NationalLibraryOfScotland/medical-history-of-british-india huggingface dataset:
48
Pleuropneumonia in Goats
in more slender and bacillary form. Bipolar staining has been distinctly seen
in smears prepared from culture.
Cultural characters. The organism on plain agar does not grow luxuri-
antly, but when growth occurs at 24 hours incubation the colonies appear
greyish, raised, circular and 1 to 2 mm. in diameter. A characteristic feature
is the margin of the growth which is heaped up for a greater part of the
colony, but a zone of ¼ to ½ mm. in the periphery is distinctly thin and glisten-
ing in appearance, showing distinct radiation with the naked eyes. Under
strict anaerobic conditions the growth is visible on agar plates, but is poor
as compared with aerobic growth. In bouillon, the growth is characterized by
the turbidity of the medium at 12 hours incubation. At 24 hours of incuba-
tion, sedimentation of the growth takes place. As the growth becomes
older, the sediment becomes heavy and appears stringy when the culture tube
containing the growth is shaken up slightly. On gelatin media at 22°C.,
only a slight growth was observed on the surface but no liquefaction of the
media occurred. The colour of the litmus milk was changed to pink, but co-
agulation did not occur. In peptone media, the organism produced indol.
The organism did not grow on bile-salt media. The production of H 2 S on
lead acetate medium was faint. Nitrate was reduced to nitrite. The
organism was slightly positive for M. B. reduction test and strongly so for
catalase test. Voges-Proskauer and methyl red tests were negative.
Fermentation reactions were obtained with different carbohydrates in
bouillon made of meat extract with the addition of 0.5 per cent of the res-
pective sugars and 1 per cent peptone using neutral red and Andrade's
solution as indicators. The reactions obtained with both the indicators were
observed to be constant as seen in the following table :—
Sugars
Saccharose . .
Lactose . .
Glucose . .
Dextrose . .
Salicin . .
Arabinose . .
Xylose . .
Neutral
red
A
O
A
A
O
A
A
Andrade's
solution
A
O
A
A
O
A
A
Sugars
Dulcite . .
Adonite . .
Laevuloso . .
Maltose . .
Glycerine . .
Mannitol . .
Inulin . .
Neutral
red
O
O
A
A
O
A
O
Andrade's
solution
O
O
A
A
O
A
O
A = Acid and no gas
O
=Negative
The various carbohydrates are fermented with different rapidity. The
first to be fermented is saccharose which is fermented within 24 hours. The
next is glucose which is fermented usually within 48 hours. All sugars fer-
mented within the first five days, and if the incubation was prolonged, say for a
week, the remaining sugars failed to exhibit any change.
Resistance of the organism. The optimum temperature at which growth
usually occurs is from 35° to 40°C., but for growth at a lower temperature,
a prolonged incubation is necessary. The organism has been observed to grow
at laboratory room temperature (30°C.), colonies appearing in 24 hours.
The organism was found quite resistant to heat; a 24 hours old bouillon
culture remained viable when heated to 58°C. in a water-bath for ten minutes.
50
Pleuropneumonia in Goats
monia, pericarditis, and also those of generalized septicaemia. The organism
could be recovered from the heart blood, the lungs and the inflammatory
effusion of the throat. About 3 c.c. of this effusion, when given intratracheally,
produced pleuropneumonia in a goat which proved fatal within five days. The
organism recovered from the buffalo calves and the goat infected with the
throat effusion of the former resembled in every detail that isolated from the
natural cases of the disease.
TRANSMISSION EXPERIMENTS
By filtered lung material
These experiments were conducted with a view to ascertaining whether
the conditon was primarily associated with a filtrable virus. In these ex-
periments the hepatized lungs from goats 571 and 596 (natural cases) were
thoroughly triturated with sand and the supernatant fluid was filtered through
Chamberland filter candle 5 bis. L. 3. The filtrate thus obtained was inject-
ed intravenously and intranasally to goats. The results are tabulated below.
Table showing the results of filtration experiments
Source of
material
goat No.
571
596
Expt.
goat
No.
