NationalLibraryOfScotland/medical-history-of-british-india huggingface dataset:
140
Contagious Pleuro-Pneumonia of Goats
Experiment 4.—Confirmation of the local reaction following the inoculation of
pneumonic lung emulsion
On the 19th of December 1938, an experimental replica of Experiment 2
was made.
Donor.—Goat No. 16 of the next consignment of suspected cases died at 12
noon having been five days on the College premises. The typical pneumonia
was found at autopsy, involving both lungs at all stages of hepatization, with
diffuse pleuritis, and 300 c.c. of serous effusion.
Procedure.—This was the same as in the preceding experiment except
that the intra-venous inoculation was with 1 c.c. of 1 : 2 emulsion in 9 c.c.
of saline, which was very well tolerated, and the control emulsion was heated
at 55°-60°C. for 45 minutes. Four experimental and four control goats were
used. As mentioned in Experiment 2, advantage was taken of the availability
of presumably virulent inoculum to test the two survivors of the control
recipients of the heated emulsion in that experiment, for any resistance
to the infection. 2.5 c.c. of the pneumonic lung emulsion was administered
subcutaneously, and the same dose intra-muscularly, both on the thigh.
Cultural tests from the lesions were made according to routine and proved
sterile on all media. Each emulsion was inoculated at the conclusion of the
goat inoculations to four guinea-pigs, 1 c.c. intra-muscularly and each to one
rabbit, 2.5 c.c. intra-muscularly, with no effect.
The details of the goat inoculations and the results are tabulated below :
TABLE VI
Experimental
Group. Fresh
P. L. E. ex-
Goat 16.
Control Group
Heated P.L.E.
ex-Goat 16.
Immunity
Test. Fresh
P. L. E. as
above.
Date
19-12-38
„
„
„
19-12-38
„
„
„
19-12-88
„
Goat
No.
48
49
50
51
52
53
54
55
37
41
I/V
1 c.c. in 10
c. c. saline.
Do.
Do.
Do.
1 c.c. in
10 c.c.
Saline.
Do.
Do.
Do.
I/Tr
1 c.c.
Do.
Do.
Do.
1 c.c.
Do.
Do.
Do.
I/M
5 c.c.
Do.
Do.
Do.
5 c.c.
Do.
Do.
Do.
2.5 c.c.
Do.
S/C
5 c.c.
Do.
Do.
Do.
5 c.c.
Do.
Do.
Do.
2.5 c.c.
Do.
I/Nas
2 c.c.
Do.
Do.
Do.
2 c.c.
Do.
Do.
Do.
Result
+ + + +
+ + + +
+ + + +
±
—
—
—
+ + + +
+ + + +
Remarks
Died on
23-12-38
Slaughter-
ed on
24-12-38
Do.
Recovered
Died on
23-12-38
Destroy-
ed on
23-12-38
Survival
days
p. ins.
4
5
5
26
4
4
The residue of untreated emulsion was split into two portions and bottled,
one with the addition of an equal volume of 50 per cent glycerine-saline
and placed in cold storage for observations on viability. (Experiment 18).
Ind. J. Vet. Sci. & Anim. Husb., Vol. X, Part II ]
PLATE
VI
FIG. 1.
FIG. 2.
FIGS. 1 and 2 show typical clinical appearance at 3 and 4 days p.i.
Ind. J. Vet. Sci. & Anim. Husb., Vol. X, Part
II]
PLATE
VII
Showing shoulder reaction following subcutaneous inoculation at the 6th day p. ins.
E. O. LONGLEY 141
Results.—As shown in Table VI above, the result confirmed the previous
experiment. Exactly the same syndrome followed in three of the four experi-
mental subjects, one of which (No. 48) died from the condition at the 4th day
p. ins. ; two being slaughtered for transmission purposes (Nos. 49 and 50) ; and
the 4th recovering after manifesting a moderate reaction. This latter died 26
days later from coccidiosis, when, at autopsy, the local lesions were found
to have resolved completely. No reaction of any kind resulted in the four
controls.
