numerous macrophages showing a marked phagocytic activity.
There is some tendency toward organization of the exudate.

Spleen — Slightly congested. The peritoneum presents
the same condition as described with the liver.

Kidney - Shows a well marked cloudy'swelling with some
congestion in the cortex and medulla. The capsule presents
the same condition as the peritoneal covering of the liver.

Adrenal glands - Show an acute inflammatory process with

 

congestion and edema. Large masses and long tangled chains
of gram staining streptococci are found throughout the cor-
tex and medulla. The cellular exudate is entirely monocytic

and shows a well marked phagocytic activity.

27

Stomach — Apparently normal except for peritoneal in-
volvement as above described.

Small intestine - Scattered throughout the small intes—

 

tines are foci of infection. These show superficial necrosis
of the epithelium, numerous streptococci in the stroma, and
inflammatory edema of the submucosa. The lumen of the intes-
tine near many of these feci shows an abnormally large number
of streptococci and rod forms. The JeJunum.shows an extensive
mucoid degeneration of the epithelium in addition to numerous
foci of infection as described. In all cases there is a con-
spicuous absence of congestion of the capillaries of the
stroma of the mucous membrane and submucosa.with a marked
dilitation of the lymph.ducts of the latter. An inflammatory
process as described above involves the serosa together with
a very marked engorgenent of the blood vessels.

Colon - The epithelium shows a marked desquanation and
mucoid degeneration with edema of the mucosa and submucosa.
Occasionally a few streptococci are found scattered through-
out the submucosa.

Uterus - Moderately congested.

Precrural_lymph glands - The left shows an acute in—

 

flammatory reaction. The gland is swollen, edematous, and
congested, and contains many streptococci, fibrin, and mono-
cytes, with very little evidence of phagocytic activity on
the part of the leucocytes and macrophages. The marginal
sinuses are dilated and packed with streptococci. The cap—
sule is of normal thickness.

Anatomical diagnosis; Streptococcus septicaemia with sero-

fibrinous peritonitis, focal hepatitis, acute focal osteo—

28

myelitis, acute lymphadenitis, acute focal enteritis, and

acute diffuse adrenalitis.

EEPWRIHENT III

Conjunctival Inoculation

Two guinea pigs, 205 and 204, weighing 345 and 465
grams respectively were inoculated by instilling a drop of
pus suspension in the right conjunctival sac. The pus sus-
pension was of the same source as used in the subcutaneous
inoculation experiment.

Guinea pig 203 developed conjunctivitis in the right
eye in 72 hours and died of septicaemia in eight days. a
pure culture of hemolytic streptococcus was isolated from
the right eye, the cephalic and cervical lymph glands, liver,
spleen, kidney, heart blood, and lung.

Guinea pig 204 developed conjunctivitis after an incu-
bation period of six days. This disappeared after a few
days. The animal remained tell and was killed after 36
days. A pure culture of hemolytic streptococcus was isolated
from the right eye, but no growth could.be obtained from.any
other organ. Unfortunately no tissues could be saved for
section from the conjunctiva as the frontal bones were so

badly crushed that they could not be cut for fixation.

Protocols

 

Guinea pig 203.
Sex — male.
Weight at time of exposure — 345 grams.

History - Swelling of the conjunctiva with lacrymation was

29

detected 72 hours after inoculation. This continued until
death. The animal was found dead on the eighth day. It had
lost 50 grams in weight.

Autopsyrfindings:

Considerable gummy exudate adheres to the lids of the eyes
and to the anterior nares.

Conjunctiva - Congested slightly and presents numerous

 

grayish white, dull, opaque foci one fourth to one mm. in
diameter.

Lacrymal gland - Somewhat enlarged and presents a few

 

greyish white, dull, opaque foci.

Pleura - Apparently normal.

‘Lungg — Occasional petechial subpleural hemorrhages.
There is very moderate congestion.througmout.

Bronchial glands -Slightly enlarged.

 

Bronchi and trachea.- Contain a small amount of floccu—

 

lent fluid.
Heart - Apparently normal.

Peritoneal cavity - Contains no excess of fluid. The

 

surfaces are smooth.
nggg — Dark reddish chocolate color. Nonnal size.
The cut surface is slightly convex. Pin point areas of
hemorrhage are found at the periphery of the lobules.
Spleen .- Apparently normal.
Kidney - There is some congestion in the cortex.
Stomach — Contains about 15 cc. of water and mucus.

Small intestine - Apparently normal.

 

Colon — Apparently normal.

Testicles - Apparently normal.

3O

Cervical lymph glands - moderately congested but show
no appreciable enlargement.
Histological examination:

Right conjunctiva - The mucous membrane is completely

 

eroded. There is a.marked thickening of the submucosa with
congestion of the capillaries, engorgement of the small
vessels,and infiltration.with small round cells and macro-
‘phages. The latter show very well marked phagocytic acti-
vity for streptococci. The entire submucosa is moderately
heavily invaded with streptococci growing mainly singly but
often forming chains of two to twelve. Humerous small foci
of early necrosis are found in which polymorphonuclear cells
are plentiful in the exudate. In the margins of these feci
and scattered throughout the submucosa are a few fibroblasts.

Left conjunctiva - Apparently normal.

Posterior gages — No evidence of inflammatory reaction
of the mucous membrane but a large quantity of exudate con-
taining a few streptococci are found adhering to the surface.
This probably comes from.the facial sinuses.

Pleura - Apparently normal.

Lung — Extensive congestion and numerous patches of red
hepatization with congestion of the alveolar walls. There
is some fibrin, a few erythrocytes, and a few streptococci
in the alveoli. There are some patches showing well marked
edema.

Bronchi - Moderate amount of éesquamation of the mucous
membrane. The walls of the smaller bronchi are edematous
and slightly infiltrated with small round cells.

Bronchial lymph glands — The marginal sinuses are dilated,

31

edematous, and Show slight hyperplasia of the endothelial
cells.

EEEEE - Slight congestion in the myocardium. The blood
shows evidence of leucocytosis.

{Eipgp - tell marked, extensive congestion and hydropic
changes are seen.

Spleen - Apparently normal.

Kidney - Congestion of the capillaries of the cortex
and medulla with tubular cloudy swelling.

Stomach - Apparently normal.

Small intestine - Apparently normal.

 

_C£>_l_o_n - Apparently normal.

Testicles - Considerable pyknosis of the spermatocytes
and desquamation of the spermatids. A few live spermatozoa
are found in the tubules of the epididymis.