88
108
83
Control
107
109
99
Control
Date of
inocula-
tion
12-9-40
„
„
„
„
„
Route of inoculation
Intravenous . .
Intranasal . .
Intratracheal .
Intravenous . .
Intranasal . .
Intratracheal .
Material
Filtrate
„
Lung
emulsion
Filtrate
„
Lung
emulsion
Dose
10 c.c.
„
5 c.c.
10 c. c.
„
5 c.c.
Remarks
The animal was kept under
observation for about a month
but showed no reaction.
Ditto.
The animal developed lesions
and symptoms of pleuropneu-
monia 3 days after inoculation.
The animal was kept for 3 month
under observation but showed
no reaction.
Ditto.
The animal died of pleuropneu-
monia 5 days after inocula-
tion.
The results of these experiments suggest that the condition is not primarily
associated with a filtrable virus.
By unfiltered infective materials and culture
These experiments were conducted with a view to testing the infectivity
of different infective materials including cultures by various routes of inocu-
lation. The materials were (1) the saline emulsion of the hepatized lung, (2)
the saline emulsion of 24 hours old growths on blood-agar media, (3) 24 hours
old growths in bouillon, (4) the pleural effusion, and (5) the blood of
experimentally infected goats. The artificial infections were obtained by
various routes of inoculation, the most ideal route being the intratracheal
one. The details of some of these experiments are given below.
Experiment No. 1—12 September 1940. Saline emulsion of the hepatized
lung tissue was injected intratracheally in 5 c. c. dose to goat 83. On
September 13, the animal showed rise in temperature to 105.0° F., discharge
from eyes and nose and loss of appetite ; on the following day it had difficulty
in breathing and occasional coughs. On September 15, the animal died and
P. G. PANDE 51
revealed consolidations of the apical, cardiac and a part of the diaphragmatic
lobes of the right lung, and pericarditis on post-mortem examination.
Experiment No. 2—12 September 1940. Goat 85 was inoculated intra-
venously with 5 c. c. of pleural effusion from goat 571 (a natural case).
The following morning the animal had a rise of temperature to 106.0° F.,
discharge from eyes and nose and succumbed in the evening. Post-mortem
examination revealed lesions of septicaemia, patches of congestion and
consolidation in both the lungs and pericarditis.
Experiment No. 3—12 September 1940. Saline lung emulsion of the
naturally affected goats 571 and 596 was injected subcutaneously in 5 c.c.
dose to goat 86. On September 14, the animal developed typical symptoms.
Two days later the animal was destroyed and the post-mortem examination
revealed lesions of septicaemia, scattered areas of consolidation in both lungs,
haemorrhagic infarcts and pericarditis.
Experiment No. 4—13 September 1940. Saline emulsion of the blood
agar culture prepared from the lung tissue of the natural case 571 was injected
intrapulmonally in 3 c. c. dose to goat 82. The animal showed all typical
symptoms of the disease on September 15 and died on the following day.
Autopsy revealed typically consolidated areas, haemorrhagic infarcts in the
lungs and pericarditis.
Experiment No. 5—15 September 1940. 10 c. c. of blood drawn from
the jugular vein of goat 82 was, after defibrinating, inoculated intravenously
to goat 105. On September 17, the animal developed slight rise in tempera-
ture and dullness persisting for two days. The animal was later killed and
post-mortem examination revealed nothing unusual. Cultures from the heart
blood remained sterile.
Experiment No. 6—18 September 1940. Saline emulsion of blood-agar cul-
ture prepared from the lung tissue of goat 82 was injected intratracheally
in 3 c.c. dose to goat 87. After exhibiting all typical symptoms the animal
died on September 22. Autopsy revealed consolidation of the right apical
and the cardiac lobes, the left apical and a part of the right diaphragmatic
lobe.
Experiment No. 7—24 September 1940. Bouillon culture prepared from
heart blood of goat 87 was injected subcutaneoulsy to goat 89 in 3 c.c. dose.
