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BULLETIN No. 8

Use aR T MENT “Or AGRICULTURE.
BUREAU OF ANIMAL INDUSTRY. a,

A

INVESTIGATIONS
INFROTIOUS DISEASES AMONG POULTRY.

THEOBALD SMITH, Ph. B., M. D., and VERANUS A. MOORE, B, 8., M. D.,

UNDER THE DIRECTIGN OF

Dr. D. E. SALMON,

‘
CHIEF OF THE BUREAU OF ANIMAL INDUSTRY.

Wyn ’ Y
= Til, \

WASHINGTON:
GOVERNMENT PRINTING OFFICE.
1895,

A

Copy um
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VY

Monogrash

ae

LETTER OF TRANSMITTAL.

U. S, DEPARTMENT OF AGRICULTURE,
BUREAU OF ANIMAL INDUSTRY,
Washington, D. C., June 29, 1895.

Str: I have the honor to transmit herewith a report of Drs. Theo-
yald Smith and Veranus A. Moore concerning investigations of certain
nfectious diseases of poultry. This subject is one of great interest and
mportance to most people of this country who are engaged in agricul-
ural occupations, and particularly to all who make a specialty of some
yranch of the poultry industry. There are a vast number of our citi-
ens engaged in the production of poultry and eggs, either for their
ywn use or for sale, and most of these at one time or auother suffer
osses and have their operations interfered with by outbreaks of infec-
ious diseases. Such diseases are as destructive with birds as with
ther kinds of animals, and until recently they have received little
systematic study from competent scientific men.

The investigations, of which an account is presented in this bulletin,
ypen up a new field for scientific inquiry. While the work which has
een accomplished is but the first step, it reveals how little has been
mown and how much is to be learned in regard to the diseases of
rds. The greater part of the losses from infectious diseases in the
youltry yard may probably be prevented by the intelligent application
f proper sanitary measures. Until the nature of these diseases is
Jearly understood, however, it is impossible to intelligently formulate
reventive measures or to prescribe successful methods of treatment.

The financial investment in domesticated birds is of such magnitude,
und the products are marketed in such vast quantities, and are so
ssential to the well being of every citizen that the problems affecting
his industry are deserving of the most careful study and investiga-
ion. The researches herein reported have been made in ihe hope and
xpectation that they would be continued until practical means of pre-
renting the common infectious diseases are placed within the reach of
il engaged in the poultry industry.

Very respectfully,
D. E. SALMON,
Chief of Bureau of Animal Industry.
Hon, J. StERLING MoRTON,
Secretary of Agriculture,

. t=

LETTER OF SUBMITTAL.

U.S. DEPARTMENT OF AGRICULTURE,
BUREAU OF ANIMAL INDUSTRY,
Washington, D. C., April 2, 1895.
Str: [ have the honor to submit herewith some preliminary studies
of infectious diseases among poultry, including fowls, turkeys, and
pigeons. Though these studies have not yet reached that state which
permits us to recommend any well-defined preventive or curative meas-
ures, it is to be hoped that they will give those immediately interested
a better insight into the nature of some of these diseases, besides fur-
nishing investigators throughout our country a more secure basis for
further experimentation.
Very respectfully,
THEOBALD SMITH,
Chief of Division of Pathology.
Dr. D. E. SALMON,
Chief of Bureau of Animal Industry.
4

TABLE OF CONTENTS.

AN INFECTIOUS DISEASE AMONG TURKEYS CAUSED BY PROTOZOA (infectious
entero-hepatitis). By THEOBALD SMITH........---..-.--.-----.------
Bpechaulechakachers Ol the diseas@s= 22.c-- so aac cece coon) s-= See soe e seas
The micro-organism (Amoeba meleagridis n. sp., 1895) -.....---.------------
The relation of bacteria to the disease...--..-..-----...---.------------
The relation of this disease to certain other diseases of poultry ..--------
GeneragconehislOnsee macs tae wees atelseica nie seieineb ciate Semen ccemlecs sce cece
PASTS I) TS CI Nae ene epee tare ne iors aim aerate Aa Sale mien Se cisicis ecccs coe ee Bcltbw mere

COWWUOTERIRS .345.0n coon eeSbod SOU EOD OE aR DE STORED Cn nee See eon eee
Bactexlaassociabted. withsthe. lesions s... 2... .25..20c. poss oc- ces sen sces
Deseription of the nonmotile bacillus......-...-. 2.---26-5---2+-e----.---
LOVE nistOnndman (re abiMnertorsseloe eee eas Soares ce cebieke ne sehccciaeccinecce
The relation of diphtheria in fowls to public health..........-.--..------
(QUA OITIE MOMS Bei te ee des AERA ee 2 ieee tye Me Syme Rens anne ema ee

A Stupy or A BACILLUS OBTAINED FROM THREE OUTBREAKS OF FowL
CHOLERA. By VERANUS A. MOORE....--..-...-----: Soehercteias esac
EIRSTOMy Ole bl OrOULDTOMER =. ec osn Sees steeiceek ees ec ceieecicccisetion dane ceeeiss
Mescuiptionvot bhelbacwhwie. 2 os. J526 esos seen cece s sce bance s-cewe ees ssc.
iemaAliceO ri tne inoculation GISCASG.--aas--c ce ssc saceessese.ccceneccne

On A PATHOGENIC BACILLUS OF THE HOG—CHOLERA GROUP ASSOCIATED WITH
A FaTal DISEASE IN PIGEONS. By VERANUS A. MOORE....-.-.--------

UB LOL ye Oleulle CISGaAkOa ae seme acele cae eisce faces f= Decent sie sacs ecee ooeecc
WCSETID NOMEOr UNO MAGUS ee. = cals a saeco clsiaten/o<taidin a cieislaiea.cicine sceteejeimic'a aisle

Page.

LIST OF ILLUSTRATIONS.

Facing
page.

Puate I. Photographic reproduction of the cecum and the spleen from a
turkey affected with the protozoan disease. .-......-.---------- 78
II. Drawings of normal and diseased ceca and of the spots on the liver. &0
III. Recent and advanced stages of the liver disease ......-.--------- 82

IV. Microscopic appearance of the diseased cxecum and liver with the
mMicroparasibes Shown. =~ see cee = eee ee ae ee eee 84

V. Drawings showing microparasites, giant cells, and flagellates from
the Jblood 5-8... 252c0-sisdeeeeee nee eee eee eee eee 86

VI. Drawings showing the diphtheritic lesions in the mouth and, on
the cornea, of adtowl: Js. 2c ce see ea seen e ee eee 88

INVESTIGATIONS CONCERNING INFECTIOUS DISEASES AMONG
POULTRY.

AN INFECTIOUS DISEASE AMONG TURKEYS CAUSED BY PROTOZOA (INFECTIOUS
ENTERO-HEPATITIS).

By THEOBALD SMITH.

[Plates I to V, inclusive. ]

In the fall of 1893 the writer received from Mr. Cushman, of the
Rhode Island Experiment Station, the organs of an adult turkey in
which the liver and the ceca were very extensively destroyed by a
peculiar disease. A second case was sent by Mr. Cushman in April
of 1894, in which the same disease was recognized. In June of 1894
the writer found the same affection in a very young turkey in western
Massachusetts. The changes induced by this affection were so pecul-
iar and yet so uniform in these three cases that a specific infectious
disease could not but suggest itself on even superficial examination.
A careful study of this material convinced the writer that the cause of
this disease was a protozoan parasite not hitherto recognized. It was,
therefore, deemed best to investigate the disease more thoroughly, and
for this purpose the writer spent three weeks at the Rhode Island Sta-
tion during the month of August. The turkeys were obtained by vol-
untary contribution and by purchase, mostly from localities within
5 to 10 miles of Kingston, R. I. Some came from a greater distance.
In this work I was greatly assisted by Mr. Cushman, in charge of the
poultry work, and by Professor Flagg, director of the station. The
chemical apparatus of the station was placed at my disposal by
Professor Wheeler. |

With the aid of the apparatus brought from Washington and that
on hand I was enabled to make a careful examination of about 50 tur-
keys. Of these, 18 were affected with the specific protozoan disease,
and many of the rest were affected with other diseases, briefly referred
to under each case.

The short space of time which could be devoted to this subject did
not enable the writer to do more than to make a preliminary study

of the disease and its cause and to collect material for further study.
7

8 INFECTIOUS DISEASES AMONG POULTRY.

Now that a firm basis for the investigation of the practical problems
has been secured, it is to be hoped that the investigation will be prose-
cuted during the coming summer, both at the experiment station of
the Bureau and elsewhere. The publication of the work thus far done
will also enable the various experiment stations to make observations
on the communicability of the disease and on means for its eradication.

The external appearances of the turkeys affected with the disease do
not seem to be constant, and this is not to be wondered at in view of
the varying intensity of the changes found in the internal organs. In
Rhode Island the disease is known as “ blackhead,” owing to certain
peculiar discolorations which take place at the height of the disease.
While it might be well to retain this as a popular name, we do not
believe that all cases of blackhead have the specific disease herein
described, nor do all turkeys affected with this disease manifest the
appearances of blackhead. More extended study is needed before we
may rely upon symptoms as a means of recognizing this disease during
life, especially as there are several other affections of turkeys quite
distinet from this concerning which there exists as yet very little defi-
nite information.

As brought out more fully further on, the disease may be recognized
by anyone who has made himself familiar with the salient facts in this
article by an examination of the organs of the diseased turkey after
death.

Among the symptoms which may be expected to appear sooner or
later, diarrhea probably occupies a prominent place. The disease of the
ceca is responsible for this. Yet we may have diarrhea with at least
one other disease of the ceca, and the presence of tapeworms may also
cause this disturbance. Emaciation was not constantly present in the
cases dissected by me. As it may accompany other wasting diseases, it
can not be depended upon as a sign of this affection.

Leaving this part of the subject we shall turn to those characters of
which the investigations have given some definite information.

SPECIAL CHARACTERS OF THE DISEASE.

The disease seems to attack turkeys when quite young. In early
June I found a turkey about 3 weeks old (No. 3) in which the dis-
ease had already made considerable progress. It seems, moreover, as
if the disease were contracted only by the young, because in the
examination of turkeys of different ages the oldest showed lesions of
the longest standing; that is, such as had undergone the most exten-
Sive transformation. In general it may be said that the age of the
turkey corresponded with the age of the disease process. The most
Serious and extensive destruction of tissue was noticed in the turkeys
in fall. In midsummer the disease was making most progress; that is,
it was freshest and the microparasites present in great numbers. It is
probable that the delicate tissues of the young are best adapted for

SPECIAL CHARACTER OF THE DISEASE. 9

the temporary habitat and rapid multiplication of this parasite. In
this regard it simply follows the rule observed by large numbers of
parasites, whose most destructive action is visited upon the young.

The primary seat of the disease are the ceca. From these the liver
is invaded. Other organs are not attacked.

The ceca of the turkey in the normal condition are represented by
two blind tubes opening into the intestine at a point about 6 inches
from the cloaca (Pl. I, fig. 1; Pl. I, fig. 1). They are directed toward
the head, the blind end being nearer the gizzard than the opening into
the intestine. They lie on either side of the latter, closely bound to it
by mesenteries. The tips only are free. The length of the ceca varies
according to age. In turkeys 3 months old they are about 8 inches
long, while in those nearly a year old they are from 10 to 12 inches
long.!

The cea are of nearly the same diameter as the intestine. When
only slightly filled they are 4 to 5 mm. (one-sixth inch) in diameter.
They may, however, be distended with feces to three times this diam-
eter. The structure of the walls seems to be identical with that of the
intestine to which they are attached. The communication between
ceca and intestine is somewhat contracted. Near it in each tube there
is a convex elevation about one-half of an inch long which is composed
of lymphatic tissue. Its surface is usually deeply pigmented. Other
lymphoid structures are represented by small elevations only three-
sixteenths of an inch in diameter, usually pigmented and scattered
sparsely over the mucous membrane. The mucosa itself is folded in
longitudinal ridges, which are frequently pigmented.

The ceca usually contain feces in greater or less abundance which
are pasty, homogeneous in texture, and greenish in color. Under the
microscope they appear to be composed mainly of bacteria. When the
feces have a softer consistency numerous flagellates of several distinct
forms may be detected in active motion in the feces of the recently
killed bird. They appear to increase in number with the increase in
the watery constituents of the feces.

Coccidia have also been detected. They were present in No. 3 and
in another young turkey of the same flock which died from some
unknown cause. In the cecal contents of No. 17, which was free from
disease, similar bodies were found, the largest measuring 32 by 18 y.
They were in form and structure like Coccidium oviforme of the rabbit’s
liver, and perhaps identical with the form seen in the ceca of fowls by
Rivolta and Silvestrini and named C. tenellum by Railliet and Lucet.?
It is very improbable that these bodies stand in any genetic relation to
the true microparasites of the disease herein described.

The lesions of the ceca are in substance a thickening of the wall,

1 The length of the intestine from gizzard to opening into ceca is about 5 feet for
turkeys 3 months old and 8 feet for those nearly a year old.
2 Zoologie Medicale, second edition.

10 INFECTIOUS DISEASES AMONG POULTRY.

followed in most cases by a destruction of the epithelium and deeper
portions of the mucous membrane. This destruction results in the
outpouring of a coagulable fluid into the tube (PI. 1, fig. 1; Pl. I, figs.
2,3, and 4). The thickening of the wall may vary considerably in
extent from case to case. It may be uniform over the greater portion
of the tube or it may be limited to cireumscribed patches. The com-
monest seat of these lesions is near the blind end of the tube, where it
evidently starts and from whence it spreads to other portions. Not
infrequently only one cecum was found diseased, the other normal.
This exemption was not due to mildness of the attack, for the other
cecum may, in such cases, be the seat of very extensive changes. Thus
of 18 cases only 11 had both ceca involved.

The affection of the cecum is due primarily to the multiplication of
the microorganism to be described below, within the walls of the
eexcum. The multiplication may take place chiefly in the mucous mem-
brane, or in the submucous tissue, or it may, though rarely, extend into
the muscular coat. The thickening of the wall is the result of several
processes—the multiplication of the parasites, the increase of the normal
tissue elements, and later on the accumulation of masses of small cells
and some giant cells.

In the early stages of the invasion the adenoid tissue between the
tubules and in the submucosa becomes greatly increased, owing to the
presence of large numbers of microparasites of round or slightly oval
outline and from 6 to 10 in diameter which stimulate the proliferation
of the tissue cells. Numerous mitoses have been seen in this stage.
The parasites seem to occupy the meshes of the adenoid tissue either
Singly or in groups or nests (PI. IV, figs. 1,2). In these meshes they
are soon enveloped in cells acting as phagocytes, so that the appearance
of. an intracellular habitat of the parasites is suggested, which is, how-
ever, only an appearance and not demonstrated by further study.

The presence of the parasites in this reticulum probably stimulates
also the accumulation of lymph cells within the spaces, in virtue of
which the mucous membrane is thickened. In this early stage of inva-
sion the epithelium, both of the tubules and of the surface, remains
unaffected. The parasites do not invade the epithelium at any time. °

As the disease progresses there is a continued increase in the cellu-
lar elements of the mucous and submucous coats and a gradual invasion
of the muscular coats. Here the bundles of fibers of the circular coat
are thrust apart by masses of cells, so that this coat also becomes
greatly thickened. The inflammation finally extends to the serous
covering, where the blood vessels become greatly dilated and give the
cxcum a congested appearance. In cases of ordinary severity the wall
of the cecum, not more than 0.2 to 0.5 mm. (one one hundred and
twenty-fifth to one-fiftieth inch) thick normally, according to distention,
becomes 2 to 3 mm. (one-twelfth to one-eighth inch) thick.

SPECIAL CHARACTER OF THE DISEASE. i ail

With the progress of the disease the mucous membrane may be shed
and a coagulable fluid poured out into the cecum. In some cases it
appears in isolated masses, which adhere to certain spots of the mucous
membrane. In others, this exudate fills the entire tube with a yellowish-
white mold (PI. I, figs. 2, 3). This is built up in concentric layers,
and consists of a mixture of blood corpuscles, fibrin, and masses of
small round cells, in variable proportion.

In the further progress of the local disease it is not improbable that
bacteria are also concerned. ‘The exudate contains immense numbers
of them and the denuded mucosa furnishes a favorable place of entry.
It is otherwise difficult to explain the continued increase in thickness
of the walls of the cecum after the mucous membrane has been shed.
This continued increase in thickness is due to an extensive infiltration
of small round cells and the presence of some giant cells. Parasites
in this advanced stage are scarce, and usually recognizable only as
vacuole-like bodies within the giant cells.

The thickening of the wall is associated in some cases with an exten-
sion of the inflammation to the contiguous wall of the intestine, which is
thereby firmly attached to the cecum. Yellowish exudates are some-
times found outside of the diseased cecum on its serous covering and
they bind it inextricably to the other cecum or to the intestine or at-
tach it to the abdominal wal]. In these stages the microparasite is not
found. It seems to have done its work by destroying the mucous
membrane and to have left the field for miscellaneous bacteria.

In no case were ovher regions of the digestive tract affected. Though
the inflammation set up in the affected cecum may have extended
through the wall to the contiguous intestine and caused the formation of
a large amount of neoplastic tissue between the adjoining tubes, the
disease did not invade the submucous or mucous membrane and the
specific parasites were absent.

Associated with this peculiar disease of the ceca is a more or less
Serious disease of the liver. In the 18 cases of protozoan disease the
liver was very extensively diseased in 16. In two cases only small
sclerotic foci were present and in these the lesions of the ceca were
evidently at a standstill, or in a condition of active cicatrization. The
affection of the liver is in tact a valuable means of recognizing this
disease, for wherever it is present we may safely infer that we have
the protozoan disease before us. The reverse is, of course, not neces-
sarily true. The disease of the ceca may be so restricted that the
secondary liver disease does not appear. Nevertheless, the value of the
liver affection is indicated by the fact that in the few cases of cecal
affection in which the disease was of a different character no changes
in the liver were manifest (Nos. 25, 41).

The appearance of the liver in the average case of this disease is
very striking.” The organitself is enlarged in proportion to the amount
of tissue involved. Twice the normal size is probably near the truth,

12 INFECTIOUS DISEASES AMONG POULTRY.

although but one was weighed to accurately determine this point. In
this case (No. 52) the liver weighed 10.7 ounces.

Over the surface of the organ are distributed roundish, discolored
spots, distinctly demarcated from the surrounding tissue. These spots
vary much in numbers, in size, and in appearance. They may be dis-
tributed uniformly over the whole surface of the liver or they may be
limited in number to a few. They vary from 3 to 15 mm. (one-eighth to
two-thirds inch) in diameter. Inasmuch as a detailed description of
the different cases examined is given in the appendix, to which the
reader is referred, only a general description will be attempted here.

In endeavoring to group these spots according to their general appear-
ance we find several types, which, as will be seen below, correspond to
different conditions of the diseased tissue. We have in the early most
active disease process, spots like those photographed and figured in Pls.
Land II. They are sharply defined circular areas of a lemon yellow or an
ocher yellow color. A careful Scrutiny shows that the spot is not homo-
geneous in structure, but made up of a delicate network of yellow, as
shown in the plates. This yellow substance represents dead tissue.

In another class of spots we have in place of the yellow meshwork an
entirely different appearance. The spot is of a mottled brownish color
and contrasts only slightly from the surrounding liver tissue by its
darker color. It may containa central yellow nucleus of dead tissue and
a narrow outer border of the same character, or the border may be a
dark brownish circular line. The entire spot has an indistinct appear-
ance and is flattened or even slightly depressed below the surface. In
some cases the spots assume still another aspect. They are uniformly
whitish and shade off somewhat gradually into the surrounding tissue.

If sections be made of the affected organ it will be noticed that these
surface spots represent masses of liver tissue in the same condition.
The spots are simply the places where these disease foci intersect the
surface. Some are found deeply embedded in the liver tissue, and
therefore not visible on the surface. The lesion of the liver is thus
represented by few or many foci of disease having in general a spheri-
cal form and appearing on the surface of the organ as round spots (PI.
III, fig.1). Occasionally the lesions become more extensive and the
death of large portions of liver tissue follows. This is well shown in
fig. 2 of Pl. III, where a large volume of liver tissue is transformed into
a yellow cheesy substance.

The changes in the liver are most easily explained by assuming that
the microparasites are conveyed by the blood directly from the diseased
ceca into the liver and there deposited in different places, where they
begin to multiply and spread in all directions, thereby forming the
spherical foci of disease which appear as circles on the surface of the
liver. This theory is borne out by the results of the microscopic exam-
ination.

SPECIAL CHARACTER OF THE DISEASE. 13

In sections of hardened tissue from the liver in which the disease has
but recently begun the affected regions are invaded by large numbers
of protozoa, which occupy a kind of reticulum formed probably from the
connective tissue stroma. The liver cells have partially or wholly dis-
appeared from these foci. The parasites occupy the meshes of the tis-
sue either singly or in groups. The reticulum is provided with a small
number of nuclei, some of which are closely applied to and curved
partly around the parasite. The blood vessels are usually much dilated
and filled with red corpuscles.

The yellow masses observed with the naked eye in the surface spots
in many cases are Shown to be patches of an amorphous substance
which takes nuclear stains very feebly, the aniline colors not at all. It
may be described as a coarse network in the meshes of which small cells,
and very rarely parasites, are seen. This substance I assume to be the
result of coagulation necrosis of the liver cells by which they have lost
their nuclei and have become fused into a formless mass. I at first
was inclined to look upon it as fibrinous in character, but the discovery
of transitional stages confirms the view that it is made up of dead liver
cells. It is probable that the plugging of blood vessels in the liver by
parasites carried from the cecum is the cause of the necrosis, since
such plugs or thrombi are not uncommon in sections of the diseased
spots.

With the appearance of the microparasites reactive changes begin
at once which complicate the process. We have at the outset an
active multiplication of the microparasites which take the place of the
original liver tissue, and a process of coagulation necrosis going on at
the same time. Soon multinucleated (or giant) cells appear which not
only take in the microparasites but which are also engaged in remov-
ing dead tissue. At least their collection in groups around and within
the necrotic areas lead us to assume this. Not infrequently they are
grouped around what appears to be a plugged vessel, or else they
occupy the lumen of the vessel itself. The presence of round or oval
vacuoles within these large cells indicates that parasites have been
taken up (PI. V, fig. 3).

In still older cases the diseased spots are found more or less filled
with small round cells which may have passed into the dead regions
from the blood vessels. in all cases the latter are more or less enlarged,
and they seem to encroach upon the liver tissue, thus filling in part
the void produced by the cell death and giving the surface of the liver
a brownish mottled appearance wherever the disease spots are.

The processes of advancing disease and necrosis or death of tissue
on the one hand, of repair on the other, seem to go on side by side, now
one now the other predominating. Of the process of repair case 16 is
a very good illustration. In this bird the liver tissue was undergoing
sclerosis associated with the true healing process, which is referred to
more in detail in the appendix,

14 INFECTIOUS DISEASES AMONG POULTRY.

Besides the lesions directly due to the microparasites in the ceca and
in the liver, there were no changes observed to indicate that the parasite
was multiplying in other parts of the body. Even where the disease
was extending locally from cecum to adjoining intestine, the process
was not due to any advance or invasion of the parasites themselves.

Regularly recurring changes in other organs due indirectly to the
destruction of liver tissue, and to the loss of the ceca as important
aids in the process of absorption were not manifest.

The blood of many of the eases was examined before the bird was
killed in order to detect any existing blood infection. No disease of
the blood was observed, however, excepting in No?16, in which a pro-
nounced leucocytosis was present. In this case the disease was disap-
pearing and the liver undergoing sclerosis, as stated above. In a
small number of cases, some diseased, some healthy (Nos. 5, 6, 12, 15,
and 33) flagellates were detected in variable numbers. They were
evidently not related in any way to the protozoan disease. These
bodies (Pl. V, figs. 6 and 7) are spindle-shaped and made up of an oval,
feebly granular, central body (nucleus) around which the cell body
forms two lateral wings and two tapering extremities finely granular in
structure. Motion was not detected at the temperature of the sur-
rounding air. Higher temperatures were not applied. The nucleus of
this flagellate is about 25 4. long and 6 4 broad. The entire organism
is 65 to 70 «long. When the blood is dried in thin films upon cover-
glasses and stained, these bodies appear in many instances much
broader than in the fresh condition. This would suggest that the
organism may be a flattish body rolled up so as to bring the two lat-
eral margins near together.

The results of the examination of turkeys made thus far indicate that
the disease may follow several courses:

(1) After a certain period of disease regenerative processes begin
which tend toward a permanent recovery.

(2) The disease may proceed so rapidly from the very start that the
affected turkeys die early in life.

(3) The disease may come to a standstill, but the amount of dead
tissue in the ceca and liver may be so great as to favor the entrance of
bacteria which are responsible directly for the death of the bird late in
the summer or fall.

The direct action of the microparasites upon the body seems to lead
to a mechanical destruction of tissue. In this respect they differ from
most bacteria, which may combine a locally destructive process with a
general poisoning of the body. The extensive destruction of liver
tissue combined with the loss of the use of one or both eeca in the
digestive process must of necessity lead eventually to weakness and
emaciation. The feebler resistance of the affected animals was brought
to our notice last summer by their speedy death after a lon g ride or a
few days of continement.

THE MICROORGANISM. 15

As regards the spontaneous cure of infected birds we may regard it
as probable that only those in which the disease comes to a standstill
before it has made much headway are likely to recover. In those in
which much destruction of tissue has taken place complete recovery is
impossible. The dead masses embedded in the cxca or the liver, or both,
form a nidus for bacteria of certain kinds. These penetrate into the
pleuroperitoneal cavity and there set up inflammation and finally lead
to a general septic infection of the body. ‘The immediate effect of the
disease may thus be very slight at the outset and the symptoms develop
slowly. The great increase in the size of the liver indicates a certain
amount of compensation for those portions which have been destroyed.

The effect of the disease on the flesh of turkeys killed in the early
stages is not recognizable. In a few cases which died from a severe
form of the disease the flesh had a mahogany color, probably due to
retained biliary substances.

THE MICROORGANISM.
(Ameba meleagridis n. sp. 1895.)

In the 18 cases of this disease which came under observation either
the liver or the ceca, or both organs, were examined microscopically in
14, In these organs certain microorganisms were detected in all but
one case and associated directly with the peculiar disease process
already described. In those cases in which the disease was recent, or
at its height, the parasites were very numerous in the affected tissues,
while in those in which the disease process was far advanced, and
associated with degenerative or regenerative changes, the parasites
were found with difficulty. These facts are summarized in the follow-
ing table:

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THE MICROORGANISM. 17

In the examination of the parasite within the tissues minute particles
of the latter were crushed in iodized amniotic fluid or in normal salt
solution. Sections were also made. Unfortunately, a freezing micro-
tome was not at hand, nor any warm stage for determining the effect of
higher temperatures on these bodies.

