NationalLibraryOfScotland/medical-history-of-british-india huggingface dataset:
288 THE INDIAN JOURNAL OF VETERINARY SCIENCE AND ANIMAL HUSBANDRY [ I, IV
into the field of experimental therapy of trypanosomiasis, so that, hitherto, the
bulk of the work with this drug has been carried out under laboratory conditions,
without the results having been sufficiently tested as to their value in the actual
prevention of the disease under natural conditions.
Of the recorded observations upon the prophylactic action of " Bayer 205 "
against outbreaks of natural trypanosomiasis in animals, the only ones that
would seem to deserve any special mention are those of Bakker [1925], and
Yakimoff and his collaborators [1927], all of whom employed the drug (for-
tunately for workers in India) against natural surra. Bakker's observations,
which were all made in Padang Sidempoean, were on horses, whilst Yakimoff and
his collaborators used the drug against camel trypanosomiasis in the Ural Region,
the parasite concerned being designated by them as Trypanosoma ninae-kohl-
iakimoff, which, however, as the authors themselves remark, probably represents
a race of T. evansi.
Bakker [1925] used the drug intravenously at the rate of 1 gramme in 5 per
cent. solution and altogether he inoculated 321 horses which were divided into
7 batches, each batch consisting of from 26 to 71 animals. In one batch of 51
animals, the inoculation was repeated after a month, but all the other animals
would appear to have received a single injection of the drug. The horses were
from places where cases of surra were actually present at the time the inoculations
were performed, or, at any rate, 10 to 12 days previous to the inoculations. None
of the treated horses developed surra or any other complication (information as to
the post-treatment observation period, however, is not available).
The observations of Yakimoff and his collaborators [1927], although made on
camels, would seem to bear quotation at some length, in view of the many points
of interest which these observations incidentally furnish in regard to the question
of the infectivity of grazing lands and transmission by vectors, both of which
factors, it is needless to say, will have to be given due consideration in order to
obtain the full measure of benefit from any scheme of drug prophylaxis against
equine surra.
From 1923 to 1927, a total of about 11,052 camels were examined in the
Ural Region, and the examination showed that on an average 25 per cent. of
the camels were infected, the infection in some districts being as high as 40 per
cent. Treatment of infected camels failed to prevent the spread of the disease,
and Yakimoff and his collaborators [1927] therefore tested the prophylactic value
of " Bayer 205 ". The test was carried out on four groups of camels. In the
first group of 20 camels, 12 received a subcutaneous injection of 5 grammes
each and the remaining 8 were kept as controls ; in the second group there were 24
PROPHYLAXIS AGAINST EQUINE SURRA
289
camels, and of these, 14 received intravenously 6 grammes of the drug each, the
other 10 being left as controls, together with a 1½ year diseased foal, which was left
without treatment for supply of the " virus" ("zur Aufbewahrung des virus");
the third group comprised 17 camels, out of which 10 received 8 grammes of the
drug subcutaneously, and the remaining 7 were kept as controls ; in the fourth
group there were 30 camels, of which 17 were given a subcutaneous dose of 10
grammes each, the remaining 13 being left as controls. All the inoculations
were performed during the first week of July 1926. The camels that were pro-
phylactically treated were branded with a special brand on the haunch. The
camels comprising the four groups were now left loose on four different sets of
premises ("rayons"), where, as will be described later, the natural conditions
varied. At this time, 564 camels were examined in other parts of the district, and
about 10 per cent. of these were found infected.
During July, large numbers of Tabanidae were noticed in the locality
where the experiments were initiated, and these were collected from all the
premises except the one where the second group of camels was kept.
The examination of blood smears from both the treated and. the untreated
animals was carried out from 1st to 25th October 1926. There were at this time
53 treated animals and 33 controls (no reference occurs in the article about the
remaining 5 controls).
The results of blood smear examination and also subinoculations made, from
1st to 7th October 1926, into guinea-pigs showed that all the 53 treated amimals
were free from infection. As to the controls, those of the first, third and fourth
groups were grazed, along with the treated animals of these groups, in elevated
("hochgelagenen") areas, where there were present large numbers of Tabanidae
and an abundance of gnats in the vicinity of large collections of water (" quellen¬
tränke "). None of the controls of those three groups was infected.
The camels of the second group (14 treated and 10 controls) were kept in
a low-lying (" niedriggelegenen") steppe, where the ground was flooded during
the spring by snow-water from the elevated areas, with weeds (" schilfrohr")
and other morassy plants, and where large numbers of gad-flies were also to be
observed. In this sector, a considerable number of diseased animals was observed
in 1925. In the course of the present experiments, the first occasion when the
camels of the second group were examined was from 8th to 10th July, and the
Naganol injections were also given at this time. The second examination was
made on 3rd August, when neither the treated animals nor the controls were
found affected. The third examination was made on 7th October, when 7
out of the 10 controls (70 per cent.) were found infected, but, as already men-
290 THE INDIAN JOURNAL OF VETERINARY SCIENCE AND ANIMAL HUSBANDRY [ I, IV
tioned, all the treated animals were found free from infection. Concurrently,
280 camels from the adjoining areas were also examined, and of these 79 were
found affected. From these observations, the authors conclude that under
natural conditions Naganol protects for at least 3 months.
The findings of Yakimoff and his collaborators, as reproduced above,
strikingly illustrate the extent to which the natural conditions of a locality
contribute towards the success of any operations designed to combat natural surra
of animals by means of Naganol prophylaxis.
