NationalLibraryOfScotland/medical-history-of-british-india huggingface dataset:
A STUDY ON DESICCATED GOAT SPLEEN VACCINE AS
AN IMMUNIZING AGENT AGAINST RINDERPEST
BY
K. S. NAIR, G.B.V.C., M.R.C.V.S., D.T.V.M.
Superintendent
R. KRISHNAMURTI, G.M.V.C.
AND
G. S. KALYANASUNDARAM, G.M.V.C.
Veterinary Assistant Surgeons, Serum Institute, Madras
(Received for publication on 14 October 1941)
T ISSUE vaccines are now widely used in combating rinderpest and
several workers have employed, in different ways, rinderpest goat spleen
in the preparation of such vaccines. In Burma, a dried goat spleen
vaccine is being used on an extensive scale and is reported to be safe, effective
and cheap. A series of experiments were conducted at the Serum Institute,
Madras, with desiccated goat spleen vaccine; the product was also tried in
selected areas of the province. The object of this paper is to record the
results obtained.
TECHNIQUE OF PREPARATION OF THE VACCINE
A private communication from Mr G. Pfaff of Burma and his [1938]
article ' Immunization against rinderpest, with special reference to the use
of dried goat spleen' gave full details of the preparation of this vaccine.
The method followed for making it was similar to that adopted by him.
Briefly stated, the mode of preparation is as follows :—
Spleens from three reacting goats were cut into pieces and a few pieces
of each spleen were passed through a tissue mincer. The resulting fine pooled
pulp was placed in a desiccator and dried in vacuo over calcium chloride,
keeping the desiccator in cold storage. The quantity of dry vaccine ob-
tained was roughly one-fourth by weight of the pulp used for desiccation.
DETAILS OF EXPERIMENTS
Experiments with the first batch of vaccine
The first batch of vaccine was prepared on 13 June 1939, using the
pooled spleen pulp from three goats belonging to the three different strains, viz
Mukteswar strain No. 1, Mukteswar strain No. 2 and Madras strain, which
were maintained at the Serum Institute. The spleens were removed on
different dates and kept in cold storage before pooling. Fresh vaccine as
305
306
A Study on Desiccated Goat Spleen Vaccine
well as vaccine kept at room temperature for one to seven days was tried.
The temperature of the room varied from 92.6° to 101.8°F. Further, to
serve as control for the material used in the preparation of the vaccine, two
buffalo-calves were each injected with 1 c.c. of one per cent emulsion, in normal
saline, of the pooled spleen pulp, taken just before putting it in the desiccator.
In all, 18 calves were used for this batch of experiments. The dose of
the desiccated vaccine given was 0.0025 gram in 1 c.c. of normal saline, and
all the calves were retested with virulent bull virus.
It was found from this experiment that:—
(a) the spleen pulp used was infective;
(b) the fresh desiccated vaccine, as also the vaccine exposed to room
temperature for one day, provoked good rinderpest reaction ;
(c) the vaccine exposed to room temperature for two days produced a
mild reaction;
(d) the vaccine exposed to room temperature for three to seven days
produced no reaction;
(e) all the calves used in (a), (6) and (c) withstood the retest with
5 c.c. of virulent bull virus, while in (d) no protection could be
detected.
There was no mortality among the calves (Table I).
It was considered that the potency of the vaccine fell off after two days'
exposure to room temperature, as spleens of different strains—with the
possibility of varying viral content—were used in this experiment. Further,
this was the first time that a satisfactory desiccated vaccine was prepared
at this Institute and it was possible that the technique might not have been
as perfect as it was in later work. However, studies were continued with
pooled spleens of a single strain of virus and yielded better results.
Experiments with the second batch of vaccine
The second batch of vaccine was prepared on 18 July 1939, using
pooled spleen pulp from three goats of the 329th passage of Mukteswar strain
No. 1. In order to test the utility of the product for field use, the vaccine
was kept under conditions simulating those under which it is despatched to
the field and then tested on buffalo-calves at varying periods of exposure
to room temperature. For this purpose, the tube of vaccine, was placed for
three days in a thermos jar with ice; it was now removed and exposed to
room temperature, which varied from 96.6°F. to 97.8°F. Trials were con-
ducted with the freshly prepared vaccine, vaccine just removed from the
thermos jar and vaccine exposed to room temperature for one to four days.