Both the goats tested for immunity reacted fully to the inoculation, one
No. 37, dying at the 4th day p. ins; the other being sacrificed that day for
purposes of transmission. No pulmonary lesions were present post mortem
in any of the 10 subjects. No immunity tests were possible later in the con-
trols to this experiment, as all died of debility, etc. before suitable test material
could be obtained.
Plate VI, figs. 1 and 2 depict typical reactions in the hind leg at the 3rd
and 4th days p. ins. Plate VII, shows a shoulder reaction at the 6th day.
Results.—The result conforms the previous result in Experiment 2, and
this finding has since been repeatedly and amply confirmed.
Experiment 5.—A second small scale attempt to effect passage of the
infection by inoculation of material from these local reactions was made on the
23rd of December 1938 on the lines of Experiment 3, and with similar incon-
clusive results.
Experiment 6.—Serial transmission through muscle lesions, 2nd passage
Advantage was taken of a severe reaction in Experiment 19 to make a
third attempt at transmitting the disease by the inoculation of effusion from
the local lesions.*
Donor.—Goat No. 95 of that experiment, slaughtered in extremis on
the 12th of February 1939, 5 days p. i., with a severe local reaction from the
injection of mixed lung emulsion and effusion from a natural case after 48
hours cold storage, provided material for the inoculation of four goats, the
first pair receiving 5 c. c. intra-muscularly of the whole effusion aspirated
direct into the syringe from the lesion, and the last pair 10 c. c. intra-
muscularly similarly collected. Two guinea-pigs were injected intra-
muscularly with 2 c. c. of the same effusion, and sterility tests on all media
were made, with no growth.
Results.—As shown above, one of each group reacted fully and one of
each incompletely. One reactor, No. 113, was slaughtered in extremis for
further transmission, as described in the following experiment.
Experiment 7.—Serial transmission through muscle lesions, 3rd passage.
Muscle effusion from goat No. 113 of the previous experiment, slaughtered
in extremis on the 20th of February 1939, was inoculated direct from the
lesions, in 10 c. c. doses, into two goats Nos. 100 and 101, the only healthy
animals then available.
*Throughout this paper, experiments are recorded not in their actual chronological
sequence, but in accordance with their logical relationship.
142
Contagious Pleuro-Pneumonia of Goats
TABLE VII
Date
12 February 1939 .
„
„
„
Goat No.
112
113
114
115
Route
I/M
„
„
„
Inoculum
Muscle effusion ex
goat 95.5 c. c.
„
Do. 10 c. c.
„
Result
+ +
+ + + +
+ + + +
+ +
Remarks
Receded by 21
February 1939.
Destroyed on 20
February 1939 for
transmission pur-
poses.
Died on 16 Febru-
ary 1939.
Receded by 20
February 1939.
Survival
days
p. ins.
8
4
Of these, one reacted fully with death at the 7th day p. ins; its fellow re-
acting incompletely with recovery.
No further passage could be attempted owing to the over-night death of
the only reactor.
Summary.—The agent in lung emulsion after 48 hours cold storage
caused full reactions in both recipient goats at the 1st intra-muscular pas-
sage ; in two of four goats at the 2nd intra-muscular passage ; and in one of
two goats at the 3rd intra-muscular passage.
Conclusions.—The experiments indicate that the local reactions can be
induced by passage from goat to goat.
Experiment 8.—Pulmonary lesions by direct spread from reacting local tissues.
It was thought that pulmonary or pleuritic lesions might possibly be
induced by inoculating virulent material intra-muscularly over the thorax
into the longissimus dorsi and longissimus costarum muscles at about the
7th rib, as a result of direct spread to the pleura and lungs from the reacting
muscles and fasciæ (' supra-costal ' inoculation). Two small experiments
to test this issue were made.
On the 14th of February 1939, a pneumonic lung emulsion pooled from
two field donor goats, Nos. 42 and 46, which was used in the test for the sus-
ceptibility of other species (Experiment 21) was administered intra-muscularly
at the above site in 10 c. c. doses to two goats.
All the four controls to the Experiment 21 reacted to the usual intra¬
musular inoculation in the thigh, in three cases with full lesions. The re-
sults in the two experimental goats are shown in Table VIII.
