NationalLibraryOfScotland/medical-history-of-british-india huggingface dataset:
ON THE TREATMENT OF BABESIA BIGEMINA INFEC-
TION IN CATTLE IN INDIA
BY
J. A. IDNANI, G.B.V.C.
Imperial Veterinary Research Institute, Mukteswar
(Received for publication on 6th July 1937)
THE introduction of trypanblue in the field of therapy of red-water in cattle,
due to Babesia bigemina infection in cattle, was the result of pioneer work carried
out by Nuttall and Hadwen [ 1909 ]. Trypanblue has ever since been used almost
universally as a remedy against this infection. In India, cattle are known to be
infected ubiquitously with various protozoan parasites which ordinarily reside in a
latent state in the tissues of the hosts and cause no manifest disturbance. It is when
the restrain on such parasites is relaxed by the onset of some febrile diseases
such as rinderpest that they multiply at such a rate as to cause serious disease
and frequently death in the affected animals [ Edwards, 1927 ]. Trypanblue,
as would appear, is rarely called into use in clinical cases of piroplasmosis in cattle
as a primary affection in this country, due to the fact that most of the animals
are ' carriers ', and it is usually in cases of resuscitation that treatment with
drugs is resorted to.
In this Institute where a large amount of work on rinderpest is carried out,
the control of Babesia bigemina with trypanblue presented serious obstacles in
recent years. The rinderpest bull virus, which is always maintained in the Insti-
tute in successive batches in hill bulls, was found to be more often than not, con-
taminated with the red-water parasites rendering it unsafe for issue in the field.
The mortality in such complicated cases occurred to the extent of 59.3 per cent
in 1933, 55.5 per cent in 1934 and 40.7 per cent in 1935, as compared to 25.6
per cent, 16.8 per cent and 23.2 per cent during the same period respectively in
uncomplicated cases. Apart from this, in the tests carried out to evaluate the
potency of anti-rinderpest serum that is manufactured at this Institute, very
misleading thermal reactions were set up due to the resuscitation of Babesia
bigemina. In such cases the usual dose of ½ to ¾ grm. of trypanblue was adminis-
tered intravenously in a 1 per cent solution per an approximate weight of 100
kg., but in a number of cases the parasites were found to be persisting, resulting
in death in several cases. Due to a large number of such failures encountered
( 273 )
274 THE INDIAN JOURNAL OF VETERINARY SCIENCE AND ANIMAL HUSBANDRY [VII, IV
in the treatment of B. bigemina infection, with the popular drug trypanblue, work
was taken in hand early in 1934 to evolve more effective measures for the control
of these organisms. A series of controlled experiments were carried out to test
the efficacy of trypanblue in larger doses on the laboratory strain of B. bigemina.
The strain was isolated and maintained in successive batches in hill bulls, but
as would appear from the data collected, the drug failed to check the multiplication
of the parasites. On perusal of the original work of Nuttall and Hadwen [ 1909 ],
it would appear that although trypanblue produced a marked amelioration of the
symptoms and checked the severity of the disease, yet in two out of the five treated
cases, the parasites reappeared in the peripheral blood on the 7th and the 8th day
respectively after the treatment. Theiler [ 1912 ] working in Pretoria on the same
lines infected twelve susceptible cattle with the local strain of B. bigemina and
treated them with trypanblue. Except one animal that died, all recovered from
the disease. In three of his treated cases, however, microscopic examination
of the peripheral blood revealed the presence of the parasites on 14th, 18th and
21st day respectively after the treatment.
A fresh stock of trypanblue was obtained from the manufacturers, E. Merck
& Co., Germany, and comparative potency tests were carried out along with the
old stock but in the experiments carried out no evidence could be obtained of the
superiority of the new stock as compared to the old. The drug was subsequently
replaced by another proprietary preparation ' Akiron R ', manufactured by Bayer
& Co., with very successful results.