Twelve calves, in all, were used for this experiment.
It was observed that the fresh vaccine, as well as the vaccine just removed
from thermos jar and the vaccine kept exposed to room temperature for one
to four days, produced moderate reactions. The immunity value was satis-
factory, as there was no reaction in any of the calves after retest. There
was no mortality in the calves (Table II).
Experiments with the third batch of vaccine
The third batch of vaccine was prepared on 1 August 1939, from the
pooled spleen pulp of three goats of the 386th passage of the Madras strain
K. S. NAIR, R. KRISHNAMURTI & G. S. KALYANASUNDARAM 307
The plan adopted for the experiment with the second batch of vaccine was
repeated here, but the period of exposure to room temperature after removal
from thermos jar was extended to seven days to determine the viability
of the vaccine at room temperature, after removal from the jar. The room
temperature varied from 89.2°F. to 97.9°F.
Eighteen buffalo-calves were used in this experiment. From Table III,
which gives details of the reactions, the following points may be noted :—
(a) Freshly prepared vaccine, the one taken out of the jar of ice after
three days' storage, and the vaccines exposed to room tempera-
ture for one to three days after removal from the jar, produced
reactions in buffalo-calves varying in intensity from a mild
thermal rise to a moderate reaction with thermal rise and
diarrhœa. At the same time, the immunity set up in all these
cases, was sufficient to withstand the subsequent test with
virulent bull virus.
(b) The vaccine exposed to room temperature for four days might be
said to have a lower potency, as a thermal rise was observed
after retest in one of the calves.
(c) The vaccines exposed to room temperature for five to seven days,
after removal from the thermos jar, produced no reactions
and conferred no immunity, as subsequent test with virulent-
bull virus set up reactions.
(d) One calf which was given the vaccine kept in room temperature
for five days and showed no reactions, died after retest.
Experiments with the fourth batch of vaccine
On 20 August 1939, the fourth batch of vaccine was prepared from the
pooled spleen pulp of three goats of the 63rd passage of the Mukteswar strain
No. 2. The plan adopted for the third batch of vaccine was repeated here,
with the addition that an emulsion prepared with the vaccine was exposed
to room temperature for three hours and six hours respectively and injected
into two buffalo-calves in each case. This was done to find out the keeping-
quality of the emulsion as such, once prepared.
The room temperature varied from 96.5°F. to 101.7°F. In all, 22
calves were used and results obtained may be stated as follows (Table
IV):
The freshly made vaccine and vaccines exposed to room temperature
for one to three days, after removal from thermos jar, were found to produce
a mild reaction and confer immunity as judged by retest with virulent blood
virus.
The vaccine exposed to room temperature for four days was erratic
in behaviour, since one of the calves showed a suspicious rinderpest reaction
after retest with bull virus. The fourth day exposure has probably brought
about a loss of potency as was observed in the third batch of vaccine pre-
pared.
The vaccines exposed to room temperature for five to seven days produced
no reactions and all the calves showed rinderpest reaction to the retest with
virulent bull virus; there was a mortality of three calves after retest.
308
A Study on Desiccated Goat Spleen Vaccine-
Emulsion kept exposed to room temperature for three and six hours
A saline emulsion of the vaccine prepared immediately after removal
from thermos jar and kept at room temperature for three hours and six hours,
was found to be active and produced moderate reactions and conferred im-
munity (Table V).
Experiments with the vaccine received from Burma
It was thought desirable to get a sample of desiccated goat spleen vaccine
from Burma ; the Director of Veterinary Services, Madras, was good enough
to get a small quantity for trial. The tubes of vaccine were received in a
sealed tin container, which was placed in a thermos jar packed with ice,
although at the time of opening the jar, the ice had melted during the journey
of five days, and the tin was found floating in water.