In the evening of the same day the animal had pyrexia (106.0° F.),
loss of appetite, lameness, discharge from the eyes and succumbed on the
following morning. Autopsy revealed patches of pneumonia, pleuritis and
lesions of septicaemia including the presence of patechae marks on the endo-
thelial lining of the heart.
Experiment No. 8—30 September 1940. 5 c.c. of the pleural effusion
from goat 89 was injected intratracheally to goat 95. The animal developed
heavy breathing accompanied with pyrexia (105.8° F.) and was destroyed on
1 October 1940. Lesions of pleuropneumonia and pericarditis were found on
post-mortem examination.
By feeding
The materials used in these experiments were the emulsified lung tissue
and 24 hours old growths of the organism on blood-agar media, fed separately,
mixed with bran.
P. G. PANDE 53
symptoms and further rise in temperature. From October 30, the symptoms
began to subside, and on November 2 the animal appeared to be normal.
VACCINATION EXPERIMENTS
A fortnight after their recovery, goats 193 and 124 were bled and their
sera contained agglutinins which caused clumping of the organism (made in an
emulsion standardized to the opacity of Brown tube No. 3) in a litre of 1:
1,000 and 1: 1,200 respectively. The sera from normal goats did not cause
this phenomenon even in a dilution as low as 1 : 20. After about a week
these animals were inoculated, one subcutaneously with 3 c.c. and the other
intravenously with 4 c.c. of 72 hours old broth culture without any ill effects.
These observations suggest that animals that have recovered from the
disease acquire antibodies in their blood in sufficient concentration to protect
them from further infection. These observations, however, formed the basis
for devising experiments on active immunization of animals with vaccines
prepared from the culture and the hepatized lung emulsion. The vaccines
prepared on the following lines were tried.
1. A 24 hours old blood-agar culture heated at 58.0°C. for 30 minutes.
2. A 24 hours old blood-agar culture formalized to the extent of 2.5
per cent.
3. A 24 hours old blood-agar culture formalized to the extent of 0.5
per cent.
4. Lung emulsion of an experimental case of pleuropneumonia forma-
lized to the extent of 0.5 per cent.
Each of these vaccines prior to its use on goats was tested for its inertness
on culture media and on rabbits, and kept for 48 hours in the refrigerator.
The heat-killed vaccine when tested on five goats was found to have no
protective value as the vaccinated animals showed no reactions and they all
died when tested for immunity with virulent lung emulsion. The vaccine
prepared from culture emulsion formalized to the extent of 2.5 per cent was
also found to be ineffective. In this experiment 20 goats were used, but all
died when tested for immunity 14 days after vaccination. It may, however,
be concluded that in case of the first vaccine the temperature at which the
culture was heated had also destroyed the antigenic property of the vaccine,
and in the case of the second, the concentration of the formalin had the same
deleterious effect. The last two vaccines were found to be successful as will
be seen from Tables I and II.
TABLE I
Vaccination experiments with 24 hours old blood-agar culture vaccine formalized
to the extent of 0.5 per cent. The dose was 5 c.c. given subcutaneously and
immunity test was performed 14 days after vaccination
Brand
Nos.
219
217
215
206
Vaccination reactions.
Dull and off feed . . .
Do.
...
Slight . . . .
104. 6° F. off feed and dull.
Immunity test
Test material
Fresh lung emulsion .
Do. . .
Do. . .
Do. . .
Dose
1 c.c.
2 c.c.
3 c.c.
5 c.c.
Route
I/t .
Do. .
Do. .
Do. .
Reactions
None
None
Slightly off feed and dull
Do.
54
Pleuropneumonia in Goats
Brand
Nos.
207
208
218
220
221
222
223
224
225
226
227
228
Con-
trol.
229
Con-
trol
Vaccination reactions.
Dull and off feed . .
Do.
...
Lame, dull and off feed .
Do.
...
104.4°F., lachrymatic, dull
and off feed
Dull and off feed . .
Do.
...
Slight
....
105.0° F., lachrymation,
nasal discharge and heavy
breathing
Dull and off feed
Slight
....
...
...
Immunity test
Test material
Fresh lung emulsion
Do. . .
Do. . .