The most frequent appearance presented by the parasites under these
circumstances was that of round homogeneous bodies with a sharply
defined, single-contoured outline (PI. V, fig. 1). Their appearance sug-
gests that of the myelin bodies found in the cells of the pulmonary
alveoli, not refrangent enough to represent oil globules, but too homo-
geneous to represent the ordinary protoplasm. Within these bodies
and situated somewhat eccentrically is a group of very minute granules,
probably representing a nuclear structure. These forms were encoun-
tered in those cases in which the parasites were found most abundant
in sections of hardened material (Nos. 5, 24, 26); also in one case in
which repair was evidently going on (No. 16). They vary somewhat in
size. In Nos.5 and 16 those that were measured were 8 to 10 jin
diameter. In No. 26 they were 12 to 14 in diameter. In No. 34 some
of the bodies were oval, having diameters of 12 and 15 jy, respectively.
They are thus distinctly larger than the parasites within the tissues,
which have undergone the hardening process. The latter are from 6 to
10 win diameter. This difference may be due to shrinkage, on the one
hand, and on the other to a slight flattening of the bodies by pressure
in the fresh preparations. These peculiar homogeneous bodies were
found, as a rule, free in the crushed preparations, although occasionally
giant cells were detected which contained a number of them. The cell
nuclei of the giant cell were not visible in the fresh condition. Numer-
ous coarse granules were embedded in its protoplasm, less frequently
fat globules.

Besides the homogeneous bodies, organisms of nearly the same size
but with uniformly granular protoplasm were also found, chiefly within
giant cells. These were not infrequently broken and rims or fragments
of protoplasm were left attached to the parasite (PI. V, fig. 2).

For the study of hardened material the tissues were removed in most
cases immediately after the bird had been killed, and placed in one or
more of the following fixing and hardening agents: 95 per cent alcohol,
saturated solution of corrosive sublimate, Foa’s solution (equal volumes
of the preceding solution and a 5 per cent solution of bichromate of
potash), Flemming’s solution (undiluted). The tissues were fixed in the
three last mentioned during one day, then washed in running water
during another day and treated with ascending strengths of alcohol,
according to the usually prescribed rules. The tissues were then passed
through absolute alcohol and chloroform, infiltrated in paraffin, and the
sections cut dry. Various staining reagents were employed, including
Delafield’s hematoxylin and eosin, alum carmine, Biondi’s triple stain,
saffranin, Gram and Gram-Weigert’s stain. Of the fixing and hard-

17897—No, 8-

9
od

18 INFECTIOUS DISEASES AMONG POULTRY.

ening agents I am unable to consider any one as superior to the
other in bringing out the parasites in the tissues when subsequently
stained. By this I do not mean to infer that there was no preference
when their action upon the tissues is considered, for here they mani-
fested their respective peculiarities already well known. But the fact
remains that now the one, now the other, tissue demonstrated the para-
sites best. In general, hardening in corrosive sublimate and alcohol and
staining in Delafield’s hematoxylin and in eosin proved the most suc-
cessful. The bodies were, however, brought out with other dyes, such as ~
methylene-blue, with variable distinctness.

The microparasites as found in sections of hardened tissue are spher-
ical or slightly oval bodies from 6 to 10 j« in diameter. In sections
stained with hematoxylin and eosin they have a homogeneous bluish-
red tint, feebler than that of the tissue nuclei of the host. They are,
however, distinctly seen, and the advantage of a more intense stain is
questionable. In most of these bodies a minute, distinctly blue ring is
seen situated centrally or somewhat eccentrically and representing the
nucleus. Thisis about2 «indiameter. With high powers a very minute
(nucleolar?) point may be seen in some parasites within the nucleus.
Other differentiations of structure were not noticed.

The presence within the connective tissue spaces of parasites in
groups of two or more individuals, as well as of single individuals,
makes it highly probable that at least in the early stages of the disease
there is active multiplication of these bodies. This appears to go on
as a Simple process of division, if we are to judge from the grouping
within the tissues. The variation in the number of individuals com-
posing such groups, as well as the: absence of any common membrane
inclosing each group, indicate at least that the multiplication is not an
endogenous segmentation such as belongs to the sporozoa.

This brings us to a consideration of the nature of these protozoa.
Their simple structure would lead us to class them with the ameebe, of
which one form is now known to produce a similar disease in the human
subject (amcebie dysentery). Amocebe are also not uncommon inhabit-
ants of the large intestine of man and certain animals. The peculiar
homogeneous structure and rigid outline of the organisms before us as
they appear when examined in tissues directly from recently killed
animals may represent a kind of encystment in the presence of the
adverse forces of the animal tissues. In the sharply defined vacuoles
within the giant cells this rigid form seems to have left a permanent
impression (PI. V, fig. 3).

The relation which the parasite bears to the microparasites found in
external tumors or warts (Jolluscum contagiosum) of fowls, or to those
flagellates which are associated with diphtheria in fowls, or finally to
the diphtheria referred to in text-books as due to gregarinosis may be
avery distant one. Yet, in the present state of our information con-
cerning these parasites, it is worth while to be on our guard before

THE MICROORGANISM. 19

making positive assertions, and leave it to future work to decide if any
relationship exists.!

We are naturally led, in a consideration of this microorganism, to
inquire what relation it bears to the tissue cells of the host. Is it an
intracellular parasite duriug any portion of its parasitic existence?
The evidence brought forward here would lead me to state that it lives
in the interstices and lymph spaces of the tissue, but not within cells.
This seems certainly true of the cecum. In the liver the liver cells
seem to become necrotic or else disappear so rapidly that it is impos-
sible to determine just where the parasites begin to multiply. They do
not live within the blood vessels, as they are not found within them
excepting perhaps in a thrombosed vessel. They must, therefore,
occupy the place of the liver cells. It is probable that they begin to
multiply in the connective tissue adjoining the blood vessel, and simply
crowd out the liver cells, leaving the connective tissue stroma of the
lobules in whose meshes they are found.

Their presence within giant cells is seen in almost every infected
organ subject to examination. In teased preparations of the fresh tis-
sues they are frequently found with remnants of the inclosing cells
still attached. This intracellular condition is, however, a purely pas-
sive one so far as the parasite is concerned.

The fate of the microparasite within the tissues of the host seems to
tend toward destruction. Both the death of the tissue itself and the
repair seem to lead to the disappearance of the parasites. In most
cases there may be seen in the same section a partial dissolution of
some of the bodies, while others are still in good preservation. Evi-
dently their life within the tissues is not very long. The attack upon
them by large giant cells in the later stages of the disease has already
been mentioned.

A discharge of the microparasites which escape destruction probably
takes place from the walls of the cecum, when these break down, into
the contents, in which they are carried outward. <A similar discharge
may take place from the liver through the bile ducts into the intestine.
Another way of dissemination would be in the death of the diseased
turkey and the dissolution of its body whereby the organisms are set
free. Speculations in this direction are, however, of little value until
experiment shall have determined whether turkeys are infected directly
from sick turkeys, from healthy, immune or recovered turkeys, in whose
ceca the parasite may perhaps vegetate indefinitely, or from some
external source.

The close analogy existing between this disease and that of amcebic
dysentery in the human being deserves at least a brief statement in
these pages. The occurrence of amcebe in intestinal affections of man
was noticed as far back as 1875 by Lisch. Since that date it has been

'The presence of numerous flagellates in the ceca of healthy turkeys has already
been pointed out on page 9,

20 INFECTIOUS DISEASES AMONG POULTRY.

the subject of many investigations.! Two of the more exhaustive
investigations which have appeared are those of Councilman and
Lafleur? and of Kruse and Pasquale.*

The points of analogy between the avian and the human disease are
that in both there is an affection of the intestine (large intestine in
man, ceca in turkeys) associated with liver disease due to amebe.

The intestinal wall in amcebic dysentery is greatly thickened, owing
to an cedematous condition. It is also thickened in circumscribed
areas and contains cavities filled with gelatinous-looking pus. The
amcebee vary much in size and contain vacuoles. They are found in
variable numbers in the bottom of the ulcers and in the discharges.
The large numbers of amcebee found in the intestinal contents led Coun-
cilman and Lafleur to infer an active multiplication therein. The pres-
ence of the parasites within the submucosa is described by these
authors in one case‘ only.

In the turkey we have seen that the parasites are always present in
the connective tissue spaces of the mucous and submucous membrane.
Their presence in the contents of the cecum is highly probable from
examinations made last summer but not definitely settled, as it is diffi-
cult to recognize the parasites in the feces.

The microparasite of the turkey disease differs from the Ameba
dysenterie in being quite uniform in its appearance, varying but
slightly in size (from 6 to 10 , in diameter) and in being free from
vacuoles. Movements characterized as amcboid have not yet been
demonstrated.

The liver affection in man appears usually as an abscess. In turkeys
it appears as a variable number of foci in which the microparasites may
be present in great abundance. The difference in the nature of the
lesions must be largely attributed to the different reaction of the tis-
sues of birds toward injuries. This brief statement must suffice to
point out the analogy between the human and the avian disease, and
the tracing of any further relationships must be left to those specially
interested in this subject.

Before concluding the discussion of the microparasite of this disease
I desire to refer briefly to peculiar organisms found in the lumen of the
tubules of the ceca. Fig. 5 of Pl. V depicts these bodies very well.
The figure represents a cross section of one of the tubules from the
diseased cecum of No. 26. It will be noticed that the tube is packed
with minute organisms of a roundish or oval outline provided with a
more deeply stained body, probably a nucleus. The organisms, after

‘For a good résumé, with citations of the literature, the interested reader is referred
to Schuberg: Centralblatt fiir Bakteriologie, XIII (1893), Nos. 18 to 22 inclusive.

°W. 'T. Councilman and H. A. Lafleur. Amebic Dysentery. The Johns Hopkins
Hospital Reports, II (1890-91), pp. 395-548.

°W. Kruse u. A. Pasquale, Untersuchungen iiber Dysenterie und Leberabscess,
Zeitschrift f. Hygiene, XVI (1894), pp. 1-149,

4 Loe, cit., p. 490,

RELATION OF BACTERIA TO THE DISEASE. Hk

being fixed in corrosive sublimate and hardened in alcohol, measure
from 4 to 5 «in diameter. The nuclear body, though it appears round
or oval in the figure, is actually more complex in form. Careful focus-
ing with a high power (x 1,000) shows a spindle-shaped or oval body
which may be bent like the are of a circle. Its shortest diameter is
about 1 yz. At first I was inclined to look upon these bodies as blood
corpuscles which had escaped into the tubules during the disease
process from the cavity of the cecum. This view is made very improb-
able by a number of facts. The red corpuscles in the capillaries of the
Same section are quite different from these bodies, both as regards the
color of the hemoglobin, and the form, size, and structural appearance
of the nucleus. These problematical objects assume a bluish-pink color
when stained in hematoxylin, and the nucleus stains quite deeply and
solidly. Again, it would be difficult to understand why all the tubules
Should become packed with these bodies if they were blood corpuscles.
Another supposition, that they might be some stage of the microparasite
of this disease does not harmonize with the fact that they are found
within the tubules of the healthy cecum (No. 47). They may be
absent in cases where the disease is most active (No. 3). Onthe other
hand, they were very abundant in No. 26, and less so in No. 16.

The most satisfactory assumption is that these organisms represent
one of the unicellular animals, probably a flagellate. It is interesting
to note that flagellates were found in large numbers in the fresh con-
tents of the cecum of No. 47,in which these bodies were found within
the tubules in sections of hardened tissue.

There is probably no genetic relation between this hypothetical .
organism and the true parasite of the disease under consideration.
The latter is larger, its nucleus is smaller and simpler in form, and its
behavior toward staining reagents is different both as regards the
cell body and the nucleus. It would seem, on the whole, that we have
here a microorganism which inhabits the cecum and by preference the
depths of the tubules. Its effect upon the health of the turkey can, of
course, be only conjectured at present.

THE RELATION OF BACTERIA TO THE DISEASE.

From what has been stated concerning the presence of the protozoan
parasite in the diseased tissues, there can be little doubt that they
must be regarded as the cause of the disease. In the course of the
investigations the possible bearing of bacteria upon the disease was
not entirely overlooked and cultures mainly upon agar were made from
the blood and the liver of a considerable percentage of the infected
turkeys. Usually, bits of liver tissue were transferred to the culture
tubes. Of the heart’s blood, one or two loops were inoculated. The
results obtained varied from case to case. As a general rule, tubes
inoculated from birds that had just been killed remained free from
growth, whereas the cultures of those which had died during the night

22 INFECTIOUS DISEASES AMONG POULTRY.

and were examined next morning were quite regularly fertile. The
bacteria isolated from these cultures were examined by Dr. C. I*. Daw-
son, assistant in the laboratory, and most of them found to be B. colt
communis. Several of these were afterward reexamined by me and his
results confirmed.

The absence of any uniformity in the bacteriological results as well as
the appearance of B. coli in the organs of dead turkeys indicates that
pathogenic bacteria were not associated with the protozoa in the cases
examined. It furthermore warns us to be careful not to attribute to
such bacteria as may be found the power of producing disease, as has
been lately done by Ligniéres,' who found JB. coli in the organs of a
small number of dead fowls and claimed, without positive proof, that
they were the cause of a septiczemia.

THE RELATION OF THIS DISEASE TO CERTAIN OTHER DISEASES OF
POULTRY.

(1) To other diseases of the ceca in turkeys.—In the investigations
made in Rhode Island three cases came under my observation which
make it certain that not all diseases affecting the ceca of turkeys are
due to the protozoa described.

Of one case (No. 7) only the intestines were obtained. One of the
attached ceca was greatly distended by an exudate, similar to that
found in some cases of the protozoan disease. At about 1 inch from
the blind end the cecum abruptly swelled out to a diameter of 14 inch.
The enlarged portion was about 2 inches long and very firm to the
touch. The whole mass was placed unopened in alcohol. When it
was subsequently opened the enlargement was found due to a firm
exudate with a narrow canal passing through the center. The exudate
readily peeled away from the wall, which was but slightly thickened.

The second case (No. 25) was similar to this. Only one of the ceca
was affected. ‘The notes on this case are as follows:

One cecum slightly distended by a rather firm cylindrical mass of feces encased in
a whitish leathery exudate made up of concentric layers. The whole is not adher-
ent to the mucosa. This membrane is dull, slightly congested. The wall is very
slightly thickened. The liver is free from disease.

In the intestines are numerous tapeworms. Two cultures were made by placing
bits of liver tissue in tubes of inclined agar. In both, colonies appeared. These
were subsequently found to be closely related to B. coli communis.

In the third case (No. 41) both cxeca were diseased, the liver normal.

One cecum was very much distended over a distance 3 inches long, attaining a
diameter of three-fourths of aninch. From the serous aspect the wall appeared very
hyperemic. When the tube was slit open the distension was found due to a cylin-
drical mass of exudate, very firm, yellowish white. The exudate appears deposited
in concentric layers. The lumen of the tube within this exudate was almost
occluded. Odor strongly fecal. The exudate easily peeled away from the wall in a
single mass. The wall was smooth and hyperemic and but slightly thickened.

'Septicemie 4 coli bacille chez la poule. Compt. rend. Soc. Biol. 1894, p. 135.

RELATION TO OTHER POULTRY DISEASES. 23

This exudate was hardened in alcohol and transverse sections cut from it. These
show that it is composed almost exclusively of a meshwork of amorphous material,
prebably fibrin. Cell groups were found in but one locality of the exudate.

The other cecum is only slightly distended. It contains a mass of exudate about
1 inch long and one-third of an inch in diameter lying loose in the cavity. The
mucosa is roughened and covered with a thin slough or exudate.

Sections of the wall of this cecum show the mucous membrane replaced by a
layer of young connective tissue cells in which tubules are absent excepting in cir-
cumscribed regions in which the remnants of the tubules are still recognizable in the
amorphous mass adhering to the submucosa, The muscular coats are not penetrated
by cell masses, but the vessels of these and the subserous membrane are greatly
distended with corpuscles.

The small intestine of this turkey contained numerous small tapeworms.

Two agar cultures were made from heart’s blood and one with a particle of liver
tissue. In one of the blood cultures a number of similar colonies developed. The
bacilli composing one of them had all the cultural characters of B. coli communis.

These three cases are of interest as indicating the existence of some
other agency, probably bacterial in nature, which affects the ceca of
turkeys. It differs from the pretozoan disease in the entire absence of
any liver affection and in the absence of any marked thickening of the
walls of the ceca. The exudate is much more abundant and obstruct-
ive than in the protozoan disease. Further investigations are neces-
sary to define more clearly the nature of this affection and its cause.
A sinilar affection of the ceca was found in two turkeys by Von Ratz
in Budapest.!

He found much thick mucus in the mouth and esophagus. The mucous membrane
of the small intestine was studded with hemorrhages and small, removable pseudo-
membranes as large as lentils. Both ceca of one turkey contained a partly dried,
cylindrical mass, and many minute worms 8 to 14 mm.long. In one excum of the
other turkey the same dry, cylindrical mass, easily removable. The mucous mem-
brane itself was covered with layers of pseudomembrane. The other sx:cum was
distended with gases and a grayish, semifluid mass. Both contained many round-
worms.

The author does not hesitate to attribute the cecal disease to the
roundworms. This conclusion is, however, merely an inference based
on the presence of worms and exudate. It is more than probable that
the disease is bacterial and secondary to the catarrhal affection of the
mouth.

(2) To disease of the cwca in fowls.—This is by no means uncommon.
Ziirn’? considers it a form of diphtheria which may affect the various
parts of the head, the digestive and respiratory organs. Hestates that
the disease of the cecum may appear as an independent affection in
water fowls and turkeys. The disease is accompanied with great
depression and weakness. The diarrhea which appears is at first semi-
fluid and mucous, later it becomes fluid and is mixed with blood. The
foul-smelling discharges may be followed by constipation, and then the

1 Blinddarmentziindungen bei Truthiihnern durch Ascariden verursacht. Deutsche
Zeitschrift f. Thiermed., XX (1893), p. 74.
* Deutsche Zeitschrift f. Thiermed., X (1883), p. 189.

24 INFECTIOUS DISEASES AMONG POULTRY.

ezeea and the rectum are found after death filled with yellow croupous
masses in layers upon the frequently ulcerated mucous membrane.
The disease lasts fourteen to twenty days or even two to three months.
Often a relapse follows an apparent cure. He states that it frequently
happens that the croupous or diphtheritic affection of the throat, nose,
and eyes appears healed, when suddenly the intestinal disease breaks
out and usually ends fatally.

The writer has seen two instances of disease of the cecum in fowls.
In one case one cecum had its walls very much thickened. The same
was true of the second case. In this the thickening was due to a
reparative inflammation following the loss of a greater part of the
mucous membrane. In sections of hardened tissue protozoa were not
recognized. It is probable that in this.fowl the disease was secondary
to a dipththeritie affection of the head.

Siedamgrotzki! describes a disease of both ceca in a hen. They
were very much enlarged. In one cecum the wall was thickened by
inflammation, but the mucous membrane was intact. In the other the
wall was also thickened and adherent to neighboring organs. The
mucosa was covered with a yellowish-brown, thin, pseadomembrane.

Disease of the ceca in poultry seems to be a widely distributed
affection, both primary and secondary to other diseases, and evidently
due to more than one ageney. Itis to behoped that more will be known
concerning these affections in the near future, and that the demonstra-
tion of one specific cause in the form of protozoa will pave the way for
the determination of the others.

GENERAL CONCLUSIONS.

In the preceding pages we have shown that there prevails a disease
of turkeys which attacks the young by preference, which runs no reg-
ular course, but varies from bird to bird in severity, duration, and
termination. It begins in the ceca and thence invades the liver
through the blood. Though restricted to these two organs, its action
is Severe enough to prove fatal to many affected turkeys. We have
seen that the disease process is always associated with a protozoan
parasite of very minute size, whose destructive action is due in the
main to its powers of rapid multiplication within the tissues. It is
probably discharged with the droppings of the diseased birds. These
are the main facts learned by the brief investigations of last summer.
While we may make certain conjectures as to the modes of transmis-
sion of the infectious agent based on these investigations, we are at
present unable to go further than this until experiments have been ear-
ried forward in other directions.

From our present standpoint I think it safe to assume that the micro-
parasite is transmitted from bird to bird without passing through

1 Bericht ii. ah Potenimeneresen « im iRcainen Sucmeeet 1872, p. 84.

CONCLUSIONS. 25

any intermediate host. The microparasite, discharged perhaps in an
encysted stage from the sick bird, is taken up with the food and water
by others and sets up disease directly.

If the parasites were taken up with insects, for example, we should
expect to find the disease diffused through all flocks. But even the lim-
ited experience of last suminer leads me to believe that certain flocks
only are infected and that by uninterrupted transmission the disease
becomes perpetuated and diffused among neighboring flocks. The
perpetuation of the parasites I am inclined to think is to be sought for
in the older turkeys, which carry them in their body, most likely in
the digestive tract during the winter. This view is supported by the
fact that turkeys not infrequently recover from the disease. This re-
covery does not mean, however, the destruction of the parasite within
the body, for we have now information concerning the persistence of a
variety of parasites—bacteria and protozoa—within the body long after
recovery.

The evidence which I have presented in this report seems to show
that the turkey is infected early in life and that infection does not take
place later on. If this be so, the transmission from the old to the young
is easily understood. If the susceptibility to infection remained for any
length of time we should expect to find the two cvca infected in every
case. Butin 7 out of 18 cases only one cecum was found diseased,
the other normal. It is true that other explanations of this condition
referable to acquired immunity, might be given, but the one cited seems
to be the most acceptable.

If the theory that the disease is transmitted more or less directly
from old to young should prove to be true, the remedy for such a state
of affairs would appear to lie in two directions:

(1) The disease might be allowed to go on and some remedy found
which will check it and lead to a cure; or

(2) The diseased flocks might be entirely destroyed and new birds
obtained elsewhere, after a thorough cleansing and disinfection of the
territory formerly occupied by the flocks.

The latter alternative, though very heroic and severe at first, seems
to me the best in the end, especially as we might search in vain for a
remedy which will check the disease in the young. Anticipating the
investigations somewhat in this direction, I had hoped to determine
the distribution of this disease in the country at large by an appeal for
the shipment to this Department of diseased turkeys or organs there-
from. Up to the present no material has been received, so that the
study of distribution has made no progress.' It is evident that if the
disease must be weeded out by a destruction of whole flocks it is highly
important to find some locality free from such disease from whence new
stock may be obtained. To obtain such information is by no means an

1The ceca of about 30 turkeys from the Washington markets were examined
during February of 1895, and found free from disease.

>.

26 INFECTIOUS DISEASES AMONG POULTRY.

easy task, and may be made still more difficult by the infusion of a
certain amount of local pride and self-interest. The disease is, how-
ever, readily recognized in most cases, so that anyene who has examined
the plates of this article and read certain portions of the text may be
able to detect it. The peculiar circular spots on the liver, coupled in
later stages of the disease with the presence of yellowish cheesy masses
or whitish scars in this organ, are very characteristic and probably
peculiar to this affection. Disease of the ceca, unless accompanied with
the spots on the liver, should not for the present be regarded as belong-
ing to this affection.

When we come to the subject of treatment some difficulties arise.
Though turkeys may appear drooping, unable to keep up with the move-
ments of the flock, and even have diarrhea, the precise nature of the
disease is not made clear by these symptoms. Among the 50 turkeys
examined many were taken alive as suspicious cases. When killed the
protozoan disease was not found. They were suffering from a variety
of other affections, the causes of which, mentioned in the order of fre-
quency, were lice, tapeworms, gapeworms, ticks, injury, and a diphthe-
ritic disease of the cea likely to be mistaken for the genuine protozoan
disease. The same treatment applied to such a variety of disorders can
hardly be looked upon very hopefully. Still, the subject merits attention.
The success which quinine has had in combating malaria leads me to
suggest its use to those who are in a position to experiment with this
disease. Since the microparasites are embedded in the walls of the
exca and in the liver the remedy to be tried must do more than act
locally in the intestines. It must act upon the parasites through the
blood, and its effect on the diseased turkey should be carefully watched
to note any poisonous action.

The disinfection of the coops and other structures designed to give
shelter to turkeys, and other poultry as well, should be carried on as for
bacterial diseases until further investigations shall have been made.
The following disinfectants are strong enough to kill spores of bacteria,
and will probably destroy the various stages of the protozoa:

(a) Corrosive sublimate (mercuric chloride), 1 ounce in about 8 gallons
of water (one-tenth of 1 per cent). Thewater should be put into wooden
tubs or barrels and the powdered sublimate added to it. The whole
must be allowed to stand for twenty-four hours, so as to give the subli-
mate an opportunity to become entirely dissolved. Since this solution
is poisonous, it Should be kept covered up and well guarded. Itmay be
applied with a broom or mop, and used freely on all woodwork. Since
it loses its virtue in proportion to the amount of dirt present, all manure
and other dirt should be first removed before applying the disinfectant.
The manure should be covered with lime.

(b) Chloride of lime, 5 ounces to a gallon of water (4 per cent). This
should be applied in the same way. ,

APPENDIX. aa

(c) The following disinfectant is very serviceable. It is not poisonous,
but quite corrosive, and care should be taken to protect the eyes and

hands from accidental splashing:
Gallon.

Ormdercan pO cracidmnnerssa se seee see ceca saos = iss foeaonsses Sasccie assis 4
(Cite SUL Oe POs ie 3a 5 KEES Gace ce bn Bena Eee ee OOC ESO OSSD Sea 4

These two substances should be mixed in tubs or glass vessels. The
sulphurie acid is very slowly added to the carbolic acid. During the
mixing a large amount of heat is developed. The disinfecting power
of the mixture is heightened if the amount of heat is kept down by
placing the tub or glass demijohn containing the carbolic acid in cold
water while the sulphuric acid is being added. The resulting mixture
is added to water in the ratio of 1 to 20. One gallon of mixed acids
will thus furnish 20 gallons of a strong disinfecting solution, having a
slightly milky appearance.

(d) Ordinary slaked lime, though it does not possess the disinfecting
power of the substances given above, is nevertheless very useful, and
should be used more particularly on infected soil.

APPENDIX.

In the following pages is given the history of the turkeys which have -
been examined up to the time of the completion of this bulletin. It was
thought best to include the brief notes of those turkeys affected with
maladies other than those of the infectious disease under consideration,
to illustrate the existence of such affections and the necessity for fur-
ther investigations.

Turkey No. 1.—Portions of the liver, czeca, and lungs received in weak alcohol
about the middle of October, 1893, from Mr. Cushman. Age of fowl not known.
Lungs dark, congested. The wall of the cecum is fully 5 mm. thick. In the liver
are disseminated firm yellowish masses simulating the cheesy matter in tuberculosis,
Tissues not in very good condition for microscopic examination. They were, how-
ever, placed in absolute alcohol and subsequently cut in paraffin. Sections of lung
tissue show all vessels densely packed with red corpuscles. No other changes.

Sections of the thickened wall of cecum stained in alum carmine, hematoxylin,
and eosin, methyl violet, according to Gram, and in Weigert’s fibrin stain. The
hematoxylin proved to be the best.