The trial consisted in inoculating two goats and two buffalo-calves with
the vaccine as soon as it was received and again into batches of two buffalo-
calves with the vaccine kept in room temperature for one to four days. The
dose of the vaccine given was 0.0025 gram in 1 c.c. of normal saline solution,
the emulsion being prepared just before inoculation in each case. Though
the dose of the vaccine prescribed for buffaloes was only one-sixth the dose
actually used in the experiment, yet inoculations were commenced with
the dose advocated for cattle, viz. 0.0025 gram, as viability of the agent was
alone looked for in this experiment. All the calves were tested for immunity
with virulent bull virus, after the reaction to vaccination had subsided.
The result of this experiment may briefly be described as follows (Table
VI):-
(a) The two goats that received fresh vaccine developed severe rinder-
pest and died.
(b) All the buffalo-calves had mild thermal reaction and at the same
time acquired immunity as judged by subsequent tests.
(c) There was no mortality among the calves.
Experiments conducted on adult he-buffaloes with varying doses
A batch of vaccine was prepared on 3 October 1939, from pooled spleen
pulp of three goats of the 342nd passage of Mukteswar strain No. 1 and stored
in Frigidaire.
In this experiment, three sets of adult he-buffaloes (serum-makers) were
selected. The first two sets, consisting of 16 and 15 animals respectively,
were injected on 16 October 1939 with the vaccine that had been
in the Frigidaire for 13 days; the animals of the first set received a
dose of 0.0025 gram each and the buffaloes of the second set received
0.00125 gram. The third set consisting of 15 buffaloes was injected on
17 October 1939 with the same vaccine stored in the cold store for 14
days, each animal receiving a dose of 0.000625 gram.
Two buffalo-calves were used as controls for this batch of vaccine and
were given 0.0025 gram each, on 16 October 1939. Both developed severe
rinderpest and died.
K. S. NAIR, R. KRISHNAMURTI & G. S. KALYANASUNDARAM 309
The result of this experiment can be summarized as follows (Table VII):—
The vaccine in doses ranging from 0.000625 gram to 0.0025 gram were
found to provoke reaction and confer immunity on adult buffaloes.
The reactions ranged from mild to moderately severe, and one buffalo of the
second batch and one of the third batch, which received 0.00125 and
0.000625 gram respectively, died of rinderpest.
In these three lots, making up a total of 46 animals, 22 had mild
reactions, 19 showed severe reactions and two of these died. All the
survivors in this experiment withstood the usual retest. It may be
noted that there was no response in five animals either to the vaccine or to
the retest. Most of the animals that had diarrhœa were found to harbour
coccidia.
Experiments with varying doses in buffalo-calves
Four pairs of buffalo-calves were given varying doses of the vaccine
which was stored in Frigidaire for 25 days. The first pair received each
0.01 gram, the second 0.005 gram, the third 0.00125 gram and the
fourth 0.000625 gram.
The first and second pairs had mild reactions and they withstood the
retest with virulent bull virus satisfactorily. One calf of the third pair had
late thermal rise, developed severe reaction and died—probably a case of
natural infection. The other calf of the third pair and one of the fourth
pair were non-reactors throughout. One calf of the fourth pair, which
showed no reaction to the vaccination, developed severe rinderpest on retest
with virulent bull virus and died (Table VIII).
In all the experiments already performed, the dose 0.0025 gram was
uniformly used for buffalo-calves and that dose was found to set up satis-
factory reaction and confer immunity. It may be said that the dose below
0.0025 gram is not satisfactory for buffalo-calves, although in adult buffaloes,
even one-fourth of this dose (viz. 0.000625 gram) was found to set up re-
action.
The viability of the vaccine in cold storage
Experiments on the viability of desiccated goat spleen vaccine, so far
performed, showed that the vaccine was viable up to 65 days in cold store
at Frigidaire temperature, but the experiments were not conclusive.
Experiments in the field
Five batches of the vaccine were manufactured on different dates and
were sent to selected areas for trial and report. The vaccine was supplied
in sealed tubes in thermos jar packed with ice, with instructions to use the
product immediately, or if that could not be done, to renew the ice in the
flask on receipt and thereafter on every third day, unless the flask could be
placed in a cold store. Each tube contained 100 doses of the vaccine
at 0.0025 gram per dose. Instructions were given to empty the tube of
powder in a sterile mortar, emulsify it with a small quantity of normal saline ;
pour the emulsion in a graduated measuring jar, add normal saline and agitate
the solution to make up 100 c.c. The dose was 1 c.c. per animal. The vacci-
nations were all done in infected areas.