METHODS, MATERIAL AND OBSERVATIONS
At the outset work was undertaken to determine the maximum tolerable dose
of trypanblue in the hope of combating the infection due to B. bigemina with
a higher dose rate. Sergent and co-workers [ 1927 ] advocated small doses in
proportion to the number of parasites present in individual cases, as they were
of opinion that one grm. of trypanblue per 100 kg. is liable to cause alarming-
symptoms due to the liberation of toxins from the destroyed parasites. But
the treatment carried out at this Institute, with 50-75 c. c. of a 1 per cent solu-
tion given intravenously, failed to cleanse the peripheral circulation in several
cases.
For the purpose of M. T. D. tests of trypanblue, fourteen animals were in-
jected intravenously at doses varying from 1.2 to 4 grms. in a 2 per cent solu-
tion at an average body-weight of 220 lb. Very interesting observations, some
of which have been recorded by Stirling [ 1927 ], were made which are worthy
of mention.
In almost all the cases, the inoculation of trypanblue was followed within a
short period, depending on the degree of the dose used, by accelerated breathing,
filling in of the folds of the skin round the eyes and the neck, lachrymation and
276
THE INDIAN JOURNAL OF VETERINARY SCIENCE AND ANIMAL HUSBANDRY [VII, IV
In a further series of experiments the effect of administering trypanblue
immediately before a combined infection with rinderpest and B. bigemina was
tested. Two healthy bulls were given 3 grms. of trypanblue each, in a 2 per
cent solution intravenously followed by B. bigemina infected rinderpest virus
a little later. Two controls were inoculated with the virus only. All the four
animals showed organisms ten to twelve days after inoculation as trypanblue
produced no effect whatever in the treated cases. All four animals, however,
lived, and were discontinued when the parasites had disappeared from their cir-
culation.
Notwithstanding the disappointing results obtained in the foregoing ex-
periments in the treatment of the tropical red-water parasites by means of try-
panblue, it was thought that these failures may be attributed to a deterioration
in the potency of the drug used owing to long storage, or to the possibility of the
particular strain of the parasites that was being used, having developed a tolerance
to trypanblue as a result of continuous passaging through animals treated with this
drug. The strain of this piroplasm was isolated and maintained in a series of
successive passages in hill bulls that had been discontinued from rinderpest
work. Only such animals were selected for this purpose as had not exhibited
B. bigemina previously. The strain was successfully established in these animals
throughout nine passages, using two animals each time. The sub-inoculations
were done with blood drawn during paroxysms, about six to seven days after in-
fection. Although none of the animals developed clinical symptoms or showed
marked thermal reaction inspite of about thirty to forty per cent of the erythro-
cytes having been infected, yet only one animal proved refractory. Three animals
showed the parasites seventy-two hours after sub-inoculation, eight animals on
the 4th day, three on the 5th, one on the 7th and one on the 11th day. In each
case the number of the parasites gradually decreased after a phase of increased
multiplication without producing any ill effects.
Treatment with a fresh stock of trypanblue, obtained from the manufac-
turers, E. Merck & Co., Germany, was now taken up with a view to test its com-
parative potency with the old stock.
In all, eleven animals were treated as
tabulated below:—
TABLE I
Table showing the results of treatment of B. bigemina with fresh and old trypanblue
No. of animal
Passage X.
Hill
Bull
484.
341
276
Date of
Infection
16th April '35
16th April '35
16th April '35
Date of
treatment
20th April '35
22nd April '35
Untreated Con-
trol.
Stock of
Trypanblue
Fresh trypan-
blue.
Old trypanblue
. . .
Results
Showed parasites for 15
days after treatment.
Showed parasites on 4th
day after treatment.
Showed parasites through-
out the period of obser-
vation.
Remarks
Severe toxic symp-
toms exhibited.
No toxic effects
produced.
. . .
TREATMENT OF BABESIA BIGEMINA INFECTION IN CATTLE
277
TABLE I—contd.
No. of animal
Passage
XI.
{
Passage
XII.
{
Passage
XIII.
{
Passage
XIV. {
Passage
XV. {
Hill
bull
111
386
213
699
647
166
198
517
623
591
594
295
584
334
407
Date of
infection
23rd April '35
23rd April '35
23rd April '35
30th April '35
30th April '35
30th April '35
8th May '35
8th May '35
8th May '35
15th May '35
15th May '35
15th May '35
24th May '35
24th May '35
24th May '35
Date of
treatment
28th April '35
28th April '35
Untreated
Control.