Do. . .
Do. . .
Fresh lung emulsion .
Culture emulsion .
Do. . .
Do. . .
Do. . .
Do. . .
Fresh lung emulsion .
Culture emulsion . .
Dose
5 c.c.
4 c.c.
3 c.c.
4 c.c.
5 c.c.
3 c.c.
1 c. c.
2 c.c.
3 c.c.
3 c.c.
2 c.c.
3 c.c.
3 c.c.
Route
I/t .
Do. .
Do. .
Do. .
Do. .
Do. .
S/c
.
Do. .
Do. .
S/c .
Do .
I/t .
S/c .
Reactions
Slightly off feed and dull
Do.
Do.
Do.
Do.
Do.
Do.
Slightly dull
None
Do.
Slightly dull
Severe reactions followed
by death
Do.
It will be seen from Table I that out of 15 animals 12 showed some degree
of reaction, and only in 3 out of 12 these may be said to have been well mark-
ed. Further, it may be noticed that the three vaccinated animals, in which
the reaction was only slight, withstood the dose of test materials as effectively
as the others.
TABLE II
Vaccination experiments with the hepatized lung emulsion formalized to the
extent of 0.5 per cent. The dose was 5 c.c. given subcutaneously and
immunity test was performed 14 days after vaccination
Brand
Nos.
129
130
134
135
131
132
128
130
127
126
138
Con-
trol
Vaccination reactions
Dull and lame . . .
Do.
...
103.8° F., off feed and dull
Slightly off feed and dull .
103.0° F., off feed and lame
104. 0° F., off feed and lame
103.4° F., off feed and lame
104.0° F., off feed and lach-
rymation
Dull and lame . . .
Dull and off feed . .
......
Immunity test
Test material
Fresh lung emulsion .
Do.
Do.
Do.
Do.
Do.
Do.
Do.
Do.
Do.
Do.
Dose
2 c.c.
Do.
Do.
Do
Do.
Do.
Do.
Do.
Do.
Do.
5 c.c.
Route
S/c
.
Do. .
Do. .
Do. .
Do. .
Do. .
Do. .
Do. .
Do. .
Do. .
Do. .
Reactions
Swelling at the seat of
injection, no other re-
action
None
Do.
Do.
Do.
Swelling at the seat of
injection, no other re-
actions
Do.
Do.
Do.
Do.
Severe reaction followed
by death
58
Pleuropneumonia in Goats
4. Experimental evidence was obtained on successful transmission of the
disease from affected to healthy goats by contact and also by feeding of in-
fective lung materials and cultures to healthy animals. It is suggested that
outbreaks in nature may be precipitated by a sudden change in weather
conditions, fatigue and other predisposing factors.
5. Preliminary attempts at vaccination with the formalized vaccines
prepared from the hepatized lungs and the blood-agar culture have given
encouraging results.
ACKNOWLEDGEMENTS
The author wishes to express his thanks to Mr T. J. Egan, I. V. S., Director,
Veterinary Services, United Provinces, for providing facilities for this work
at the Provincial Veterinary Laboratory, Lucknow.
REFERENCES
Dungal, N. (1931). J. Comp. Path. and Ther. 44, 126
Hutyra, F., Marek J. and Manninger R. (1938). Sp. Path. and Ther. of Diseases of
Domestic Animals, 4th edition, Vol. 1, Bailliere, Tindall and Cox, London
Jones, F. S., and Little, R. B. (1921). J. Exp. Med. 34, 541
Jones, F. S., and Little, R. B. (1922). J. Exp. Med. 36, 273
Kolayli, C. and Raif, M. (1935). Rec. Med. 111, 38
Montgomerie, R. F., Bosworth, T. J., and Glover, R. E. (1938). J. Comp. Path. and
Ther. 51, 87
Naik, R. N. (1939). Ann. Administration Rep. of the Vet. Invest. Officer, Province of
Bombay, 13
Newsom, I. E., and Cross, F. (1923). J. Amer. Vet. Med. Ass. 62, 759
Shirlaw, J. F. (1939). Proceedings, 3rd Anim. Husb. Wing Meeting, Feb., New Delhi