Cervical lymph_glands - Swollen, congested, and infil-
trated by inflammatory edema, polymorphonuclear leucocytes,
and finely granular material involving most of the glands.
This condition involves a large part of the peripheral
sinuses and to a great extent the capsule. The marginal
sinuses also contain a variable number of proliferating
endothelial cells. A diffuse infection with Streptococcus
is found throughout the gland, growing singly and also
forming chains of from two to twenty. very little evidence
of phagocytosis can be found. ‘

Anatomical diagnosis: Acute conjunctivitis with acute

 

lymphadenitis and early bronchopneumonia.

Guinea pl; 204

Egg - female.

Height at time.of exposure - 465 grams.

Tistogy - Lacrymation with swelling of the conjunctiva was
noticed after 24 hours. This gradually subsided after six
days. The animal at no time appeared to be sick. It was

killed on the 56th day. The gain in Wei ht was 140 grams.

Autopsy findings:

 

The animal is in.good condition.

..

Conjunctiva - Apparently normal. lo tissues can be

 

saved for section as the skull is too badly crushed in
killir; the animal.

Yasal passages — Apparently normal.

 

Pleura — Apparently normal.

Luggs — Apparently normal.

Bronchi - Some thickeninr of the bronchial walls is found.
Trachea - Apparently normal.

Eeart — Apparently normal.

Peritoneal cavity - Contains no excess of fluid. The

 

surfaces are smooth.
l££§£_- Chocolate brown color. Apparently normal.
Spleen - Apparently normal.
Kidnevs - Apparently normal.
Stomach — Apparently normal.

Small intestine - Apparently normal.

 

Colon — Apparently normal.
Uterus - Apparently normal.
Ovaries — Apparently iormal.

Cervical lymph glands _ Apparently normal.

33

Histological‘examination:

 

 

Pleura - Normal.

Lupgs — There are a few scattered foci surrounding the
bronchi shoring marked thickening of the alveolar mall due
to fibrosis and, to a lesser extent, endothelial prolifera-
tion.with a few monocytes in the alveoli.

Bronchi — Koderate annunt of desquamation of the mucosa.
The exudate in the lumen is made up chiefly of desquamated
cells with an occasional leucocyte. The bronchial rall is
somewhat thickened by fibrosis. This condition extends out
into the lung tissue for a distance of from .5 mm. to szn.
as above described.

Bronchial glands - Apparently normal.

 

Heart - The myocardium presents a vell marked cloudy
swelling with some evidence of fatty changes.

Liver — Very hydropic. Some cells have degenerated so
badly that the nuclei have become shrunken.

Spleen — Apparently normal.

Kidney - There is a very extensive well marked tubular
aloudy swelling.

Stomach - Apparently normal.

Small intestine — Apparently normal.

 

Colon — Apparently normal.
Uterus — Apparently normal.

Cervical lymph ngmds — Apparently normal.

__L—.

*J‘J'fifi‘ffi "1 T‘ Pm? “T IV
arLL‘CJ J . u-.'.f.J.‘.~: .

Inoculation via nasal passases

 

Two guinea.pigs, 205 and 206, weighing 470 and 220 grwus
respectively were exposed by instilling one drop of normal
salt solution pus suspension into the right nostril. This
was accomplished by holding the animal in one hand with the
nostrils in a vertical position and using a medicine dropper
to make the instillation.

Both animals died of septicaemia in eight day

A pure culture of hemolytic streptococcus was obtained
from.the liver, spleen, heart blood, and cervical lymph
glands of both pigs. The organism was obtained also in

culture from the lungs of guinea pi: 205.

Protocols

 

Guineappig 205.

 

Sex - male.
Teight at time of exposure — 470 grams.
History - Ho clinical symptoms were observed until the eidhth
day when a slight nasal discharge developed and the animal
showed signs of general malaise. The animal died eight days
after exposure. The loss of weight was twenty grams.
Autopsy findings:
The condition of the animal is good. There is a small amount
of dryed nasal discharge adherent to the nostrils.

Pleura — Apparently normal.

Lupa - A well marked patchy congestion with numerous

scattered petechial hemorrhages is observed. The accessory

lobe is completely atelectatic.

55

Bronchi and trachea - Filled with.a frothy serous exu-
date.

Bronchial lygpp‘glapdg - Apparently normal.

£3333 - Apparently normal.

Peritoneal cavity — Contains no excess of fluid. The
surfaces are smooth.

£2123.“ Hedium chocolate brown color. It appears nor—
mal except for a grayish white dull opaque focus one cm. in
diameter on the left lobe of the liver.

Spleen - Apparently normal.

Kidney — Iarked congestion.

Stomach - Apparently normal.

Small intestine — Apparently normal.

.99lpp - Apparently normal. I

Testicle — Apparently normal.

Cervical lymph glands - Ioderately congested, pliable
and firm in consistency, and moderately enlarged averaging
5 mm, in diameter.

Nasal passages - Kucous membrane of nasal cavity and
turbinate bones is very congested, somewhat thickened, and
covered with a thin layer of mucus.

Facial sinuses - Frontal, palatine, and maxillary show
a well marked congestion of the mucosa and contain a large
amount of thin, White, opaque, granular fluid.

Histological examination:

Pleura — Apparently normal.

Lppg - Well marked and extensively congested. Patchy
areas of red hepatization in which the alveoli are loaded

with streptococci, fibrin, and a few erythrocytes are found

36

throughout both right and left lunfis. hunerous patches of
edema are also found scattered throujhout the lung. The
condition is essentially what is usually described as acute
patchy bronchopneumonia with variable recognizable stages
from congestion and edema to red hepatization.

Bronchi and pgachea — The bronchi are partly filled

 

with debris consistin: mainly of desquamated cells. T.e
bronchial mucous membrane is almost completely eroded. The
bronchial wall appears to be thickened and moderately infil-
trated by mononuclear cells.

Bronchial lymph glands — Congested and edematous.

 

Seart - Shows a slight cloudy swelling.

Liver - Voderate congestion. A slifiht amount of fatty
degeneration is found in the central portion of the lobules.
There is a necrotic focus one cm. in diameter just beneath
Glisson's capsule. This is loaded with streptococci growing
singly and in long chains. N serous leucocytes are found
infiltrating the periphery of the lesion but show no phago-
cytic activity for the bacteria. A zone surrounding the
lesions contains numerous fibroblasts, macrophages, and
polymorphonuclear leucocytes, but no evidence of phagocytosis
is recognizable. The surface of the liver at this point con-
tains a larg deposit of fibrin and polymorphonuclear leuco-
cytes.

Spleen — Apparently normal.

Kidney — Ioderate cloudy swelling of the tubular epi—
thelium. Slight congestion of the capillaries of the medulla.

Stomach — Apparently normal.

Small iptestine — Apparently nomnal.

 

37

Colon — Apparently normal.

Testicle — Apparently nonail.