The thickening of the wall is due in the main to cell infiltration of the submucous
tissue. The entire mucous membrane is sloughed away, and the submucosa presents
a ragged appearance along the exposed border. Throughout the section, and more
particularly near the serous aspect, there are many distended capillaries filled with
blood corpuscles. The nature of the cellular infiltration not recognizable, owing to
imperfect hardening. In certain areas there may be seen, however, a peculiar col-
lection of cells having several nuclei and inclosing large vacuole-like spaces. These
spaces were subsequently recognized as having been occupied by protozoa. These
phagocytes, with several nuclei shaded off into exceedingly large giant cells, over
80 in diameter, often forming large patches by collecting into groups. They are
quite uniformly dotted with nuclei, and contain relatively very few parasites.

Sections of liver tissue showed an extensive substitution of the parenchyma by
numerous and greatly dilated blood vessels within newly formed connective tissue.

28 INFECTIOUS DISEASES AMONG POULTRY.

In such areas only a few islands of liver cells are recognizable. Parasites not seen.
Giant cells rare.

Turkey No. 2.—lLiver and a portion of the intestine, with ceca attached, sent by Mr.
Cushman from Rhode Island on April 2, 1894, and received April 5. Turkey probably
a large one.

Post-mortem changes well under way when organs were received. It was, how-
ever, not difficult to recognize the disease. Only one cxum affected. About one-
half of the tube, beginning with the blind end, is almost completely occluded with a
firm, pale-yellowish exudate having the appearance of concentric layers when cut
across, like the rings of a tree. The occlusion is complete at the blind end. The
tube, usually only 0.5 to 1 em. (two-fifths inch) across, is distended to 2 em.
(four-fifths inch). The exudate is firmly attached to the greatly thickened walls.
Sections of hardened tissue show that the entire wall of the cecum, or what is
left of it, is densely infiltrated with round cells. The lesions characteristic of the
earlier stage of parasitic invasion no longer recognizable.

The liver is beset with a considerable number of circular, slightly depressed areas,
which have a pale, mottled appearance. ‘They vary in diameter from 5 to 10 mm.
(one-fifth to two-fifths inch). When incised the same appearance is presented within
the liver tissue, showing that these circular spots simply represent the surface of
roundish masses of tissue which have undergone the same change.

In sections of hardened tissue each disease focus is shown to be the scene of a com-
plex process of destruction. ‘The periphery consists of normal liver tissue, the capil-
laries of which are distended, and gorged with red corpuscles. Within the diseased
focus there is disseminated a peculiar meshwork of amorphous, homogeneous matter
which stains but feebly in hematoxylin. Its meshes are large enough to hold one
or more tissue cells. In some places it resembles more closely mammalian fibrin in
its appearance. Liver cells are either entirely absent or present in isolated numbers
where this substance appears. Besides this there are areas of considerable cell infil-
tration and, scattered about without regularity, giant cells. Parasites are present,
but in small numbers.

From the liver bits of tissue were placed in three agar and three gelatine tubes.
At the same time bits were placed under the skin of arabbit and a guinea pig. The
cultures remained sterile and the animals well.

Turkey No. 3.—Small specimen; probably 3 or 4 weeks old. Died in the morning of
June 6, 1894, on a farm in Berkshire County, Mass. Examined several hours after
death. Weather quite cold. In this case both ceca and the liver were involved.
In the former the walls were considerably thickened, but there was no exudate
within the tube and the mucous membrane appeared intact.

In sections of the ceca hardened in alcohol and in corrosive sublimate the wall
was found of variable thickness. In some regions it was nearly norma! on one side
and considerably enlarged on the opposite side. In others the entire cross section
was uniformly thickened. The mucosa is still in position and but slightly altered.
The cells of the crypts show extensive mucous metamorphosis. The enlargement of
the wall is due mainly to an extensive infiltration of the submucosa and of thecircular
band of muscular fibers with the protozoa. The muscular fibers are pushed apart into
strands, or else entirely obliterated. , These microparasites are found throughout the
entire section in uniform abundance. They are comparatively rare in the reticular
tissue of the mucosa between the crypts. In the fresh contents of the ceca were
found coccidia with distinctly double-contoured wall. The long axis measured 15
to 20 4, the short 10 to 13 x.

The liver of this bird was dotted with about a dozen circular spots, varying in
size, some 7 mm. in diameter. These spots consist in the main of minute yellowish
lines embedded in the liver tissue and loosely interlacing to form a circular wheel-
like area. They thus differ from some other cases to be recorded in less destruction
of liver tissue within the circular area, for the yellowish lines represent necrosis of
liver tissue.

APPENDIX. 29

In sections of liver hardened in corrosive sublimate and alcohol the foci of disease
no longer contained any liver tissue. This is replaced by numerous microparasites
surrounded and enveloped in a meshwork of reticulated tissue rich in nuclei. Giant
cells are present in moderate numbers.

At the time this young turkey died two others were found dead and were exam-
ined. Neither was affected with this disease. ;

Turkey No. 4.—Has been dead for two days. Decomposition well under way.
Obtained from a dealer. Extensive spotted disease of the liver and thickening of
walls of ceca with exudation into the tube.

Turkey No. 5 (Farm A).—Probably 8 to 9 weeks old. Received alive August 3.
Blood from vein of the skin examined. A few fusiform flagellates seen. Nothing
abnormal with the corpuscles. Killed. On the neck a small tick. Mouth, trachea,
and lungs normal. The digestive tract normal with exception of the duodenum and
the ceca described below. The former shows marked pigmentation in points and
strice (vill). In the rectum and cloaca contents pale-yellow, liquid, containing urate
spheres in abundance and numerous flagellates.

Both cxea are diseased. They are unusually short, only one-half the length as
usually found in turkeys of the same age (PI. I, fig. 2). Both are distended from one-
half to three-fourths of an inch in diameter, and very firm, asif filled up. Vessels
on the serous surface injected.

~The condition of one of them is shown in PI. II, fig. 2. The cecum is slit open
longitudinally, and the cut surface exposed to view. The lower two-thirds of the
tube is completely occluded by a firm exudate. On the cut surface the wall of the
cecum is shown as an irregular line, indicating marked thickening along the whole
length of the tube. The exudate is pale yellowish-white in color, excepting below,
where it is largely made up of a blood clot. The main mass contains centrally an
irregular cavity. Above, the tube is filled with small round stones, probably dis-
charged from the gizzard. The other cecum has its wall very much thickened, as
shownin PI. II, fig. 3. The exudate is also present, but not attached to the walls in
the alcoholic preparation.

The liver, which is enlarged slightly, is spotted on all surfaces with isolated and
confluent areas of a circular outline and pale yellow in color. They rarely exceed
4 mm. (one-sixth inch) in diameter. On closer scrutiny the yellowish color is seen
to appear as a network. These disks correspond to masses of diseased tissue within
the substance of the liver. ;

In crushed, fresh tissue are found isolated and agglomerated bodies of a round
form, with a homogeneous colorless disk and a single-contoured outline (Pl. V,
fig. 1). Near the center of these bodies is seen a small mass of very fine refrangent
granules. They are 8 to 10” in diameter. One per cent acetic acid will not affect
their appearance.

Three cultures on inclined agar were made with a large and two small bits of liver
tissue and a large bit of spleen tissue (about the size of a pea). Furthermore, tubes
were inoculated with a loop of blood and with a loop driven into the liver tissue.
The six tubes remained free from growth.

Turkey No. 6 (Farm B).—Selected because of slightly reduced condition. Cooped
for two days. Found dead August 5.

In abdomen some blood-stained serum. Considerable hyperemia of serous cover-
ing of gizzard and of mesenteries. The blind ends of ceca knotted inextricably
together, enveloped in a mass of yellow exudate, and attached by means of it to
neighboring coils of the intestine and to the abdominal wall. The latter is infil-
trated and discolored at this point. After some dissection it was found that only
one cecum was primarily diseased. This was filled with an exudate at the blind
end for a distance of three-fourths of an inch.

The liver is somewhat enlarged, and on its surface are a small number of pale
yellowish areas fully one-half of an inch in diameter, They correspond to masses of
similarly affected tissue in the liver,

wif”

30 INFECTIOUS DISEASES AMONG POULTRY.

Sections of liver tissue hardened in alcohol were made through these large pale
spots. In these only small fragments of recognizable liver tissue were found inter-
spersed in the diseased focus. Much of this was occupied by a meshwork represent-
ing necrosed liver cells fused together. Small necrotic foci were also detected outside
of the main focus, embedded in still unchanged tissue. The necrotic meshwork is
arranged in spots generally connected, the interspaces being filled up with groups
of minute cells and occasional patches of microparasites. Giant cells are common,
usually within or on the circumference of the necrotic areas. The microparasites
are also inclosed in these cells. The scarcity of these bodies makes it probable that
the disease is not longer progressing.

Cultures were made by transferring a loop of heart’s blood and a loop which had
been forced into the liver through a scorched area to inclined agar.

The blood culture remained sterile. The liver culture contains manifestly several
species, only one of which was studied, and found to resemble B. coli.

Turkey No. 7.—Received from a dealer the intestines of 16 turkeys for examination.
In one of these there was disease limited to one cecum. (See page 22.)

Turkey No. 8 (Farm C).—Received about August 3 and cooped temporarily.
Found dead August 6; alive the day before.

The lesions in this animal are restricted almost entirely to the cca. In one
exeum the mucosa is beset with about eight peculiar elevations, which have a central
yellowish slough and a surrounding infiltrated zone, making the whole crater-like
body about three-eighths of an inch in diameter. On the serous surface some of
these necroses show as whitish, discolored areas, with injected border.

The other cecum contains but one ulcer. In the liver several small areas are
found which appear necrotic, but whose nature is doubtful.

Trausverse sections of a portion of the wall of the cecum hardened in alcohol
were made through one of the ulcers. The mucous membrane had sloughed away
over the center of the ulcer, and the submucous tissue and the muscular coat were
extensively infiltrated with masses of cells.

Two cultures were made on inclined agar, one with a loop of heart’s blood, the
other with a bit of liver tissue. The latter remained sterile. From the former two
bacilli were isolated, one corresponding to B. lactis aérogenes, the other to B. coli.

Turkey No. 9 (Farm D).—Brought to laboratory dead, Probably 38 months old.
The disease is restricted to the ceca and the liver.

In one cecum there are three, in the other four, thickenings of the wall, which
appear as opaque yellowish spots under the serous covering. Two thickenings are
situated near the blind end, the rest near the attached end. From the mucous sur-
face they appear as yellowish spots from one-eighth to three-sixteenths of an inch
thick in the center and thinning out from that point.

The liver is beset with large, isolated and confluent roundish patches of a grayish
color, mottled with yellowish specks. The same appearance is presented by the cut
surface.

In crushed preparations of the liver a few round, uniformly granular bodies, some
within what appear to be the remnants of cells, detected.

Sections of liver tissue hardened in alcohol present the usual appearances charac-
teristic of the disease. Sections passing through the cireular spots show that a
considerable area of such spots consists of the homogeneous meshwork resulting
from necrosis and fusion of the liver cells. In some of these, giant cells are situ-
ated, singly or in groups, many of them inclosing vacuoles which represent the
former seat of microparasites. The regions not occupied by necrotic tissue are
largely made up of cells which have a small, round, densely-stained nucleus, envel-
oped in considerable protoplasm. The nature of these cells is unknown, although
it is probable that they correspond to the round cells in the inflammatory processes
of mammalia. In the liver tissue beyond the diseased circle isolated foci are pres-
ent, which are made up mainly of the cells described and some interspersed giant

APPENDIX. 31

cells. Throughout the diseased area the various cell forms are undergoing degener-
ation. This manifests itself by a breaking up of the nuclear substance into stained
granules of limited number. This phenomenon is especially frequent in the epithe-
lium of the bile ducts, and more rarely the giant cells themselves show this degener-
ation. :

Sections were also made through one of the thickenings in the wall of the exea.
These showed that the thickening was due in the main to a very extensive cell
infiltration of the submucosa and of the muscular portion of the wall. Under the
serosa the blood vessels are much dilated, and gorged with red corpuscles. The
mucosa is not thickened, but is more or less infiltrated at the base. Much of the
epithelium is gone, both of the surface and of the tubules.

The microparasites are poorly preserved, but a little scrutiny shows them to be
present in certain regions of the mucosa, submucosa, and rarely in the cell masses
between the displaced muscular bundles. In the mucosa and submucosa they have
invaded large patches, and here they are ranged side by side and are inelosed in a
reticular tissue, each parasite by itself. Giant cells are also present in groups,
usually consisting of little else than a mass of parasites enveloped in a nucleated
network of protoplasm.

Turkey No. 10 (Farm E).—Probably 8 to 9 weeks old. Supposed to be infected
because unthrifty. Cooped for a few days. When killed no lesions found. On the
skin were some ticks and lice: in the intestines a few tapeworms.

Turkey No. 11.—From the same flock and in the same condition as No. 10. No
lesions found when animal was killed. A few ticks on border of meatus of ear.
Lice abundant.

Turkey No. 12 (Farm C).—Probably 3 weeks old. Cooped for a week before it was
killed. A few flagellates in blood from veins of skin.

Tapeworms in upper small intestines. In soft contents of cecum immense num-
bers of flagellates. No disease of cxecum or liver,

Two cultures made with bits of liver added to agar remained sterile.

Turkey No. 13 (Farm F).—Brought alive. because suspected of being diseased. No
lesions discovered on dissection. Many small tapeworms in duodenum. In eeeal
contents many flagellates.

Turkey No. 14 (Farm G).—About 3 months old. Taken from a flock August 8 be-
cause of lack of strength to keep up with the rest when driven. Indications of diar-
rhea. Placed in a coop, where it died during the night. Examined next morning.

Slight odor of decomposition. A few small warts on skin of neck. The various
organs were found normal, with the following exceptions:

Mucosa of duodenum’ almost blackish, from intense injection and pigmentation of
villi.

Both cieca diseased. The left is slightly distended. Onserous aspect two yellow-
ish spots, with markedly injected borders, corresponding to thickenings of the walls
near the blind end of tube. The mucous surface of one is smooth; to the other an
exudate isattached. Besides the thickening at these spots, the free half of this cecum
is somewhat thickened uniformly.

The right cecum is very much distended over two-thirds of its length. From the
serous surface local thickenings are recognizable, which have a yellowish, mottled
appearance. The small intestine is firmly attached to one of these. ‘The disease has,
however, not invaded the wall of the latter. The border of these spots is intensely
hyperemic. When the cecum is slit open its width is three to four times that of the
undistended tube, and the thickness of the wall varies from one-eighth to one-half
of an inch, being not less than one-eighth of an inch over three-fourths of the entire
length. When the brownish feces were washed away the increased local thicken-
ings were found covered with firm exudates, usually attached in but one spot.

Sections were examined of that portion of the cecal wall which was very much
thickened, and to which. the contiguous small intestine was inseparably attached by
the new growth, . ‘

32 INFECTIOUS DISEASES AMONG POULTRY.

The mucosa of the cecal portion had sloughed away, while that of the embedded
small intestine was intact. The neoplastic tissue between cecum and intestine
was fully 1 em. (two-fifths inch) thick. Inasmuch as the infiltration probably fol-
lowed the narrow mesentery between cecum and intestine the original boundary
lines of the execal wall are no longer recognizable. The muscular coat of the cecum-
may be traced for only a short distance into the neoplasm, when it disappears. Micro
parasites were not seen distinctly in the diseased tissue.

The liver is very much enlarged, and dotted everywhere with roundish spots of
varying appearance. The majority are from 5 to 12 mm. in diameter, round or slightly
oval. The center of each is usually occupied by a group of yellowish dots and the
circle is bounded by a narrow yellowish ring. The space of the circle is mottled
brownish. Aiong these spots there are also circles of a completely yellowish color
On the convex surface of the left lobe there is a very firm, ring-like, yellowish mass,
cutting like firm cheese.

In crushed preparations of fresh liver tissue from within the brownish circles many
giant cells are seen. They consist of a meshwork of protoplasm of a rather coarsely
granular character inclosing spheres which appear homogeneous. The giant cells
are up to 30 /c in diameter.

Sections of liver tissue hardened in alcohol and in Foa’s solution were also exam-
ined. The foci of disease contain necrotic areas in which are numerous giant cells
each inclosing a number of microparasites. In some portions there is much cell
infiltration in the interlobular tissue around the portal vessels. Among the cells the
protozoa are recognizable.

Turkey No. 15 (Farm H).—Obtained alive and killed. About 3 months old. No
cecal or liver disease detected. In duodenum numerous tapeworms. This turkey
had probably been injured, for blood extravasation was found under skin of skull.

Turkey No. 16.—About 3 months old. Obtained with No. 14 from the same flock
(Farm G). Suffering with diarrhea. After being cooped for two days it was killed
August 10. The blood taken from a cutaneous vein of the breast under the wing
showed extensive leucocytosis.

The digestive tract is normal, with following exceptions: The mucosa of duodenum
ishyperemic. Tapeworms absent. The rectum, i. e., the bowel from the ceca to
cloaca, is also hyperemic. It contains yellowish masses made up of microscopic
spherical crystals (urates).

Both exca are affected. Contents of a pasty, brownish (nearly normal) character,
containing a considerable number of microparasites. Flagellates not seen.

The disease is manifested in both ceca symmetrically by a thickening of the walls
of the distal half of each tube. There are a few yellowish spots noticed under the
serous covering of the thickened region. These correspond to spots of maximum
thickness. The mucous membrane appears intact. The slight pigmentation noticed
on the mucous folds is frequently present in cca otherwise normal.

In tangential sections of the thickened wall in the fresh condition considerable
numbers of round microparasites, of a homogeneous.appearance, and about 8 to 10 u
in diameter are seen. ‘They are not bound to cells, but appear to be displaceable
under the cover glass independently.

In transverse sections of one cecum hardened in corrosive sublimate and alcohol
the increase in thickness of the wall was found to be due to hyperplasia of the sub-
mucous tissue and cellular infiltration of the muscular coat. The mucosa itself was
intact, the epithelium in place. Interspersed in the hyperplastic supmucosa are
small nests of giant cells, some containing well-defined microparasites, others only
vacuoles, somewhat larger than the parasites. The latter are not diffusely scattered
through the tissue, but restricted to these and a few other foci in which they exist
free in the meshes of the tissue. The masses of cells between the bundles of muscular
fibers resemble those of the submucosa and are free from parasites.

The liver is considerably enlarged. The surface is not smooth but slightly rough-
ened, Scattered over the various surfaces there are in all six or seven yellowish-white

APPENDIX. 3

homogeneous sclerotic areas which correspond to masses in the liver tissue. In cut-

ting into the liver these foci are found to sheath the hepatic veins. Outside of these

sclerotic foci the liver tissue is mottled with gray. In general, the entire liver
appears to have passed through a process of sclerosis.

Jn sections hardened in Foa’s solution and in corrosive sublimate the pathological
change going on is shown to be essentially a sclerosis following the protozoan infec-
tion. Within the disease focus the liver tissue, recognizable as such, is present only
in irregular patches of variable extent. The remainder has been replaced by an
actively developing connective tissue, still rich in nuclei. Within these areas there
is an extensive formation of bile ducts. Parasites were seen in but one spot, where
anumber of multinucleated cells had ranged themselves around what appeared to be
an old thrombus. Here, beyond the encircling giant cells, a group of similar con-
tiguous cells contained asmall number of protozoa. The encircling cells themselves
also contamed a few, and one parasite was recognizable within the thrombus itself.
Outside of these spots undergoing repair, the blood vessels are everywhere inclosed
in sheaths of new connective tissue very rich in nuclei, encroaching slightly upon
the liver tissue itself.

Turkey No. 17 (Farm I).—About 3 months old. Found August 11 in a dying condi-
tion in a flock in which a considerable number had died during the past month. The
only lesions found were a rather pale condition of the organs. In the small intestine
numerous tapeworms. No liver or cecal disease. In the ceca feces are rather dry,
and contain a considerable number of coccidia, varying slightly in size, the largest
32 uw long and 18 « broad.

Turkey No. 18.—From the same flock as No. 17, and quite sick when found. Killed
and placed on ice over night. :

In this bird no marked lesions of any kind were noted. The duodenum was

hyperemic and contained more or Jess mucus. The contents of ceca are fluid and
contain numerous flagellates. Cultures on agar from blood and liver remain sterile.

Turkey No. 19 (Farm J).—About3 to 4 months old. Taken August 11 from a large
flock, because suspected of being diseased. Died shortly after being caught. Placed
in refrigerator over night. In the pleuroperitoneal cavity a dark clot and much
blood-stained fluid; ecchymosis of the left abdominal wali. Source of hemorrhage
not detected; probably brought on while being caught.

Organs very pale but not diseased, with exception of liver and the right cecum.
In the latter there are two diseased areas. Near the blind end there is a ring-like
thickening of the wal], which has a yellowish color from the serous aspect. The
other area is covered with a small mass of exudate, and the thickening of the wall
beneath it covers an area about one-half of an inch square. The remainder of the
excum pigmented in spots and lines.

The liver is very much larger than in the normal condition and covered every-

~where with circular spots of varying appearance. ‘There are:

(1) Two completely necrotic, yellow masses of cheesy character, one in the base
of each lobe.

(2) Circles of a grayish-yellowish color throughout.

(3) Spots of a mottled brownish appearance. The mottling represents distended
vessels. 2

(4) Spots like the preceding with a necrotic mass in the center. These spots vary
in size up to a diameter of three-fourths of an inch.

In crushed preparations of the fresh liver the microparasites are distinctly rec-
ognizable. Some appear as free granular bodies, others are inclosed, either singly
in a cell, or in groups in large cells. These inclosed bodies are usually free from
granules and may show a minute nucleus. They measure from 10 to 13 1 in diameter,

In sections of liver tissue from this case hardened in alcohol the lesions were found
-to correspond to those in other cases. Sections through a brownish spot showed
‘everywhere marked distension of the vascular channels, These were filled with

17697-No, $—_3

o4 INFECTIOUS DISEASES AMONG POULTRY.

corpuscles. The parenchyma has largely disappeared and small cells have taken
its place. In a few places of the section microparasites are found collected together
in large patches. In these, giant cells are occasionally seen.

Turkey No. 20 (Farm I, see Nos. 17 and 18).—Found in field resting on keel and
moving along ground with aid of wings. Obtained August 11. Killed and exam-
ined August 12.

In trachea six pairs of gapeworms attached. In cecum and rectum a yellowish
finid (urates) containing many flagellates. No disease of liver or ceca,

Turkey No. 21 (Farm K).—Picked out of a large flock August 11, 1894, because it
was much weaker than the rest. No distinct appearances of disease. When killed the
organs were found normal, with the exception of the intestines. A mass of exudate
lying adjacent to rectum in abdominal cavity binds together rectum, ceca, and
neighboring coils. This mass has acylindrical shape and is about 2 inches long and
three-fourths of an inch at greatest diameter. It consists of spongy yellowish
exudate, incased in a smooth, leathery capsule. There is no disease of the mucous
or submucous tissue in any part of the intestines. The exudate is evidently the
result of a tear in the rectum which permitted the escape of feces into the abdo-
men. These became subsequently permeated with and encased in fibrin while the
tear healed up. Odor of this mass strongly fecal. A bit of liver tissue is placed on
inclined agar. The latter remains sterile.

Turkey No. 22.—Large female, over 1 year old. Died on Block Island during the
night of August 12. Examined on the farm next day. No evidence of liver or
cecal disease. In abdomen a partially smashed eggshell attached lightly to mes-
enteries.

Turkey No. 23 (Farm L).—About 3 months old. Received, dead, by express August
14. Probably died two days ago. Slight odor of decomposition.

Duodenum very much discolored by pigmentation. Both ceca considerably dis-
tended. The left is normal, the right diseased. Fully one-half to two-thirds of its
entire length beginning with the blind end is thickened. There are a number of
centers where the thickness is increased. At one of these there has been an out-
pouring of exudate on the serous membrane. The mucous surface of these local
thickenings is covered with firm masses of exudate.

The liver contains one sclerotic focus which has a whitish mottled appearance.
Itis otherwise normal. Three adult pairs of syngames in the trachea.

In sections of the wall of the greatly enlarged cecum hardened in alcohol the
lesions were of an advanced character. The mucosa was almost entirely sloughed
away. Only a few remnants of the slough remained, containing some tubules recog-
nizable as such. The submucous tissue was greatly thickened by cell infiltration,
and the muscular coat was divided into sections by cell masses which extended to
the serous membrane. Under the latter the blood vessels appeared greatly dilated
and filled with corpuscles. The microparasites are still present, but in small num-
bers. They appear in the neoplastic tissue in small patches inclosed by a fringe of
round cells. Giant cells are numerous and arranged in groups. Only in one group
were microparasites inclosed.

Turkey No. 24.—Taken August 13 from a flock on Block Island, because it was sus-
pected of being diseased. Killed August 14. No lesions were discovered in the
organs. One gapeworm found in the trachea and several minute tapeworms in the
intestine.

Turkey No. 25.—Taken from the same flock. When killed disease was found in
one of the ceca. (See page 22.)

Turkey No. 26.—About 3 months old. Brought August 15 from a large herd
(Farm G), and reported as having been feverish for a few days past. The bird is
killed and found in well-nourished condition.

Disease limited to one cecum and to the liver. Fully one-third of the cecum has
its wall thickened, The thickening is most pronounced in spots, to some of which a

APPENDIX. 35

firm, pale-yellowish exudate is attached. A portion of the diseased tube is placed
unopened in hardening fluids for further study.
The liver is enlarged to two or three times its normal size, and uniformly per-

‘meated with closely set, but usually discrete, spots, varying in size, a few having a

diameter of one-fourth of an inch. They are of a yellowish color, this being due toa

fine yellowish network, of which the spot is in fact made up. The details are well

shown in the photographic reproduction and in the colored drawing. (Pl. I,
fies. 1, 2.)

Tn teased and crushed preparations of the fresh liver a large number of micro-
parasites are detected. They appear as round, pale, homogeneous bodies, with a

sharp line as a border. They are less refrangent than fat globules, and suggest
somewhat the color and refrangibility of myelin or Buhl’s bodies as they are found

in the alveolar epithelium of the lungs of cattle. Their diameter ranges from 12 to
14 w. Some are surrounded by a narrow rim of proteplasm, containing one or more
nuclei, evidently the remains of a cell.

Transverse sections of the diseased cecum show a mass of exudate oceupying a
portion of the lumen. In the exudate are embedded strips of perfectly preserved
epithelium. The exudate appears amorphous and contains small foei and even large
patches of cell masses, probably originating from the intlammatory processes going on.

The thickening of the wall iS due in this case chiefly to infiltration of the sub-
mucous tissue. In one portion of the sections the museular portion of the wall is
infiltrated to the serous covering. Parasites are, however, not detected in the sub-
mucous or muscular cell masses, but are situated exclusively and in very large num-
bers in the reticular tissue of the mucosa, between the erypts. In this tissue, as
well as in the epithelium of the crypts, numerous mitotic figures are recognizable,
Giant cells are also present, but in small numbers. The microparasites are lodged,
generally singly, in cavities surrornded by one or more nuclei. In some places few
or many may be detected, packed away in a space of the reticular tissue.

The parasites are well brought out in the tissue hardened in corrosive sublimate
and alcohol, and stained in hematoxylin and eosin, They appear as round or
slightly oval bodies, with a faint reddish tinge of the body and a‘minute blue, ring-
like, central or eccentric body, probably the nucleus. They vary from 6 to 8 “ in
diameter.