3rd May '35 .
3rd May '35 .
Untreated
Control.
17th May '35
Untreated .
Untreated
Control.
21st May '35
20th May '35
Untreated
Control.
28th May '35
28th May '35
Untreated Con-
trol.
Stock of
Trypanblue
Fresh trypan¬
blue.
Old trypan-
blue.
...
Fresh trypan-
blue.
Old trypan¬
blue.
...
Fresh trypan¬
blue.
...
...
Fresh trypan¬
blue.
Old trypan¬
blue.
...
Fresh trypan¬
blue.
Old trypan¬
blue.
...
Results
Showed parasites for 5
days after treatment,
reappeared on 15th and
17th day.
Showed parasites occa-
sionally during the period
of observation.
Showed parasites through-
out the period of obser-
vation.
Showed parasites through-
out the period of obser-
vation.
Ditto .
Showed parasites through-
out the period of obser-
vation.
Showed parasites for 3
days after treatment.
Reacted late (15th day).
Showed parasites through-
out the period of obser-
vation.
Showed parasites through-
out the period of obser-
vation.
Ditto
Showed parasites through-
out the period of obser-
vation.
Showed parasites on the
2nd day after treatment.
Showed parasites through
out the period of obser-
vation.
Showed parasites through
out the period of obser¬
vation.
Remarks
No toxic
effects
produced.
Severe toxic symp-
toms exhibited.
...
Slight toxic symp-
toms exhibited.
Ditto.
...
No toxic effects
produced.
...
...
No toxic effects
produced.
Ditto.
...
Slight toxic symp-
toms exhibited.
Ditto.
...
TREATMENT OF BABESIA BIGEMINA INFECTION IN CATTLE
279
About this time a communication was received from Havero Trading Com-
pany, Calcutta, offering samples of a proprietary preparation, ' Akiron R ', manu
factured by Bayer & Co., for trial in ' Babesiosis' and ' Theilerioses' of cattle.
At that time no reference could be obtained in available literature regarding the
therapeutics of this drug, on which experiments could be based, but the oppor-
tunity was taken and the drug obtained free of charge. It was received in 5 and
10 c.c. ampoules of a 5 per cent solution, the fluid being of a light yellow colour
and ready for use. The original laboratory strain of B. bigemina was maintained
and in the subsequent series of experiments the efficacy of ' Akiron R ' was tested
with very successful results. These have been tabulated below:—(Tables II-A
and II-B).
TABLE II-A
Showing results of treatment of B. bigemina infection with ' Akiron R .'
No. of animal
Passage
XVI.
{
Passage
XVII.
{
Passage
XVIII.
{
Hill
bulls
246
219
1094
571
58
55
639
500
370
Date of
infection
31st May '35
31st May '35
31st May '35
7th June '35
7th June '35
7th June '35
13th June '35
13th June '35
13th June '35
Date on which
parasites
appeared
12th June '35
∆ B. big.
7th June '35
1st June '35 to
5th June '35.
11th June '35
∆
B. big.
7th June '35
10th June '35
16th June '35
∆
B. big.
16th June '35
∆
B. big.
16th June '35
Date of treat-
ment and dose
12th June '35. 10
c.c. sub-cutane-
ously.
Untreated . .
Do. .
11th
June '35.
10 c.c.
intra-
venously.
Untreated . .
Do. .
16th
June '35
5 c.c. sub-cuta¬
neously.
16th June '35
5 c.c. intramus-
cularly.
Untreated Control
Results of blood
smears
Negative following
day till the period
of observation.
Showed parasites
for 5 days after
appearance.
...
...
...
...
Negative following
day till the period
of observation.
Parasites not seen
again as above.
...
Remarks
Thermal reaction for
48 hours, inappe¬
tence, local swelling
lasting for a week,
increased salivation
for 2 hours.
...
'Carrier' animal para-
sites
persisted
throughout the pe-
riod of
observa-
tion.
Died soon after the
injection of ' Akiron
R.' poisoning.
Parasites
persisted
throughout the pe-
riod of observation.