Cervical lymph glands - doth middle cervical lymph
glands are enlarged about 7 mm. in diameter due mainly to
hyperplasia of the lymphoid cells. A few foci of early
infection, apparently of pyemic origin, are found near the
periphery of the gland containing numerous streptococci.

The lymphoid cells in these foci are entirely replaced by
large mononuclear cells which have great phagocytic power
for streptococci. in occasional polymorphonuclear leucocyte
showing very little phagpcytic activity for bacteria of this
species is found scattered throughout these lesions. At the
peripheral zone of these lesions numerous fibroblasts and a
few vascular endothelial cells can be found trying to incap—
sulate the developing lesion. The marginal sinuses are
partly filled in some places with proliferating endothelial
cells. The capsule is thickened sli:htly by fibroblastic
hyperplasia.

Nasal cavit - A large quantity of pus is found on the

 

surface containing many degenerated leucocytes and a large
number of chains of streptococci. The mucosa is congested,
edematous, and in nany places shows desquamation of the
superficial epithelial cells. The edema of the connective
tissue stroma is of an inflammatory nature containing some
fibrin and an occasional leucocyteo.

Facial sinuses - Present the same condition as that

 

described for the nasal cavity.
Anatomical diagnosis: Acute mucOpurulent rhinitis and

sinusitis with septicaemia.

38

Guinea pig 206.

gpg - male.

Weight at time of exposure - 226 grams.

history — On the third day the animal develOped a cepious
nasal discharge and made slight rattling noises from the
throat when breathing. After eight days the animal vas
found dead. The loss in weight was ten grams.

Autopsy findings:

The animal is in fair condition.

Pleural cavity - Contains no excess of fluid. The

 

pleural surfaces are smooth.

Lupgs - hoderate congestion.

Bronchi - Some of the bronchi show some thickening of
the alveolar malls involvin: the lnn: tissue in a narrow

surrounding zone.

Bronchial lvmnh glands — Apparentlyinormal.

 

Heart - Apparently normal.

Liver - Koderately congested.

Spleen — Apparently normal.

Kidney - Apparently normal.

Stomach - Apparently normal.

Small intestine - Apparently normal.
Colon - Apparently normal.

Testicles - Apparently normal.

 

sublaxillary and retropharyngeal lymph glands - Slightly

 

enlarged and congested.

Nasal passages and facial sinuses - All contain a copious

 

quantity of thin, white, creamy pus. The mucous membranes are

hyperemic and appear slightly thickened.

59

Histological examination:

 

Pleura - Apparently normal.

Lungs - Moderate to slifhtly contested, with small foci
of atelectasis. Several small foci of consolidation
surrounding the bronchi with lymphocytic infiltration of the
alveolar walls and an exudate in the alveoli made up of leuco-
cytes and lymphocytes. ho evidence of streptococci can be
found in the sections.

Sronchi — A rather marked desquamation of the epithelium
of the bronchi is dominant throughout the lung in some places
completely plugging the lumen. A lymphoid infiltration
involves the wall of the small bronchi, peribronchial tissues,
and in some places the surrounding lung tissues to a variable
extent.

Bronchial lymph glands - Apparently normal.

Heart — Apparently normal.

'Liver - Very hydropic and moderately congested.

Spleen - Apparently normal.

Kidney'- Tell marked tubular cloudy swelling and a
moderate congestion in the cortex and redulla. The subcap-
sular spaces and tubules contain numerous shadow cells Which
are possibly desquamated and degenerated cells from the
glomerular epithelium.

Stomach — Apparently normal.

Small intestines - Apparently normal.

 

Colon — Apparently normal.

Testicles - Apparently normal.

 

Submaxillary and retrophapyngeal lymph glands — The

glands are swollen, congested, and edematous. There are a‘

40

few scattered foci of infection, some of them.involving the
marginal sinuses and others located in.the deeper structures
of the glands. The cellular content of these foci is made
up mainly of polymorphonuclear leucocytes showing a marked
phagocytic activity for streptococci and a few mononuclear
cells showing very active phagocytosis. The marginal zones
of these foci contain numerous proliferating fibroblasts.
There is a slight dilatation of the marginal sinuses and a
slight proliferation of the endothelial lining. The cap—
sules are thickened moderately by edema, infiltration.with
lymphocytes, and proliferation of fibroblasts.

Basal passages — The surface of the rmcous membrane is

 

covered with a large amount of exudate made up mainly of
polymorphonuclear leucocytes and numerous streptococci
growing singly and in short chains of from two to six. There
is slight desquamation of the superficial epithelial cells

of the mucosa.rith some edema an congestion.in.the connec-
tive tissue stroma.

Anatomical diagnosis: Acute rhinitis with acute lymphaden—

113180

EXP RIMENT V

L‘J

Skin Inoculatign
Two guinea pigs, 20? and 208, were prepared by clipping
the hair over a small region on the left lateral wall of the
abdomen and scraping the skin with the blade of a scalpel
over an area about 5 cm. in diameter until it appeared

slightly hemorrhagic. Next a drOp of the pus suspension of

41

of the same source as used in the other experiments was
lightly massaged into the denuded area.

Guinea pig 20? died of streptococcic septicaemia in
eight days. A pure culture of hemolytic streptococcus was
recovered from peritoneal fluid, liver, spleen, kidney,
uterus, pleural fluid, heart blood, lungs, cervical lymph
glands, and the subcutaneous tissues at the point of inocu—
lation.

Guinea pig 208 was killed by a blow after 55 days. A
pure culture of hemolytic streptococcus was isolated from
abscesses in the precrural and cervical regions. To cultures
could be obtained from any other organ.

Another guinea pig, 301, neixhing 310 grams was pre-
pared as above and inoculated with a freehly isolated eighteen
hour nutrient agar culture of hemolytic streptococcus taken
from guinea pig 207. A drop of physiological NaCl suspension
of this culture was massaged into the abraded area of skin.
The animal Showed no clinical symptoms of acute lymphadenitis
and was killed after 43 days. Abscesses from Which pure cul-
tures of hemolytic streptococcus were isolated were found
involving the precrural, cervical, and submaxillary lymph
glands. Cultures from all other organs were negative. The
results of this experiment might sugrest that the resistance
of the individual animal has a greater influence on the
course of the streptococcic infection than either the viru-
lence of the organism or aqe of the animal. Guinea pig 20?
died of a fulminatin: type of infection while streptococci
from this case produced chronic lymphadenitis when subse—

quently inoculated in a similar manner into another animal

42

of about the same age and the same sex.

Protocols

 

Guinea pig 207.

‘ggx — female.

