NationalLibraryOfScotland/medical-history-of-british-india huggingface dataset:
174
Contagious Pleuro-Pneumonia of Goats
It is thus evident that the morbidity in the controls under discussion
(73.3 per cent) was indicative of an average virulence of the test strains and
average susceptibility in the particular groups. The 17.6 per cent morbidity
in the vaccinated groups is clearly significant (3.2 σ approximately) and indi
cates a considerable acquisition of immunity.
With the intra-venous inoculation the result obtained was the four out
of five morbidity in the controls and a morbidity of two in the eight vaccinated
goats, a reduction of 55 per cent but this result, while strongly suggestive,
requires larger number of animals to be conclusive. It is noteworthy that the
two reactors had received the larger dose of 10 c.c. intra-venously.
Considering the results with all vaccinated and control animals, the result
obtained is :
TABLE XXXVI
+
—
Total
Vaccinated
7
34
41
17 per cent
83 per cent
Control
14
11
25
56 per cent
44 per cent
Total
66
The standard error here is 12 per cent and the observed difference 39
per cent or 3.25σ. This is clearly significant, and relates to sufficient num
bers of animals to justify a positive conclusion in respect of formalised prepara
tions in general.
Experiment 31.—Active immunisation with a carbolised vaccine
At the same time as the formalised vaccines used in Experiments 29 and
30 were prepared, two carbolised vaccines (0.5 per cent phenol) were prepared
each from the same material, and administered after 48 hours cold storage
i.e. on consecutive days, to a group of seven healthy goats (virulence controls
as in Experiment 29). These were tested at the same time as the groups in
the two preceding experiments, with the results shown in Table XXXVII.
As shown, one of the seven goats died of the infection at the 20th day
p. ins., and another positive reactor was destroyed in extremis at the 18th
day p. ins. The remaining five were negative at slaughter 23 days p. ins.
Discussion.—As already described, the test material employed here in
fected four of the five healthy controls, with natural death in two at the 13th
and 6th days p. ins., and extensive lesions in a third slaughtered for material
at the 14th day p. ins. The virulence of the test material is thus established
and some significance definitely attaches to these results, with no lesions in
five of the seven vaccinated goats. It will be recalled that the same materials,
formalised, when administered subcutaneously, protected all the seven goats,
and the 2nd preparation administered alone intra-venously, protected six out
of eight goats.
E.
O.
LONGLEY
175
TABLE XXXVII
Goat
No.
556
557
558
559
560
561
562
1st vaccination
Date
29
July
1939
Do
Do
Do
Do
Do
Do
Material
Phenolised
vaccine
(0.5 per cent)
ex
goats 461, 462 and
467 in cold store for
48 hours.
10 c.c. S/C.
„
„
„
„
„
„
2nd vaccination
Date
30
July
1939
Do.
Do.
Do.
Do.
Do.
Do.
Material
Phenolised
vaccine
(0.5 per cent) ex
goats 463, 466, 468
and 470 in cold store
for 48 hours.
10 c.c. S/C.
„
„
„
„
„
„
Immunity test
Date
12
August
1939
Do.
Do.
Do.
Do.
Do.
Do.
Material
P. L. E. in Tyrodes
(1:1) ex goats 505,
529 and 531.
15 c.c. I/Nasal.
„
„
„
„
„
„
Results
+ + + +
—
—
—
+ + + +
—
—
Remarks
Destroyed on
1
September 1939.
Destroyed
on
4 September 1939.
Do.
Do.
Destroyed
in
extremis
30
August 1939.
Destroyed
on
4 September 1939.
Do.
Survival
days p. ins.
20
23
23
23
18
23
23
176
Contagious Pleuro-Pneumonia of Goats
B. Active immunisation with living ' Virus.'
Experiment 32.—Active immunisation by the subcutaneous inoculation of virulent
pneumonic lung emulsion.