D
310
A Study on Desiccated Goat Spleen Vaccine
From the reports received, the following observations were made :—
So far, 1,988 cattle and 686 buffaloes were vaccinated, of which 1,878
cattle and 666 buffaloes were available for observation. Of these, 984 cattle
and 491 buffaloes had reactions with a mortality of 10 cattle (0.5 per cent)
and 11 buffaloes (1.7 per cent). The reactions may be classified under two
headings, viz. mild and severe. 47.1 per cent of the cattle and 59.4 per
cent of the buffaloes had mild reaction and 5.3 per cent of the cattle and
14.3 per cent of the buffaloes had severe reaction. 47.6 per cent of the
cattle and 26.3 per cent of the buffaloes showed no reaction. Amongst the
severe reactors some animals were rendered unfit for work for two or three
months after vaccination, some of the milch cattle had dried up and two she-
buffaloes aborted. It was found necessary to give anti-rinderpest serum to
35 cattle and 91 buffaloes to check the severity of reaction.
DISCUSSION
Experiments were conducted at the Serum Institute, Madras with desic-
cated goat spleen vaccine, on susceptible young buffalo-calves and adult
he-buffaloes. While in young calves the reactions observed were not of a
severe nature, in adult buffaloes rather severe reactions were set up. The
vaccine was also sent to selected areas in this province for trial, and the ex-
perience was such that it had to be discontinued. It was reported to have
set up severe reactions resulting in death of some animals. Many of the
cattle were said to have lost condition, while some milch cattle became dry.
In Burma, this vaccine is widely used and very good results are reported,
though there is a note of warning in the annual report of the Civil Veterinary
Department, Burma, for the year 1939-1940 : ' it would seem inadvisable
to inoculate buffaloes, unless they can be well fed and well cared for and given
at least 30 days' rest after inoculation.' It was also stated ' the inoculated
buffaloes lost a lot of condition during the reaction and this notwithstanding
the fact that they were well fed and kept under ideal conditions.'
SUMMARY
Desiccated goat spleen vaccine was prepared at the Serum Institute,
Madras, with pooled spleen pulp from at least three goats killed on the fourth
day after yielding rinderpest goat blood virus.
2. Experiments were conducted with this vaccine on susceptible young
buffalo-calves and adult buffaloes and it was found that the vaccine provoked
reactions and conferred immunity.
3. The vaccine can safely be said to be potent for three days when kept
at room temperature.
4. It is suggested that the vaccine has a lesser potency when exposed
to conditions prevailing in the plains for four days.
5. The vaccine has no immunizing value after four days' exposure to
temperature of 97°F. and upwards.
6. No appreciable difference in the nature of reactions set up was ob-
served in buffalo-calves when doses varying from 0.0025 gm. to 0.01 gm.
were given. Doses below 0.0025 gm., set up no reactions and conferred
K. S. NAIR, R. KRISHNAMURTI & G. S. KALYANASUNDARAM 311
no immunity, although in adult buffaloes, doses varying from 0.000625 gm.
to 0.0025 gm. produced satisfactory reactions and conferred immunity.
7. It is suggested that the desiccated rinderpest goat spleen would
produce a reaction to confer immunity.
8. In the field, in some places, the vaccine was found to have set up severe
reactions and, from the experience gained, it cannot be said that the vaccine
is a safe one.
REFERENCES
Burma, Ann. Rept. C. V. D., 1939-40
Pfaff, G. (1938). Onderstepoort J. Vet. Sci. & Ani. Ind. 11, 2
TABLE I
To determine the viability at room temperature of dried goat spleen vaccine—
Batch No. I of Madras
Dose—0.0025 gm.
Serial
No.
1
1
2
3
4
5
6
7
8
Period
of
storage
2
Nil . .