The lumen of the mucous gland is densely packed with bodies roundish in outline
and about 4 « in diameter. The cell body is stained a homogeneous red with eosin,
the nucleus bluish with hematoxylin. (See page 20.)

Sections of the liver were made from material hardened in alcohol and in corrosive
sublimate and aleohol. They were stained in various ways, the most satisfactory
stain being hematoxylin and eosin, by which the microparasite was most clearly
brought out. Methylene blue was also satisfactory in bringing the microparasites
into relief.

The sections show within some of the areas indicated by the yellowish spots
extensive disappearance of the liver cells proper and the presence of a meshwork or
network of a homogeneotis substance, which suggests a fibrinous exudate, although
it has not the fibrillated strneture of mammalian fibrin. This meshwork is not uni-
formly distributed over the affected area, but appears m foci which are connected
with one another. The space left is occupied by numerous parasites and giant cells
and an occasional group of liver cells. In other toci the liver cells are replaced by
areticulum containing numerous parasites. Usually but one parasite is contained
in a mesh and is closely surrounded by one or more nuclei.

Turkey No. 27.—Taken from another Block Island flock as a suspicious case.
About 3 months old.

No lesions found in this bird. In the trachea three gapeworms; in the intestine
many large tapeworms.

Turkeys Nos. 28 and 29.—Brought from Fishers Island, New York, No lesions iy
these cases, Many large tapeworms in the intestine,

36 INFECTIOUS DISEASES AMONG POULTRY.

Turkeys Nos. 30 and 81 (Farm J).—About 3 to 4 weeks old. Brought because
they were not doing well. Killed. Intestine found nearly occluded with small
and large tapeworms.

Turkey No. 32 (Farm C).—About 3 months old. Killed August 17. No lesions
found. In duodenum several tapeworms. In cecum flagellates.

Turkey No. 83 (Farm B).—About3 months old. In the blood from a subeutaneous
vein some flagellates. No lesions in the internal organs. In duodenum some tape-
worms.

Turkey No. 34.—Obtained from Farm K August 18. Died onthe way to laboratory.

Bird about 10 weeks old. In body cavity blood-stained fluid. One-half of the
left cxeecum, beginning at the blind end, has its wall thickened. The serous covering
j8 reddened and shows a few yellow patches which correspond to certain specially
thickened regions of the wall whose mucous covering is necrosed.

The liver is large and covered everywhere with rather large, round, dark-brownish
patches having a narrow, yellowish boundary line. These correspond to diseased
foci within the liver.

In fresh liver tissue, crushed or cut in sections, oval bodies 12 to 15 «4 in diame-
ter are present in considerable numbers. They appear homogeneous; contents not
differentiated. Similar bodies, inclosed in groups of two or three in large cells, are
also present. They are homogeneous, round in outline, from 8 to 12 « in diameter.

In sections of liver tissue hardened in alcohol, when examined with a low power,
an immense number of microparasites appear embedded in the tissue. These vary
but slightly in size (8 to 9). Throughout the invaded spot the blood vessels, large
and small, are irregularly dilated and filled with red corpuscles, among which the
parasites are not seen. The tissue between the blood vessels is no longer recogniza-
ble as hepatic tissue. In some parts of the section it consists in the main of the
round parasites, single or insmall groups, each individual or group inclosed ina rim
of protoplasm provided with one or more nuclei. There is thus inclosing the para-
sites an imperfect meshwork present whose origin and nature is not evident. In
some places it would appear that the parasites are embedded in the liver cell. The
nuclei of such cells have undergone proliferation and some of these proliferated cells
have degenerated into groups of minute, deeply-stained granules. The impression
conveyed by such cells is that they are giant cells. They differ, however, from
those giant cells which appear in a later stage of the disease. The meshwork of
tissue which has undergone necrosis is present in but a few small areas.

Turkey No. 85.—From the same flock as No. 34 (and No, 21). Obtained August 18.
Died on the way.

This case closely resembles the preceding. One-half the length of both ceca
involved. The walls are thickened uniformly and in this portion are some small
excrescences easily removed in toto, leaving only a slight depression.

Liver spotted on all surfaces as in preceding case. Duodenum blackish from
injection and pigmentation. Muscular tissue has a yellowish tinge.

In the trachea one gapeworm, and some roundworms in the duodenum,

Turkey No. 36 (Farm N).—About 3 months old. Obtained, just dead, on August 18.

In this case the liver and ceca are diseased. The liver is spotted on all surfaces.
The spots vary in appearance. A few have a mottled, pale-yellowish aspect and
are fully 1.5 em. (three-fifths inch) in diameter. The remainder are round blotches
of a mottled, dark-brownish appearance. Many of these have coalesced into small
groups.

The wall of the left cecum is very slightly thickened and stillsmooth. The right
isin thesame condition. At the middle it has a circular ring-like thickening, about
2 em. (four-fifths inch) wide, of a yellowish appearance on section. Sections were
prepared of this region after hardening the entire cecum in alcohol.

The thickened wall, about 8 mm. in diameter at the thickest portion, consists
entirely of neoplastic tissue. The mucous layer is gone and the muscular layer is

APPENDIX. 37

no longer recognizable. ‘The bulk of the enlarged wall is composed of small cells, in
which are interspersed in round and elongated patches large numbers of multinu-
cleated cells, much smaller in size than the giant cells usually encountered. The
cells do not distinctly show the microparasites within their substance except in a
few cases, probably owing to the time and manner of hardening.

Turkey No. 37.—Obtained August 18 from Farm O. About 4 weeks old. Died the
following night. No lesions found, although intestine coutained many tapeworms.

Turkey No. 38.—From the same flock as preceding. About 3 months old. Died
during the following night. No lesions. One gapeworm in trachea, and very many
small tapeworms in intestine.

Turkey No. 39.—From the same flock. About 9 weeks old. Died the following
night. Numerous tapeworms in intestine.

Turkey No. 40.—From the same flock August 18. Cooped until August 20. On
that day the bird was resting on its breast with eyes closed. Killed. Many small
tapeworms in intestine.

Turkey No. 41.—Probably 8 weeks old. Brought to laboratory August 18, 1894.
Cooped temporarily and found dead August 20. Diseasein this case limited to both
ceca, (See page 22.)

Turkey No. 42.—Obtained August 18 from Farm I. About 3 months old. Sus-
pected of being diseased. When killed no lesions were detected.

Turkey No. 43.—Obtained August 18 from Farm P. Cooped. Found dead August
20. The only discoverable lesions were a puffing out of the tissue below one eye,
due to accumulation of a glairy secretion (roup). The same exudate found in the
trachea. Agar tubes inoculated with tracheal exudate remained sterile. Numerous
tapeworms in intestine.

Turkey No. 44.—Obtained from Farm Q August 18. Many turkeys are reported to
have died on this place. Bird about 3 months old. No lesions found when it was
killed. Many tapeworms in intestine.

Turkeys Nos. 45, 46, and 47.—Obtained from Farm R August 18 and cooped for
several days. When killed nothing abnormal detected. A few tapeworms in the
small intestine of No. 45 and No. 47, and many in No. 46. In the semiliquid con-
tents of the ceca of No. 47 very many flagellates.

Turkey No. 48 (Farm 8).—Obtained August 18. Cooped until August 21. Bird
appears well. When killed a few tapeworms were found in the intestine.

Turkey No. 49.—Received August 18, from a dealer, the intestines of nine turkeys,
In only one of these slight traces of cecal disease. This is manifested by a nodu-
lar, roughened condition of the mucosa of one cecum near the attached end. In
all intestines tapeworms present.

Turkey No. 50.—Obtained September 26 from a farm in the District of Columbia.
Slight odor of decomposition. The internal organs are free from disease. In the
trachea one pair of gapeworms. The head is badly diseased. On the root of the
bill a projecting tumor overlapping the nares, which is about two-thirds of an inch
in diameter. It is of a soft, medullary character and has a wart-like structure.
Besides this large tumor there is a small nodule on the lid of left eye. Right eye
completely closed by excrescences on the lid and cheesy masses in conjunctival sac.
Cornea opaque. In onenasal passage a large cheesy mass. No liver or cecal disease.

Turkey No. 51 (Farm B).—Received from Mr. Cushman, Kingston, R. I., October 8,
1894. Died on the way. Turkey quite large; probably 5 months old. Decompo-
sition well advanced. ‘The disease in this case was limited to the cieca, the liver, and
the peritoneum. The intestine was covered in places by a firm, yellow pseudomem-
brane fully 1 mm. thick, and the coils were glued to each other so that they could
be separated only after much dissection. The ceca appeared as an irregular mass
partly covered with the psendomembrane. Attempts to dissect out both ceca were
fruitless, as they were firmly attached to each other. Only one was diseased, how-
ever. This was much enlarged, its walls thickened, and the mucous membrane

38 INFECTIOUS DISEASES AMONG POULTRY.

beset with large, firm masses of exudate. In the liver were two foci of disease,
each about 1 em. in diameter. One is made up of firm, pale-grayish tissue, probably
largely cicatricial; the other is similar in appearance, but has small, yellow, cheesy
masses disseminated through it.

Owing to the very advanced stage of decomposition, cultures were not made. It
was evident, however, that the cause of death was the septic peritonitis, which was
secondary to the cecal disease.

Turkey No. 52.—Received, dead, with No, 51. Large male bird; probably 5 months
old. Very little decomposition.

Head free from disease. Heart and lungs normal. The pleuroperitoneal cavity
contains a large amount of blood-stained serum. Serous coat of gizzard very much
injected.

Serous coat of proventriculus dotted with elevated yellowish points and lines
where the diseased liver lay in contact with it. Spleen small, contains a minute
yellow focus; probably also the result of neighboring disease of the liver.

Duodenum, and a portion of the intestine below it, intensely hyperemic and pig-
mented. A few small tapeworms present. Pigmentation and hyperemia gradually
become less and disappear near openings of ceca. Below these the intestine is
coated with a yellowish liquid, largely composed of urates.

Both ceca contain pigment spots, and the longitudinal folds are pigmented. In
one cecum, near the blind end, there is a thickening of the wall about half an
inch across, covered with a brownish-yellow slough on the mucous side. The serous
aspect is discolored, in part yellowish. This is the only lesion found in the e:ea,

The liver is very large, weighing 304 grams (10.7 ounces). It is extensively dis-
eased. ‘The lesions are best described by grouping them under several heads:

(1) Necrosis of the liver tissue is present in both lobes in the form of large and
small masses of yellow, firm, cheesy masses (PI. III, fig. 2). In the left lobe fully
one-third of the lobe is converted into asingle, yellowish, homogeneous mass, situated
at the base or attached portion. The necrosed tissue is sharply defined from the
adjoining living tissue by an irregular line. The main branch of the hepatic vein
is bounded by the necrosed tissue, and the branches from the dead mass are occluded
with pale, friable thrombi. In the right lobe, at the base or attached portion, there
are several quite large cheesy masses of very irregular outline embedded in the living
tissue and in part visible on the surface. Besides these large masses there are dis-
seminated through the liver numerous smaller masses of cheesy tissue, either singly
or in groups. These small foci are usually associated with reparative processes.

(2) Foci which have healed and cicatriced wholly or in part are represented by
isolated regions, varying in size and attaining a diameter in one case of three-fourths
of an inch. The appearance of these foci varies considerably. They may show as
pale pinkish-gray spots penetrated by a network of vessels corresponding to the
interlobular markings. Others may be the seat of scars and slight depressions char-
acterized by increased vascularity and a pale pinkish-gray appearance on the liver
tissue. Finally, they may be intermingled with small cheesy foci, which are located
either centrally in the healed region or in numerous foci around it.

Over the entire surface of the liver patches containing networks of enlarged ves-
sels having a blackish color give the liver an injected appearance. Thereis nowhere
an indication that the disease as such is still going on. The extensive destruction
of liver tissue, as well as the repair, appear to be the result of an invasion of ‘proto-
zoa now completely checked.

The false membranes which cover some of the healing spots on the liver and the
abundant fluid in the abdominal cavity indicate a secondary infection with bacteria
through the necrosed tissue.

ph

regi

here tye

A PRELIMINARY INVESTIGATION OF DIPHTHERIA IN FOWLS.
By VeRranus A. MOORE.
(With Pl. VI.)

It is well known that a disease which is characterized by the forma-
tion of an exudate on one or more of the mucous membranes of the
head is widely distributed among the fowls of this country. Although
its existence is of long duration its cause and the character of the
exudate have not been satisfactorily determined. In Europe outbreaks
of a malady comparable to this in their manifestations have been
described as epizootics of fowl diphtheria. There are in this country
a few popular articles on diphtheria in fowls, but, generally speaking,
the term most frequently employed to designate a disease on the heads
of poultry is “roup.”! The range of morbid anatomy which this term
originally indicated is not known, but its present conception is exceed-
ingly broad, including, as shown by tle American literature,’ nearly,
if not all, the noticeable disorders of the heads of poultry.

Writers differ in reference to the nature of this disease. Many
poultry raisers consider it a local affection, having its origin in the
improper care of the fowls, while others insist that it is a highly con-
tagious disease. The latter class strengthen their position by clinical
history, in which they show that fowls which had previously been free
from this disease soon became infected after the introduction of one or
more “roupy” fowls into their midst. Reports show that it is the
usual experience that when the disease is introduced it almost invari-
ably remains in a more or less chronic form for an indefinite period,
notwithstanding the adoption of the best known methods of caring
for the fowls. It is stated by Bennett that “it is undoubtedly trans-

!The origin of this term is somewhat obscure, but it is supposed to be a corruption
of croup, and its application explained on account of a peculiar hoarseness accom-
panying the respiration of the affected birds.

2This literature is confined for the greater part to articles in poultry and agri-
cultural papers and journals. They are written by poultry fanciers or farmers, who
record their experience with the various diseases of fowls, and, as would be expected,
they deal more with the treatment than etiology, and while suggestive are of little
value in the study of the cause and nature of the maladies of which they treat.

39

40 INFECTIOUS DISEASES AMONG POULTRY.

mitted from the sick to the well fowls by their drinking out of the same
vessel, as the discharges from the nostrils of the sick birds contami-
nate the water as they drink.”

The lesions and symptoms attributed to this disease are exceedingly
variable. Among the more careful writers we find ‘“‘roup” defined as
“a purulent catarrhal affection of the air passages.” The localization
of the lesions, however, in the air passages is not universally sup-
ported. It is usually stated to affect, to a greater or less extent, the
conjunctiva and the mucosa of the mouth, pharynx, and larynx, as
well as that of the nares. The first symptoms are said to be ‘dullness
and loss of appetite, discharge from the nostrils, loud breathing, and a
dry cough.” ‘This is followed by the appearance of an exudate on the
mucosa of the nares, which extends in all directions, often covering
the eyes with a cheesy substance. The secretions are said to ocecasion-
ally accumulate within the sides of the face, causing them to swell to
an extensive degree. The appetite is either very poor, or ravenous, in
which case the food is not digested. Emaciation follows and death
may occur in from three to eight days, or, as is usually the case, the
disease becomes chronic and the fowis eventually recover.

The economic importance of fowl diphtheria appears to be greater
than that of any of the other poultry diseases. ‘There are no statistics,
however, by which the total amount of loss from this cause can be even
approximately obtained, but the numerous reports of its occurrence
are sufficient to show that it is very large. Unlike a rapidly fatal
disease such as cholera, the loss is not confined to the fowls which die,
but in addition it includes a heavy shrinkage in the poultry products;
due to the chronic course of the disease in large numbers of fowls
which eventually recover. The small value of the individual and the
fact that only a small percentage of those affected die has evidently
caused the importance of the disease to be underestimated. It is evi-
dent, however, that the frequency of its occurrence and its wide
geographical distribution render this affection a serious obstacle to
the poultry industry! of this country. In addition to the usual chronic
form, there are,if reports are true, frequent epizootics in which thou-
sands of dollars’ worth of poultry are destroyed.

In addition to the theory of contagion, there are many conditions
mentioned in poultry literature under which this disease may be
expected to occur. Thus, it is affirmed that fowls exposed to a draft
at night will ordinarily become affected. The nature of the food, venti-
lation, and cleanliness are also considered important factors in pro-
ducing or preventing this malady. The exudates are described as
both catarrhal and diphtheritic, but the description of the lesions is
exceedingly indefinite.

‘The economic importance of this industry is little understood. ‘The Eleventh
Census shows that in 1890 there were in the United States 258,472,155 barnyard
fowls (exclusive of ducks, turkeys, and geese), and that the egg product for 1889
was 817,211,146 dozens with a money value of $98,000,000.

PRELIMINARY INVESTIGATIONS, Al

The foregoing résumé of the present knowledge of the disease gener-
ally designated as “roup” is important in showing the various condi-
tions recognized and included under that term. There are other so-
called fowl diseases, such as influenza and distemper, reported to
be characterized by somewhat similar symptoms and lesions. Their
descriptions are so indefinite, however, that it is impossible to differen-
tiate them from certain forms of the disease more frequently called
“roup,” but which, on account of the nature of the lesions, will be con-
sidered as diphtheria in this preliminary report.

I:

During the past year numerous reports of epizootics among poultry!
have been received, and several sick and dead fowls have been sent to
this laboratory and to the experiment station of this Bureau for exam-
ination and diagnosis. Certain of the sick fowls improved rapidly
and soon fully recovered. Others were dead and far advanced in post-
mortem changes when received, while a few were in a condition more
suitable for bacteriological and pathological study.

In all, about 36 fowls came under observation. Several of them
subsequently recovered; a few died with tumors; in one case death was
undoubtedly due to a traumatism;? 2 died of fowl cholera, and the
remainder, 18 in number, were affected with somewhat similar lesions of
the mucous membranes of the head, although they came from reported
epizootics of cholera, diphtheria, and “‘roup.” The examination of
these fowls revealed the interesting fact that the lesions corresponded
very closely to those described by Loeffler,’ Klammer,t Babes and
Puscariu,’ Eberlin,® Loir and Ducloux,’ and others, as diphtheria im
pigeons, fowls, and other birds.

As the disease was not studied in the field it is impossible to define
the conditions under which it occurred, or to estimate the full extent
of the loss it occasioned. The fowls examined and here reported came
from five different flocks, in some of which there is a history of conta-
gion. Unfortunately, the fowls sent to us were usually among the last
to be affected.

The methods employed have varied according to the exigencies of the

1In addition to the fowl disease, a similar affection of pigeons, more particularly
of young squabs, was reported from Waynesboro, Pa., and several of the affected
birds were sent to this laboratory for examination. The lesions found in these
birds resemble somewhat closely those found in the mouths of fowls.

2It is not uncommon to find pieces of wire or nails in the crop and proventriculus.
In one case, two eight-penny nails and a piece of wire about 2 inches long were
found piercing the walls of the proventriculus and penetrating the liver and lungs.

’Mittheilungen aus dem Kaiserlichen Gesundheitsamte, Bd. II (1884), S. 421.

4 Berliner thieriirzt. Wochenschrift, 1890, No. 18, S. 138.

5 Zeitschrift f. Hygiene, Bd. VIII (1890), S. 374.

® Monatshefte f. Thierheilkunde, Bd. V (1894), S. 433.

7Ann. de l’Inst. Pasteur, Tome VIII (1894), p. 599.

42 INFECTIOUS DISEASES AMONG POULTRY.

different cases. In order, therefore, to present the facts obtained in
these preliminary studies, it seems best to record somewhat briefly the
history of each outbreak, so far as determined, together with the notes
on the individual fowls.

OUTBREAKS.
It

Late in December, 1893, Mr. W., of Middleport, N. Y., sent several
adult fowls! affected with “roup” to the experiment station of this
Bureau. Mr. W. stated that he had lost a considerable number of
fowls from this disease during the past two or three years. He
believed that the disease was introduced into his flock by the purchase
of some choice birds about three years before. Since its first appear-
ance it had continued to exist in his flock, usually running a chronic
course, but occasionally breaking out in a more acute and destructive
form. He had tried many methods of treatment without success.

The majority of the fowls sent by Mr. W. appeared to be well upon
their arrival at the experiment station. A few of them were killed
for examination and found to be normal throughout. Five fowls, how-
ever, were suffering with a slight or more extensive exudate on the
nasal mucosa, or conjunctiva. These were carefully watched and
examined from time to time with the results indicated in the appended
notes.

Fowl No. 1.—This fow] was suffering from an exudate in the left eye. December 25
Dr. Smith made several cultures from this exudate. From these two nonpathogenic
bacilli were isolated.

December 28: A portion of the exudate was removed from the eye of fowl No.1
and inserted beneath the skin, over the abdomen, of arabbit. On the following day
there was considerable swelling at the point of inoculation. This continued for sev-
eral days, when it subsided. The rabbit died February 10,1894. It was very much
emaciated, but no bacteria were found in the organs, either in cover-glass prepara-
tions or in cultures.

January 25, 1894: Fowl No 1 was found dead; very much emaciated. The left
eye was covered with a thin layer of a friable, yellowish exudate, composed of
broken-down cells and bacteria. No fibrin discovered. The nictitating membrane
thickened; cornea opalescent. The mucosa of the nares, mouth, and pharynx were
apparently normal. No lesions were found in the internal organs. Culture media
inoculated with small pieces of the liver remained clear. :

Two white mice were inoculated subcutaneously with bits of the exudate from the
eye. They died on the fourth and sixth days, respectively, after the inoculation.

From the organs of the mouse which died on the sixth day a nonmotile bacillus,
resembling morphologically and in its cultural characters the bacillus of rabbit sep-
ticeemia, was isolated. Experimental animals inoculated with small quantities of
culture remained well. This bacillus was carefully studied and retained in subeul-
tures until the fall of 1894, when a rabbit inoculated intravenously with a moderate
quantity of pure culture died on the second day with lesions similar to those pro-
duced by the slightly attenuated bacillus of rabbit septiczemia.

1The term fowl is used in this article in its restricted sense, referring to Gallus
domesticus oniy. This disease, however, is said to affect several other species.

OUTBREAK I. 43

The mouse which died on the fourth day exhibited an enlarged and discolored
spleen. The other organs were normal in appearance. A cover-glass preparation
showed it to contain a large number of bacilli. Tubes of agar and bouillon inocu-
lated with bits of the spleen pulp developed into pure cultures of a motile bacillus,
which was subsequently found to be possessed of marked pathogenic properties.
Although fatal to such a large number of experimental animals, experiments to pro-
duce a disease in fowls similar to the one from which it was obtained gave negative
results.

Notwithstanding the marked pathogenesis of this organism, its cultural and physio-
logical manifestations showed it to be closely related to Bacillus coli communis,' though
differing from it in several important particulars.

Fowl No. 2.—January 4, 1894: This fowl appeared to be well excepting a protu-
berance beneath the left eye. In order to examine this thoroughly the fowl was
killed with chloroform. The autopsy showed much emaciation. The protuberance
consisted of an encysted tumor lying immediately beneath the conjunctiva. It was
about 2 em. long and about one-half as thick. Upon section it was found to contain
a yellowish, caseous substance, which emitted a peculiar, penetrating, and repulsive
odor. The nasal duct was clear. The eye itself was not visibly affected. The liver
was unusually fatty. In the left ling were a few hepatized areas. Other organs
appeared to be normal.

Cover-glass preparations made from the caseous substance contained many bac-
teria. Those from the other organs showed no microorganisms. From the caseous
substance a large number of cultures were made in agar, bouillon, glycerin-agar,

‘On account of the importance of considering all bacteria possessed of pathogenic
properties, of however slight degree of virulence, the more significant characters
and properties of this bacillus are appended.

Morphology.—A small, actively motile bacillus, varying in length from 1.2 to 2 y,
ends rounded. It is appreciably larger in bouillon than in agar cultures. Flagella
‘easily demonstrated. Stains readily with the ordinary aniline dyes.

Culture characters.—On agar it develops in twenty-four hours into a moderately
vigorous, moist-appearing, glistening growth of aneutral grayish color. The growth
is not viscid. When isolated the colonies are round, slightly convex, varying from
1 to 3 mm. in diameter. In gelatin the growth is very feeble in stick cultures.
After forty-eight to seventy-two hours a delicate grayish growth appears along the
needle track. The colonies in plate cultures appear on the second day as minute
grayish points. Onpotato the day after inoculation at 36° C. a light, lemon-yel-
low colored growth appears. At the end of from four to six days the growth has
increased slightly in quantity; the color is somewhat darker. When cultivated in
peptonized bouillon the fluid becomes heavily clouded in twenty-four hours. After
about three days a thin, friable, grayish membrane forms over the surface of the
liquid. The reaction, which at first is faintly alkaline, becomes slightly acid, but
subsequently strongly alkaline. It does not grow in acid bouillon,

The casein of milk is precipitated in from two to three days. It is faintly acid in
reaction. In about seven days the casein becomes firm and covered with a clear
serum.

Its action upon sugars is very constant. Dextrose and lactose are fermented with
the formation of gas. In both cases 46 per cent of the capacity of the closed branch
of the fermentation tube is filled with gas. The proportion of CO, to the H in the
gas varies slightly with the two sugars; in the dextrose it is 3:11, in the lactose
4:11. The production of gas is completed in from three to four days. Fermenta-
tion tubes containing saccharose bouillon become uniformly clouded throughout,
strongly alkaline in reaction, but no gas is formed. {

A marked indol reaction was obtained.

Pathogenesis.—When isolated this organism was fatal to white mice in from
twenty-four to forty-eight hours when inoculated subcutaneously with from one to

44 INFECTIOUS DISEASES AMONG POULTRY.

gelatin, blood serum, bouillon containing glucose in fermentation tubes, and anaéro-
bic-agar cultures. From these two aérobic bacilli and one streptococcus were iso-
lated, all of which were harmless for experimental animals.

Fowls Nos. 3, 4, and5.—When received these fowls exhibited a very slight quan-
tity of a mucus-like exudate in the nasal cavities. They were thought at the time
to be the best cases for an experiment to test the contagiousness of the disease, and
very soon after their arrival they were penned with six healthy fowls.

February 21, 1894: About seven weeks later, rabbit No. 144 was inoculated subeu-
taneously with a loop of the nasal discharge of fowl No. 3. February 23 the rabbit
was found dead. . The autopsy showed aslight purulent infiltration into the subcu-
taneous tissues at the point of inoculation. The spleen was slightly enlarged and
of a dark, purplish color. The liver was hyperemic, and the cortex of the kidneys
deeply reddened, due to injected blood vessels. There were several punctiform hem-
orrhages beneath the serosa of ileum and lower colon, The lungs were markedly
emphysematous. Stained cover glass preparations made from the blood, spleen, and
liver showed innumerable short rod-shaped bacteria with ends rounded, usually
exhibiting light or unstained centers. Pure cultures of this bacillus were obtained
from the spleen and blood.

February 24: A second rabbit (No. 145) was inoculated subcutaneously with the
nasal discharge. It was found dead at 4 p.m. February 25, The autopsy and bac-
teriological examination revealed the same condition and results as those described
for rabbit No. 144.