Eglght at time of exposure - 2%“ grams.

history — hfter five days a circumscribed, firm, hot, pain-
ful smelling develOped at the point of inoculation. This
rapidly became necrotic and a scab developed over the sur—
face which remained until death. The inflammatory swelling
spread out peripherally involving mammary glands and the
left precrural lymph gland, whidh'became enlarged and hard.
The animal was found dead eight days after exposure.
autopsy findi If s:

The hair coat is dull and rough. The loss in neizht is 65
grams. a necrotic area, 5 mm. in diameter, covered Yith a
thick scab is found on the skin over the abdominal wall 5
mm. to the left of the median line and 1 cm. in front of
the pubis.

Pleura — The thoracic cavity contains aheut 5 cc. of
serosanjuinous fluid containing a brick dust-like sediment.
The pleural surfaces are dull and rough, but no great amount
of fibrin deposit can be found.

Lunas — The right apical, about three fourths of the
right cardiac, and a small area on the ventral forder of the
right diaphragmatic, and all of the left apical lobes are
consolidated and show a variation in color from.a dark red

to a dull grey. These areas cut with increased resistance

and have a dull nodular cut surface from.which no creamy

fluid can be expressed. Extensive congestion and some edema

43

are found in regions net stoninj consolidation. ihe lung is
about 25 f air containing.

Pronchi — Cn section the'walls appear somerhat thick—
ened. The lumen contains a spell quantity of serosanfuinous
frothy fluid.

fironchial lynnh_;l3gd§ - Enlarged an? sometJat edematous.

heart - ; small amount of serosanjuinous fluid is found
in the pericardial sac. the epicardium shows a few pateclial
hemorrhages on the auricular walls and around the base of the

pulmonary arteries and aorta.

Peritoneal ca1:tv — Contains ahout 10 cc. of serosan-

"—‘p A

In
I

guinous brownish fluid containing a la fie quantity oi fine

brick dust sediment.

lgggg_- Dark red in color. It is about nornal in size.
There are about twelve white translucent glistening areas and
a few dull opaque grayish thite areas from 1 to 3 mm. in
diameter on the surface of the liver just beneath Glisson's
capsule and extending to a depth of about 1 mm. in the liver
tissue. Lhe cut surface is convex and bleeds sliflhtly. a
few dull opaque grayish White foci averafiin: about 1 mm. in
diameter are found scattered throughout the orfian.

Spleen - Slijhtly contested.

Kidneys - Capsule strips easily leaving a smoofli alis—
tenin; surface of a chocolate brorn color. The cut surface
is convex and shows slijht congestion in the capillaries of
the medulla.

£23ch .. Apparently noraal .

Small intestine — Apparently normal except for 1 moderate

 

congestion of tne serous coat.

44

Colon - Apparently normal.
Uterus - Apparently nonmil.

Precrural lymph glands - The right gland is apparently

 

normal. The left is slightly enlarged and firm in consis-
tency.

Histological examination:

 

Pleura - Markedly'thickened and covered with a thin
layer of fibrin, rich in mononuclear cells and containing a
few chains of streptococci.

lungs — About three fourths of the right cardiac and a
small area on.the ventral border of the right diaphragmatic
lobes are completely consolidated. Fibrin, a few erythro-
cytes, desquamated epithelial cells, and long tangled chains
of streptococci are found in the alveoli. This condition is
associated with a general pulmon ry hyperemia accompanied by
edem . The edema is confined to those areas in which there
is congestion but no hepatization. The edematous material
is finely granular, acidophilic, and almost hyaline in
character. Scattered throughout the consolidated regions
are patchy areas one fourth to one half mm. in diameter,
surrounding the terminal bronchioles ard smaller brondui.
The exudate is chiefly leucocytic in character with a few
desquamated epithelial cells and macrophages. Numerous
streptococci are found in these areas, a few of which are
phagocytosed by the leucocytes and macrophages.

Bronchi - The bronchi are the seat of an acute catarrhal
inflammation with a distinctly purulent exudate in the
lumen. There is a W611 marked and extensive desquamation of

the epitheliuu of the bronchi and bronchioles with exudation

45

of polymorphonuclear leucocytes into the walls. Kany strep-
tococci are present in the exudate, and occasionally a few
may be found in the bronchial walls.

a
l

Bronchial lymp1_glands — Very hyperemic and edematous.

 

Several foci of liquefaction necrosis averaging 250 u. in
diameter involve the germinal centers. There is no cellular
exudation involved in these foci, but the lymphocytes and
endothelial cells are apparently very active.

;ea£t — Occasionally a small hemorrhage of about 1 mm.
in diameter involves the connective tissue stroma of the
auricular epicardium. The epicardium is very hyperemic and
shows a rather marked thickening due to edema. The myocar—
dium shows distinct degenerative changes evidenced by cloudy
swelling and fatty degeneration of the muscle cells.

Liver - federately congested and presents a slitht
fatty degeneration in the central portion of the lobules.
One focus of early necrosis 1 mm. in.diameter is found con-
taining numerous streptococci growing singly and in chains
of two to twel e. There is a slight exudation of polymor-
phonuclear leucocytes and a few macrOphages at the margin
of this lesion. Not much phagpcytosis of bacteria is seen
by either of these cells. Glisson's capsule is swollen
slightly and covered with a thin layer of fibrin containing
numerous erythrocytes.

Spleen - The organ is engorged with blood and edematous.
The architecture is very indistinct due to a ride spread and
scattered necrosis of the ymphoid cells. humerous bacteria
are found scattered throughout the spleen pulp and in.the

lymph sinuses. The capsule is thickened slightly. Numerous

46

large masses of streptococci are found on its surface. ho
where tlroughout the virole orjan is there any evidence of
leucocyte exudation.

Kidney _ Presents a *ell rarked tu ular cloudy swelling
with necrosis of the epithelium. There is a finely granular,
acidophilic, alhuninous material in the tubules., also in the
subcapsular spaces of the ’thl eruli. The capillaries of the
medulla and cortex are moderately contested. The glomerular
loops are engorged rith blood. Kany of the glomeruli show
Show deg generative chan es vith escape of erythrocytes into
the subcapsular Spaces.

Stomach - Apparently nornal except for a moderate con—

"i

gestion and thickening 01 the serosa uith numerous erythro-
cytes and large masses of bacteria on the surface.

Small intestine — Apparently normal. The serosa shore

 

the same condition as that of the stomach.

Colon — Apparently normal. The serosa shove the same
condition as that of the stomach.

Uterus - Apparently normal. Cerosa greatly hick ened,
edematous and congested. Iumerous streptococci are found
on the surface together with a few erythrocytes and a small
amount of fibrin.