As is well known, the common method of vaccination against bovine
pleuro-pneumonia is to inoculate virulent lymph from the pulmonary lesions,
or virulent culture, subcutaneously into the tip of the tail. An examination
of the value of this, or a similar procedure, was clearly indicated in the caprine
disease. Three points are noteworthy in this connection :
(1) That whereas inoculations of bovine material subcutaneously on
the trunk in bovines give very regular results, ordinarily fatal,
our experience with inoculations of caprine material, though all
inocula have invariably been fresh (1-2 hours) and the materials
used in their preparation most carefully selected for virulence,
has been that reactions are much less regular and fatal, in toto
in less than 50 per cent of cases.
(2) That full local reactions follow minute doses (0.1—0.5 c.c.) of
exudate, in bovines, where we have on several occasions failed
to induce reactions in goats with volumes of apparently typical
concentrated inocula as great as 20 c.c., and in one case shown to
be fully virulent at insufflation.
(3) That the factors limiting the local reaction in the bovine tail are
believed to be (i) the reduced temperature at this site which
enforces propagation of the organism at other than its optimum
temperature, and so reduces its virulence, and (ii) the absence
of loose vascular connective tissue favourable to the local re-
action.
Item (1) above raises an important question. Is the ' virus ', injected
as potent antigenically when avirulent, or only slightly virulent, as when fully
lethal on injection ? If not, biological assay of virulence would have
to precede any issue of vaccine and only lethal inocula employed—an
important point, as it would entail discarding a high percentage of preparations
and involve considerable trouble and expense.
Regarding item (3), if the local temperature of the inoculated tissue were
the prime factor, full and fatal reactions could be anticipated following inocu-
lation at any site in goats in Madras, where the temperature of the atmosphere
is, over the period available for this research, approximately that of the body.
To this may be added that the tail of the goat is not strictly comparable with
tip of the tail of the ox, as it is much shorter, and provides no such effective
barrier to the spread of a reaction.
It was, therefore, decided to conduct experiments to study these issues,
and in view of item (2), to use the maximum convenient dosage of virulent,
concentrated pneumonic lung emulsion, viz. 5 c.c.
Scheme of experiment.—The plan followed is a little complicated. It was
proposed to select special sites (as the tail) for the inoculation of pneumonic
lung emulsion, and to contrast the local reactions at these sites with those at
sites on the trunk in controls. For a satisfactory conclusion of the safety of
the inoculatio sites, highly virulent test material was obviously required.
E.O.LONGLEY
177
It was further proposed in a proportion of goats to reinforce the inoculations
with a second subcutaneous inoculation on the trunk, which would test for any
immunity to inoculation at the same time as it reinforced any acquired im-
munity, as in Experiment 27.
The sites selected for the inoculations were :
Group A.—the tail
Group B.—the ear
Group C.—the metacarpal region
Group D.—control inoculations subcutaneously over the shoulder
Of these, (A) needs no explanation. The ear inoculations, (B) were sug-
gested by the lack of loose vascular connective tissue in the ear ; the obvious
suceptibility of the ear to any reduced external temperature, in which
respect it is far more in line with the bovine tail as an inoculation site than
is the goat's tail ; and to the fact that the ears, being pendulous, should not
favour gravitational spread to the body. Furthermore, inoculations at this
site are most convenient, the part being immediately accessible in the standing
posture with the minimum of control, and requiring no preliminary clipping
of hair. The metacarpal region (C) was selected as a control to the gravita-
tional limitation of the spread of infection and as having little loose fascia.
In practice this site would be unsuitable, as it necessarily induces temporary
lameness, and could not be called convenient.
The actual procedure was as follows:
(1) Each group, A—C and controls (Group D) comprised ten goats,
which were inoculated as above.
(2) 14 days p. ins., five goats (Group E) were separated from these, one
from each and an additional one from Group A, leaving eight goats
to each group A—D (three intercurrent deaths had occurred)
(3) Each such group was split into two groups of four each.
(A 1 —D 1
and A 2 —D 2 ).
(4) A 1 —D 1 were then (at 14 days) tested for immunity by insufflation,
together with ten healthy controls (Group F).
(5) A 2 —D 2 were simultaneously inoculated subcutaneously on the
shoulder with the same emulsion as used for the above test and
five healthy controls were treated similarly (Group G).
(6) 15 days later these (A 2 —D 2 ) were tested for immunity by insuffla-
tion, together with Group E, Group G (the healthy controls to
the ' reinforcing ' inoculation), and fresh healthy controls
(Group H).