Do.
Thermal reaction last-
ing 48 hours ; local
swelling.
...
Parasites
persisted
throughout the pe-
riod of observation.
D
280
THE INDIAN JOURNAL OF VETERINARY SCIENCE AND ANIMAL HUSBANDRY [VII, IV
TABLE II-A—contd.
No. of animal
Passage
XIX. {
Hill
bull
712
341
707
Date of
infection
19th June '35
19th June '35
19th June '35
Date on which
parasites
appeared
22nd June '35
∆ B. big.
22nd June '35
∆ B. big.
22nd June '35
Date of treat-
ment
and dose
22nd June
'35
5c.c. sub-cuta¬
neously.
22nd
June '35
5 c.c. intramus-
cularly.
Untreated Control.
Results of blood
smears
Negative the fol-
lowing day till the
period of observa¬
vation.
Parasites not seen
again as above.
...
Remarks
Slight thermal and
local reaction.
Thermal reaction last-
ing
48
hours.
Slight lameness.
Parasites
persisted
throughout
the
period of observa-
tion.
∆ = Very rare. ∆ = Rare.
TABLE II-B.
No. of animal
Passage
XX. {
Pasage
XXI. {
Passage
XXII. {
Hill
bull
715
686
638
730
305
669
465
41
558
Date of
infection
28th June '35
28th June '35
28th June '35
4th July '35
4th July '35
4th July '35
12th July '35
12th July '35
12th July '35
Date on which
parasites
appeared
1st July '35
4th July '35
2nd July '35
5th July '35
to 8th July '35
9th July '35
5th July '35
15th July '35
15th July '35
16th July '35
Date of treatment
and dose
4th
July '35
+ + B. big. 5 c.c.
sub-cutaneously.
6th
July
'35
+ B. big. 5 c.c. in-
tramuscularly.
Untreated Control
Untreated . .
Do. .
Do. .
18th July
'35
+ + + B. big. 5
c.c. sub-cutane¬
ously.
18th
July '35
+ + B. big. 5 c.c.
intramuscularly.
Untreated Control
Result of blood
smears
24 hours after treat-
ment parasites ap-
peared degenerated
and decreased; at
48 hours smears
negative till the
period of observa-
tion.
5 hours after treat-
ment parasites ap-
peared degenerated,
smears negative at
24 hours and after.
...
∆ B. big . .
∆ B.big . .
Control . .
24 hours after treat-
ment a small num-
ber of degenerated
parasites seen. Ne-
gative at 48 hours
and thereafter.
Do. .
...
Remarks
Appreciable thermal
reaction for
48
hours; local swel-
ling.
Increased
saliva-
tion noticed for
about one hour
after treatment.
Parasites persisted.
'Carrier,' parasites
did not increase.
Parasites did not
increase.
Parasites persisted.
Marked thermal re-
action for
24
hours after treat-
ment.
Slight thermal re-
action for
24
hours with inap¬
petence.
Parasites persisted.
+ = few.
+ + = frequent.
+ + + = numerous.
TREATMENT OF BABESIA BIGEMINA INFECTION IN CATTLE
281
TABLE II-B—contd.
No. of animal
Passage
XXIII.
{
Passage
XXIV.
{
Passage
XXV.
{
Passage
XXVI.
{
Hill
Bull
674
721
681
28
726
42
41
709
10
4
12
644
Date of
infection
18th July '35
18th July '35
18th July '35
25th July '35
25th July '35
25th July '35
31st July '35
31st July '35
31st July '35
6th Aug. '35
6th Aug. '35
6th Aug. '35
Date on which
parasites
appeared
19th July '35
24th July '35
24th July '35
29th July '35
29th July '35
29th July '35
4th Aug. '35
3rd Aug. '35
2nd Aug. '35
9th Aug. '35
6th Aug. '35 &
11th Aug. '35.
11th Aug. '35
Date of treatment
and dose
25th July '35
+ + B. big. 5 c.c.
intramuscularly.
25th
July '35
+ + B. big. 5 c.c.
sub-cutaneously.
Untreated Control
31st
July '35
++ B. big. 5 c.c.
sub-cutaneously.