{in a_ region of in oculation — Presents an area.of

 

 

necrosis in v. hich lar rge masses of strepmococci are found.
The lesion involves the stratum lucidun, stratum germina—
tivum, of the epidermis and the pars papillaris and pars

reticularis of the derma. The subcutaneous tissues are

greatly thickened due to engorgcmelt cf tie blood vessels,

edema, and numerous variable sized he :::orrha3es. Dense

47

masses and tangled chains of streptococci are found scattered
throughout the subcutaneous tissues in many places so numer-
ous as to obscure the other structures. The veins and arteries
show enormous thickening of their walls due to acute inflamma—
tory processes. Kany of the veins present thr mbi on their
internal layers. No where throughout the entire inflamed

area is there any evidence of leucocytic or monocytic exuda-
tion.

Precrural lymph glands - Left gland is apparently normal.

 

The right gland is increased to about 1 cm. in length by 5
mm. in Width. It is very edemic and heavily loaded vith
streptococci throughout. The structure is so indistinct as
to be unrecognizable due to the great pressure of the fluid
contained causing a spreading apart of the cellular struc-
tures. There is no evidence of leucocytic exudation in any

part of the gland.

 

Cervical lymph glands — Present the same condition as
the precrurals.

Anatomical diagnosis: Streptococcus septicaemia.

 

Guinea pig 208.

.§§§’- male.

Weight at time of exposure - 245 grams.

History - Seven days after exposure a slight thickening of
the skin with a superficial area of necrosis 5 mm. in dia-
meter was found at the point of inoculation. This was
entirely healed after fourteen days, and at that tine the
left precrural lymph gland became palpable as a hard shot-

like body about the size of a small pea. This slovly

48

developed into an abscess which became .5 cm. in size at th
time the animal was killed. On the 28th day the middle cer-
vical lymph glands became slightly enlarged and more firm
than normal. They gradually increased in size until they
became 1 cm. in diameter. The aninnl was killed on the 53rd
day following exposure.

Autopsy findings:

 

The animal is in fair condition. It has :ained 175 grams
since exposure. The hair coat is smooth. There is a group
of circumscribed swellinqs in the cervical region and one
in front of he right shoulder joint.

Pleura - There is no excess of fluid in the pleural
cavity. The surfaces are smooth.

Lunns - A few very small irregular areas of congestion
are found. The cut surface is regular except for a few
raised thickened areas surroumding the bronchi that cut with
increased resistance.

Bronchi — The lumen contains a srall amount of purulent
fluid. The walls are thickened and stand out as raised
branching areas on cut surface.

Bronchial lymph glands - Appear slightly enlarged and
edematous.

Heart - Apparently normal.

Peritoneal cavipy -. Contains no excess of fluid. The
surfaces are smooth.

Liver - It is light greyiSh brown color.

Spleen — Apparently normal.

Kidnexs - The capsule is tense and strips with ease

leaving a smooth surface of a light areyish brovn color.

49

The organ cuts without increased resistance and bleeds
slightly. The cut surface is raised slightly and shows a
fairly well marked hyperemia in the cortex and medulla.

Stomach - Apparently normal.

Small intestine - Apparently normal.

gglgn - Apparently normal.

Testicles - Apparently normal.

 

Precrural limph glands — The left gland is .5 cm. in

 

size, firm.in consistency, and cuts with increased resistance.

Cervical and cephalic lymph Egands — Both middle cervical

 

and submaxillary lymph glands average 1 cm. in diameter and
are converted into abscesses vith a thick dense fibrous wall
which cuts with increased resistance allowing a thick granu—
lar greyish white pus to ooze out.
Histological examination:

Pleura - Apparently nornal.

Lungs — There are no lesions that are attributed to
streptococcus infection.

Bronchi - There is a mild catarrhal degeneration with
some desquamation of epitheliun.in some bronchi. The alveoli
contain some granular degris and a few desquaiated cells.

Pronchial glands - Apparently normal.

 

Heart - Apparently normal.

Liver - Shows a well marked and extensive hydropic
degeneration of the liver cords.

Spleen - A hyalin necrosis involves a part of the peri-
phery of the Halpighian corpuscles.

Kidney - Very marked cloudy swelling of the tubules

with.sore fine granular albuminous material.in their lumina.

50

There is a.moderate hyperemia of the capillaries in the cor-
tex and medulla.
Etomach — Apparently normal.

.Small intestine — Apparently normal.

 

Colon — Apparently normal.

Precrural lymph gland — The left gland presents an

 

advanced chronic inflamnatory condition. The capsule is
thickened about ten times by fibroblastic hyperplasia vhich
envelops the marginal sinuses completely obliterating them.
There is a well marked increase in connective tissue frate-
work throughout the gland with numerous endothelial cells
and macrophages completely replacing the lymphoid cells.
humerous foci of infection containi 3 many streptococci and
and a collection of leucocytes are found throughout the
cortical portion of the gland. These show a tendency to
necrosis in their centers. Those in the more central pore
tion of the gland are entirely necrotic and converge to form
a suppuratin: mass about two mm. in diameter.

Cervical and. submaxillary lyngph glands — Chow a condi—

 

tion similar to that of the precrural gland described above
only apparently more rapid in abscess formation, The capsule
is thickened about fifteem.times, completely obliterating

the marginal sinuses. Just beneath the capsule are a number
of individual abscesses, completely incapsulated, measuring
from .5 mm. to 1 mm. in diameter. Towards the center of the
gland these converge to form large abscesses averaging about
5 mm. in diameter.

Anatomical diagnosis: Chronic streptococcus lyn hadenitis.

 

51

Guinea.plg'301.

§p§ - female.

Weight at time of exposure - 510 grams.

Eistopy - A slight thickening of the skin was noticed 48
hours after inoculation. This healed promptly, and no other
clinical symptoms could be observed until the fifteenth day
when a slight enlargement becane palpable in the left pre—
crural and submaxillary lymph glands. These gradually in—
creased in size and averated about 1 cm. in diameter at the
time the animal was killed 43 days after inoculation.

Autopsy_findings:

 

Condition.good. A group of circumscribed swellings like
marbles are found in the cervical and submaxillary regions
and in.the left flank.

Pleura - The cavity contains no excess of fluid. The
surfaces are smooth.

Agppg§’- A few small slightly congested areas are found.
The anterior borders of both diaphragmatic, all of the left
cephalic, and about one half of the right cardiac lobes show
partial atelectasis. The accessary lobe is completely

atelectatic.

Bronchi and trachea - Apparently normal.

 

Bronchial glands - Apparently normal.

'Epgpp - Apparently normal.

Abdominal cavity - Contains no excess of fluid. The
surfaces are smooth.

Lizpp.- Iedium chocolate brown in color. Apparently
normal.

Spleen - Apparently normal.