Fowl No. 3.—February 25: This fowl was founddead. It was very much emaciated.
The nasal cavities were occluded with a thick muco-purulent substance. At the
external nares it had a yellowish color and was dried down into a firm, crust-like
mass. Throughout the nasal passages to the posterior opening the substance was
lighter in color and very viscid. The eyes were not affected. The internal organs
appeared to be normal. Tubes of culture media inoculated from the liver and blood
remained clear. The fact that two rabbits had been recently inoculated from the
nasal discharge with like fatal results precluded the necessity of further inocula--

two drops of a pure bouillon culture. The lesions were characterized by edema
about the place of injection, and an enlarged and discolored spleen. Gray mice
remained well after a similar injection.

A short series of inoculations were made in white mice, and gray mice succumbed
to the inoculation with a culture from the spleen of the second white mouse that
died. ‘The lesion found was a much enlarged spleen as in white mice.

A guinea-pig inoculated in the abdominal cavity with 0.3 cc. of a bouillon
culture was found dead on the following morning. The abdominal organs were
covered with a grayish exudate, which was in the form of shreds over the intestines.
A few ecchymoses beneath the pleura and peritoneum. The peritoneal exudate
ontained innumerable bacteria.

Rabbits are not susceptible to subcutaneous or small intravenous injections of
pure cultures.

A fowl inoculated in the wing vein with 0.3 cc. of a bouillon culture died on the
second day. The organs contained very few bacilli. Tubes of culture media inocu-
lated with bits of the liver developed into pure cultures of this bacillus.

A fowl inoculated subcutaneously with 0.6 cc. of a bouillon culture died in forty-
four days. No bacteria were found in the organs and no lesions were discovered.
The cause of death is questionable.

A large number of inoculations were made beneath the conjunctiva of fowls, but
no lesions were produced. Loops of the growth from agar cultures were rubbed
cover the conjunctiva and into the nasal cavities with negative results.
| Four fowls were fed with 240 cc. of bouillon cultures of this bacillus mixed with
alittle mill food. They remained perfectly well.

OUTBREAK II. 45

tions at this time. The bacillus obtained from rabbits Nos. 144 and 145 was exceed-
ingly abundant in the nasal discharge, and especially so in preparations made from
scrapings of the nasal mucosa.

Fowl No. 4.—This fowl had presented external appearances similar to those of fowl
No. 3.

February 24: Rabbit No. 97 was inoculated subcutaneously with a loop of the
nasal discharge. It was found dead on the following morning. The examination
showed its organs to contain innumerable bacteria identical with those found in
rabbits inoculated from fowl No. 3.

Fowl No. 5.—Like Nos. 3 and 4, this fowl suffered apparently from a nasal catarrh.
The eyes and mouth were normal.

February 28: Rabbit No. 149 was inoculated with the nasal discharge. The rab-
bit died in less than twenty-four hours from septiczemia, the same as the rabbits
inoculated from fowls Nos. 3 and 4.

Fowls Nos. 4 and 5 died soon after the inoculations into rabbits were made. They
were not brought to the laboratory, but examined at the experiment station by Dr.
F. L. Kilborne, who reported them to be much emaciated and the nasal cavities filled
with a muco-purulent substance. The bacteria obtained in pure cultures from the
rabbits inoculated from fowls Nos. 3, 4, and 5 were carefully studied and compared
and found to be identical in morphology and cultural characters and to resemble
the bacillus of rabbit septicemia. The bacillus obtained from fowl No.1 was not
distinguishable from it except in its degree of virulence. a

The six fowls penned with fowls Nos. 3, 4, and 5 from the latter part of December,
1893, until the middle of February, 1894, remained perfectly well. The disease in the
affected ones remained confined to the nasal cavities. Inthis experiment all the fowls
drank from a small basin, and their food was kept in adish arranged so that they
were all compelled to eat from it. The disease was not contracted by the exposed
fowls.

II.

In October, 1894, four fowls were received from Mr. W., of Newbern,
N.C. In an accompanying letter he stated that they were suffering
from a disease which was called cholera, and which had killed hundreds
of fowls about Newbern during the summer and early fall. ;

All of the fowls (Nos. 65, 66, 67, and 68) were adults of a mixed breed.
Upon inspection two of them were found to be apparently well and the
other two were suffering from an exudate over the conjunctiva of one
eye and the mucosa of the nasal passages. As this disease was re-
ported to be contagious it was thought that the apparently well fowls
(Nos. 66 and 67) had simply been exposed, and consequently they were
kept for a considerable time waiting for the disease to develop. They
were subsequently chloroformed and carefully examined, but no lesions
were found in either case, and culture media inoculated with pieces of
the organs remained clear.

Fowl No. 65 was chloroformed soon after its arrival and carefully examined. It
was much emaciated. The left eye was covered with a thick yellowish exudate,
which was somewhat elastic. The microscopic examination showed small and large
round cells and bacteria. A fibrin-like substance was present in moderate quanti-
ties. The exudate was firmly attached to the cornea but not to the conjunctiva
covering the inside of the lids. The mucosa of the nares and mouth appeared to be
normal, The intestines were slightly reddened, due to injection of blood vessels.
They contained a large number of roundworms and several tapeworms attached to the

46 INFECTIOUS DISEASES AMONG POULTRY.

mucosa. In the intestinal wall were several nodules.! The other organs appeared
to be normal.

Tubes of culture media inoculated with the liver remained clear. A tube of agar
inoculated with the exudate from the eye developed into a pure culture of a motile
bacillus belonging to the colon group of bacteria.

Fowl No. 50 was inoculated in both eyes with bits of the exudate from the eye of
fowl No, 65. The mucosa of the conjunctiva over the lower lids were scraped and
the exudate thoroughly rubbed into the irritated surface. The inoculation wound
healed rapidly and the eyes subsequently remained well.

Fowl No. 68 was chloroformed and examined November 9. Fowl very much emaci-
ated. The left eye was covered with a cheesy, yellowish, rather firm exudate, which
formed a cap over the eye and which was easily removed. It emitted a strong putrid
odor. The eye was much flattened and the cornea had an opalescent appearance.
The posterior portion of the eye was normal. The saucer-shaped exudate was
loosely attached to the conjunctiva over the lids and cornea, ‘There was a small
amount of firm cheesy exudate in the nasal cavities. The mucosa of the mouth and
pharynx was normal in appearance. ‘The liver, spleen, kidneys, intestines, and lungs
also appeared to be normal. Although the fowl was emaciated, it is highly proba-
ble that the exudate in the eye would soon have sloughed and that recovery would
have followed. The appearance of the nasal mucosa indicated that the cheesy sub-
stance in the nasal passages had come from the eye, having coagulated after passing
through the lachrymal duct. The disease appeared to be localized in the left eye.

Fowls Nos. 54 and 55 were inoculated by scraping the mucosa of the conjunctiva
and pharynx and thoroughly rubbing the irritated surface with the exudate from
the eye of fowl No. 68. The inoculation wound healed and the fowls remained well.
About ten days later they were reinoculated in a similar manner with the growth
from an agar culture made from the eye of fowl No. 68. No lesions were produced.

III.

In the fall of 1894 several chickens suffering from this disease were
examined from the flock belonging to Mr. T., who resided near Alexan-
dria, Va. The insidious nature of certain forms of this malady is well
illustrated by this outbreak. Several healthy young chickens were
desired for experimental purposes, and Mr. T. reported that he possessed
some very thrifty chickens of the desired age, and his account of the
history of his fowls showed that the place had been free from fowl dis-
eases for several years. Nine of the chickens were secured and placed
on the experiment station. A few days later two of them were brought
to the laboratory, where, upon inspection prior to their use, they were
found to be suffering with an exudate which covered the greater part
of the mucosa of the mouth and pharynx. The other seven chickens
were examined by Dr. Schroeder, director of the station, who found a
few of them to be similarly affected. The remainder appeared to be well,
but were subsequently attacked. Some of these were allowed to lie,
and others were killed in the earlier alae of the disease for exam-

sane fe summer of f 1894 numerous roe i were aaa to be affected with a
disease of the intestines, due to the presence of tapeworms. This disease is espe-
cially interesting, owing to the resemblance of its lesions to those of tuberculosis.
Ihave described it somewhat fully in an article entitled ‘A Nodular T:eniasis of
Chickens,” to appear in a forthcoming report of this Bureau,

OUTBREAK III. 47

ination. Mr. T. states that later in the fall he lost a few chickens
from this disease, and a large number were affected which recovered.

Fowl No. 60 was brought to the laboratory October 17. On the following day it
was found sitting in a crouched position, feathers ruffled, and the head drawn close
to the body. It breathed through the mouth. The eyes were usually closed; no
exudate on the conjunctiva; temperature 108° F. Thesymptomsapparently improved
during the next few days, but on the morning of October 22 it was found dead.
This chicken was about 2 months old and was much emaciated. On the lower
side of the cornea of the right eye there was an area about 2 by 1 mm. covered with
a grayish exudate loosely attached to the cornea. The mucosa over the entire left
side of the mouth was covered with a yellowish exudate one-fourth to 24 mm. in
thickness. It was easily removed and exceedingly friable. The mucosa of the
pharyax, larynx, and nares was normal. The internal organs appeared to be nor-
mal. Cultures made from the exudate, liver, and kidneys contained a variety of
bacteria, which, however, were not isolated and studied in pure culture.

Fowl No. 59 was received at the laboratory at the same time, and it manifested
symptoms similar to those described for No. 60. After a few days the symptoms
showed a general improvement, which continued until October 20, when it was
chloroformed for examination.

The autopsy showed the fowl to be very much emaciated. The mucosa of the
nares and the conjunctiva was normal. On the floor of the mouth, on each side of
the tongue and extending back on either side of the larynx, there was a band of
grayish exudate about 2 mm. in width, firmly adhering to the mucosa. In micro-
scopic section the exudate was found to be composed for the greater part of round
cells and bacteria. (See Pl. VI, fre. 3.)

The internal organs were normal in appearance. Tubes of agar inoculated with
bits of the liver and kidney developed into cultures containing several forms of bac-
teria, including a micrococcus frequently encountered and Bacillus coli communis.

A guinea-pig was inoculated subcutaneously with a small piece of the exudate,
including subjacent muscle. A small swelling developed at the place of inoculation.
Five days after inoculation the guinea-pig was chloroformed and examined. At the
place of injection the subcutis was infiltrated with a pasty, purulent substance over
an area of 5 by 3 cm. The adjacent muscle necrosed. Thoracic and abdominal
organs normal in appearance. No bacteria were found in cover-glass preparations
and subsequently in cultures made from the organs. The pus in the local abscess
contained innumerable bacteria, apparently of the same species. Bouillon and agar
inoculated from the infiltrated substance developed into pure cultures of a nonmotile
bacillus, which was carefully studied and found to be similar in its pathogenesis and
cultural characters to that of slightly attenuated rabbit septiczemia.

Fowl No. 69.—In the latter part of October this fowl was attacked, and a thick
exudate formed on the larynx. November 2 it was chloroformed. The eyes were
covered with exudate which agglutinated the lids. Upon opening the lids the exu-
date was easily removed from the conjunctiva covering the lids, but it was firmly
attached to the cornea. The exudate formed sancer-shaped caps which covered the
eyes, being 2 mm. thick over the cornea, gradually thinning out to a sharp margin.
About the margin of the firm exudate was a small quantity of partially coagulated
mucous substance. The mucosa of nares normal. On both the anterior and poste-
rior sides of the laryngeal opening there was an area about 3 mm. in width covered
with a grayish exudate adhering firmly to the mucosa. It was removed with diffi-
culty, leaving araw surface. The kidneys were mottled with grayish lines, due to
large quantities of urates in certain of the tubules. Other organs appeared to be
normal.

November 2: Rabbit No. 215 was inoculated subcutaneously with a small quantity
of the coagulated mucus from the eyes,

48 INFECTIOUS DISEASES AMONG POULTRY.

November 5: Rabbit found dead this morning. At the point of inoculation the
subcutis was infiltrated with a purulent substance over an area 2 by 5em. The
pleura over the Jungs and parietes was covered with a thin cellular exudate.
Lungs hyperemic. Spleen much enlarged and dark colored. Kidneys hyperemic.
The liver mottled with areas covered with a delicate grayish exudate. The mucosa
of the lower colon sprinkled with punctiform hemorrhages. Cover-glass prepara-
tions from the organs contained innumerable short bacilli with ends rounded. Pola-
stain not marked.

Rabbit No. 225 was inoculated with 0.3 ce. of a bouillon culture obtained from the
spleen of rabbit No. 215. It was found dead on the fourth day with lesions similar
to those found in rabbit No. 215. Stained cover-glass preparations made from the
liver and spleen contained innumerable polar-stained bacteria. Pure cultures or
this organism were obtained from the liver. A guinea-pig inoculated into the abdom-
inal cavity with 0.3 ce. of a bouillon culture from rabbit 215 died within twenty
four hours. A subcutaneous inoculation with a similar quantity produced death in
six days.

Fowl No. 70 was received at the same time as No.69 and chloroformed for exami-
nation. Much emaciated. The left eye was closed, the lids agglutinated together;
the eye was covered with a yellowish cheesy exudate adhering quite firmly to the
cornea. It gave off a disagreeable odor, The exudate had sloughed from the con-
junctiva of the lids. In the nasal passages there was a small amount of cheesy exu-
date. Mucosa of mouth normal. The internal organs appeared to be normal. No
cultures were made.

Fowl No. 71 was found dead November 5. This fowl was much emaciated. The
left conjunctival sac contained considerable semiliquid substance. The mucosa
was slightly reddened and the cornea had an opalescent appearance. No other
lesions were found. Cultures made from the exudate in the eye contained several
forms of micrococci and a short nonmotile bacillus.

Rabbit No. 225 inoculated subcutaneously with the liquid from the eye.

November 12 the rabbit was found dead. The autopsy showed extensive purulent
infiltration into the subcutaneous tissue about the point of inoculation. The tho-
racic and abdominal organs were covered with a thin grayish exudate composed of
cells and bacteria. Spleen enlarged; pulp soft. No bacteria were found in cover-
glass preparations from the organs, but in those from the exudate there were innumer-
able short bacilli with ends rounded, frequently exhibiting a polar*stain. Pure cul-
tures of this bacillus were obtained from the spleen and liver.

November 14: Fowl No. 64 was inoculated in the conjunctiva with 0.2 cc. of a
bouillon culture of the bacillus obtained from fowl No.71. On the following day
there was much swelling of the eyelid. The swelling increased, with accompanying
reddening, for about two days, when it began to subside and finally entirely disap-
peared. The fowl continued well.

November 14: Fowl No. 65 (previously inoculated on the mucosa of head) was
inoculated in the wing vein with 0.3 ce. of a fresh bouillon culture of this bacillus.
The fowl remained well.

Two small chickens were inoculated beneath the mucosa of the mouth and also
the conjunctiva with several drops of a pure culture of this bacillus without pro-
ducing any appreciable effect.

Fowl No. 72 was found dead November 7. It was much emaciated. A thin gray-
ish substance covered the cornea from which a nonpathogenic bacillus was iso-
lated. No lesions could be found, and culture media inoculated from the organs
remained clear.

Fowl No. 73 was found dead November 9. It was much emaciated. The conjunctiva
ana mucosa of the nares were normal, The roof of the mouth and the pharynx
were covered witha thick grayish exudate, which was removed with difficulty, leav-
ing araw surface. The entire tongue was surrounded with a thick exudate, which

rl eae A eee

OUTBREAK Ivy. 49

was easily removed. The examination of sections showed a cell infiltration extend-
ing through the mucosa into the subjacent muscles. No other lesions were found.
On account of a scarcity of rabbits no inoculations were made from this fowl.

Fowl No. 75 was found dead November 22. Fowl considerably emaciated. The left
eye was covered with a thick necrotic exudate, which was easily removed from both
the eye and conjunctiva of the lid. The exudate extended through the nasal duct
intothe nares, In the posterior nares the exudate was bordered by a layer of a gray-
ish, mucus-like substance. Small areas on the roof of the mouth and about the
glottis were covered with a grayish, firmly-adhering exudate, which left a raw
surface upon removal. No other lesion discovered.

November 22: Rabbit No. 226 inoculated subcutaneously with the mucus-like
substance from the nares.

November 23: Rabbit found dead this morning. Owing to the large amount of
exudate injected beneath the skin the local lesion was unusually severe. The
abdominal viscera were hyperemic. Cover-glass preparations made from the organs
contained many elongated bacteria. A pure culture of a nonmotile bacillus was
obtained from the liver. A rabbit inoculated subcutaneously with 0.3 cc. of this
culture died of septicemia in less than twenty-four hours. This bacillus was iden-
tical in its morphology and cultural characters with those obtained from fowls Nos,
59, 69, and 71,

IV.

In October, 1894, a severe epizootic was reported to have occurred
among fowls belonging to Mr. P., residing on Bennings road, in the
District of Columbia. Mr. P. stated that a considerable number of his
fowls had died from what he thought to be diphtheria, on account of
the grayish membrane observed on the mucosa of the mouth and
pharynx. Unfortunately, he did not notify us of this disease until it
was nearly over, so that we had an opportunity of examining only the
three last fowls affected. Two of these were not received until some
time after death, and the other died on the third day after its arrival.
The results of these examinations are appended:

Fowl No. 58.—This fowl was received at the experiment station October 9. It was
found dead on the morning of October 12. Fowl very much emaciated. The mucosa
of nares, mouth, and pharynx appeared to be normal. No lesions were found in the
internal organs excepting a few nodules due to certain forms of tapeworms in the
intestine. Cover-glass preparations of the blood and liver showed no bacteria. In
those from the kidney a very few short bacilli with rounded ends were observed.
Tubes of bouillon were inoculated with bits of the blood, liver, and kidney. The
tubes inoculated with the blood and liver developed into impure cultures of ordinary
saprophytic bacteria. The culturefrom the kidney contained a short motile bacillus
belonging to the colon group.

Fowl No. 61.—A small chicken, about 10 weeks old. It had been dead about
twenty-four hours when received. Much emaciated. Along the cleft in the hard
palate there was a narrow strip covered with a thick grayish exudate, which was
firmly adherent to the mucosa. On the posterior surface of the pharynx and also
about the glottis were small areas covered with a similar exudate. The lefteye was
covered with a firm, yeliowish mass of necrotic exudate, which was easily removed.
The color of the puteeiall organs was slightly altered, ane to post-mortem changes;
otherwise they appeared to be normal.

A guinea-pig was inoculated subcutaneously with a bit of the exudate, including
the subjacent tissue from the roof of the mouth. A slight swelling occurred at the
point of inoculation. Four days after the inoculation the guinea-pig was chloro-

17897—No. 5—-4

50 INFECTIOUS DISEASES AMONG POULTRY.

formed and culture media inoculated from the local lesion. These developed into
pure cultures of a motile bacillus belonging to the colon group.

Fowl No. 62.—This fowl was received at the same time as No. 61. The lesions were
confined to the nasal passages, which were occluded with a yellowish cheesy sub-
stance, and the left eye, which was covered with a quite firm, easily removable
necrotic exudate. The conjunctiva of the lids were not appreciably affected. The
cornea was opaque.

A guinea-pig was inoculated with a portion of the exudate from the nasal cavities.
It was chloroformed six days later for examination, The subcutaneous tissue about
the point of inoculation was infiltrated with a purulent substance over an area of
about 1 cm. in diameter. No other lesions. From the local lesions pure cultures
of a motile bacillus belonging to the colon group were obtained.

¥.

This outbreak occurred among the fowls belonging to Mr. H., who
resides at Brookland, D.C. He states that he had about 75 fowls, all
of which were in excellent condition up to the purchase of a cock, which
was subsequently found to be diseased. A large number of his fowls
became infected. They usually recovered, although several of the badly
affected ones were killed. Late in October Mr. H. brought a hen which
was suffering from the disease to this laboratory. This was one of the
last of his fowls to beattacked. At the time it came under my observa-
tion the disease was confined to the left eye and an area of firmly attached
exudate on theroofof the mouth. This fowl was kept under observation
for several weeks, and cultures and animal inoculations were made from
the exudate as indicated in the appended notes.

Fowl No. 63.—Received October 20. The left eye was considerably swollen, the two
lids being agglutinated together. The cornea had an opalescent appearance. Res-
piration apparently normal. Temperature 107° F. Fowl sits with the head drawn
close to the body. Tubes of agar and bouillon inoculated with the exudate developed
into pure cultures of a nonpathogenic micrococcus.

A white mouse was inoculated subcutaneously with a loop of the exudate from the
eye. It remained well for about three weeks, when it died. As it was badly decom-
posed when examined no cultures were made.

Fowl No. 64 was again inoculated in both eyes with the exudate from the eye of
this fowl. The exudate was thoroughly rubbed on the conjunctiva. Four days later
the inoculation was repeated. The eyes remained apparently perfectly well.

October 22: A guinea-pig was inoculated into the abdominal cavity with 0.4 cc. of
a bouillon culture from the eye exudate. It remained well.

October 24: The left eye of fowl No. 63 was covered with a quite firm, nearly
white exudate, which causes it to protrude considerably. The lids were firmly agglu-
tinated together, being separated with much difficulty. The lid was lined with a
thick, quite firm, whitish exudate. Nictitating membrane much thickened. There
were several areas of a grayish exudate in the roof of the mouth. Cultures in agar
and bouillon made from the eye contained two species of micrococci. The growth
in the agar consisted of a few colonies only.

A whitemouse inoculated with the exudate was found dead on the third day. From
the liver a culture of a micrococcus was obtained. This organism did not prove
fatal in subsequent inoculations.

October 27 the eye was entirely closed. The agglutinated lids were separated and
a wedge-shaped mass of necrotic exudate was easily removed. This covered the
entire inner surface of the lower lid, being thicker at the exposed or free margin.

VARIETIES OF LESIONS. | 51

In section it was found to be composed very largely of broken-down cells. The
cornea had an opalescent appearance and was decidedly flattened. A nonpatho-
genic bacillus was obtained in agar cultures from the eye.

November 9, the fowl appeared to be improved. The eye was covered with a
yellowish exudate which emitted a peculiar, offensive odor. It was easily removed
leaving a roughened, nearly healed surface, The cornea was opaque and depressed.
The exudate from the mucosa of the mouth had sloughed, leaving a healed surface.

The fowl was chloroformed for further examination. It was much emaciated,
The internal organs appeared to be normal. Tubes of bouillon inoculated from the
liver, spleen, and blood remained clear.

1h

A review of the disease found in the 18 fowls shows that while there
are many dissimilarities in the lesions in the different fowls they can not
be easily construed to represent different diseases. Although much of
the confusion which characterizes the popular conception of ‘ roup” is
apparent in thefragmentary history obtained of the outbreaks, a more
careful study of the individual cases will show that there is a striking
similarity between them. The lesions in the fowls examined were for
the greater part localized on the mucosa of the organs in the head.
Although all of the fowls which died were much emaciated, there were,
with few exceptions, no lesions or evidence of organic disease in the
internal organs. The cause of death and the extreme emaciation is
difficult to explain in those cases where the lesions were confined to
one eye or to the mucosa of the nares, excepting on the supposition
that some poisonous or toxic substance was absorbed from the seat of
the disease. In those cases where the lesions were in both eyes, or in
the mouth and throat, difficulty in finding or swallowing food affords a
rational explanation.

A study of the lesions revealed the interesting fact that in some cases
the exudate was of acroupous character, and in others of a diphtheritic
nature. In a considerable number of the fowls the exudate was more or
less sloughed from the underlying tissue. The hypothesis is suggested
by these varied conditions that perhaps the apparent croupous exudate
is the first stage of the diphtheritic condition, and that the sloughed
mass of decomposing exudate so frequently found is the last or advanced
stage. There is a certain amount of evidence to support such an hypoth-
esis, both in the description of fowl diphtheria in Europe and in the
cases here recorded. The three stages or varieties of lesions which
represent the types of this disease as encountered in these investiga-
tions may be more definitely defined as follows:

(1) Anexudate of a serous or muco-purulent character in the conjune-
tiva or nasal cavities. Ordinarily this condition can not be recognized
in the mouth. The mucosa in these cases is apparently but slightly
altered. (Fowls Nos. 3,4, 5, Outbreak I, and No, 71, Outbreak III.)

(2) The mucosa over a smnall or larger area is covered with a spread-
ing exudate of a grayish or yellow color, It is firmly attached to the

52 INFECTIOUS DISEASES AMONG POULTRY.

mucous membrane, and when removed leaves a raw, bleeding surface.
Sections through this exudate and the subjacent tissues show that the
epithelial layer is destroyed, and the underlying tissue infiltrated
with cells. The extent of the infiltration varies in different individuals.
(Fowl 59, Outbreak I.)

(3) The mucosa is covered with a thick mass of exudate, varying in
color from a milky white to a lemon yellow or brown. It is easily
removed, leaving a more or less granular and healed surface. This
sloughed mass is frequently dried at its margins to the adjacent tissue.
It emits a strong putrid odor, due to decomposition. The drying of the
margins prevents the fowl from expelling the exudate after it becomes
separated from the underlying tissue. (Fowl 63, Outbreak V.)

The evidence to support the supposition that the three forms or types
of exudate described are different stages in the same morbid process,
as gathered from the post-mortem notes and bacteriological study of the
cases here reported, may be summarized as follows:

(a) Abnormal conditions, representing the intermediate and con-
necting links between the specific types of lesions, were frequently
encountered.

(b) Although but one form of exudate was usually present in a single
fowl, there were marked exceptions, in which two, and in one case the
three, forms were coincident. Thus in fowl No. 75 (Outbreak IIT) the
eye was covered with a sloughed exudate. In the posterior nares there
was a layer of muco-purulent substance, and on the mucosa of the
mouth were areas of a diphtheritic exudate. Fowls Nos. 69 and 73
furnish examples where the diphtheritic process and the sloughed exu-
dates were present.

(c) The same species of a pathogenic bacillus was associated with
each form of exudate. It was almost invariably found with the first
and second, but rarely with the third.

In the fowls which died, the exudates were for the greater part in the
advaneed stage, although there were several fatal cases in which the
lesions were restricted to an abnormal quantity of a serous or muco-
purulent, more or less viscid, exudate in the conjunctiva or nasal cavi-
ties. The best illustration of the diphtheritic process was found in
certain of the fowls killed for examination.

The distribution of the lesions shows that the conjunctiva was most
frequently affected. The exudate in the nasal cavities was in a few
cases undoubtedly the result of the coagulation of the liquid which had —
passed during the course of the first stage from the conjunctiva through
the lachrymal duct into the nares. In certain of the other cases, how-
ever, the lesions appeared in the nares only (fowls Nos. 3,4, and 5).
Inadvertently, in these cases no material for sectioning was saved from
the mucosa of the nares or conjunctiva. Sections of the exudate, with
subjacent tissue from the cornea and the mouth, show that there is a cell

BACTERIA ASSOCIATED WITH THE LESIONS. 53

infiltration into the mucosa which destroys the epithelial layer and
frequently the submucous tissues to a considerable depth. (See P1. VI.)

The fact should not be overlooked, as pointed out in the post-mortem
notes, that the disease in the eye was confined to the conjunctiva and the
cornea, the posterior portion remaining apparently normal. Although
the lesions were found more frequently in the eye, the number of fowls
examined was not large enough to admit of the conclusion that this is
generally the case.