31st
July '35
+ + B. big. 5 c.c.
intramuscularly.
Untreated Control
6th
Aug. '35
+ + + B. big.
5 c.c. sub-cuta-
neously.
6th
Aug. '35
+ + + B. big.
5 c.c. intramus-
cularly.
Untreated Control
Untreated . .
Do. .
Do. .
Results of blood
Smears
24 hours after treat-
ment a small num-
ber of degenerated
parasites seen. Ne-
gative thereafter.
Do. .
...
24 hours after treat-
ment a small num-
ber of degenerated
parasites seen, nega-
tive thereafter.
24 hours after treat-
ment parasites ap-
peared degenerated
and decreased, 48
hours a very small
number of degener-
ated parasites seen
and negative there-
after.
...
Negative at 24 hours
after treatment till
the period of ob-
servation.
24 hours after treat-
ment a small num-
ber of degenerated
parasites seen, nega-
tive thereafter.
...
...
...
...
Remarks.
Marked
thermal
reaction for 48
hours after treat-
ment.
Slight thermal re-
action, increased
salivation
for
about an hour,
inappetence for
2 days.
Parasites persisted.
...
Inappetence for 2
days.
Parasites persisted.
Thermal reaction
for 24 hours after
treatment.
...
Parasites persisted.
Parasites did not
increase.
Do.
Do.
+ = few.
+ + = frequent.
+ + + = numerous.
In connection with the work of testing the curative effect of ' Akiron R'
upon B. bigemina, it would appear from the foregoing tables that in all thirty-three
bulls were artificially infected in a series of eleven batches, each comprising three
animals. Of these, sixteen animals were subjected to treatment and the remaining
served as controls. The first animal handled was No. 571 (passage XVII, Table
II-A.) which died as a result of intravenous injection of 10 c.c. ' Akiron R.' The
second animal No. 246 (passage XVI) received the same dose of the drug sub¬
cutaneously on the day that parasites were first observed, but it exhibited a severe
TREATMENT OF BABESIA BIGEMINA INFECTION IN CATTLE
283
TABLE III
Results of treatment of B. bigemina with ' Akiron R ' at 1 c.c. dose
No. of animal
Passage
XXVII. {
Passage
XXVIII.{
Passage
XXIX. {
Hill
bull
87
65
704
34
733
486
14
546
Date of
infection
15th Aug.'35
15th Aug.'35
20th Aug.'35
20th Aug.'35
20th Aug.'35
26th Aug.'35
26th Aug.'35
26th Aug.'35
Date of treatment
and dose
20th Aug. '35 : + B. big.
1 c.c. intramuscularly.
20th Aug. '35: + B.big.
1 c. c. subcutaneous-
ly.
Untreated Control .
......
26thAug.'35 + + + B.
big. 1 c. c. subcutane¬
ously.
Untreated .
Ditto
31st Aug.'35: + + + B.
big. 1 c.c. spontane-
ously.
Results
Parasites
persisted
throughout the period of
observation.
Parasites seen for 48 hours
after treatment.
......
......
Parasites persisted for 4
days after treatment.
......
......
Parasites
persisted
throughout the period
of observation.
Remarks
......
......
Parasites
persisted
throughout the period
of observation.
Did not react.
......
Parasites
persisted
throughout the period
of observation.
Ditto
......
+ = few.
+ + + = fairly numerous.
In further trials the dose was raised to 2 c.c. per 100 kg. where by a total
of five animals were treated at the height of infection as indicated below in Table
IV-A.
TABLE IV-A
Results of treatment of B. bigemina with ' Akiron R ' at 2 c.c. dose
No. of animal
Passage
XXX.
{
Passage
XXXI. {
Passage
XXXII.{
Hill
bull
58
88
645
46
72
73
1
447
118
Date of
infection
2nd Sept.'35
2nd Sept.'35
2nd Sept.'35
9th Sept.'35
9th Sept.'35
9th Sept.'35
17th Sept.'35
17th Sept.'35
17th Sept'35
Date of treatment
and dose
9th Sept.'35 : + + + B.
big. 2 c.c. intramus-
cularly.