(7) On the following day half of each group A 2 —D 2 received a second
insufflation test, together with fresh healthy controls. (Group
I).
Summarising, the experiment was designed to examine the following
issues, with proper controls :
(1) The possibility of avoiding fatal local reactions at the inoculation
of virulent material by employing special sites.
(2) The immunising value of such inoculations, alone and reinforced
at 15 days, against tests by one and two insufflations.
(3) The immunising value of such inoculations at 14 and at 29 days
p. ins., i.e. the time factor.
178
Contagious Pleuro-Pneumonia of Goats
Procedure.—The three groups A—C received on the 11th of July 1939,
pneumonic lung emulsion pooled from three goats at the 12th passage of strain
42/46, in 5 c.c. doses subcutaneously at the sites named. A control group of
ten goats (Group D) received the same dose subcutaneously on the shoulder.
The inoculum was the same as that employed for the 1st test of a formalised
vaccine (Experiment 27).
The reactions were similar at all the three experimental sites, i.e. fairly
marked swelling with slight tenderness, complicated with lamensess in
Group C, inoculated on the leg. All reactions except in the latter had
almost completely receded by the 3rd day p. ins.
The controls reacted moderately and in one of these (No. 412) there was
the typical progressive reaction resulting in death on the 7th day p. ins.
It is evident from this, (and the low morbidity in the insufflation controls
in Experiment 27 mentioned above, viz. three out of ten, where the same
material was administered by insufflation) that the preparation was not very
virulent, though the danger of the trunk inoculation is demonstrated by the
one fatality.
Fourteen days later the groups were split as described. Pneumonic
lung emulsion pooled from three reactors at the 13th passage of the same
strain was prepared, and administered intra-nasally to half the experimental
and control animals (A 1 —D 1 ) and to ten healthy controls. The balance
of the pneumonic lung emulsion was given to the remaining goats
(A 2 —D 2 ) in 5 c.c. doses subcutaneously on the shoulder, and to five healthy
controls similarly (Group G). All goats inoculated showed reactions ; slight
in those vaccinated, where they had receded by the 3rd day ; marked in three
of the five controls ; but in all receding within a week. This inoculum was
thus also not virulent at inoculation, though as below, where the 1st immunity
test is recorded, virulent by insufflation.
Considering the first test ; one of the vaccinated goats died from inter-
current infection at the 20th day p. ins., the balance being slaughtered at 28
days. One goat only, No. 417 of Group D, had the pneumonia—a small
patch of consolidation in the cardiac lobe of the right lung. Of the controls,
five developed pneumonia with natural deaths in three cases at the 15th,
16th and 18th days p. ins., respectively.
As intended, 15 days after their 2nd inoculation Groups A 2 —D 2 together
with the control Groups E and G., and six healthy controls, (Group H) received
a test by insufflation. The material used was pooled from six goats slaughtered
on the 9th of August 1939. Three days later, half of Groups A 2 —D 2 i.e. 2 goats
from each, received a second test insufflation with material ex the 14th passage
together with five fresh healthy controls. The test dose in all cases was
15 c.c. of 1 : 1 emulsion in Tyrode's solution.
The final results are shown in Tables XL—XLV.
E.O.
LONGLEY
179
TABLE XXXVIII
Immunity test after one inoculation of pneumonic lung emulsion.
(14 days p. ins.)
Group
A 1
B 1
C 1
D 1
Goat
No.
453
454
455
456
431
432
433
434
443
444
445
446
411
413
416
417
Date
11 July
1939.
„
„
„
„
„
„
„
„
„
„
„
„
„
„
„
Inoculum
P. L. E. in
Tyrode's (1:
1) ex goats
381, 383 and
384.
5 c. c.
„
„
„
„
„
„
„
„
„
„
„
„
„
„
„
Seat
S/C Tail
S/C Ear.
S/C Leg
S/C
Shoulder
Test
Date
25 July
1939.
„
„
„
„
„
„
„
„
„
„
„
„
„
„
Material
and route
P. L. E. in
Tyrode's (1:
1) ex goats
311,424 and
426. 10 c.c.
I/Nasal.