Kidneys - The cut :urface is raised slightly, dull, pale,
pinkish red color, suggestive of cloudy swelling.

Stomach — Apparently normal.

Small iptestine - Apparently nonial.

Colon — apparently normal.

Uterus — Tight horn is 2 m3. in diameter and non preg—
nant. ghe left horn is 6 my. in diameter and apparently in
a state of early pregnancyw

Ovaries - Present no unusual condition.

Precrural lymph glands - The right gland is apparently

 

normal. The left gland is 6 31-11. in diet-deter, presents a
thick tough capsule which cuts with increased resistance
allowing a small quantity of thick granular grayish white
pus to escape.

Iiddle cervical lynrh glands - Average 12 cm. in diaa

 

meter and are abscessed the same as the precrurals.

Eubmaxillapy_and sublin;pal‘;lands — Average 9 mm. in

 

diameter and are all abscessed.

Yistolonioal examination:

—

 

 

Pleura - A parently normal.

Lungs _ Apparently normal except for a few small areas

of consolidation from .1 to .5 an. in diameter located around
the bronchioles and larfier bronchi. These are oh acterized
as a low grade inflammatory process, apparently of long
standing, with marked thickening of the alveolar walls due

to fibroblastic and endothelial proliferation. The exudate
in the alveoli is scanty in amount and made up of macrophages
with a few eosinOphiles and occasionally a few polymorpho-

nucl ear leucocytes .

55

Bronchi — Show a mild catarrhal inflammation with some
desquamation of the epithelium and a little leucocytic exuda-
tion. The bronchial walls are thickened slithtly.

Bronchial lymph_glands — Apparently normal.

M - Apparently normal.

‘Eiggp - Moderate cloudy swelling of the epithelium.

Spleen - Apparently normal.

Kidney - Bell marked tuhular cloudy smelling.

Stomach - Apparently normal.

Small intestine — Apparently normal.

 

Colon - Apparently normal.

Uterus - The left horn is pregnant. (Fetus is not
found in section.) ‘fhe fetal nenbranes and all coats of
the uterine wall are apparently normal. Tie mucous mem.
brane is 1.5 mm. thick. The mucous membrane of the riaht
horn is .4 mm. hick and shows no evidence of injury.

Lymph glands - They are described collectively as all

 

the glands recorded above in the autopsy protocol show the
same condition. They are ermirely corNerted intc>abscesses
and bear no resemblance to lymph glands. ri‘he capsules are
greatly thickened, one of them.l/4 mm. thick. The subcap-
sular areas show considerable fibroblastic hyperplasia,
numerous endothelial cells, leucocytes, and streptococci.
Towards the center the mass becomes necrotic.

Anatomical diagnosis: Chronic suppurative streptococcus

lymph adeniti s .

DISCUSSION

Both acute lymphadenitis pith septicaemia and the

54

chronic form with abscessation of the lymph glands were seen
in this guinea pig epizootic. Out of 82 infected guinea
pigs examined, 26 died of acute lymphadenitis with septi—
caemia. These animals were apparently nornal until a few
hours before death. Host of them were found dead nithout
any clinical symptoms being noticed. Upon histoleical
examination no evidence of chronic disease process attribu-
table to streptococcus could be found. There was also a
great tendency for chronic cases suddenly to become acute,
resulting in septicaemia and death of the animal. Fifty two
guinea pigs were examined which developed chronic lymphaden-
itis. Eighteen of these animals died of septicaemia.

The most frequent point of localization was in the car-
vical lymph glands. Sixty five guinea pigs showed involve-
ment of these glands. Next in frequency are the submaxillary
glands with 42 animals involved. Others are as follows:
prefemoral in fourteen animals, sublingual in twenty, pre-
scapular in six, posterior scapular abscesses in two, the
internal illiac in.three, and the spleen in one case.

Among the septicaemic cases bronchopneumonia was the
most frequent complication. Forty four cases of pneumonia
were found, and hemolytic streptococci were found in each.
Twenty seven cases were complicated with streptococcus
pleuritis, nine with focal infection in the liver, eight with
focal infection in the heart, seven.with streptococcus peri—
tonitis, and four each pith focal infection involving the
spleen, intestine, and uterus.

Due to the insiduous nature of his disease at the onset

and the apparent contagiousness, together with a lack of

efficient diagnostic methods, the control of this infection,
once it has hecome estahlished in a flock of guinea piss, is
iifficult. Tiseased pigs_can tn readily located by palpation
of the superficial lymph glands when the disease has become
far enouah advanced to produce an appreciable increase in the
size of these rgans. However, this method alone does not
serve to locate the cases at the beainning of infection.
Indeed, some of the cases of acute lymphadenitis pith septi—
caemia showed no enlargement of the lymph glands palpable
through the skin even though they showed, upon histological
examination, sone smelling with inflamratory edema and were
teeming pith streptococci.

With these facts in mind then, it would appear that
either a more efficient method of diatnosis is necessary,
some method of immunization, or a more effective rethod of
sanitation. I am.not in.a position to offer any suzaestions
for improving the diagnosis or for immunization. Herbert
and Neyer (15) were unable to produce local immunization in
guinea pigs against streptococcus infection by means of a
Besredka filtrate. Single or multiple doses of heat killed
faccines gave no protection against subsequent inoculations,
and no local non specific resistance could be induced by
injecting sterile broth or salt solution intradermally.

Where one is able to maintain a small colony of guinea
pigs, it would appear that the spread of infection could be
best prevented by separating the animals into individual
cages, thereby eliminating the possibility of a carrier
coming in contact with unaffected animals and using precau-

tions in handling the feed, feed dishes, and litter of these

animals.

3

6.

CO

C‘ 'T "5 4.
5.) AIAIA‘ELRY

Clinical, bacteriolotical, and pathological aspects of a
guinea pig epizootic are described.

B. type of hemolytic streptococcus is found to be the
etiological factor, affecting principally the peripheral
lymph g ands.

Roth acute septicaemic forms and chronic forms with
abscesses in the lymph glands are found in spontaneous
and artificially infected ruinea pims.

Cases of chronic lymphadenitis frequently terminate in
septicaemia.

The gland lesions in acute lymphadenitis coreist of in—
flammatory edema with numerous streptococci with or
Without leucocytic exudation.

The gland lesions in chronic lymphadenitis begin as focal
lesions containing numerous macrOphages, usually a few
streptococci, and occasionally a few leucocytes and
develop into abscesses through central necrosis and peri-
pheral extension of the lesion.

Pyemic focal streptococcus lesions occur in liver, heart,
uterus, spleen, small intestine, bone marrow, thyroid
glands, and adrenal glam-.3.

a simple method of demonstrating gram positive bacteria
in tissue sections, which at the same time prahlces sec—
tions stained excellently for histolorical study, is

described.