As shown, both reacted fully at the site of inoculation. In No. 128
there was an extensive oedema of the entire side of the trunk from about the
4th rib to the point of the hip. In the thoracic cavity there was 100 c. c.
of serious effusion with typical pleuritis, and adhesion between the apical and
cardiac lobes of the right lung and the pleura. Portions of the cardiac lobe
of this side presented typical early lesions with hepatisation and oedema.
E. O. LONGLEY 143
TABLE VIII
Goat
No.
128
129
Date
14 February
1939.
Do.
Inoculum
P. L. E. in
Tyrodes
(1:1) ex 42
and 46.
Do.
Route
I/M
Supracostally.
Do.
Dose
10 c. c.
„
Lesions
Local
+ + + +
+ + + +
Thoracic
Unilateral
pleuritis
and pneu-
monia.
—
Remarks
Died on 20
February
1939.
Died on 19
February
1939.
Survival
days
p. ins.
6
5
In the other case, an extensive local reaction was present, but with no ex-
tension to the thoracic viscera.
Experiment 9.
The second experiment, also in two goats, was made with pneumonic
lung emulsion from a natural case, 5 c. c. being similarly administered on the
right side. Two healthy control goats were inoculated with the same dose
intra-muscularly in the thigh. One died 5 days p. ins. with a severe reaction,
the other reacted moderately and recovered.
Of the ' supra costals ', one reacted only slightly, with early recession of
the lesions. The other died 6 days p. i. with oedema of the thoracic wall
and an early and typical pleuritis with marked congestion of the capillaries
of the parietal pleura of that side. The visceral pleura and lungs were normal.
Summary.—(1) Of four goats receiving intra muscular inoculations in the
back over the thorax, three died with severe local reactions in the muscles
and connective tissues.
(2) In one goat, a pleurisy developed adjoining the site of reaction ;
in the other there was a similar pleurisy with pneumonic involvement of the
contiguous cardiac lobe.
Experiment 10.—Transmission by means of intra-venous inoculations of virulent
pneumonic lung emulsion, with aprevious induction of
experimental pulmonary infarcts.
Daubney [1935] has recorded the experimental induction of the pul-
monary lesions of bovine pleuro-pneumonia by injecting into the jugular
veins of cattle fragments of agar impregnated with the pleuro-pneumonia
organism from culture, and this has since been confirmed [Mettam and Ford,
1939]. It is supposed that the resultant pulmonary infarcts provide a nidus
suitable for the localisation of the organism in the lungs.
An experiment on similar lines was undertaken, the procedure being to
introduce fragments of sterile lung into the jugular veins by means of a bleed-
ing canula attached to a 20 c. c. syringe charged with saline, and 24 hours
later, to inoculate 5 c. c. of virulent pneumonic lung emulsion intra-venously.
E. O. LONGLEY 145
TABLE IX
Goat
No.
124
125
126
127
122
123
Date
15
February
1939.
Do.
Do.
Do.
Do.
Do.
Inoculum
P. I. E. ex 42/46 at
24 hours cold stor-
age. Dose 5 c. c.
„
„
„
P. L. E. ex 42/46 at
24 hours cold stor-
age. Dose 2.5 c.c.
„
Route
I/Nasal.
„
„
„
I/M
„
Results
Lungs
+ + + +
+ + + +
+ + + +
±
—
—
Local
reaction
—
—
—
—
+ + + +
++++
Remarks
Destroyed on 3
March 1939.
Died
on
2
February 1939.
Do. .
Destroyed on 3
March 1939.
Destroyed in ex-
tremis on 21
February 1939.
Destroyed in ex-
tremis on 23
February 1939.
Survival
days
p. ins.
16
15
15
16
6
8
lesions. On the left side extensive completely organised adhesions were
present over the entire pleura, and the entire cardiac and apical lobes of the
lung presented lesions of a chronic interstitial pneumonia. No lesions re-
sembling active caprine pleuro-pneumonia were present, appearances suggest-
ing rather a post-pneumonic fibrosis. Whether this animal could have re-
covered from a previous attack of the specific disease can only be conjectured.
In the light of subsequent experience this is probable.