AN OUTBREAK OF BOVINE ENCEPHALOMYELITIS
BY
S. R. HASSAN, B.A., M.R.C.V.S., L.M.
AND
J. A. IDNANI, G.B.V.C.
Imperial Veterinary Research Institute, Izatnagar
(Received for publication on 23 December 1941)
THE object of this article is to record an outbreak of encephalomyelitis
which occurred among the cultivation and dairy animals of the Institute
at Izatnagar during the winter of 1938-39 and which lasted for about three
months. The dairy herd consisted of 43 cows, three stud bulls and 45 young
stock. These animals were accommodated in temporary thatched sheds at
a distance of about one hundred yards from 50 bullocks and 35 buffaloes
used for cultivation work.
Prior to the outbreak, no case of encephalomyelitis at this Institute
is on record. With regard to the origin of the outbreak, it may be stated
that, on account of constructional work then in progress on the estate, a
number of outside horses and cattle had access to the Institute. Altogether
20 animals, viz. five bullocks, one buffalo bull, 12 cows, one buffalo calf and
a pony contracted the disease and died.
The first case, bullock 1320, was seen to be ill on 5 December 1938
and died three days later. It had previously been working on the estate
area where it had contact with the outside animals. It was housed in a
shed along with other bullocks and buffalo bulls near the dairy animals.
The two dairy byres, a furlong from each other, had the same attendants,
who may have carried the infection from one place to the other. Various
groups of animals also came into contact with one another whilst watering
and grazing. The disease was mainly confined to the vicinity where it had
originally started. Table I shows the time between the onset of the disease
and death in individual cases.
TABLE I
Animals involved in the outbreak
Animal
Bullock 1320
Bullock 1543
Cow 100
H ifer 172
Cow 91
Bull-calf 3
Bullock 136
Cow 80
Age
in years
9
8
5
2
6
1
8½
7
Onset
of illness
5-12-38
7-12-38
8-12-38
9-12-38
10-12-38
11-12-38
12-12-38
11-12-38
Duration
in days
3
3
7
2
4
4
3
6
59
60
An Outbreak of Bovine Encephalomyelitis
TABLE I—(contd)
Animal
Cow 66 . . . . .
Cow 106
.
.
.
.
.
Bullock 40 . . . . .
Cow 82
..
.
.
.
Cow 142 . . . .
Bullock 114 . . .
.
Cow 74 . . . . .
Cow 153 .. . . .
Buffalo bull 296 . . . .
Cow 56 . . . . .
Cow 40 . . . ..
Pony 5B . . . . .
Age in
years
7½ 5
2½ 7
3
14
7
3
7
8
10
7
Onset of
illness
13-12-38
14-12-38
16-12-38
25-12-38
25-12-38
30-12-38
30-12-38
31-12-38
3-1-39
18-1-39
2-2-39
25-2-39
Duration
in days
6
5
5
7
5
3
7
6
2
7
7
3
NOTE. All the above animals died of the disease
It would appear from Table I that the epidemic, in spite of the severity
of the clinical cases, showed no tendency to extensive spread in the form of
clinically apparent cases among in-contact cattle. At any one time, there
were never more than 2 or 3 obviously affected animals.
It is interesting to note that, both among the dairy cows and the culti-
vation bullocks, the early cases lasted for a shorter time than the later ones
though all ended fatally.
SUSCEPTIBILITY
No animal less than one year old contracted the disease although young
calves were kept quite near the sheds where the outbreak started. The
animals affected were mostly adults, buffaloes appearing to be less susceptible
than cattle.
PERIOD OF INCUBATION
The incubation period in natural cases could not be definitely determined.
For what the information is worth, however, it may be noted that the inter-
val between the disease of cow 153 and the onset of symptoms in the dairy
animal to be affected next, viz. cow 56 was 12 days.