From the observations thus far made the provisional theory is enter-
tained that the three forms of the exudate—serous or muco-purulent,
diphtheritic, and sloughed mass—represent three stages in the course of
the same disease. It is easily understood that fowls examined in the
first stage would be said to be affected with a catarrhal condition of the
mucosa of the eyes or nares. It is highly probable that in many cases
the disease never reaches the second stage (see fowls Nos. 3, 4, 5, and
71), and if these cases alone were examined the diphtheritie condition
would not be suspected. It appears, however, that in the majority of
cases the disease runs its course, and membranes are formed, slough,
and recovery follows. It is further presumable that the disease in
question appears sometimes in a virulent and destructive form. I am
in possession of statements from poultry raisers which show that there
are occasionally epizooties of a disease characterized by exudates in
the eyes, nose, or mouth, which runs a rapidly fatal course. It appears
that it is such outbreaks which have been reported in Europe as diph-
theria, and not the low form of chronic disease which I have encountered.

Itis highly probable that there are many affections of fowls resembling
somewhat closely the lesions found in the cases examined which are
etiologically different. There is much doubt respecting the cause of
the apparently sporadic cases of this disease. Whether they differ
other than in the degree of virulence of the specific organism from the
rapidly fatal disease sometimes reported can not be determined until
such epizootics can be thoroughly studied. It is hoped that the investi-
gation of this disease or class of diseases may be continued until the
doubtful points concerning their nature and cause are clearly explained,
and efficient methods of prevention and treatment are determined,

BACTERIA ASSOCIATED WITH THE LESIONS.

It has already been shown that bacteria were not found in the internal
organs or blood of the affected fowls when they were examined before
post-mortem changes occurred. It has also been stated that a non-
motile, pathogenic bacillus was found associated with the lesions, espe-
cially in the first and second stages, in a considerable number of the
fowls.

The examination of the exudates for bacteria was attended with
many difficulties, owing to the presence of a large number of sapro-

5A INFECTIOUS DISEASES AMONG POULTRY.

phytes.' The method found to be the most trustworthy for the isola-
tion of the pathogenic bacteria was the subcutaneous inoculation of
experimental animals with the exudates. If the inoculated animals
died, pure cultures of the pathogenic organism were obtained from the
blood, liver, or spleen. If they survived, they were subsequently chloro-
formed and cultures of the parasitic bacteria obtained from the tissue
at the point of inoculation.” By this process the saprophytes were
destroyed in the tissues and only those organisms encountered which
were suspected to be of more or less etiological importance.

The success of the animal inoculation over the culture methods in iso-
lating the nonmotile pathogenic bacillus is explained from the fact that
it is not vigorous in its growth on culture media and that it was ordina-
rily crowded out by more vigorously growing saprophytes. It isfurther
presumable that in the older exudates its virulence is gradually lost
(see fowls Nos. 1 and 59), so that its presence could not be detected
readily by animal inoculation. If rabbits had been inoculated, however,
with large quantities of the exudate from the more advanced disease a
few additional cases containing the pathogenic—but attenuated—bacil-
lus would undoubtedly have been found. The severe local lesions pro-
duced in rabbits by the injection of such material caused them, with
few exceptions, to be abandoned. It is evident, in view of the results
obtained, that rabbits were the most efficient experimental animals to
use in this work, and that fowls were the mostrefractory. The scarcity
of rabbits, however, led to the use of other animals in a few cases.

Cover-glass preparations made from the lesions of the affected fowls
were carefully stained and examined.’ Those prepared from the surface
of the exudate and from the necrotic masses generally contained innu-
merable bacteria, but no predominating species was observed. Similar
preparations from the base of the exudate in the more recent stages con-
tained a large number of short bacilli with the ends rounded. These
forms frequently exhibited a polar stain. Several preparations were

1A large number of species of bacteria were isolated and studied somewhat
carefully. Among them are several quite interesting forms, some of which were
suspected to be of more or less economic importance. Prominent among these was a
bacillus which resembled morphologically and in certain of its cultural characters
the bacillus of tuberculosis. It was not pathogenic for guinea-pigs or fowls, and
did not take the Koch stain.

2It is of interest to note that several varieties of colon bacteria were isolated in
this way. One of these possessed such variations from the colon bacillus in its cul-
tural manifestations and it was possessed of such marked pathogenesis that for a
time 1t was suspected of bearing some causal relation to the disease. It was briefly
described in a footnote on page 43. :

2Tn both fresh and stained preparations a long spirillum was found to be quite
abundant. It wasfrom2to4 win length. Thus far I have been unable to detect it in
artificial cultivations,

a

en

i

stained for tubercle bacilli, but with negative results.

SUMMARY OF

INOCULATIONS.

5

5

A summary of

the results obtained from animal inoculations is appended in tabulated

form
Animals inoculated with the exudate from affected fowls.
Animals Inoculated with | Date of Result Raman
inoculated. exudate from—_ CSUN: vers?

| inoculation.

Rabbit No. 117...

Eye, fowl No. 1

| Dec. 28, 1893 |

Died Feb. 10, 189+.

Rabbit very much emaci-
ated; no bacteria found
in its organs.

White mouse....).-..-. Goteer aos ase ee | Jan. 23,1894) Died Jan. 27, 1894..| Enlarged and discolored
| spleen; a pathogenic mo-
| _ tile bacillus.

10 SE See mee Meee CER aaa eae Re eee doi =. Died Jan, 29, 1894.-| Spleen enlarged; nonmo-
| — tile pathogenic bacillus.

Rabbit No. 144...) Nares, fowl No. 3-... Feb. 4, 1894 | Died Feb. 28, 1894.) Septicemia; nonmotile

: | pathogenic bacillus.

Rabbit No, 145.:.)..... dO So 34- sewses= Feb. 24,1894 | Died Feb. 25, 1894. Do.

Rabbit No. 97....| Nares, fowl No. 4.-..|..-.. GO ascent eee QGr ease nee Do.

Rabbit No. 149...| Nares, fowl No. 5- 4 Feb. 29,1894 | Died Mar. 1, 1894..| Do.

Guinea-pig 222...

| Chloroformed Oct.

Local lesion contained Ba-

; i 13, 1894. cillus coli communis.
Guinea-pig 223...| Nares, fowl No. 62..|....- Cleaner Chlorotormed Oct. Do.
15, 1894.

Guinea-pig 209...) Mouth, fow] No.59..| Oct. 20,1894 Chloroformed Oct. | Local lesion contained non-
| 25, 1894, motile pathogenic bacil-
lus. F

Fowl No. 50..-... Eye, fowl No. 65....| Oct. 24,1894 | Remained well....| No local reaction.

Fowl No. 54.....- Eye, fowl No. 68..--.| Oct. 27, 1894 |..-..- GO:22 saeoeac Soe Do.

Fowl! No. 55......|-- REG 0) Pie seer poeeee Seer (Oe eee deren: Os Sas= shite Do.

Rabbit No. 215...) Eye, fowl No. 69....| Nov. 2,1894| Found dead Nov Pleuritis; hemorrhages in

5, 1894. the intestines; nonmo-

| tile pathogenic bacillus.

Rabbit No. 223...| Eyes, fowl No. 71...) Nov. 5,1894) Found dead Dec. | Pleuritis and peritonitis;

19, 1894. nonmotile pathogenic
bacillus.
Rabbit No. 226...| Nares, fowl No. 75..| Nov. 22,1894 Found dead Noy. | Septicemia; nonmotile
23, 1894. pathogenic bacillus.

The table shows that the pathogenic bacillus was obtained from the
lesions of fowls Nos. 1, 3, 4,5, 59, 69, 71, and 75, or nearly 50 per cent of
those examined. It is a significant fact that the bacillus was obtained
from all the fowls but one (No. 61) where’ the lesions either in the
eye, nares, or mouth were not far advanced. In one case (No. 73) no
inoculations were made. The almost constant appearance of this bacil-
lus in the more recent lesions renders the apparent small percentage of
cases from which it was isolated of much greater importance than it
would at first be considered. A comparative study of the bacteria from
the different fowls showed that those obtained from fowls Nos. 1, 59, 69,
and 71 were attenuated in varying degrees, while those from the other
cases were sufficiently virulent to destroy rabbits within twenty-four
hours. This variation in the degree of virulence of the cultures obtained
from different fowls is interesting and important. There was no differ-
ence in their cultural characters. A glance at the post-mortem notes
will show that in the fowls from which the attenuated forms were
obtained the lesions were apparently older or more advanced than in
the other cases.

In previous investigations! rabbits were inoculated with the nasal
and pharyngeal secretions of several healthy fowls. These rabbits

1 Bulletin No. 3, Bureau of Animal Industry, 1893, p. 47.

56 INFECTIOUS DISEASES AMONG POULTRY.

remained perfectly well. Although septic bacteria are known to be
common in the secretions covering the normal mucosa of the upper air
passages of several species of domesticated animals, such organisms
have not been reported from the mucosa of healthy fowls.

In 1890 two fowls! which died very suddenly at the experiment sta-
tion of this Bureau were examined. Oneof them exhibited a croupous
exudate extending from the larynx into the trachea, and the other a
swollen condition of the mucosa of the mouth and m@sophagus. No
pathogenic bacteria were obtained from the fowl] suffering from the exu-
date, but from the other were obtained pure cultures of a bacillus not
distinguishable from that of swine plague.

It is impossible to positively identify the pathogenic bacillus found
associated with the lesions in this disease with the one described by
European writers as the cause of fowl diphtheria. Morphologically it
is similar to the one described by Loeffler, but its pathogenesis is differ-
ent. This may be due, however, to difference in the degree of virulence.
There is much obscurity in the reference to the properties of the bacilli
found by other investigators as the probable cause of diphtheritic affee-
tions of fowls and birds, as their descriptions are exceedingly meager,
being often limited to the morphology and possibly the character of the
growth on one or two of the more commonly used culture media.

In those species of pathogenic bacteria which have been more fully
described a more accurate comparison is possible. Thus, the bacilli of
fowl cholera, swine plague, and rabbit septicemia are found to be com-
parable with the bacillus about to be described, and found in the lesions
of this disease of fowls. While it is not my purpose to discuss at this
time the identity or relationship of these bacteria, it is important to
know that a bacillus associated with the peculiar lesions of this disease
should be so similar to the one described by European writers as the
cause of fowl cholera. The more essential properties of the bacillus
found, apparently the etiological factor, in the lesions of the diphtheritic
disease of poultry which I have studied are appended :

DESCRIPTION OF THE NONMOTILE PATHOGENIC BACILLUS.

Morphology.—A nonmotile, rod-shaped organism 0.8 to 1.5 “4 long and from 0.8 to
1.2 4 thick. The ends are oval, and the shorter forms appear to be nearly spherical.
In bouillon they are frequently in short chains and in clumps. When stained with
the aniline dyes in cover-glass preparations made directly from animal tissues they
exhibit a light center, occasionally showing deeply stained poles. In preparations
from cultures this character is much less marked. No capsule has been positively
demonstrated, although certain preparations suggest its existence. It stains readily
with the aniline dyes ordinarily used. It does not retain the coloring matter when
treated after the Gram method. :

Culture characters.—On agar at 36° C. the growth is not vigorous. It is of a neu-
tral gray color, with a glistening, moist-appearing surface. The growth is slightly
viscid, and adheres quite firmly to theagarsurface. The condensation water becomes

' Special Report on Swine Plague, Bureau of Animal Industry, Department of Agri-
culture, 1891, p. 158.

DESCRIPTION OF THE BACILLUS. 57

faintly clouded. Within agar the colonies appear as minute grayish dots. The
growth is completed in about forty-eight hours. In agar plates (double disks) it
emits a disagreeable, pungent odor, similar to that observed in cultures of the
swine-plague bacillus.

It does not grow in alkaline, peptone gelatin. It does not develop on potato.

Milk remains unchanged in appearance for six weeks. Cultures which had grown
for several days were boiled. No change was produced.

Alkaline bouillon at 36° C. becomes uniformly clouded in twenty-four hours. The
reaction becomes acid in from one to two days. It does not grow in acid bouillon.

In bouillon containing sugar the growth was slightly more vigorous. In the fer-
mentation tubes the liquid in both branches becomes cloudy. Gas is not produced.
In bouillon containing dextrose and saccharose the alkaline reaction is changed to a
strongly acid one in twenty-four hours ; the bouillon containing lactose remained
alkaline,

In a weak solution of peptone containing salt it grew feebly, but gave a decided’
indol reaction. The phenol reaction was not detected.

Thermal death point.—This organism is destroyed in bouillon cultures heated in a:
water bath for five minutes at 58° C. Anexposure of ten minutes to a temperature
of 54° C. was not fatal.

Effect of drying.—The bacillus is destroyed in twenty-four hours by dryimg on
cover-glasses at the temperature of the laboratory.

Effect of tow temperature.—Dry cover-glass preparations were exposed in a sterile
test tube to a temperature varying from zero to a few degrees above for a period of
seventeen hours. At the end of that time they were placed in tubes of bouillon and
put in the incubator. These tubes contained pure cultures of this bacillus on the
following day.

Disinfectants.—A few preliminary tests showed that this organism was very sensi-
tive to ordinary disinfectants. To determine more accurately the strength of the
solution and the time necessary for a fatal exposure, Dr. C. F. Dawson kindly ecar-
ried out an extensive series of experiments, the more important results of which are
appended:

Commercial sulphuric acid in one-eighth of 1 per cent solution destroys in thirty
minutes; in one-fourth of 1 per cent it kills in ten minutes.

Lime water destroys the organism in one minute. When diluted four times (0.015
per cent lime) it requires thirty minutes. Carbolic acid in strengths of 0.5, 1, and 2
per cent destroys this bacillus in sixty, five, and one minutes, respectively.

The frequency with which sulphur fumes are used to disinfect chicken houses led
to the testing of their efficiency on this organism. The methods and results reported
by Dr. Dawson are as follows:

**A drop of bouillon culture was placed upon a sterile cover-glass inside a large
glass jar which had been specially prepared for the test. About a half teaspoonful
of sulphur was placed upon a sheet of asbestus and ignited. The lid of the jar was
put on and in a short time the jar was filled with the white fumes. In three hours
the cover-glasses were carefully removed and dropped into a tube of sterile bouillon
and placed in an incubator. They invariably remained clear.”

Pathogenesis.—Rabbits inoculated subcutaneously with 0.1 cc. of the virulent
bouillon cultures died in from eighteen to thirty-six hours with lesions similar to
those produced by swine-plague bacteria of a virulent type. The organs contained
large numbers of oval bacteria exhibiting a polar arrangement of the protoplasm.
Those inoculated in a similar manner with the attenuated bacilli lived several days.
They exhibited more or less severe local lesions and exudates on one or more of the
serous membranes. Adult fowls inoculated subcutaneously with 0.5 cc. of a fresh
bouillon culture of the virulent form remained well. A young fow] (about 6 weeks
old) inoculated in a similar manner with 0.3 ce. died on the fourth day. The kid-
neys were abnormally yellow in color and contained large quantities of urates.

58 INFECTIOUS DISEASES AMONG POULTRY.

Other organs were apparently normal. No bacteria were found in cover-glass prepa-
rations from the organs, but tubes inoculated with the blood and liver developed
into pure cultures of the bacillus injected.

Several white and gray mice were inoculated subcutaneously with pure cultures.
One of the white mice died. The others remained well.

Several efforts were made to produce the local disease by injecting pure cultures
of this bacillus into the nasal cavity of fowls. These gave invariably negative
results. Cultures were also fed in large quantities to several fowls without produc-
ing the disease.

The fact that diphtheritic lesions were not produced in fowls with
this bacillus appears to be an argument against its specific relation-
ship to the disease. Loeffler produced the disease in pigeons with the
bacillus he isolated. In these investigations the conditions under which
the inoculation experiments were made were presumably quite different
from those surrounding the fowls which developed the disease. The
variations in the degree of virulence of the organism from the different
fowls renders it difficult to determine whether the infection depends on
the virulence of the bacillus, or a predisposing condition on the part of
the fowls. The etiological relation of this bacillus to the disease in
question, however, is neither demonstrated nor affirmed, although the
evidence in hand is quite enough to warrant the working hypothesis
that it is, to a certain extent at least, an exciting agent in the produe-
tion of the accompanying lesion.

PREVENTION AND TREATMENT.

The nature of this disease, as determined by the foregoing investiga-
tions and the published statements of a large number of poultry raisers,
indicates that it can be prevented, and if introduced it can be cured.
Although it will be necessary to determine experimentally the efficiency
of curative agents on a considerable number of fowls before formulas
for treatment can be laid down, several important suggestions present
themselves at this time, especially in reference to preventive measures.

In order to prevent this disease it is evident that many conditions
must be strictly complied with. The character of the food and the
general sanitary conditions, including cleanliness, ventilation, and the
temperature of the poultry houses, must be considered. Undoubtedly
there is much to be learned in connection with the proper care of poul-
try, especially in rural districts where it is probable that many methods
now in use will, after a careful investigation, be found defective. The
determination of these details must necessarily be made by those
actually engaged in the raising of poultry. In addition to the general
sanitary methods, the following rules should be observed:

(1) Fowls which have an exudate on any of the mucous membranes
of the head, or which have come from flocks in which such a disease
exists or has recently existed, should not be introduced among other
poultry.

—s

——

PREVENTION AND TREATMENT. 59

(2) If the disease appears in one or more fowls of a flock they should
be immediately separated from the well ones. If possible, the source
of the infection should be determined and removed.

(3) The quite common practice of allowing fowls from different flocks
to run together during the day should be discouraged.

(4) Care should be taken to avoid the possibility of bringing the virus
of the disease from affected flocks in the dirt or excrement which natu-
rally adheres to the shoes in walking through an infected chicken yard.
The same care is necessary in the interchange of working implements,
such as shovels, hoes, ete.

It is evident to any careful observer that the fact is too often over-
looked that fowls, owing to their method of living, are more liable to
infection than other farm animals. This is especially true when they
are allowed to run at random, as they too frequently are, picking their
living from the garbage pile and barnyards, or securing even more
unwholesome food. There is little doubt that many so-called outbreaks
of contagious disease among fowls are simply enzooties brought about
by improper care. The efficiency of these few suggestions in reference
to the prevention of this disease is demonstrated by the success of
certain poultry raisers who adhere strictly to the teachings of sanitary
methods.

The wide distribution, the large number of fowls affected, and the
usual chronic course of this disease render it one of the few poultry
affections for which curative measures promise to be of practical value.
Although prevention is the safest of cures, when the disease is once
introduced as it is in a very large number of flocks, the necessity for
remedial treatment is apparent, and where economy is to be considered
should be recommended. The practice sometimes followed of destroy-
ing all of the affected birds should be discouraged. Although experi-
ments have not been made to test the efficiency of remedies already
recommended, or to investigate the practicability of others, the testi-
mony of many practical poultry raisers is, as previously stated, to the
effect that the disease is amenable to treatment.

The most certain of the known methods of treatment is the local
application of certain disinfectants, among which a weak solution of
carbolic acid appears to be the most satisfactory. The fact that the
lesions are so much exposed renders the disease especially favorable for
topical applications. The administration of mild stimulants has also
been recommended. In addition to the medicinal treatment, it is of
much importance that the affected fowls be provided with proper food
and kept in dry, warm,and well-ventilated apartments. In the course
of this study a considerable number of badly affected fowls fully
recovered in the animal room of this laboratory with no treatment other
than a warm, dry atmosphere.

If the disease has reached its third stage, it is frequently necessary
to remove the sloughed exudate, which is retained on account of the

60 INFECTIOUS DISEASES AMONG POULTRY.

attachment of its margins, before the application of the disinfectant.
A few fowls have been received where it was evident that death would
soon have followed from starvation or a closure of the respiratory pas-
sages, and which were immediately relieved and cured by removing the
obstructing exudate. There are certain disinfectants and stimulants
recommended, which I am advised will give good results, if adminis-
tered early in the course of the disease, by mixing them with the food
and thus eliminating the necessity for individual application.

THE RELATION OF DIPHTHERIA IN FOWLS TO PUBLIC HEALTH.

A comparison of the bacillus of diphtheiia in man (Klebs-Loeffler)
with the one described by Loeffler as the cause of diphtheria in fowls,
shows that morphologically and in their pathogenesis for experimental
animals the two organisms are in no way alike. ‘There is also a marked
difference in the nature of the exudates in fowls and in man. The non-
identity of these diseases has been clearly pointed out by Ménard.!
Although these diseases are shown by several observations to be unlike
in their etiology and the character of their lesions, the transmission of
fowl diphtheria to the human species, and vice versa, is affirmed by
several writers.

Gerhardt? reports 4 cases of diphtheria among 6 workmen who had
charge of several thousand fowls, many of which died of diphtheria,
in Wesselhausen, Baden. There were no other eases of diphtheria in
the neighborhood, and the evidence was quite conclusive that the dis-
ease was contracted from the affected fowls. It is also stated that an
island on the northeastern coast of Greece had been free from diph-
theria for at least a third of a century, when a dozen turkeys, several
of which were diseased, were introduced. Soon afterwards diphtheria
appeared in a house near the garden where the turkeys were kept.
The disease became epidemic on the island causing the death of 36
people, or over 40 per cent of those attacked.

Debrie* reports briefly the clinical history of 6 cases of diphtheria
which occurred in the garrison of Sebdou, and states that while the
sixth case (2 were fatal) was still under treatment in the hospital 10
fowls kept in a house not far from the hospital were attacked with
diphtheria, and exhibited symptoms strikingly like those present in the
human beings. Five of the 10 fowls died, and two heads were sent to
Arloing, who confirmed the diagnosis of fowl diphtheria. The fowls
were fed by a hospital attendant, and it was ascertained that an identi-
cal outbreak had occurred among the fowls at a neighboring place
from which one of the 6 cases of human diphtheria had been brought.
Debrie is inclined to the view that human diphtheria is transmissible
to fowls and fowl diphtheria to man.

‘Revue d’ Hygiene, tome XII (1890), p. 410.
* Revue f. Thierheilkunde u. Viehzucht, Bd. VI (1883), p. 180.
5 Reviewed in Centralblatt f. Bakteriologie, Bd. XIII (1893), p. 730.

CONCLUSIONS. . 6]

Cole! reports an interesting case near Jacksonville, Ill. <A flock of
fowls became affected with a disease characterized by an exudate on
the mucosa of the head. Some of the exudates emitted a foul odor.
As the weather was cold, one of the chickens was taken into the house
where a child about 24 years old fondled it. Four days later the child
was taken sick apparently with diphtheria, from which it died. There
were no other cases in the neighborhood, and the affected chicken was
the only possible source of infection.

The diphtheritic disease of fowls reported by Loir and Ducloux (loc.
cit.) in Tunis, in 1594, spread to the people of that place, resulting in an
epidemic of serious proportions. Ménard refers to the fact that men
employed to feed young squabs contracted diphtheria by blowing the
masticated food into the mouth and crop of squabs suffering with that
disease. Schrevens® reports several cases of diphtheria in children in
which he traces the source of infection to certain poultry.

Although the number of reported cases of the transmission of fowl
diphtheria to the human species, and vice versa, is small in comparison
with the extent of the disease in poultry, the evidence that such a
transmission is possible is quite sufficient to discourage the careless
handling of diseased fowls. It is a quite common practice, especially
in the rural districts, to bring the sick chickens into the house for treat-
ment, where the children of the household are allowed to fondle them
at will. It is not improbable that when this disease is thoroughly
investigated thé number of cases of direct infection from this source
will be found to be much larger than it is at present supposed. Until
such investigations are satisfactorily completed the indiscriminate
handling of diphtheritie chickens, especially by children, and the
exposure of fowls to the infection of diphtheria in the human species,
whereby they may become carriers of the virus, should be strenuously
avoided.

CONCLUSIONS.

The preceding investigations, though preliminary, are important in
bringing together certain existing facts and in a measure correlating
certain confusing and contradictory theories in reference to a few of
the more common so-called diseases of poultry. Although many
important questions, especially concerning the etiology, are as yet unsat-
isfactorily answered, the facts elicited are of much value as a basis for
future investigation. The definite results which have been obtained,
and which mark the progress made in our knowledge of fowl pathology,
may be summarized in the following determinations:

(1) Many of the so-called diseases of fowls which are characterized

1Archives of Pediatrics, XI (1894), p. 381.

2Cole’s description of the disease shows that it was undoubtedly the same as the
one I have studied.

$ Bulletin de PAcad. Royale de Méd, de Belgique, VIII (1894), p. 380,

62 INFECTIOUS DISEASES AMONG POULTRY.

by an affection of the mucous membranes of the head, and popularly
designated as ‘“roup,” diphtheria, influenza, and sometimes cholera,
resemble each other so closely in their manifestations that they may be
considered as belonging to one and the same disease. This disease is
distinct from the rapidly fatal malady which is better known as fowl
cholera.

(2) The lesions, as usually encountered, are diphtheritic in nature.
In the advanced stages the accumulating exudates decompose and emit
a putrid odor. In the earlier stages they are frequently of a serous or
muco-purulent character, and not infrequently fowls die before the diph-
theritic condition appears.

(5) The same species of a pathogenic bacillus is associated with
apparently different forms of lesions, more particularly in the serous or
muco-purulent and diphtheritic. This bacillus is not distinguishable
from the one described by European writers as the cause of fowl cholera
and can not be differentiated in parallel cultures from the bacillus of
swine plague. It is comparable in certain respects with the supposed
specific organisms of certain diphtheritic diseases of poultry and birds.
Its causal relation to this disease, however, is not demonstrated.

(4) This disease usually runs a slow, chromic course from which the
majority of the affected fowls recover. Its long duration enables the
lesions to be more or less modified from the effect of external contami-
nation. Outbreaks in which the disease runs a rapidly fatal course are
reported, but thus far they have not been encountered in the investi-
gations of this Bureau.

(5) This disease, or, as may subsequently be found, diseases, is ame-
nable to treatment. The treatment consists largely in good hygiene,
and the removal of the accumulated exudate when the disease has
reached the advanced stage. The local application of disinfectants is
indicated.

(6) A large number of slight or more severe enzootics among fowls,
due to local unsanitary conditions, are popularly considered as outbreaks
of some contagious disease.

(7) Judging from the results of these investigations and the recorded
experience of certain poultry raisers, it is highly probable that this
malady would have been prevented in a large proportion of the flocks
in which it now exists if careful sanitary methods had been followed
and precautions taken against the introduction of the disease through
the purchase of affected fowls.

A STUDY OF A BACILLUS OBTAINED FROM THREE OUTBREAKS OF FOWL
CHOLERA.

By VERANUS A. MOORE.