„
„
„
„
„
„
„
„
„
„
„
„
„
„
„
Result
(Lungs)
—
—
—
—
—
—
—
—
— —
—
—
—
—
—
++
Remarks
Destroyed on
22 August
1939.
Destroyed in
extremis
14 August
1939.
Destroyed on
22 August
1939.
Do.
Do.
Do.
Do.
Do.
Do.
Do.
Do.
Do.
Do.
Do.
Do.
Do.
Sur-
vival
days
p. ins.
28
20
28
28
28
28
28
28
28
28
28
28
28
28
28
28
TABLE XXXIX
Controls
Group
F
Goat
No.
521
522
523
524
525
526
527
528
529
530
Date
25 July 1939
„
„
„
„
„
„
„
„
„
Route
and dose
I/Nasal 10 c.c.
„
„
„
„
„
„
„
„
„
Material
P. L. E. in Ty-
rode's ( 1 :1) ex
goats 311, 424
and 426.
„
„
„
,.
„
„
„
„
„
Results
(Lungs)
—
+ + + +
—
—
—
+ + + +
+ + + +
—
+ + + +
+ + + +
Remarks
Destroyed on 22
August 1939.
Died on 9 August
1939.
Destroyed on 22
August 1939.
Do.
Do.
Do.
Do.
Do.
Died on 12 August
1939
Died on 10 August
1939.
Survival
days p. ins.
28
15
28 28
28
28
28
28
18
16
H
180
Contagious Pleuro-Pneumonia of Goats
TABLE XL
Immunity test after two inoculations of pneumonic lung emulsion—1st insufflation
Gr oup
A 2
B 2
C 2
D 2
Goat No.
459
460
437
439
449
450
415
419
Date
11 July
1939.
„
„
„
„
„
„
First inoculum
P. L. E. in Ty-
rode's
(1:1)
ex goats 381,
383 and 384,
5 c.c.
„
„
„
„
„
„
Seat
S/C Tail
„
S/C Ear
„
S/C Leg.
„
S/C
shoulder
Second Inoculation.
Date
25 July
1939.
„
„
„
„
„
„
„
Route and
dose
S/C
Shoulder
5. c.c.
„
„
„
„
„
„
„
Inoculum
P. L. E. in Ty-
rode's
(1:1)
ex goats 311,
424, 426.
„
„
„
„
„
„
„
Test
Date
9
Aug
1939.
„
„
„
„
„
„
„
Material
P. L. E. in Ty-
rode's
(1:1)
ex goats 298.
484, 499, 522,
534 and 548.
15 c.c. I/Nasal
„
„
„
„
„
„
Result
(Lungs)
—
—
+ + + +
—
—
—
—
Remarks
Destroyed
on
30
August
1939.
Do.
Do.
Do
Do.
Do.
Destroyed
in
extremis
14
August 1939.
Destroyed
on
30
August
1939.
|
Survival days
p. ins.
21
21
21
21
21
21
5
21
Ind. J. Vet. Sci. & Anim. Husb., Vol. X, Part II ]
PLATE
VIII
FIG. 1
FIG. 2
FIG. 3
FIG. 4
FIG. 5
FIG. 1. Oedematous and
cellular infiltration of the
pleura and interlobular
septa. Alveoli hepatised
FIG. 2. Early oedema of
alveoli and pleura with
cellular infiltration, and
severe congestion of
capillaries and larger
vessels
FIG. 3. Showing
the
oedema extending into
the surrounding healthy
lung
FIG. 4. A later stage than
that depicted in Fig. 2.
All the intra-alveolar
cells are leucocytes, and
scrutiny will reveal the
persistence of the capil-
lary congestion referred
to in the text
FIG. 5. A similar stage to
the
above,
showing
severe
c a t a r r h a l
inflammation
of two
bronchioles filled with
mucopurulent exudate
All sections stained with haemotoxylin and eosin. × 70
Ind. J. Vet. Sci. & Anim. Husb., Vol. X, Part II ]
PLATE
IX
FIG. 1
FIG. 2
FIG. 3
FIG. 4
FIG. 5
FIG. 1. Alveoli in advanc-
ed grey hepatisation in
direct contiguity with
alveoli in the same
lobule in the oedematous
stage. Infiltration of
the septa is also evident
FIG. 2. Infiltrated inter-
lobular septa separat-
ing three lobules at
different stages. Note
emphysema
FIG. 3. Advanced grey
hepatisation.