Goat No. 124 had typical lesions. Both lungs presented the same pic-
ture. An area of grey hepatisation about the size of a tennis ball at the
caudal extremity of the diaphragmatic lobe, and early lesions in the apical
and attached portion of the cardiac lobes, from which fluid exuded freely
on incision—the typical early lesion. All lesions from this goat were excised
with due asepsis and emulsion prepared in the usual way for further passage.
Experiment 12.—Serial pulmonary transmission by intra-nasal insufflation of
pneumonic lung emulsion : 2nd passage
Procedure.—The above preparation was administered on the day of
preparation to 14 goats as follows :
Eight received the. fresh lung emulsion, four by intra-nasal insufflation
of 5-6 c. c. and four ' virulence ' controls by intra-muscular inoculation of 5
c. c. Six control goats received a portion of the lung preparation after
heating to 55°—60°C. for 40 minutes, four by intra-nasal insufflation and
two by intra-muscular injection in 5 c. c. doses. The technique used was
identical with that in the previous experiment. The details and the results
are shown in Table X.
146
Contagious Pleuro-Pneumonia of Goats
TABLE X
Intra-nasal insufflation
Group
I. a.
II. a.
Goat No.
187
188
189
190
183
184
185
186
Date
3 March 1939
„
„
„
Do.
„
„
„
Inoculum
Fresh P. L. E. ex
Experimental
Donor No. 124.
5-6 c. c. I/Nas.
„
„
„
Do. heated to 00°C. for
40 minutes.
5-0 c.c. I/Nas.
„
„
„
Results
(Lung)
±
+ + + +
+ + + +
+ + + +
—
—
—
—
Remarks
Slaughtered on April
1939.
Died on 30 March 1939
Slaughtered
on 24
March 1939.
Died on 28 March 1939
Slaughtered on 1 April
1939.
Do.
Died on 18 March 1939.
Slaughtered on 1 April
1939.
Survival
days
p. ins.
29
27
21
25
29
29
15
29
TABLE XI
Intra-muscular inoculation (' Virulence ' and ' Avirulence ' controls )
Group
I. b.
II. b.
Goat No.
177
178
179
180
181
182
Date
3 March 1939
„
„
„
Do.
„
Inoculum
Fresh P. L. E. ex
Experimental Donor
No. 124. 5 c. c. I/M
„
„
„
Do. heated to 55°-60°C.
for 40 minutes.
„
Results
(local
lesion)
+ + + +
+ + + +
+ + + +
+ + + +
—
—
Remarks
Slaughtered on 6 March
1939.
Died on 8 March 1939
Died on 9 March 1939
Destroyed on 8 March
1939.
Survival
days
p. ins.
3
5
6
5
E. O. LONGLEY 147
Results.—It will be seen from the table that the inoculum was virulent
on intra-muscular inoculation in the ' virulence' controls, causing full re-
actions in all the four recipients, fatal in both instances where the disease was
allowed to run its course. The heated emulsion, however, caused no reaction
whatever in either recipient.
Of the experimental groups, three of the four who received the virulent
unheated material intra-nasally developed the typical pneumonia, in two cases
with natural death at the 25th and 27th days p. i. In the remaining goat
(No. 187), slaughtered at the 29th day p. i.. small foci of congestion in the
diaphragmatic lobe of the left lung were the only lesions evident. Of the three
definite positive cases, the two which died, Nos. 188 and 190, showed lesions
as follows :
Goat No. 188—Late consolidation of both diaphragmatic lobes with
pleuritic involvement. Early lesions (hepatisation with surrounding oedema)
of both cardiac lobes.
Goat No. 190—Consolidation of the entire left lung with focal lesions in
all lobes of the right lung. Both lungs were covered with coagulated effusion
with extensive parietal pleuritis.
The remaining reactor, No. 189, which was slaughtered on the 24th of
March 1939, 21 days p. i., had complete hepatisation of both apical and cardiac
lobes and partial consolidation of the diaphragmatic. Slight costal pleuritis
was evident, but no effusion. The material from this goat was used for
transmission purposes, as detailed in the next experiment.
As shown in the table, one of the insufflation controls receiving heated
emulsion died from other causes at the 15th day p. i. No pulmonary lesions
were present in any.
Experiment 13.—Serial transmission by intra-nasal insufflation of pneumonic
lung emulsion: 3rd passage.