Although a large number of rapidly fatal outbreaks among fowls are
reported annually from the central and southern sections of the United
States, very little is positively known of the nature and cause of the
disease producing them. In 1879-80 Dr. D. E. Salmon! investigated
certain epizootics among fowls in North Carolina, in which he found a
disease comparable to the one described as cholera des poules (fowl
cholera) in France. With the exception of his investigations, which
were made before the development of bacteriological methods now
considered essential and which were restricted to an inquiry into the
nature of the disease in very few outbreaks, we have no definite knowl-
edge of the disease known as fowl cholerain America. In Europe this
disease has been extensively studied and the specific organism carefully
described by several observers; the descriptions of the specific bacillus
differ somewhat in reference to certain properties, but, without attempt-
ing a review of the voluminous literature on the subject, suffice it to say
that it is generally accepted that the bacillus of fowl cholera is very
closely related to, if not identical with, the bacillus of rabbit septica-
mia. The results recorded by Dr. Salmon show that he obtained a
similar organism from the fowls he examined, but unfortunately the
condition of bacteriological science, at that time did not permit the
determination of certain important differential biological properties.

The further inquiry into the nature and cause of this disease of fowls
has been attended with much delay on account of the difficulty in pro-
curing suitable material. Although this may. appear paradoxical in the
presence of so many reports of epizootics, a moment’s reflection upon
the existing condition will reveal its truthfulness. It was shown in the
previous article that there is a great looseness in the use of the term
‘‘fowl cholera,” so that many, if not all, of the forms of local, infectious,
and contagious diseases of poultry are frequently designated by it.
Again, there is a hesitancy among poultry raisers to report the exist-
ence of a disease in their flocks, especially before its seriousness is

' Reports of the Department of Agriculture, 1880, 1882.

. 63

64 INFECTIOUS DISEASES AMONG POULTRY.

demonstrated by the death of several individuals when it is usually
too jate for an investigation. Repeatedly, reported epizootics have
been visited by different investigators of this Bureau, but invariably
the conditions found precluded the determination of the nature of the
disease which had already run its course and disappeared.

In the summer of 1894 three serious outbreaks of fowl cholera were
reported, and from each of them a single fowl was received for exami-
nation. From the organs of each of these fowls a nonmotile patho-
genic bacillus was obtained in pure cultures. As I was unable to study
the outbreaks in the field, the investigations here recorded are neces-
sarily confined to the study of the organism obtained from these fowls
and the disease produced by feeding and inoculation experiments.
Fowls and rabbits inoculated with pure cultures of the bacillus, isolated
from each of the affected fowls, produced similar fatal results, showing
that very little if any difference existed in the degree of virulence pos-
sessed by the different cultures. A comparative study of their cultural
characters and physiological properties showed them to be identical.
The study of this organism, however, showed that it possessed certain
properties which have not heretofore been attributed to the bacillus of
fowl cholera, and which are not possessed by the bacillus of swine
plague or rabbit septiczemia, bacteria generally supposed to be iden-
tical with it. The history of the outbreaks, so far as learned, and the
conditions found in the fowls examined are appended:

April 7, 1894, a dead fowl was received from Mr. S., who resides near the city of
Washington, D.C. He had lost nearly all of his fowls (about 50) with cholera dur-
ing the preceding few weeks, the one brought to us being the last to die. He was
requested to notify us if the disease continued in his flock, and to send all the
fowls that were subsequently attacked. As no other fowls were received, it is pre-
sumable that the disease had disappeared and that the fowl examined was the last
to be affected.

The fowl received was a hen about 2 years old. She was in a good condition.
The liver, spleen, kidneys, and lungs, appeared to be normal. The crop contained a
considerable quantity of dirt, kitchen seraps, corn, and other food. In the abdomi-
nal cavity were two large blood clots and a broken egg. The mucosa of the head
normal. Comb pale.

Cover-glass preparations made from the liver showed no bacteria. Tubes of agar
anid bouillon were inoculated from the liver and blood. On the following day these
contained pure cultures of a nonmotile bacillus. It was fatal to fowls, rabbits,
guinea-pigs, and mice.

Tele

August 8, 1894, a dead fowl was received by express from Mr. 8., of Tackett’s
Mills, Va. <A postal card from him bearing date of August 8, addressed to the De-
partment of Agriculture, reads in part as follows: ‘‘I send you by Adams Express a
hen with genuine cholera. I have lost about 80 hens and pullets in the last month.”
Sal About three weeks later a letter was received from Mr. 8. in which hestated
that he had lost about 100 hens and pullets in the last two months from fowl] cholera.

DESCRIPTION OF THE BACILLUS. 65

The autopsy of the fowl received showed that death had occurred sometime before
its arrival. The organs were somewhat discolored from post-postem changes. The
liver was fatty, kidneys yellowish in color, due to an excess of uratesin the tubules.
Heart muscle pale. The contents of the intestines were greenish and semiliquid
in consistency. Mucosa of cloaca had a yellowish color. Tubes of bouillon and agar
were inoculated from the liver. The following day the tubes contained pure cul-
tures of a nonmotile bacillus, which upon subsequent study was found to be identi-
eal with the bacillus obtained from the fowl examined from the first outbreak.

088

In August, 1894, while engaged in other investigations, Dr. Theobald Smith had the
opportunity of examining a fowl which died in an outbreak of fowl cholera on Block
Island, R. I. The fowl died August 13. It was kept on ice until August 15, when it
was catefully examined. It came from a flock of about 70 fowls. The disease was
reported to have begun in June, and at the time this fowl died only about 14 or 15
were still living. The autopsy notes made by Dr. Smith show that the organs
appeared to be normal. Four cultures were made from the spleen, liver, and blood.
On the second day following these contained a moderately vigorous growth of anon-
motile bacillus.

August 31: Dr. Smith gave me these cultures for further study. ‘They were found
to be identical and pure. A series of inoculations and a careful study of the mor-
phology and physiological properties of this organism showed that it was identical
with the bacillus obtained from the other two fowls.

Although the data concerning the nature of the disease in the three
outbreaks is exceedingly meager, the facts in hand are quite enough to
show that it was practically the same in its manifestations in the three
localities. From other sources I have learned that fowl cholera, or a
rapidly fatal disease known as such, is not uncommon in the vicinities
where two of tie outbreaks occurred. No one can doubt that, if the
history given is correct, the disease in the flocks from which the fowls
-eame was of a highly contagious, or at least infectious, nature. The
presence of the same species of a pathogenic bacillus in the organs of
the three fowls is evidence supporting such a conclusion. As the history
of the disease in each flock and the examination of the fowls received
showed the absence of diphtheritic lesions, it is necessary to consider
the disease in question either fowl cholera, in the general acceptation
of that term, or to add a new disease to fowl pathology, which, from
our limited knowledge of this disease in the United States, does not
appear to be justifiable.

DESCRIPTION OF THE BACILLUS.

Morphology.—A nonmotile, rod-shaped organism, varying somewhat in size accord-
ing to the media in which it develops. In tissues of fowls or rabbits it is from 1.4
to 1.8 long an from 1 to 1.3 4 thick. No spores or vacuoles have been discovered.
The ends are rounded, so that in the shorter forms it could easily be mistaken for
a micrococcus. In the tissues it frequently appears in small clumps. Involution
forms are common. In cultures on agar it is more slender than in tissues. When
examined in a hanging drop preparation, especially at the edge, it frequently shows
a marked polar arrangement of the cellular protoplasm. In these preparations
there is observed a marked dancing motion of the organisms. In old bouillon cul-

17897—No. 8 5)

66 INFECTIOUS DISEASES AMONG POULTRY.

tures short chains composed of bacilli united end to end are sometimes observed.
It stains with the ordinary aniline dyes, but retains the coloring matter very feebly,
or not at all, when treated after the Gram method.

Cultural characters.—Agar.—On this medium at 37° C. the growth is moderately
vigorous. It has a grayish, glistening appearance. Isolated colonies are from 1
to 2 mm. in diameter, convex, with sharply defined borders. Agar plates emit a
peculiar penetrating odor.

Gelatin.—In this medium the growth is less vigorous. In stick cultures it is more
abundant along the line of inoculation than on the surface. Isolated colonies are
about 0.25 mm. in diameter, appearing as homogeneous bodies to the unaided eye, but
slightly granular under low magnification. They have a grayish-yellow color.

Potato.—On the surface of potatoes a delicate grayish-yellow growth appears after
about forty-eight hours. Frequently there is no development, owing, presumably, to
the acid reaction.

Alkaline boullion.—In twenty-four hours alkaline bouillon: becomes uniformly
clouded. Reactionacid. The bouillon becomes clear after several days. The growth
settles with the formation of a small quantity of friable sediment. The bacteria live
for at least four weeks.

Acid bouillon.'—Very faintly clouded; acid reaction. Slight sediment, which is not
viscid.

Milk.—No change is produced in the appearance of milk for about four weeks. It
then begins to change to a clear opalescent-appearing fluid. In about six weeks
the milk has a decidedly opalescent appearance. Strongly alkaline inreaction. The
addition of a few drops of acetic acid precipitates the casein. A microscopic exam-
ination shows that the fat globules are destroyed. This condition is presumably due
to the alkali produced by the growth of the bacteria. This appearance of milk is not
distinguishable from that produced by the bacillus of hog cholera.

Fermentation of sugars.—Alkaline bouillon containing 1 per cent dextrose in fer-
mentation tubes becomes uniformly cloudy throughout. Strongly acid in reaction.
Similar tubes containing saccharose and lactose become clouded throughout, but they
remain alkaline in reaction, the degree of alkalinity increasing withage. Gas isnot
produced during the growth of the bacillus in bouillon containing sugars.

Indol and phenol.—Cultures in a peptone solution give a strong indol reaction when
treated after Kitasato’s method. No phenol reaction could be obtained when cul-
tures were treated according to Weyl-Lewandowski.

Thermal death point.—Tubes of bouillon inoculated with several drops of a fresh
bouillon culture, exposed in a water bath to a temperature of 58° C. for fifteen
winutes or longer, did not develop. Those exposed for ten minutes or less developed
into pure cultures of the inoculated bacillus. An exposure at 54° C. for thirty min-
utes produced no deleterious effect.

Effect of disinfectants.—A one per cent solution of carbolic acid was fatal in five
minutes. <A one-fourth of one per cent solution of commercial sulphuric acid de-
stroyed life in ten minutes, Lime water was fatal in ten minutes, and a solution
composed of one part lime water and three parts distilled water was equally as
effective in one hour. Sulphur fumes were also fatal in the test applied by Dr. C. F.
Dawson. A drop of a bouillon culture was placed upon a sterile cover-glass inside a
large glass jar which had been specially prepared for the test. About a half tea-
spoonful of sulphur was placed upon a sheet of asbestus and ignited. The lid of the
jar was put on and in a short time the jar was filled with white fumes. In three
hours time the cover-glasses were carefully removed and dropped into tubes of sterile
bouillon. These tubes were placed in an incubator. They invariably remained
clear.

‘Bouillon prepared from beef without neutralizing or rendering alkaline by a
solution of soda or potash.

NATURE OF THE INOCULATION DISEASE. 67

Effect of low temperature.—Bacteria dried on cover-glasses were placed in sterile
test tubes and exposed in a temperature of from zero to a few degrees above for
seventeen hours. This exposure did not appreciably retard their subsequent mul-
tiplication in bouillon.

The effect of drying.—This bacillus withstood drying at the ordinary temperature
on cover-glasses for seven days,

THE NATURE OF THE INOCULATION DISEASE.

The disease produced in the experimental animals with pure cultures of this
bacillus is, when compared with the described effect of the bacillus of fowl cholera,
exceedingly interesting.

Fowls inoculated in the wing vein with 0.3 cc. of a fresh bouillon culture died in
from three to eight days, usually on the fifth or sixth day. The temperature begins
torise,on the second day after inoculation. It reaches 109- to 111° F.! the day before
death occurs. In cases where the fowls live from five to six days they appear per-
fectly well for at least three days, when the feathers begin to have a slightly ruffled
appearance. The exposed portion of the head usually, but not invariably, becomes
pale. Toward the last they sit in a crouched position, with the head drawn close to
the body. They refuse food, but usually take more or less water. There is some-
times a slight diarrhea. That portion of the excrement secreted by the kidneys
becomes yellow.

The changes observed at post-mortem are not conspicuous. The liver is moderately
enlarged, soft and fatty. The spleen is occasionally enlarged. The kidneys are
marked with yellowish lines, due to the injection of the tubules with urates. The
intestinal mucosa is sometimes hyperwmic and sprinkled with punctiform hemor-
rhages, especially in the rectum. The contents of the intestines are, as a rule,
greenish in color and normal in consistency. The mucosa of the cloaca is frequently
of a yellowish color, due to urates. The heart muscle is pale. In afew cases, where
the fowls resisted for an unusually long period, tubercle-like nodules were observed
in the heart and lungs.

The most marked pathological changes were found to occur in the blood. Within
twenty four to forty-eight hours after an inoculation into the veins a diminution in
the number of the red corpuscles and an increase in the number of leucocytes was de-
termined. This decrease in the number of the red cells and the increase in the white
ones continued until death occurred. This isillustrated by the notes from a somewhat
typical case.

Fowl No. 82.

{Inoculated February 6.]

Temper- | Number of | Number of
Date. ature. red cor- white cor- |
F.° puscles. puscles. | Remarks.
|
MAG ORAR VAG. om ae ose srk eon oes onc 107.4 3, 744, 444 22, 222 |
HEDPIALY ne tee Sa: snc eueecise™ 109 3, 417, 391 26,087 | Apparently well.
Webueany Shccces ean eS ee sc8 108. 2 2, 784, 700 55, 000 Do.
Bebrdary Oe sess ek saeceee: 108. 4 2, 807, 692 76, 925 Do.
Meprmanry, (iis. ai. ice~ 3) cases ss 107.4 3, 481,818 90,909 | Feathers ruffled; refuses food.
MEDIA. 13.6 2256 os capi is Sines 110. 2 2, 133, 333 100,000 | Very quiet; comb pale.
TOG] tye Ee) Zl SE ee eee ae 108 2, 530, 000 140, 000 | Fowl] dies later in the day.

Unfortunately the changes produced in the blood of fowls by bacterial diseases
are so little known that the extent to which this marked change in the blood is
characteristic of this disease can not be determined until further investigations with
other pathogenic bacteria are made.

1The normal temperature of fowls, as determined by taking the temperature of
the fowls used before inoculation, varies considerably, ranging from 104° to 108° F.

68 INFECTIOUS DISEASES AMONG POULTRY.

The method of destruction of the red corpuscles is not as yet satisfactorily
explained. In his report on fowl cholera Dr. Salmon (loc. cit.) Ulustrated leucocytes
surrounding the red corpuscles, but the marked diminution of the red cells was not
determined, although he speaks of the pale color of the blood. In a few prepara-
tions of fresh blood I have observed similar phenomenon, and also have seen portions
of red cells within the granular leucocytes. The determination of the extent of this
mode of destruction of the red corpuscles, however, necessitates further investiga-
tion. Itis of interest to call attention to certain other observations in connection
with their destruction.

In carefully heated cover-glass preparations of healthy fowl’s blood stained with
eosin and methylene blue the nuclei are colored a deep’ blue, and the cellular proto-
plasm surrounding the nucleus is stained with the eosin. In similar preparations
made from the blood of the affected fowls there are a greater or less number of cor-
puscles which do not take the eosin stain. In these the portion of the corpuscle
surrounding the nucleus remains unstained or becomes slightly tinted with the blue.
It occasionally contains one or more vacuoles, and the margin is frequently broken.
In some instances a considerable portion of the corpuscle has disappeared. There
are a few free nuclei.

Fowls fed with pure cultures and with the viscera of birds dying of this disease
usually succumb in from eight to fifteen days. The subcutaneous inoculation with
moderate quantities of pure cultures gave negative results.

The organs and the blood of fowls dying from the inoculation disease contained
comparatively few bacteria. This was shown in the cover-glass preparations and
also in cultures. Frequently the inclined surface of agar rubbed with a large piece
of the liver or spleen, or a clot of blood, contained on the following day isolated
colonies. In sections the bacteria were frequently found lying in clumps within the
capillaries.

In rabbits the disease resembles very closely that produced by attenuated hog-
cholera bacteria, although the fatal dose is much larger than for that organism.
Subcutaneous inoculations with 0.1 to 0.2 ce. are rarely fatal. When death follows
the lesions are characterized by slight local reactions, necrotic areas in the liver,
enlarged and discolored spleen, and an infiltration of cells into the follicles of the
vermiform appendix and glands about the ileo-cxecal valve. An intravenous inocula-
tion of 0.2 to 0.3 ee. of a fresh bouillon culture produces death in from two to three
days. In these cases the spleen is engorged with blood, the liver swollen, and the
jutestinal mucosa contains hemorrhagic areas. A larger dose produces death in from
eighteen to twenty-four hours. Ai intravenous injection of a small quantity of
culture produces lesions similar to those following the subcutaneous inoculation.
Cover-glass preparations from the liver, spleen, and blood made soon after death
show few elongated bacteria. If a rabbit is allowed to le for twenty-four hours
after death cover-glass preparations show innumerable oval bacteria, which usually
exhibit a polar stain.

A guinea-pig inoculated in the abdominal cavity with 0.2 ec. of a bouillon ecul-
ture died in seven days. The autopsy showed about 50 ce. of a clear, lemon-colored
serum in the abdominal cavity. The viscera were covered with a grayish membra-
nous exudate. The pleural cavity contained a large quantity of serum. Lungs
collapsed. Subcutaneous inoculations of 0.5 cc. proved fatal in frem six to eight
days. There was a purulent infiltration into the subeutis about the point of inocu-
lation. Spleen enlarged, dark colored, and friable. Liver pale and usually fatty. No
intestinal lesions. Comparatively few bacteria in the organs as shown in cover-
glass preparations. :

White and gray mice inocuJated subcutaneously with 1 to 2 drops of bouillon
culture died in from three to five days. The only pronounced macroscopic lesion
observed was an enlarged and discolored spleen. Mice inoculated similarly with a
much smaller quantity (a single small loop of culture) remained well.

NATURE OF THE INOCULATION DISEASE. 69

It was thought that perhaps the bacillus was attenuated because the
fowls were among the last to die in their respective outbreaks. On that
account several series of inoculations were made in different animals
for the purpose of increasing its virulence, the culture from the fowl in
the first outbreak being used. These gave negative results. Several
other experiments are now in progress for the purpose of accelerating
its pathogenesis. It seems, however, that its virulence is not easily
changed by any of the ordinary methods. After nearly a year’s pres-
ervation in subculture it is as rapidly fatal as when first isolated.

The bacillus here described differs from the description of the bacil-
lus of fowl cholera (rabbit septicemia or swine plague) in the following
particulars:

(1) Morphology.

(2) Its effect upon milk.

(3) Its effect upon sugars.

(4) Its degree of virulence for rabbits and fowls.

(5) The nature of the inoculation disease, especially in rabbits.

The morphological changes consist in a slight increase in size and
the feebleness with which it takes the polar stain. The fact is inter-
esting, however, that the polar stain is well marked when the bacillus
is allowed to multiply in the dead animal body.

The saponification of milk is very marked. This property belongs to
the hog-cholera group of bacteria, and it has not occurred in parallel
- cultures of the swine-plague bacillus or other members of that group,
in which it is generally conceded by authorities in bacteriology the
fowl-cholera bacillus belongs.

The bacilli of rabbit septicemia and swine plague produce acids in
bouillon containing dextrose and saccharose, while this bacillus changes
dextrose only. I have not had an opportunity of studying the bacillus
of the European disease, and consequently this variation is assumed on
the authority of the statements that these species are identical.

Thevirulenceof this bacillus is much less than that ordinarily recorded
for fowl cholera. Dr. Salmon produced fatal results in fowls by the
subeutaneous injection of minute quantities of a pure culture, while
this organism requires for fatal results an intravenous injection of a
much larger amount. In rabbits the dissimilarity is much greater.
The bacillus of fowl cholera is said to destroy rabbits with acute septi-
cemia within twenty-four hours after a subcutaneous inoculation with
small quantities of pure culture, but this organism produces a disease
characterized by certain definite anatomical changes.

The theory that possibly the variation in the lesions is due to an
attenuation of the bacillus or a resistance on the part of the animal
does not hold, as the disease produced in rabbits with this organism is
different from that obtained with attenuated rabbit-septiceemia bacteria.
It has frequently been pointed out in the publications from this labora-
tory that attenuated rabbit-septicemia or swine-plague bacteria pro-

70 INFECTIOUS DISEASES AMONG POULTRY.

duce local infiltrations and an inflammation followed by an exudate on
one or more of the serous membranes.

Another point of difference, taking the bacillus of fowl cholera to be
identical with that of swine plague, is the inability to render rabbits
immune to the disease by previous injections of sterilized cultures.
A carefully conducted experiment was made in which 10 cee. of steril-
ized bouillon culture was injected intravenously in repeated small
doses in two rabbits. They were subsequently inoculated with a mini-
mum fatal dose of pure culture. They died with the control rabbits.
Such negative results are invariably obtained with hog-cholera bacteria,
but many experiments with swine-plague bacteria show that such treat-
ment will quite as surely produce a pronounced resistance to that disease’

There are, however, certain similarities in the disease produced in
fowls with the one studied by Dr. Salmon in his investigations in North
Carolina. These are, for the greater part, the rise in temperature, the
period of incubation, yellowish color of the urates, paleness of the
comb, and the refusal of food. The gross pathological changes are
also comparable. The diarrhea was less marked in the disease I have
produced.

The study of this organism and its pathogenesis are as yet too
incomplete to admit of further comparisons at this time. The fact has
been shown that in the inoculation disease there is slight or no diar-
rhea, and pronounced pathological changes occur in the blood. Should
these conditions be found to exist in the disease as it occurs in epi-
zooties it is evident that the term fowl cholera is an unfortunate one.
It has further been pointed out that the bacillus obtained from three
outbreaks of reported fowl cholera differs materially from the organ-
ism generally accepted, and ofttimes described by European investiga-
tors as the specific cause of that disease. It is quite possible that
there are several varieties of fowl cholera, and that the bacillus here
described is the etiological factor of one of these forms. The extent
to which this bacillus is distributed in this country as the cause of epi-
zootics among towls can not be determined until further investigations
are made.

1 Bulletin No. 6 of the Bureau of Animal Industry, 1894, p. 65.

ON A PATHOGENIC BACILLUS OF THE HOG-CHOLERA GROUP ASSOCIATED WITH
A FATAL DISEASE IN PIGEONS.

By VERANUS A. MOORE,

HISTORY OF THE DISEASE.

In July, 1894, an investigation was made into the nature of a reported
epizootic among pigeons at Vineland, N. J. The disease was found,
however, to be restricted to the cotes belonging to Mr. A. H. Hawley,
and while it was said to be very destructive, its course was slow, and
the heavy loss which it occasioned was due to its continued existence
and spread. The only symptom observed was a general emaciation,
or “going light,” as expressed by pigeon fanciers. He had examined
post-mortem a large number of the birds, but the only apparent patho-
logical change noticed, other than the loss of flesh, was an exceed-
ingly pale heart muscle.

In addition to the affection mentioned, a few pigeons had died from
a disease known as ‘‘megrims,” which is characterized by emaciation
and a peculiar turning of the head from one side to the other, with
occasional paroxysms of flying about, apparently in an aimless man-
ner. The economic importance of this disease, however, was said to be
insignificant, and it is considered here in connection with the more
serious affection on account of the identity of the bacilli isolated from
the diseased pigeous.

At the time of my visit Mr. Hawley had a pigeon in the advanced
stage of each of these apparently different diseases, and several
others which he said showed beginning symptoms. As these diseases
appeared to have followed outbreaks of a rapidly fatal affection which
had occurred at variable intervals among his pigeons during the pre-
ceding few years, a brief historical sketch of the disease in his cotes is
of special interest.

Mr. Hawley stated that be began to raise pigeons early in 1889. At
first he kept about 160 pairs. For nearly three years his pigeons
remained well. Late in the fall of 1891 he purchased 50 pairs, aged
about 10 months, from the Fanciers’ Pigeon Club, of Philadelphia.
About four weeks later these pigeons began to die from a disease des-

: 71

12 INFECTIOUS DISEASES AMONG POULTRY.

ignated by him as cholera. About one-half of the purchased pigeons
died. The disease did not spread to the pigeons formerly kept on the
place. A second and more destructive outbreak occurred in the
spring of 1892. During the summer two other quite severe outbreaks
occurred. In August, 1893, he lost 125 birds from the same cause. At
this time the disease was unusually rapid in its course, the pigeons
being sick for only twelve to fourteen hours. The cotes in which the
disease occurred were thoroughly sprayed with kerosene emulsion and
the floors were washed with carbolic acid. They were vacated and left
unoccupied until the spring of 1894, when 75 pairs of pigeons, selected
from the other cotes, were placed in three of the apartments, 25 in each,
and 17 pairs were purchased and put in a fourth apartment of the same
house. The four apartments were arranged side by side, occupying the
entire building. The pigeons remained well for about a month, when
cholera broke out in two (first and third) of the apartments. About 15
birds died, when the disease was checked by the use of medicines.

Soon after the first outbreak of so-called cholera, late in 1891, pigeons
began to die of the disease under investigation. The number of deaths
continued to increase until the profit of the business was consumed and
the extermination of the pigeons threatened. A record of the mortality
kept during the first few months of 1894 showed that in February 83,
in March 36, and in April 66 deaths occurred. In May and June the
loss was about the same. In July a much smaller proportional number
had died up to the time of the investigation. The prevalence of the
disease among young pigeons was noted by Mr. Hawley, although old
birds were occasionally affected. <All of the affected pigeons were
examined.

Pigeon No. 1.—This pigeon was found dead on the morning of July 19. It was
about 6 weeks old. There was evidence of aslight diarrhea. It was much emaciated.
The mucosa of the eyes, nares, and mouth was normal. The liver had a peculiar
greenish color, due, presumably, to post-mortem changes. The kidneys were of a
pale yellowish color. Intestinal mucosanormal. The heart muscle was exceedingly
pale, fatty. Tubes of agar were inoculated from the liver and blood. These con-
tained on the day following pure cultures of a motile bacillus belonging to the hog-
cholera group, as will be shown in its subsequent description.

Pigeon No. 2.—This was an old pigeon. It had been sick for several days. It
turned its head continually from one side to the other and walked with difficulty.
The left eye was closed and contained a small quantity of mucus. It was killed for
examination. Pigeon much emaciated. No lesions were found in thoracic or abdom-
inalorgans. In the subarchnoid space over the cerebellum and the posterior lobes of
the cerebrum there was a grayish-yellow, friable exudate one-half to 1 mm. in thick-
ness and easily removed. Subjacent brain tissue reddened; otherwise normal in
appearance. The dura appeared to be normal. The exudate was composed for the
greater part of degenerating cells and bacteria. Tubes of agar were inoculated from
the exudate over the brain and from the heart blood. The latter remained clear;
the former developed into pure cultures of a bacillus not distinguishable in its bio-
logical character from the one obtained from the organs of pigeon No. 1.

Allof the suspected pigeons were killed and examined, but in every case the organs
appeared to be normal and the culture media inoculated remained clear.

DESCRIPTION OF THE BACILLUS. 73

DESCRIPTION OF THE BACILLUS.