N o t e
severe persistent con-
gestion of two venules
FIG. 4. Necrosis
with
fibrosis of adjacent
pleura
FIG. 5. Fibrosis of lung
around a bronchiole
E.
O.
LONGLEY
189
all resemblance to muscle is lost, only the oedematous fluid, leucocytes, and
isolated fragments of unlysed degenerated muscle fibres remaining. (Plate
X, Fig. 5).
THE ETIOLOGICAL AGENT
(i) The cultivability of the etiological agent
As already indicated, no growth has been obtained by sowing any of the
many samples of material on any of the ordinary laboratory media, nor on
the special media prepared with goat's flesh and goat blood and serum, nor
on Soparker's medium. Attempts were made to cultivate the organism on
the special ' V. F., S ' digest medium recommended and used by Turner
(1936) in his studies on the bovine organism, using bovine and goat serum,
but the results were inconclusive.
Inoculating tubes of the broth with increasing quantities of sterile exudate
resulted in 3-4 days, and in two instances in the development of a corres-
ponding turbidity in the respective tubes. Dark ground observations of the
turbid fluid revealed in both instances very numerous coccoid bodies, with
no such change in the incubated controls. The ordinary sterility of the
inoculum was established by control inoculations of the usual media. Primary
subculture was, however, negative. This was effected only at the culmina-
tion of the work, when no further attempts could be made, and no retrans-
mission to goats was practicable. It is suggested that a study of the culti-
vability of the organism on this medium, or on slight modifications of it, is
indicated.
Through the kindness of Dr C. G. Pandit, Director of the King Institute,
Madras, it was possible to inoculate a number of eggs with the fresh sterile
exudate. Reactions resulted in a proportion of cases, and passage was
attempted, but a progressively increased mortality in the embryos resulted
in failure at the 2nd and 3rd passages respectively in the two attempts made.
Bacterial sterility was in all instances properly controlled.
Macroscopically the reactions were diffuse and revealed on section
ecto-dermal necrosis, cell-nest formation, and endo-dermal proliferation
(Plate XI, Fig. 1). No quantitative work, and no reinfection of goats from
the embryos, could be attempted.
(ii) Morphology of the etiological agent
1. Tang (1935) and his co-workers, and Turner (1935) in their extensive
dark-ground studies of the morphology of the organism of bovine pleuro-
pneumonia, describe five principal phases in its life-cycle, though the classi-
fications of the two workers have slight points of divergence. For these
phases Turner has proposed the term ' genethodes '
The time available for the study of the morphology of the organism of
the caprine disease has been insufficient for an exhaustive survey, and the
study has been somewhat handicapped by the failure to effect satisfactory
cultivation on artificial media. Sufficient evidence has been adduced, how-
ever, to justify the inclusion of the caprine organism in the same genus as
Ind. J. Vet. Sci. & Anim. Husb., Vol. X, Part II ]
PLATE
X
FIG. 1
FIG. 2
FIG. 3
FIG. 4
FIG. 5
FIG. 1. Early muscle lesion
showing oedematous and
cellular infiltration of
interfascicular connec-
tive tissue
FIG. 2. Extension of the
oedema to the inter-
fibrillar
connective
tissues with ' hyaline '
degeneration of muscle
fibres
FIG. 3. Showing massive
infiltration of inter-
fascicular
connective
tissues, early inter-
fibrillar infiltration and
degeneration of muscle
fibres
FIG. 4. Later stage with
the focal accumulations
of leucocytes referred to
in the text
FIG. 5. Final stage.
Muscle fibres almost
disappeared
Ind. J. Vet. Sci. & Anim. Husb., Vol. X, Part II ]
PLATE
XI
FIG. 1.
FIG. 2.
FIG. 3.
FIG. 4.
FIG. 5.
FIG. 1. Section of chorio-
allantoic membrane of 14
day chick embryo 4 days
after inoculation of fresh
sterile pneumonic effusion.
FIG. 2.