Lung emulsion from goat No. 189, of the preceding experiment, slaughtered
on the 24th of March 1939 for the purpose, was prepared as usual and ad-
ministered intra-nasally by the same technique as previously to four
healthy goats, and also in 10 c. c. doses subcutaneously and 10 c. c. intra-
muscularly (total 20 c. c.) to two healthy goats. Guinea-pigs were inocu-
lated as usual with no reaction. The results are shown below :
TABLE XII
Goat
No.
203
204
205
206
207
208
Date
24 March 1939
„
„
„
Do.
„
Inoculum
Fresh P. L. E. ex Donor
189 8 c. c. I/Nas.
„
„
„
Do.
10 c. c. S/C
10 c. c. I/M.
Results
Lungs
+ + + +
+ + +
+ + + +
+ + + +
—
—
Local
reaction
—
—
—
—
±
±
Remarks
Died on 9 April 1939.
Destroyed on 5 April
1939.
Died on 4 April 1939
Died on 10 April 1939
Tested I/Nas. on 4
April 1939 and 5
April 1939.
Survival
days
p. ins.
15
12
11
16
Result
(Lungs)
+ + + +
—
D
148
Contagious Pleuro-Pneumonia of Goats
Results.—All the four experimental goats reacted to the insufflation with
typical and extensive pulmonary lesions, in one case as early as the 11th and
in another at the 12th days p. ins. Nos. 204 and 205 were used for the next
passage. Neither of the local inoculations produced any but insignificant
reactions which had receded completely in a week, although the dosage (20
c. c.) was eight times the smallest infecting dose used to date.
Discussion.—No further details of passages are given. Passage of the
infection has since been maintained on these lines up to the conclusion of
the work, and to the 14th passage.
The results at the 3rd passage, where the pneumonic lung emulsion was
100 per cent infective at insufflation and apparently non-infective in the
control inoculations suggests that the infectivity of pneumonic lung emulsion
by the two routes is to some extent independent.
Finally, Experiment 12 confirms the findings at inoculations, that the
infection is destroyed by exposure to heat at 55°—60°C. for 40 minutes.
Experiment 14.—The induction of pulmonary lesions by the intra-nasal
insufflation of emulsion from the local reactions.
The disease had been successfully passaged from an original pneumonic
emulsion through lungs and also through local muscle and subcutaneous re-
actions and it was clearly indicated to attempt the re-production of pul-
monary lesions from the latter, to complete the cycle of transmission
shown below
Procedure.—On the 27th of April 1939, five healthy goats (Nos. 221 to
225) had received 12 c. c. intra-muscularly of pneumonic lung emulsion pre-
pared from goats Nos. 209 and 212, these being reactors to strain 42/46 at
its 5th intra-pulmonary passage. Of these five goats, three reacted with local
lesions and were slaughtered at the 3rd day p. i. for material. The muscle
effusion was aspirated and mixed with emulsion of selected portions of re-
acting muscle tissue, the mixture strained, and administered by the usual
insufflation technique to healthy goats, to six without treatment, and four
controls after heating it at 55°—60°C. for 45 minutes. The details and the
results are tabulated in Table XXIII.
As shown, all the six experimental goats developed pneumonia from the
10th to 16th p. ins., in four cases with natural deaths, and in all cases with
severe and typical pulmonary lesions. None of the four controls had any pul-
monary lesions at ultimate autopsy.
E. O. LONGLEY 149
TABLE XIII
Intra-nasal insufflation of emulsion of reacting muscle
Goat
No.
231
232
233
234
235
236
237
238
239
240
Date
30 April 1939
Do.
Do.
Do.
Do.
Do.
Do. .
Do. .
Do. .
Do. .
Material
Muscle emulsion ex goats 222
to 224.
I/Nasal 20 c.c.
„
„
„
„
„
Do. heated 50—60 o C. for 45
minutes.
„
„
„
Result
(Lungs)
+ + + +
+ + + +
+ + + +
+ + + +
+ + + +
+ + + +
—
—
—
—
Remarks
Died on 12 May 1939
Died on 16 May 1939
Destroyed on 11 May
1939.