CLINICAL OBSERVATIONS
Illness in the first two cases, bullocks 1320 and 1543, appeared suddenly
and diagnosis was not easily established. Colic and spasms of the muscles
of the hind parts of the body were the only symptoms evinced. Both animals
died within a space of two days. In the meantime, other animals were observ-
ed to be affected and in them the symptoms assumed a more or less definite
course. Marked lassitude and dullness were common premonitory symptoms,
the animal being partly or wholly off its feed. In some cases, there was sali-
vation and champing of the jaws, with or without twitching of the super-
ficial muscles of the face and neck. As the disease progressed, incoordina-
tion and loss of power with absence of sensation in the hind limbs became
manifest. Later, paralysis supervened and the animal collapsed. The tem-
perature was either normal or subnormal, except in cases showing excitement
Repeated bellowing and attacks upon the surroundings were characteristic
of such cases. The neck became stiff and was bent backwards, symptoms
S. R. HASSAN AND J. A. IDNANI 61
which were not relieved by withdrawing 10 c.c. of cerebrospinal fluid.
The amount of this fluid and the rate of its flow appeared to be normal. When
down, the animals were seen to strain but this seemed to be largely due to
spasms of the muscles connected with the act of defecation. Towards the
end, the eyeballs became fixed and staring. Hyperaesthesia, which was
present in the early stages, appeared to pass off in one or two days and was
followed by drowsiness and finally by coma and complete paralysis.
DIFFERENTIAL DIAGNOSIS
Two possibilities, other than encephalomyelitis, could not be entirely
ruled out ; one, some form of poisoning producing nervous symptom,
the other rabies. To dispose of the former, chemical examination of fodder,
etc. was undertaken but in no case was any poison detected. Rabies was
considered unlikely as none of the affected animals was known to have been
bitten by any carnivorous animal. Moreover, Negri bodies were found in
no case examined. Further, rabies in cattle is usually of the furious type,
while in the present outbreak only three animals (cow 91, bullock 114 and
buffalo 296) exhibited maniacal symptoms.
CONTROL
Measures deemed to be suitable were taken to check the spread of the
disease, viz. segregation of animals showing symptoms, isolation of suspects
and in-contacts and thorough disinfection of the premises. The usual graz-
ing areas were closed and the animals were sent in small batches to new pas-
tures. In spite of such precautionary measures, the disease continued to
spread and eventually subsided spontaneously. Hexamime (urotropine)
was tried in several cases but without satisfactory results, injections being
given intrathecally in dosage of 2.0 c.c. of a 10 per cent solution, sometimes
repeated on two or three consecutive days.
MORBID ANATOMY
In none of the cases examined at autopsy, were gross lesions suggestive
of any specific disease seen. Although in most cases a certain amount of
gastro-enteritis was present, the principal seat of lesions was the brain and
spinal cord. The meninges were congested and pin-point hæmorrhages in
the parenchyma of the brain were more or less constant.
HISTOPATHOLOGY
Histological examination of the brain and spinal cord was undertaken
in all cases. Tissues were fixed in 10 per cent formalin or in Bouin's fluid.
The report of the pathologist on the first two cases, on which a diagnosis
of encephalomyelitis was established, states : ' Brain and spinal cord showed
marked diffuse inflammatory changes, non-suppurative in character, asso-
ciated with encephalomyelitis, consisting chiefly of necrosis and neurono-
phagia of nerve cells. The grey matter of the dorsal (posterior) horns appear-
ed to be practically devoid of nerve cells, with distinct glial proliferation
with monocytes forming perivascular cuffs. No polymorphs were seen.
The vessels of the pia arachnoid were congested and those in the meningeal
fold, like the vessels of the grey and white matters, showed distinct cuffing.
The pathological changes were most marked in the spinal cord, less in the
64
An Outbreak of Bovine Encephalomyelitis
RABBIT 3—(Vide Table II)
(Susp.=suspension)
(n. r.=no reaction)
All inoculations consisted of brain-cord mixed suspension 0.5 c.c. intranasally
PLATE VII
[ Indian J. Vet. Sci. & Anim. Husb., Vol. XIII, Part I ]
Crinum defixum Ker-Gawl (Family Amaryllidaceae)
showing the leaves, bulb and roots.