A method of eradication is recommended consisting of main-
taining a small colony of guinea pigs and isolating each

animal in an individual cage.

57

    

TRBLE I

Biochemical reactions of cultures from

5 cases.

 

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KEY T TAQLE II

Lymph glands
Early focal lesions
Acute lymphadenitis

Chronic lymphadenitis

Lungs
Congestion
Femorrhage

Pneumonia

Streptococci in section
Few, completely phagocytosed
Ioderate num er, completely phagocytosed
Kuherous, partly phagocytosed

Numerous, no phagocytosis

Streptococci in culture
Light growth
moderate growth

Heavy growth

59

++

+++

++

+++

++
+++

++++

++

+++

 

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With abscessation.

Guinéa fEUb — C ervl ca 3.3, _ 117131152, 1 :13 5 (3 w _ ‘5 :3. 1 ‘2 O I T “' : v“; :31... A... n 7 23751911113
pig LO. max1lle y “ ' ‘ * sgapular ‘ ‘W‘V' * A :;1130
1 +++
2 + “‘1‘“-‘ ‘-
5 ++
4 A ++" + ‘" ”M —.‘— ~:—+
5 +++ +++ L
7 +++ * l“
8 +++
10 +++ ++ ++ ‘w
11 +++ _‘ *7
12 %+ n- h}, '
15 w++ +++
141. +++ +++ - +++“
15 +++ + +++
17 ++ +
18 ++ +++ +++ +++ 4‘
19 +++
BC ++ ‘++
21 +++ L++
< 4 VWmvamme..- .. - - - w
25 +
24 ++#
25 ++ +++ +++
26 +
27 + + + +
28 ++
29 +++ —*¢~
30 +++ + tiim‘-‘ +++
51 +
55 ++
56 ++ +++
57 +++ +++ . -—
58 ++ ---- ~
59 +++ +++
40 +++ +++
41 + + +++
4-2 + + w
43 ++ .—
44 ++
45 +++ +++ +++
46 + ++
47 ++
i 4
48 +++
49 + +++ +++
50 +++ +++
51 +++ ++ +++
52 +++ +++ ++
55 ++ +++
54 ++ +++
56 ++
57 +++ ++ +++ +++
58 +++ ++ +++
60 +
61 ++ ++ +
62 +++
66 +++ ++
57 +++
68 ++
69 +++ ++
70 ++ ++ + ++ +++
71 + , +
72 ++
73 ++ ++ + +++ +++
74 +++
75 ++ +++
7
76 +++ +
77 +++
78 +++ ++ ++ +++
79 +++ +++ ++
80 +++ ++ +
101 +
102 +
201 +++ +++
202 +
203 +
205 +
206 + +
207 + + +
208 ++ ++ ++ +++
301 ++ +++ + +++
Key: Early focal lesions +
Acute lymphadenitis ++
Chronic lym_1adenitis +++

79

 

 

62

DESCRIPTIOE 0F PLATJS

Plate‘l
Fig. 1. Guinea p13 24, shoring enlarged cervical lymph
gland.
Plate_££
311‘: o 2 0

Section from submaxillary

13mph fland of guinea

a. Early focus of infection containini nume‘ous

macrophages, few fibroblasts, and an occasional

polymorphonuclear leucocyte. A few scatters

V—~u

streptococci can be seen in this lesion at
hither magnification.

b. Targinal sinus. Iagnification approx. 200x.

Fig. 5. Bronchial lymph :land from juinea pi: 207.

a. Focus of liquefaction necrosis showing no evi—
dence of cellular reaction.

‘ ‘ '

o. .arginal sinus.

c. Septun.

Kagnification approx. 200x.

Plate III

Lymph gland from.auinea pie 7, showing edema,
degeneration of the lymphocytes, and ClL'dpS of
streptococci in the lymph sinuse (a).

Yafinification appr V 0COX.

4.. . 2‘!

.4 K.‘

I;rmh.qland frmn‘uUJea pig 0"“7. -Knme lymph-

adenitis shoring edema and cTains of streptococci.

hagnification approx. A”Cx.

J.\

Fig. 6.

63

Plate-£3
Lymph gland from guinea pig 67. ftreptococci in
the marginal sinus, Shovine phagocytosis by macro—
phages. Kaenification approx. 400x. .
Lymph gland from guinea pig 59 showing early
abscessation at (a) with great thiekeninz of the
capsule. The great thickening of the capsule is

indicated by the distance between the two arrows.

Eagnification approx. 200x.

Plate.y
Lymph gland from same (guinea pig as figure '7,
showing an advanced abscess at (a) just prior to
rupturing. fhe almost complete erosion of the cap-
sule at (b). Compare with the thickness of capsule
in the earlier abscess shown in figure 7.
0. Blood vessel.
d. Areolar tissue.
Kagnification approx. 200x.
Section from lung of :uinea pig 74 showing early
acute bronchopneumonia with fibrinous exudate and
leucocytes in.the alveoli.
a. Bronchus shouin: sliflrt desquamation of epi—
thelium and peribronchial round cell infiltra-
tion.

,s‘

Iagnification appr x. 206x.

64

Plate _V_I

 

Fig. 10. Lung from guinea pi? 69 showing acute broncho—

pneumonia.

a. Sazfital section of a bronchus.

b. Vronchial exudate made up of desquamated epi-

helial cells and polymorphonuclear leucocytes.

The bronchial epithelium shows marked desquamation
at (c).

d. Slight hyperemia of the alveolar capillaries
with slight swelling of the alveolar epithelium.
The exudate in the alveoli is :ade up of leuco—
cytes, macrophages, and desquamated epithelial
cells. Etreptococci are visible in these alveoli
at higher majnification.

8. Area in which there is considerable leucocytic
exudate distending the alveoli and compressing
their walls.

Kagnification approx. BCCK.

Lung from guinea pig 27 shoving brondiOpneumonia.

a. Bronchus showin: desquamation of epithelium with
exudate in the lumen made up diiefly of desqua—
mated cells and mucus.

o. Yarked swelling of the epithelial lining of the
alveoli with an exudate made up of leucocytes,

‘

macrOpnages, and desquamated epithelial cells.

A few alveoli in this rejion show atelectasis.

Hagnification approx. 200x.

Fig;- 14:0

Plate VII

 

Lung from guinea pig 43.

a. A focus of early necrosis. ihe alveoli sur—
rounding this focus show pneuionia and contain
leucocytes, desquar-ated epithelial cells and
edema.

Kagnification approx. 200x.

Lung from fuinea pig 44. Pneuaonia win; a fistu-

lous tract Openin: on the pleural surface.

hagnification approx. 200x.