Photo-micrograph
of the organism in a
section of reacting muscle,
with staphylococci from
culture superimposed on
the section for compari-
son. Note non-coccoid
forms, similar to those
revealed by dark-ground.
Stained
Carbol-thionin.
×1260
FIG. 3. Cluster of the
organism in oedematons
connective tissue in
alveolar wall. St. Carbol-
thionin. ×630
FIG. 4 & 5. Typical dis-
tribution and appearance
of the organism in pul-
monary lesions.
Ind. J. Vet. Sci. & Anim. Husb., Vol. X, Part II ]
PLATE
XII
FIGS. 1-15. Dark-ground photo-micrographs of variations of the
organism in pulmonary effusions. × 1890.
Fig. 15. × 630
FIG. 16. Phase corresponding to genethode IV. × 675
Ind. J. Vet. Sci. & Anim. Husb., Vol. X, Part II
]
PLATE
XIII
FIG. 1.
FIG. 2.
FIG. 3.
FIGS. 1, 2 & 3. Typical field with
pulmonary effusion under dark-
ground illumination. The coccoid,
stellate, filamentous and many
other forms are evident. ×945,
×1890 and × 3780 respectively
Ind. J. Vet. Sci. & Anim. Husb., Vol. X, Part
II]
PLATE
XIV
Diagram 1.
Diagram 2.
Diagram 3.
Ind. J. Vet. Sci. & Anim. Husb., Vol. X, Part II ]
PLATE
XV
Diagram 4.
Diagram 5.
E.
O.
LONGLEY
195
new order, the Borrelomycetalcs, should be recognised seems fully justified.
It is accordingly suggested that the organism of caprine pleuro-pneumonia
should be grouped with that of the bovine disease and of agalactia, and
designated Borrebmyces peripneumoniae capri.
GENERAL SUMMARY
(1) Contagious pleuro-pneumonia of goats occurs as a specific epizootic in
the Madras Presidency, and is transmissible experimentally.
(2) The symptoms, course, pathology, and histopathology of the disease
are described.
(3) The etiological agent is a highly pleomorphic organism, which, in fresh
material examined by dark ground illumination, bears close resemblance to
the organism of contagious bovine pleuro-pneumonia.
(4) At least the micrococcal forms of the organism are demonstrable in
formol-fixed sections of both the local lesions arising by inoculation, and early
lung lesions. It is Gram negative, but stains well with Leishman and Geimsa,
best with carbol-thionine. Its average estimated size is 0.3—0.5 µ.
(5) The organism is unusually fragile, and at room temperature loses most
of its virulence in 24 hours, with the addition of both glycerine and Tyrode's
solution. In cold storage it is non-infective at 96 hours, but a reduced infec-
tivity has been retained to seven days, but no longer, where either Tyrode's
solution or glycerine has been added.
(6) The organism is destroyed by exposure to heat at 55°—60° C. for
40 minutes, and also by 0.25 per cent formalin and 0.5 per cent phenol, at
0°—5°C, in 48 hours.
(7) It is not cultivable on ordinary media, nor on several special media
employed. The reaction in the developing chorio-allantoic membrane of the
developing chick embryo suggests that it is cultivable by this means.
(8) In one of two experiments made, the agent passed through the Seitz
filter.
(9) The inoculation of fresh pneumonic material either subcutancously
or intra-muscularly gives rise to an acute, progressive, and usually fatal oedema
at the site of inoculation, with death at from three to eight days p. i.
(10) 'The inoculation of effusion from these lesions gives rise in a percentage
of cases to similar lesions, and the infection has by this means been carried to
the 3rd passage.
(11) Pleuritic and penumonic lesions can be induced by direct extension
from similar lesions in the musculature and connective tissues in the thoracic
wall.
(12) The pneumonia can also be induced by experimental pulmonary
infarction, combined with a subsequent intra-venous inoculation of virulent
material.
(13) The pneumonia can regularly be induced by the intra-nasal insuffla-
tion of either pneumonic long emulsion, or effusion from the local reactions
The resultant morbidity is 75 per cent with a mortality of nearly 100 per cent.
in reactors.
(14) Sheep are highly susceptible to inoculation, but do not appear to be
susceptible to insufflation.
K