Morphology.—An actively motile bacillus, varying in size from 1 to 1.6 in length
and about 1 /¢ in width. The size varies appreciably in different media. No spores
have been detected. The ends are rounded in cultures, but in tissue they appear to
be more pointed. In preparations from tissues it usually appears in pairs, united
end toend. Frequently, however, it is observed in clumps. The flagella are easily
demonstrated by Loeffler’s process. The number of filaments vary, eight being the
maximum number observed in a single bacillus. In preparations from tissue it
usually exhibits a light center with a uniformly stained periphery. In preparations
from cultures it usually shows very little, if any, unstained area. It does not take
the Gram stain.

Cultural characters.—Agar.—On the inclined surface of this medium after twenty-
four hours at 37° C.a grayish, glistening, nonviscid growth appears. When isolated
the colonies are convex, 0.5 to 1.5 mm. in diameter. Edges sharply defined. The
growth is increased slightly during the next twenty-four hours. In stick cultures
there is a grayish growth along the course of the needle track.

Gelatin.—In this medium the growth is not vigorous. Surface colonies appear as
small grayish dots. When magnified they are finely granular in appearance and of
a yellowish tint. In older colonies there is a distinct band which is less granular
in appearance about the circumference. It is not liquified.

Potato.—It forms on potato a thin, glistening growth, having a faintly yellowish
color. When the reaction of the potato is yery acid no growth appears.

Alkaline bouillon.—This medium becomes uniformly clouded after twenty-four
hours. The reaction is faintly acid, owing to the fermentation of the muscle sugar.
After a longer growth a thin, grayish membrane forms over the surface of the liquid
and a grayish band frequently appears on the sides of the tube at the surface of the
liquid. The reaction becomes strongly alkaline in old cultures. In acid bouillon the
growth is feeble and the reaction is not appreciably changed.

Milk.—When this medium is faintly acid in the beginning it slowly becomes alka-
line. In about three weeks it gradually changes. to a clear, opalescent fluid, due,
presumably, to the alkali! produced by the growth of the bacteria, as the milk at
this time is very strongly alkaline. The addition of a few drops of acetic acid pre-
cipitates the casein. If the culture is allowed to remain in the thermostat for sey-
eral weeks the volume of the milk shrinks, due to evaporation, and pine remaining
part becomes viscid and of a light brownish color.

Alkaline bouillon containing dextrose.—In this medium it produces a marked fer-
mentation of the sugar with the formation of gas. The acid resulting from the fer-
mentation gives the fluid a strong acid reaction. The gas produced (when collected)
in the fermentation tube replaced 54.8 per cent of the liquid in the closed branch.
The gas contained 35.7 per cent CO, (gas absorbed by potassium hydrate) and 64.3
per cent H (explosive gas).

In similar bouillon containing lactose and saccharose the liquid became uniformly
clouded throughout both branches of the tube. No gas was formed. The reaction
remained alkaline, and after some days the degree of alkalinity, as detected by litmus
paper, was much increased. :

A marked indol reaction was determined by Kitasato’s method. 2

Thermal death point.—This organism is destroyed in cultures by an exposure of ten

‘Tt was shown in Bulletin No. 3, 1893, p. 43, that the addition of a few drops of a
strong solution of caustic potash or soda would produce this appearance in milk.
*This consists in cultivating the bacillus at 37° C. in a sterilized solution composed
of 1 per cent peptone and 0.5 per cent sodium chloride in distilled water for from
one to three days. To these cultures 1 cc. of a 0.02 per cent solution of potassium
nitrate, freshly prepared, and about 3 drops of chemically pure sulphuric acid are
added; a violet-pink color indicates the presence of indol.

74 INFECTIOUS DISEASES AMONG POULTRY.

minutes to a temperature of 58° C.in a water bath. It resisted drying on cover-
glass preparations made from bouillon cultures at the temperature of the room for
eight days. Similar tests were not made with preparations made directly from the
organs of animals.

Disinfectants.—The effect of lime and carbolic acid upon this organism shows that
a solution of lime weaker than lime water is not effective, and that a 2 per cent solu-
tion of carbolic acid requires one hour to destroy life.

Pathogenesis.—Rabbits inoculated in the ear vein with 0.3 ce. of bouillon culture
died in thirty-six to forty-eight hours. There was a general congestion of the inter-
nal orgaus. The intestinal mucosa was especially reddened in certain areas. A sub-
cutaneous injection of a similar quantity of culture produced death in from four to
six days. In these cases there was considerable purulent infiltration into the sub-
cutaneous tissue about the place of inoculation. ‘The liver was sprinkled with
necrotic areas and the spleen much enlarged, dark colored, and friable. Whena
smaller quantity of culture was injected the local lesion was much more severe, and
the follicles of the vermiform appendix and also certain of those of Peyer’s patches
were infiltrated with cells, giving them the appearance of round, whitish dots.
White mice died in from three to six days after a subcutaneous inoculation with one
to three drops of bouillon culture. The only lesion observable was an enlarged and
discolored spleen. Guinea-pigs died in from five to ten days after subcutaneous inoc-
ulations with from 0.1 to 0.2 ce. of a fresh bouillon culture. Pigeons resisted a sub-
cutaneous inoculation with a small dose, but a larger quantity (0.2 to 0.5 cc.)
injected into the circulation or pectoral muscle produced death in from two to eight
days.

Two pigs inoculated with 5 cc. each of a bouillon culture, one subcutaneously
and one in the femoral vein, remained apparently well.

The description of this bacillus shows that it undoubtedly belongs to
the hog-cholera group of bacteria, although it differs in certain impor-
tant properties from the specific bacillus ordinarily encountered in
destructive outbreaks of hog cholera. As several varieties ! of the hog-
cholera bacillus have already been described, it is important to note the
specific variations of the bacillus in question from the virulent form
described? originally and the one which is generally meant when this
organism is mentioned. The differences are as follows:

(1) It is appreciably larger.

(2) In bouillon a delicate membrane appears on the surface of the
liquid and in old cultures a deposit is formed on the sides of the tube.

(3) There is a marked indol reaction.

(4) It is less rapidly fatal in small doses for experimental animals,
and the lesions produced in rabbits are comparable to those follow-
ing the inoculation with the more attenuated varieties. The negative
results from the pig inoculations are parallel with many similar ones
heretofore made with the virulent variety.

‘In Bulletin No. 6 of the Bureau of Animal Industry, 1894, Dr. Theobald Smith
has pointed out the difference that exists in the morphology, cultural characters, and
pathogenesis of the hog-cholera bacillus as encountered in different outbreaks of
that disease. He also found that several bacilli associated with diseases other than
hog cholera belong to the same group. These are a bacillus found by himself in a
mare after abortion, Bacillus enteriditis, and Bacillus typhi murium.

2Second Annual Report of the Bureau of Animal Industry (1885), p.212. It was
called at that time swine-plague bacterium.

DESCRIPTION OF THE BACILLUS. 75

(5) It is slightly more resistant to the action of certain disinfectants.

The variations, with one exception, have already been noted in some
one or other of the varieties described by Dr. Smith. It is of interest
to note that of the seven varieties of hog-cholera bacillus which he
has pointed out the indol reaction was markedly present in but one
variety (Bacillus cholere suis y), and that a nonvirulent form. This is
of interest, especially as the indol reaction is prominent in the colon
group of bacteria. The greater resistance to disinfectants is also inter-
esting.

The discovery of this bacillus in a disease of pigeons is of much
interest in the accumulation of data concerning the distribution of this
organism and the range of its disease-producing power. The source of
infection is somewhat obscure, but the history points very clearly to its
introduction with the pigeons purchased in Philadelphia. Accepting
that as the source of infection, it.is presumable that the acute disease
breaking out at variable intervals was due to this organism, which for
undetermined reasons became increased in virulence, or on account of
certain conditions the pigeons were rendered more susceptible to it. A
careful inquiry failed to reveal the existence of outbreaks of hog cholera
in Vineland for many years. It was also learned that the disease found
among Mr. Hawley’s pigeons had not appeared in other cotes in that
vicinity.

The conditions under which the affected pigeous were kept and the
care exercised in their management seemed to be all that could be
desired. The cotes, however, were located in a slight depression, the
land surrounding them being appreciably higher, so that drainage is
imperfect. The soil is quite sandy, however, and dries quickly after
rains.

The fact was elicited in conversation with a number of pigeon raisers
in southern New Jersey that a rapidly fatal disease, comparable to the
one described by Mr. Hawley, does occasionally occur among pigeons
where the source of infection can not be explained. I was unable to
find such an outbreak, although one gentleman told me of recently
losing 1,500 birds in one of these outbreaks. The results of the inves-
tigation at Vineland render a careful study of the so-called pigeon
cholera of much interest. It is not improbable that future inquiry will
find that these outbreaks are due to bacteria belonging to the hog-
cholera group. ;

The appearance of this bacillus in the brain of the pigeon affected
with the so-called “‘megrims” is of special interest. It was stated by
Mr. Hawley that the old birds were affected in this way. This disease
is reported by pigeon raisers to be invariably fatal and more or less
common, although comparatively few birds are affected. In this case,
however, it appears that the disease was etiologically the same as that
which caused the large number of deaths among the younger pigeons,
the lesions being localized on the brain. This fact renders the further

76 INFECTIOUS DISEASES AMONG POULTRY.

investigation of “megrims” important in order to determine whether
the single case here reported is comparable in its etiology to the dis-
ease generally. The symptoms appear to be characteristic, so that the
disease is readily recognized by pigeon fanciers.

The fact that pigeons, which are ordinarily quite refractory to hog-
cholera bacteria, should be affected with a disease due apparently to
an organism belonging to that group of bacteria is of much importance.
Although it has been shown that several varieties of the bacillus
described originally from outbreaks of hog cholera are associated with
other diseases and in different species of animals, they appear not to
have heretofore been encountered as the apparent cause of any disease
in pigeons or other birds, with the possible exception of the bacillus
described by Loeffler as the cause of pigeon diphtheria.

The importance of the results of this investigation, which was
materially restricted by the limited number of examinations possible,
is centered in a few determinations in reference to the hog-cholera
bacillus.

(1) The discovery of a member of the hog-cholera group of bacteria
apparently as the etiological factor in a chronic disease of pigeons.

(2) The appearance in pure culture of a variety of the hog-cholera
bacillus from the exudate covering a portion of the brain of a pigeon
killed in the last stages of the disease popularly known as ‘“ megrims.”

(3) The presence of a marked indol reaction in cultures ot a decidedly
pathogenic bacillus of the hog-cholera group.

(4) The discovery of a variety of the hog-cholera bacillus in which
the cultural characters and the physiological property of producing
indol tend to assimilate to a marked degree those of closely related
saprophytic forms, but which in its pathogenesis for experimental
animals falls but little below that of the virulent form originally
described. This striking commingling of saprophytic tendencies and
highly parasitic powers tends to strengthen the proposed theory that
the hog-cholera bacillus is derived from the closely related colon group
of bacteria.

DESCRIPTION OF PLATE I.

Fic. 1. Ceca of turkey No. 5 (somewhat reduced). Both are distended and filled
with exudate. They are unusually short. (See fig. 2 on P1. II.)

Fic. 2. Liver of turkey No. 26 (dorsal aspect). Reduced to seven-ninths of original.
The circular spots which are seen on the surface are shown in color in fig.
ton Pl. III. The other surfaces of this liver presented a similar appear-
ance. The liver is considerably larger than in health.

BUL. 8, BUREAU OF ANIMAL INDUSTRY. PLATE |

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DESCRIPTION OF PLATE II.

Fic. 1. Ceca of a healthy turkey, probably 9 months old. The central tube is the
small intestine, the food passing downward in the direction of the arrow.
Below, at the junction of the ceca with the intestine, the food is drawn
into the ceca by suction.

Fic. 2. One cecum of turkey No. 5 (see fig. 1, Pl. I), cut open longitudinally. The
middle portion of the tube is greatly distended and occluded with an
exudate which is firm in consistency. The upper portion contains small
stones which have passed down from the gizzard. The irregular thicken-
ing of the wall of the tube is shown by a faint line bordering the exudate.

Fic. 3. The other cecum of No, 5, cut transversely. The great thickening of the
wall is shown, together with the presence of a loose plug within the tube,
consisting of a firm exudate. Both this and the preceding figure were
drawn from the alcoholic preparation. (Natural size.)

Fic. 4, One cecum of turkey No. 23, slit open and the mucous membrane exposed.
The wall of this organ is very much thickened, as may be easily seen by
comparing with the healthy tube of a turkey fully six months older (in
fig. 1). On the mucous membrane are masses of exudate firmly attached to
certain spots of increased thickness. (From an alcoholic preparation. )

Fia. 5. Spots on the liver of turkey No, 34 (natural size). Drawn from an alcoholic
preparation.

80

PLATE II

BuL. 8, BUREAU OF ANIMAL INDUSTRY.

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DESCRIPTION OF PLATE III.

Fias. 1 and 2. Spots on the liver of turkey No. 5. (See photographic reproduction,

fig. 2of Pl. I.) The spots are shown both isolated and confluent. The
reticulated structure of these spots is better brought outin the photograph
referred to.

Fic. 3. Left lobe of liver of turkey No. 52 (convex surface). The large yellow area

82

in the upper portion of the figure represents a mass of dead tissue pene-
trating nearly through the entire thickness of the liver. On the right the
pale grayish spot represents diseased liver tissue which is undergoing
repair. Similarly, the spots in the lower portion of the figure correspond
to diseased regions partly healed. Several other spots readily detected in
the specimen could not be clearly brought out in the figure. The diffuse
change, probably reparative, is shown along the lower margin.

BUL. 8, BUREAU OF ANIMAL INDUSTRY. PLATE III

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DESCRIPTION OF PLATE IV.

Fig. 1 (X500). A-portion of the mucous membrane of a cecum of turkey No. 26,

FIG

PIG

FIG

cut transversely. The tissue was hardened in corrosive sublimate and alco-
hol, cut dry after embedding in paraffin, and stained in Delafield’s hema-
toxylin and in eosin.
(a) Nuclei of the epithelium of a crypt or tubule struck nearly tangentially
by the section.
(a’) Mucin cell represented by a cavity in the section.
(b) Spindle-shaped cells belonging to the reticulum of the tissue between the
tubules.
(ce) Protozoa, single and in compact groups, situated within the meshes of the
reticulum.
(d) Multinucleated (giant) cell which has enveloped some of the protozoa.
(x<500). From the same section.
(a) A large group of parasites inclosed in a space of the mucous membrane.
(a’) Single parasite.
(b) A tissue cell in process of division.

. 3. Section of the liver of turkey No. 34. Hardened in alcohol, cut dry in

paraffin, and stained in hematoxylin and in eosin, A large collection of
protozoa (a) occupying the space formerly occupied by the liver cells.
They are probably somewhat forced apart by artificial pressure, for in
other sections of the same liver compact masses of protozoa of equal size
were not infrequently seen. The parasites are surrounded by capillaries
(b) dilated and filled with red corpuscles; (c) round cells or leucocytes.

. 4 (x500). From the same section as fig.3. Protoza (a) embedded in a granular —

matrix containing a few nuclei. Some of them largely disintegrated (b).
(c) A capillary filled with red corpuscles. The matrix resembles the pro-
toplasm of the liver cells.

84

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PLATE IV
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DESCRIPTION OF PLATE V.

1. The protozoan parasites as they appear in crushed and teased fresh tissue

(1,000).

2. A parasite with granular body to which a fragment of a giant cell still

adheres. The bodies in this fragment are fat globules (1,000).

3. Section through the liver of No. 9. Hardened in alcohol and stained in

hematoxylin and eosin (X260). A group of indistinctly demarcated giant
cells containing a considerable number of vacuoles which indicate the
position of inclosed microparasites. This group of giant cells is situated
within what appears to be a thrombosed blood vessel. The coats of the
vessel are faintly traceable. The surrounding tissue is involved in coagu-
lation necrosis, presents a homogeneous appearance, and contains scattering
nuclei.

4 (xX500). A giant cell over 80 in diameter among a group of ten or more

slightly smaller ones found in the wall of cecum of turkey No.1. The
mucous membrane had ulcerated away, and the greatly thickened remain-
der (3 to 4mm. thick) was completely infiltrated with round cells, (a) Pro-
tozoa, two of them within the cell.

5 (xX500). Transverse section through a crypt or tubule of the diseased cecum

of turkey No. 26. The lumenof the tubule is packed with minute bodies,
probably flagellates. (a) Vacuole-like spots representing mucin cells.
(b) Mitosis.

6 (x500). Blood from healthy turkey No. 12, dried as a thin film ona cover-

glass and stained, after being fixed at 120° C., in Ehrlich’s hematoxylin.
(a) Red corpuscles, (b) leucocytes, (¢) flagellates, very much broadened
in appearance by the drying process.

7 (xX500). Flagellate as seen in the fresh blood of healthy turkey No. 33.

86

PLATE V

BUL. 8, BUREAU OF ANIMAL INDUSTRY.

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DESCRIPTION OF PLATE VI.

. The roof of the mouth of fowl No. 73, showing the position of the exudate.

(Natural size. )

. The floor of the mouth, showing the exudate over the tongue. (Natural

size.)

Section of exudate with subjacent tissues, from mouth of fowl No. 59, showing

destruction of the epithelium and cell infiltration of the underlying
tissue (X17).

. Sections of the cornea of fowl No. 65 after the removal of the mass of

88

sloughed exudate (X17).

PLATE VI

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Fia. tl

HAINES, DEL.

Fic. IV

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A. HOEN & CO.

INDEX.

Page

perme ema SPINEL YW 1.574200 aka tiea aw elbe Me alak es ce-s siavian/ouon'becd sccklc weeee 18, 19

Ameba meleagridis, appearance in giant cells........-.-...0-22 2 ee eee eeee eeee 19

TISSUOK ee aemimtiacaceteeaen oe ema ae es oe 17

effect of staining revgents... 5 sotensss ane seceeu tees. one ose 17

LU) BORE Ae Ie Sone Arras co Se eee ee ee Bee 19

itl .CONNeCHIVS TISSUE AHACOS. .osece ene Pasa care fee e 4 <i 18

is it an intracellular parasite?.......-....-.2+ sess. s-2--e 19

; multiplication ...-..---... SiGumagan teas Some aan eee § 19

LIC HIDE RS See Seton On OCS SCO Sr ab CMB Ra SaeOTOaGarbeSaae tee 18

MLO COS ie 2 crs creat Sete ets sheers obs Se teres eae ears se ers Sea 15

Animals, inoculation with exudate of fowls affected with diphtheria. .... .... 55

PBACTLUUSICOUU COMMUNES cect ees a epee ie ome He eee Menlo oe eeoeuseta ule 23, 43, 47, 49

CESEP PHOnMOEAMIBby.: a tosetesicadiscne cee nc kway eases 43

CLOG RATIIES: Gn eB coc o's SECRBONE SS SoOSISB SESS DAC ROC EEE Sa aan Ee = 74

Le DU RU peek aoe 2 een CS See aeelai tte nald oe aciehowaclee vues 74
Blackhead. (See Protozoan disease of turkeys.)

ee Cd ert ines one alas a om Shore pare hoe ese sie taleSestS sem cacicamae ance 9

“TOLDEIIN Gg EGE « Ahh Re iO aC SPE ae See hee ee ne ee 59

Chilonidexofelimeys... o-oo soc ence =~ i= ee wrafeinie slots hee ioe Renal views Sees dese sins ce 26

Cholera des poules. (See Fow] choleza.)

COED QO ONDE S456 GbE Se Gave Aas SRB ORES Ee COCO Oo DEES ee Ee nee ee nae 9

UPON 26G 8d GpcbBas OS IS On SOME IES SSID ee a a eee 9

ELD Sp ISIN PSYC 1h TO ia SS ieee a ne Se 26

CROPPESSTNAS) SUD) PUOTENTES Ses eonm Oe Soars Te SSE Es SICH Sete ete = Pie 1 i se ee 26

CouncilmaniangelatleuninvestiPablons..--+2 426 2oce2 206 o5e6scce csc sec ecee 20

Pa ee we eae Lar OUA Terres ave eran Soe Se wlele Ree Aiea ew n~ hone case wets alse 54

eRe ONE BERG ttR ES INO oo ied ere oie a= oo) aie nnn) Dnielmiaieye ald Sie a Soe wings) aq = Se Sec e.c = wis zi

Day SO les Ory emer tee nA , f oes miei al eele classe Gi icine SS S2 seeiseceicm esos 22, 57, 66, 73

LOO SEERA CT) Tim Se eee ee eC a ae 39

Conditions Havorabletaasie goes. es oc2 se so seco. eee oe cen 40

COMCACLOUSMCSS Re ae peer as 2 oes naa Sil Scie oe Se se ees 40

ECONOMIC RIN POLLAN CO mme me isee se serail eos ieee = - 5 eiaiene 40

IDEALS ots eae AE mettre aati id epee oc ae Seti ba 41

NaS HO Geese Se oad BESS SOOO COS CHE SES Cone Ee ee ae aes 51, 52

MEVHOUSTOn INVES iG AtONy cose a) sees San) o= ee nes cin2 > aon 41

OUGHTA Ks iMVeSTIS AEM pean See ee See acto aes 42, 46, 49, 50

pathogenic bacillus associated with the lesions -.-.--..... 42,

44, 45, 47, 48, 49, 53, 55, 56

prevention andi treatment)... -eeem- cscs. .csccc. ccc oie cea 58

relation toppubliguhealih) .-- seems. -- 8-02. ces eee 60

saprophytic bacteria associated with -....--.-...---..-.- 53

SY NUP COMIS ert recta teenies Seats Selo )2 ro 3) <icte nce Pciwin vnmele gions 40

IRCAREMMEDISCONS:.-\— 92 Suctiee anemone eee cla owes oa ba cise ees ces debs Hees 71

description of bacillus associated with ..--........-..----- 73

relation to Bacillus choler@ suis ...-- 73

ECONOMI GUM PoTtanCOssssaeeeseises a e+ a2 Seo ed ec comc nace esse G2

WUISDON Yasar ereiaes eters eee ec aic ce wa aicicle maid See erate 71

Ey RUDI ace 66 = sodisore te See eee ae Ses een ee eee 71

TEE CCS MOET ONG scm cetoiet esac ere i= cle cic mc ec Scie ee wie isis oseie ices: oats 10

90 INDEX.
MERAY 5 oo6bdsa6c0 c060 weossdbnSpso6 Gbdbos sasooooSed sndo ado nokoasac ane 21
laioro ME TOLESSOL, (ASSIS CBI CO ear ote meee ea renee ere lee U
Ibi ll lig SESE a6 oe oe ose noses acouieeapuoon Boeu conn Gee bSO Sons HES Oab Bane “63
nature of inoculebiom Gisease == =o. sseeeesa eee eee eee ae 67
outbreaks. 35. -¢. se Seats coe Saris oem eet emote eee eee 64, 65
pathogenic bacillus obtained from fowls............-...--.-.---- 65
its relation to rabbit septicemia and swine- |
plague bacli) 26 22'1 5.2522 5.25 eee 65, 69
(CUT CUES 9 IOOO BA SOOO OF SI05 Saco SSSq Some GOCsS6 Sass GeSSa6 cans Sben ase ase 42
Giant cells, ‘appearance of parasites =-c2=e-emes-- ae -e es ee ee eae teens 17
Hawley, A. H., experiments in-raising: pigeons). —2---- oo eons saeetseee 71
Kalborne, BF. L..,.examinationvoftowls.ssssee-ces- 6 15-6 cee eee eee 45
Kruse and Pasquale, investigations... 2.5020. 226) cole. oe cs el en 20
JGEUKKO ATMOS 55 Sooo ss SoS aS oon SS55 S500 coga gS cece ese cdddosdseduu sSosce 14, 67, 68
AY Weyer bey} spol | LOS) oo So So5s00 casoce. Secu csadonoe od sede cress AobSee cues oBoC (Oris
Microparasites. (See Amceba meleagridis. ) :
SU UTE OV: GU OUI DEIN, oa Sen Sosa aoot Bago csSce0 odee soneoy oe asongeSe esace9 Hsoc 18
Nonmotile bacillus in diphtheritic lesions of fowls. (See Diphtheria. )
etiological relation to the disease -..--....----.--,------- 58
WHOSE ease os osns see tao bo acos OSS S Some Coes SoS E SRS Cases So Olga Ses Sob sacs cece 10
Pathogenic bacillus of the hog-cholera group. (See Disease in pigeons. )
LETT a ee GTS NOUESS 225 SSeco so cstono sass cose nono enesoocasecascnsos cece 71, 72, 73
Protozoan diseaserotturkey 22: =. 222: c2e2s0e 22s dente we ete ences ee seeeeeoee 7
analogy to.amobic dysentery 222-5-2ss-5 =) 23 = ee ee = 19
appearance of liver<==- 2-2 sees Seca cceas eee eise eee iit
chanesiot ‘the liven: s.-ce. ose eas seen eee sree nee 12
extentioh cecal diseaseu--- ee = neces eee eee eee eae 16
liver Gisease::.- 2+ 22.2 3.2256 soe eee eee 16
external appearance ~---e----.---= == a SE ae ea HSa 8
Giant cells -- 5. aee se eae see See ee eee 11,13
pakasites| inytherlivertes. ss]. oe ee a ae eee eee ae 13
mw biplicabtionesce=--et--s e eeeee 13
relation'of- bacteria opto --) - se = a ee ee eee 11, 21
to certain other diseases of poultry ......-..---- 22
diseases.of- ceca in: fowls «=.----.--+----.---- 26
other cea disease in turkeys.-....-..-..-. oes 22
seat Of GIseAse = se= tees oe ee oe eee nme ee eee 9
special characters .--.------.--.---: sae. 3422-222 225-2 == 8
Eh ALONG) Se seco ocemcodeceaascmons nso Ceoccseon onsnSoroSss 8
URSIN Nine eietoosD oo GOSS MCU See AUS OSINCCaS SQSOGOUSESOu 26
von Ratz, S., investigations -...------------------ +--+ ------- +--+ ----++ +--+ 23
1 ao Ww I bates io bree moos aoemeiconoa boo cucu WsopBouooeeowanckosenseds cdsene ral
Roup. (See Diphtheria.) ;
Salmon, D. E., results of investigations ......---.--------------------++-++---- 63, 68
Schroeder, E. C., experiments. ----.---.-------------- +--+ ----------+--------+ 46
Slaiked! Mime 22s fee tase ante a a ee eh eee 27
Ube siahis Oy sgooneebenorsesees Cosa Gace cance ar cuocemsascescon besos SS50a8ce 46
MOXLC)/ SMPSLAN GO acs a a Se eee a a ae ata a ahd er ed ed ad lt 51
TPraumatisiiee see cee ces ceeee con steno te ence rreee eee eee eee era 41
Turkeys, diseases of. (See Protozoan disease.)
SKAMMUNATLONs MISTOLY cee ea ee ie ieeie ete ete ole eet eter 27
LER NOL) sib b Wee a enISa Vinee eases 5ocoss Dace Osea Scop emo Soecesss0¢ 8
Voluntary: CONG DUWbUOUS) osetia ae etal tf
Vineland, N. J., epizootic diseases among pigeons-....----------------------- 71
VE ee ee ge erie aes Se ee SOG. c hc aSOG SSIS aS0,6 OB - Bae e Seeo aS 18
Wheeler, Professor, acknowledgments.............-..--..------------------- 7

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