Plate

4
'5“

II

i—i

Liver from guinea pig 2C2. Focus of necrosis with
no evidence of cellular reaction. Iumerous pyknotic
liver cells are seen thzeughout the central part of
the lesion. The arrow points to a group of strepto-
cocci barely visible at this magnification.
a. Liver cells showing fatty degeneration.
Kagnification approx. 200x.
Same lesion as figure 14 magnified approx. 400x.
a. Streptccocci growing singly and in chains of

two to s x elements, some phagocytosed by

Kupffer cells.

b. Pat globules in liver cells.

Plate IX
Liver from guinea pig 202. Focus of infection
shoring chains of streptococci.

Kagnification approx. 400x.

Fig. 170

Fig. 18.

F180 19 o

66

Spleen from guinea pig 53. Acute spleenitis showing
edema.and phagocytosis of streptococci by spleeno—
C3rteSo

Magnification approx. 400x.

rarer
Mammary gland from.virgin guinea pig 207, showing
acute interstitial mastitis. The alveoli are dis—
tended with mucoid material. The interlobular
tissues are very edematous. The lower half of the
picture shows early necrosis. With higher magnifi-
cation numerous streptococci can be found in this
region.
Magnification approx. 200x.
Uterus from guinea pig 45. Pyendc focus in the
muscular coat.
a. Polymorphonuclear leucocytes.

b . Macrophages .

Magnification approx. 2001:.

ID/afe I

 

é?

 

63

Plate 1U

 

é?

—-———7
p
late IV
_ 70

 

 

7/

7.!

Plate V1

 

Fig. H.

 

Fla te V11

 

73

74

Plate VII]

 

[’2‘

[3.

 

FigJJ’.‘

 

 

Fn‘g . I 9.

76

ACIQTOE’LTDGlaic-

I wish to express my hearty appreciation
to Doctors E. T. Hellman and I. Forest
vHuddleson for valuable assistance and sugges-
tions in the study of this problem, to Doctor
L. B. Lholl for assisting in interpreting
some of the microscopic lesions, to Fr. W. L.
Hallmann for the use of the photomicrographic
equipment, and to Fr. J. E. Pratt for making

the photographic prints.

77

78
B BLIOGRAPHY

l. Eberth, E. J. Zwei hykosen des murschweinchens. Virchow's
Arch., 100, 15-27, 1885. (Cited by Parsons, Am. Jour.
Eyg., 8, 556, and holman, Jr. Hed. Res., 55, 15, l.)

2. Malassey, L. and h. Vignal. Tuberculose Zoogleique Forme ou
Espere de Tuberculose sans Bacilles. Arch. de Physiologic,
15, 569, 1885. (Cited by Parsons, Am. Jour. Eyg., 8, 556.)

Sur 1e micro-organisms de la tuberculose zoogleique. Ardh. de
Physiologie, 4, 51, 1884. (Cited by Parsons, Am. Jour.
Hyg., 8, 556.)

5. Chantemisse, A. La tuberculose zoogleique. Ann. de l'Inst.
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4. Dor, L. Pseudo—tuberculose bacillaire. Compt. Rendus de
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5. Weber. Inaug. Dissert. Tunich, 1901. (Cited by holman,
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6. Wagner, G. A. Puerperalerkrankung bei uprschweinchen. Cent.
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7. Boxmeyer, Chas. H. Epizootic lymphadenitis, a new disease of
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8. Teacher, John H. and J. A. G. Burton. Infectious abortion in
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9. Smith, Theobald. Sore bacteriological and environmental
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10. Holman, W. L. Spontaneous infections in the auinea pig. Jour.

Med. Res., 55, 151, 1916.

79

11. Hardenbersh, John G. Epidemic lymphadenitis with formation

12.

13.

14.

15.

16.

17.

18.

19.

20.

21.

of abscess in buinea pigs due to infections with hemolytic
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Parsons, Elizabeth I. and Poscoe R. Hyde. Spontaneous strep-
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8(5), 556, 1928.

Cunningham, J. 8. Epizootic lymphadenitis in guinea pigs due
to an incapsulated mucoid hemolytic streptococcus. Jour.
Inf. Dis., 45, 474, 1929.

V

Megrail, Emerson, and Robert H. Hoyt. Epizootic lymphadenitis
in guinea pigs. Jour. Inf. 313., 44, 245, 1929.

Hubert, L. D. and Meyers, K. F. Cutaneous immunization
against the streptococcus of guinea pig lymphadenitis.

Jour. Inf. Dis., 44, 489.

Hutyra, Franz, and Josef Karek. Special pathology and there-
peutics of the diseases of domestic animals. 1, 460, 1922.

éurgey, D. K. Differentiation of cultures of streptococcus.
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Rosenow, E. C. Transmutations with the streptococcus pneu—
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Johnson, hm. J., Jr. Influence of surface tension depressants
on the growth of streptococcus. VI Studies of the strepto—
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Gay, F. P. and L. F. Merrison. Clasmatocytes and resistance
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Beckwith, T. D. and 3. J. Rose. Virulence of hemolytic

Streptococcus. JOUI'O Info 313., 3'7, 227, 19250

80

22. Fisk, Etta and Earl L. Durky. A study of the classification
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25. Branham, Elizabeth Saier. Production of lung hemorrhages and
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24. Brown, J. Howard. The use of blood agar for the study of
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25. Talker, E. W. Ainley. The value of sugar tests for strep-
tococci. Jour. Path. and Bact., 15, 124, 1911.

26. Durke, Victor J. Notes on the gram.stain with a description
of a new method. Jour. Bact., 7, 159, 1922.

27. Palentine, Eugenia. A method of inoculation of blood plates
for the identification of streptococcus. Jour. Inf. Dis.,
45, 167, 1929.

28. Gordon, K. E. The differentiation of streptococcus. Jour.
Path. and 3act., 15, 525, 1911.

29. Branch, A. Spontaneous infections in guinea pigs, pneumoco-
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Inf. Dis., 4, 555, 1927.

50. KcLeod, J. I. The streptococci. Chapter II. A system.of
bacteriology in relation.to medicine. Vol. 2, pate 50.

51. Sabin, Florence R. and Chas. A. Doan. Relation of monocytes
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52. Jungeblut, C. W. The role of the reticulo endothelial system
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1927.

54.
55.

59.

40.
41.

42.

81

Jaffe, R. H. The reticulo endothelial system. Arch. of
Pati- o and. Exp. lied o , 4(1) , 1327 o
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Brown, J. Howard. A method for differential staining of gram
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Karsener, Howard T. Iuman Patholofy, 1926.

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Lee, Bolles, Kicrotomist's Vade-Kecum, 1928.

 

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