9th Sept.'35 : + + + B.
big. 2 c.c. subcutane¬
ously.
Untreated Control .
17th Sept.'35: + + +B.
big. 2 c.c. intramus-
cularly.
17th Sept.'35: + + + B.
big. 2 c.c. subcuta¬
neously.
Untreated Control .
24th Sept.'35: + + +B.
big. 2 c.c. intramus-
cularly.
......
Untreated Control .
Results
Cured .
Do. .
...
Cured .
Do. .
.
...
Cured .
...
...
Remarks
A few degenerated parasites seen up
to 24 hours after treatment and
negative thereafter.
Ditto
Parasites persisted throughout the
period of observation.
Same as in 58.
A few degenerated parasites seen up
to 24 hours after treatment and
negative thereafter.
Parasites persisted.
Same as in No. 58.
Did not react.
Parasites persisted.
+ + + = fairly numerous.
+ + + = numerous.
284
THE INDIAN JOURNAL OF VETERINARY SCIENCE AND ANIMAL HUSBANDRY [VII, IV
The results incorporated in Table IV-A indicate that a dose of 2 c.c. of
' Akiron R ' was sufficient and safe for successfully combating infection with
B. bigemina in bulls averaging 100 kg. in body-weight. Furthermore, a few
trials with 1.5 c.c. were also carried out but of the eight artificially infected bulls
treated with this dose one animal continued to show the parasites for seventy-
two hours after treatment, while in the others the circulation was free of parasites
within forty-eight hours. Finally more tests were executed to confirm the cura-
tive effect of ' Akiron R ' with 2 c.c. as shown in Table IV-B.
TABLE IV-B
Results of treatment of B. bigemina with ' Akiron R' at 2 c.c. dose
No. of animal
Passage
XXXVIII.
{
Passage
XXXIX.
{
Passage
XL.
{
Passage
XLI.
{
Passage
XLII.
{
Hill
bull
l49
148
158
l22
240
139
249
318
315
293
261
312
165
291
183
Date of
infection
29th Octr.'35
29th Octr.'35
29th Octr.'35
5th Novr.'35
5th Novr.'35
5th Novr.'35
13th Novr.'35
13th Novr.'35
13th Novr'35
21st Novr.'35
21st Novr.'35
21st Novr.'35
28th Novr.'35
28th Novr.'35
28th Novr.'35
Date of treatment
and dose
4th Novr.'35 : + + + B.
big. 2 c.c. subcuta¬
neously.
Ditto .
Untreated Control .
13th Novr.'35 : + B. big.
2 c.c. subcutaneously.
Ditto .
Untreated Control.
21st Novr. , 35: ++ B.big
2 c.c. subcutaneously.
21st Novr.'35: +B.big.
2 c.c. subcutaneously.
Untreated Control .
28th Novr.'35 : + + + B
big. 2 c.c. subcuta-
neously.
28th Novr.'35: + + +B.
big. 2 c.c. subcuta-
neously.
Untreated Control .
6th Decr.'35 : + + B.
big. 2 c.c. subcutane-
ously.
Ditto .
Untreated Control .
Results
Cured .
Do. .
...
Cured .
Do. .
...
Cured .
Do. .
Cured .
Do. .
...
Cured .
Do. .
...
Remarks
A few degenerated parasites seen up
to 48 hours after treatment and
negative thereafter.
A few degenerated parasites seen up
to 24 hours after treatment and
negative thereafter.
Parasites persisted throughout the
period of observation.
Negative at 24 hours after treatment
and thereafter.
Ditto.
Parasites persisted for 10 days after
appearance.
A few degenerated parasites seen up
to 24 hours after treatment and
negative thereafter.
Ditto.
Parasites persisted throughout the
period of observation.
A few degenerated parasites seen up
to 24 hours after treatment and
negative thereafter.
Ditto.
Parasites persisted for 4 days after
first appearance.
A few degenerated parasites seen up
to 24 hours after treatment and
negative thereafter.
Ditto.
Parasites persisted throughout the
period of observation.
+ =few. + + = fairly frequent. + + = frequent.
+ + + = fairly numerous. + + + = numerous.