Destroyed on 12 May
1939.
Died on 10 May 1939
Died on 12 May 1939
Died on 22 May 1939
Destroyed on 20 June
1939.
Do. .
Do.
Survival
days
p. ins.
12
16
11
12
10
12
22
51
51
51
Passage. Pneumonic lung emulsion prepared from three of these re-
actors was administered intra-nasally on three successive days to five healthy
goats, two of which developed the pneumonia, as below :
TABLE XIV
Goat
No.
246
247
248
249
250
1st insufflation
Date
10 May
1939
Do.
Do.
Do.
Do.
Material
P. L. E.
ex goat
235.
20 c. c.
I/Nas.
Do.
Do.
Do.
Do.
2nd insufflation
Date
11 May
1939.
Do.
Do.
Do.
Do.
Material
P. L. E.
ex goat
233.
20 c. c.
I/Nas.
Do.
Do.
Do.
Do.
3rd insufflation
Date
12 May
1939.
Do.
Do.
Do.
Do.
Material
P. L. E.
ex goat
234.
20 c. c.
I/Nas.
Do.
Do.
Do.
Do.
Result
(Lungs)
—
+ + + +
+ + + +
—
—
Remarks
Died on 4 June
1939.
Died on 25 May
1939.
Destroyed on 23
May 1939.
Retained
for
Hyper-immurii¬
sation. (Expt.
26).
Destroyed on 25
May 1939.
Sur-
vival
days
p. ins
25
15
13
...
15
Both these reactors furnished material for a further passage, and this
strain of virus has been maintained to date.
Experiment 15. Confirmation of the infectivity at insufflation of effusion from
the local reactions
An experiment in confirmation of the preceding experiment was made
on the 18th of July 1939, on similar lines but using the effusion only as
opposed to the musele emulsion then used.
150
Contagious Pleuro-Pneumonia of Goats
Procedure.—The donor was goat No. 412, inoculated subcutaneously
with 5 c. c. of pneumonic lung emulsion ex goats 381-383 and 384 at the 11th
passage of strain 42/46, one of the controls to the immunisation experiment
of that date (Experiment 32) and which was slaughtered in extremis on the
18th of July 1939. The effusion from the extensive local reaction was as-
pirated and defibrinated by agitation with glass beads, diluted with an equal
volume of Tyrode's solution and administered in 10 c. c. doses intra-nasally
by insufflation to six goats. The results are shown below :
TABLE XV
Goat
No.
492
493
494
495
496
497
Date
18 July 1939
Do.
Do.
Do.
Do.
Do.
Route and dose
10 c. c. I/Nasal
Do.
Do.
Do.
Do.
Do.
Material
Muscle effusion in
Tyrode's (1:1)
ex goat 412.
„
„
„
„
„
Result
(Lungs)
+ + + +
+ + + +
+ + + +
+ + + +
—
+ + + +
Remarks
Destroyed on 5 August
1939.
Do.
Do.
Died on 4 August
1939.
Destroyed
on
5
August 1939.
Do.
Survival
days
p. ins.
18
18
18
17
18
18
The pneumonia resulted in five of the six goats, in one case with natural
death 17 days p. ins., in the remainder at autopsy on slaughter 18 days p. ins.
All lesions were good and typical, with pleurisy and effusion in three cases.
Summary.—(1) In two separate experiments involving a total of 12
goats, insufflation of both emulsion and the effusion from local reactions
induced the pneumonia in 11 goats.
(2) Intra-pulmonary passage was successfully effected from these induced
pneumonias.
(3) None of the four controls receiving the emulsion after treatment with
heat at 55°—60°C. for 45 minutes had any pulmonary lesion.
(3) By contact
(a) Experiment 16. Contact with natural cases
Procedure.—Contact experiments have been in progress off and on
throughout the course of the work. The procedure has simply been to intro-
duce healthy goats into a loose box where donor or reactor goats were housed.
Over long periods the box was not cleaned out in the hope of enhancing the
risk of infection. In the current experiment as many as 12 suspected cases
from the field, of which more than 50 per cent were actual diseased subjects,
have at times been on hand, housed in one such loose box 20' × 20' in size,
and the facilities for contact infection should have been ample.