1 day
2 days
3 days
4 days
5 days
6 days
7 days
Reaction
to
vaccination
3
1. Severe
2. Nil.
1. Moderate
2. Nil .
1. Mild
2. Nil .
1. Nil.
2. Nil .
1. Nil .
2. Nil .
1. Nil .
2. Thermal*
1. Nil . .
2. Nil . .
1. Thermal*.
2. Nil .
Reaction
after
retest
4
Nil
.
Nil
.
Nil
.
Nil
.
Nil
.
Mild .
Moderate .
Moderate .
Severe .
Mild .
Moderate
Nil . .
Nil . .
Mild .
Severe
Moderate .
Interval
between
vaccination
and retest
5
Days
24
13
23
12
15
11
10
10
9
9
8
22
7
11
20
6
Remarks
6
* Presumably not
Rinderpest.
*Presumably not
Rinderpest.
NOTE.—Two calves were used for each serial number.
D 2
312
A Study on Desiccated Goat Spleen Vaccine
TABLE II
To determine the viability at room temperature of dried goat spleen vaccine
Batch No. II of Madras
Dose—0.0025 gm.
Serial
No.
1
1
2
3
4
5
6
Period
of
storage
2
Nil . .
Nil . .
1 day . .
2 days
3 days
4 days
Reaction
to
vaccination
3
1. Mild . .
2. Mild . .
1. Moderate .
2. Mild . .
1. Mild . .
2. Mild . .
1. Mild . .
2. Mild . .
1. Mild . .
2. Mild . .
1. Mild . .
2. Mild . .
Reaction
after
retest
4
Nil . . .
Nil . . .
Nil . . .
Nil. . . .
Nil . . .
Nil . . .
Nil . . .
Nil . . .
Nil . . .
Nil . . .
Nil . . .
Nil . . .
Interval
between
vaccination
and
retest
5
Days
22
26
19
19
18
18
17
17
21
21
20
20
Note:
Two calves were used for each serial number.
In serial numbers two to six, the vaccine was kept in thermos jar with ice packing
for three days and then kept at room temperature for the number of days specified
against each.
K. S. NAIR, R. KRISHNAMURTI & G. S. KALYANASUNDARAM 313
TABLE III
To determine the viability at room temperature of dried goat spleen vaccine—
Batch No. III of Madras
Dose—0.0025 gm.
Serial
No.
1
1
2
3
4
5
6
7
8
9
Period
of
storage
2
Nil . .
Nil . .
1 day .
2 days .
3 days .
4 days .
5 days .
6 days .
7 days .
Reaction
to
vaccination
3
1. Mild
2. Moderate
1. Mild
2. Mild
1. Moderate
2. Severe and
died.
1. Mild
2. Nil
1. Moderate
2. Moderate
(late).
1. Nil .
2. Nil .
1. Nil .
2. Moderate*
1. Nil . .
2. Nil. .
1. Nil . .
2. Nil . .
Reaction
after
retest
4
Nil . .
Nil . .
Nil . .
Nil . .
Nil . .
. .
Nil . .
Mild . .
Nil . .
Nil . .
Mild . .
Nil . .
Severe and
died.
Nil
Severe .
Mild . .
Mild . .
Nil . .
Interval
between
vaccination
and
retest
5
Days
18
21
15
18
19
..
16
12
17
27
10
10
13
28
12
9
8
8
Remarks
6
* Probably a
case
of
natural
in-
fection.
Note:
Two calves were used for each serial number.
In serial numbers two to nine, the vaccine was kept in thermos jar with ice packing
for three days and then kept at room temperature for the number of days specified
against each.
314
A
Study on Desiccated Goat Spleen Vaccine
TABLE IV
To determine the viability at room temperature of dried goat spleen vaccine—
Batch No. IV of Madras
Dose—0.0025 gm.
Serial
No.
1
1
2
3
4
5
6
7
8
9
Period
of
storage
2
Nil . .
Nil . .
1 day. .
2 days . .
3 days . .
4 days . .
5 days . .
6 days . .
7 days . .
Reaction
to
vaccination
3
1. Mild . . .
2. Mild . . .
1. Nil . . .
2. Nil . . .
1. Mild . . .
2. Mild . . .
1. Mild . . .
2. Mild . . .
1. Mild . . .
2. Mild . . .
1. Nil . . .
2. Nil . . .
1. Nil . . .
2. Nil . . .
1. Nil . . .
2. Nil . . .
1. Nil . . .
2. Nil . . .
Reaction
after
retest
4
Nil . . .
Nil . . .
Nil . . .
Nil . . .
Nil . . .
Nil . . .
Nil . . .
Nil . . .
Nil . . .
Nil . . .
Nil . . .
Moderate .
Moderate .
Moderate .
Severe and died
1 Severe and died
Mild . .
Severe and died
Intorval
between
vaccination
and
retest
5
Days
14
14
7
7
21
19
15
18
17
17
7
7
7
7
7
7
7
7
Note:
Two calves were used for each serial number.
In serial numbers two to nine, the vaccine was kept in thermos jar with ice packing
for three days and then kept at room temperature for the number of days specified
against, each.
K. S. NAIR, R. KRISHNAMURTI & G. S. KALYANASUNDARAM 315
TABLE V
To determine the viability of the emulsion in normal saline of dried goat spleen
vaccine, at room, temperature for (1) 3 hours and (2) 6 hours
Dose—0.0025 gm.
Serial
No.
1
1
2
3
Period
;
Of
storage
'
2
Nil . . .
3 hours . . .
6 hours . . .
Reaction
to
vaccination
3
1. Nil . .
2. Nil . .
1. Mild . .
2. Nil . .
1. Moderate .
2. Nil . .
Reaction
after
retest
4
Nil . . .
Nil . . .
Nil . . .
Nil . . .
Nil . . .
Nil . . .
Interval
between
vaccination
and
retest
5
Days
7
7
20
7
17
7
Note.—Two calves were used for each serial number.
TABLE VI
To determine the viability at room temperature of dried goat spleen vaccine of
Burma
Dose—0.0025 gm.
Serial .
No.
1
1
2
Period
of
storage
2
Nil
.
.
Nil . .
Reaction to
vaccination
3
Goat No.1—Severe and died .
Goat No 2—Severe and died .
Calf No.1— Nil . . .
Calf No.2—Nil . . .
Reaction
after
retest
4
..
..
Nil . .
Nil . .
Interval
between
vaccination
and
retest
5
Days
..
..
6
6
316
A Study on Desiccated Goat Spleen Vaccine
TABLE VI—contd.
Serial
No.
1
3
4
5
6
Period
of
storage
2
1 day .
2 days .
3 days .
4 days .
Reaction to
vaccination
3
Calf No. 1—Nil . . .
Calf No. 2—Nil . . .
Calf No. 1—Mild . .
Calf No. 2—Nil . .
Calf No. 1—Mild . .
Calf No. 2—Mild . .
Calf No. 1—Mild . .
Calf No. 2—Mild . .
Reaction
after
retest
4
Mild . .
Mild . .
Nil . .
Nil . .
Nil . .
Nil . .
Nil . .
Nil . .
Interval
between
vaccination
and
retest
5
5
5
16
4
15
15
16
16
Note.—Two calves were used for each serial number (2 to 6).
TABLE VII
To determine the immunizing properties of the dried goat spleen vaccine in
adult buffaloes and the effect of using varying doses of the vaccine
Serial
No.
1
1
2
3
4
Dose
2
gm.
0.0025
0.00125
0.000325
0.0025
Number 1
of animals
vacci-
nated
3
16
15
15
Two
calves as
controls
Reaction to vaccination
Mild
4
10
9
3
...
Severe
5
3
4
12
2
Mortality
6
Nil
1
1
2
Reaction
after
retest
7
Nil
Nil
Nil
...
Interval
between
vaccination
and retest
8
Interval rang-
ing from 8
days to 21
days.
Do.
Interval rang-
ing from 7
days to 15
days.
...
Remarks
9
Vaccine kept in
cold storage for
13 days was used.
Three
non-re-
actors throughout.
Vaccine kept in
cold storage for
13 days was used.
Two non-reactorg
throughout.
Vaccine kept in cold
storage for 14
days was used.
Vaccine kept in
cold storage for
13 days was used.