A mention should be made here of a very recent paper by Bubberman
[ 1931 ], where reference is made to certain experimental observations indicating
that in the horse a prophylactic dose of 1 gramme of Nagonol per 150 kg. body
weight will produce in the majority of cases " a condition of non-susceptibility "
to natural surra which will last for a month or more.
A brief reference may also be made to the experiments carried out by Berg
[ 1925 ] upon the prophylactic action of " Bayer 205 " against cattle trypanoso-
miasis in South Africa. As one would expect from what has been said earlier in
regard to the action of this drug on T. congolense infection, the results obtained by
Berg were very disappointing, the drug having conferred practically no protection,
even when employed in doses of 10-25 grammes, for cattle exposed to natural
infection in fly belts 2 days after the inoculation.
From the records summarized above, it would appear that except for the
congolense group of trypanosomes, " Bayer 205 " has been more or less successfully
used for prophylactic inoculation against all of the common species of trypanosomes
affecting domesticated animals. The failure of the drug to affect trypanosomes of
certain species, while it is endowed with marked trypanocidal properties for others, is
a point of importance for workers in India, inasmuch as this might be held to explain
any failure on their part to use the drug successfully against individual cases of
animal trypanosomiasis, which, although apparently due to T. evansi, might, in
reality, be due to some other undetermined species of "small" trypanosomes
analogous to one of the species of the congolense group of trypanosomes. It would
be useful to remember this possibility in dealing with outbreaks or sporadic cases
of " surra " in India, and to endeavour definitely to establish the identity of the
causative parasite whenever cases are encountered which appear markedly
resistant to the prophylactic or therapeutic action of " Bayer 205 ".
RECOMMENDATIONS, WITH A DISCUSSION ON
PROPHYLAXIS AGAINST EQUINE SURRA BY ME
A perusal of the available literature shows that few workers have made actual
recommendations in regard to the employment of "Bayer 205 " for prophylactic
PROPHYLAXIS AGAINST EQUINE SURRA
291
purposes under field conditions, although the records of their own work, as
summarized above, provide useful indications as to the manner in which the drug
might be successfully used for such purposes.
For prophylaxis against dourine, Pfeiler (cited by Ruppert and Edwards),
recommends two doses of 3 grammes each, to be administered at an interval of eight
days during the covering season.
For Mal do Caderas, Migone and Osuna [ 1922 ] recommend a dose of 2
grammes of " Bayer 205 " to be given in physiological salt solution.
Writing about T. venezuelense infection which is reported to kill hundreds of
horses every year in Venezuela, Tejera [ 1924 ] recommends that for prophylaxis
against this disease, 2 grammes of "Bayer 205'' should be injected every six
months.
For prophylaxis against the form of camel trypanosomiasis known as
"su-auru", Emelin and Zeiss [1928] employed a dose of 5 to 10 grammes of
" Bayer 205 ", and they consider doses above 10 grammes as unnecessary. It is of
interest that for curative purposes they employed only 4 grammes, the drug being
injected into the large abdominal vein.
Coming to surra, Yakimoff and his collaborators (loc. cit.) consider that for
prophylaxis against the disease in camels in the Ural Region during the summer
period, a dose of 6 grammes of " Bayer 205 " should be effective.
Rodenwaldt and Douwes [ 1922 ], speaking of equine surra, recommend that
all animals " which are suspected of being infected " with surra should receive a
prophylactic injection of 1 gramme for every 150 kilogramme body weight, this dose
being repeated every 4 weeks, for the authors believe that by injection of non-toxic
doses of the drug during the first days of the incubation period of surra, it is
possible to inhibit outbreaks of the disease. According to Kligler and Weitzman
[ 1924 ], however, the injection of the drug during incubation cannot be depended
upon for preventing the disease from developing.
According to Bakker [ 1925 ], all horses in the district should be injected with
1 gramme of " Bayer 205" when surra appears amongst these animals and
horses from outside the district should not be admitted for a time unless they also
are inoculated. A single injection should protect for a month and the injected
animals could still be used, so that ordinary business did not suffer.
The results of experiments carried out by Edwards [1926] in India (ante) appeared
to him to warrant the conclusion that " for horses continually exposed to the risk of
natural infection in surra districts the prophylactic administration of the drug
intravenously at the rate of 1 gramme per 1,000 lb. body weight at intervals of a
D
292 THE INDIAN JOURNAL OF VETERINARY SCIENCE AND ANIMAL HUSBANDRY [ I, IV
fortnight during the period of risk should be capable of warding off the disease ".
More recently, Allen [ 1930 ], in India, has also made a recommendation which is
essentially to the same effect, namely, that for prophylaxis against equine surra,
the drug should be used at the rate of 10 c. c. of a 10 per cent. solution per 1,000 lb.
body weight.
In the rationale of successful prophylaxis, it would seem necessary that the drug
should be maintained in a sufficiently trypanocidal concentration in the circulation
for the desired period. This brings us to the much disputed question of the
mechanism of the trypanocidal action of " Bayer 205 ". In regard to this, Kligler
and Weitzman [ 1924-25 ] would appear to have adduced sufficiently convincing
evidence (based on the results of experiments mentioned earlier in this paper) in
support of the contention that the drug confers protection so long as it remains in a
concentration sufficient to affect the parasites, so that it would seem unnecessary,
as some workers have done, to invoke the aid of certain hypothetical substances
produced, as they believe, as a result of the interaction between the drug on the one
hand and the tissues of the host on the other. It has been urged in support of the
latter contention that the drug behaves very differently when tested against the
parasites in vitro and in vivo. As to this, Kligler and Weitzman [ 1925 ] have
demonstrated that such differences are more apparent than real: if the amount of
dilution undergone by the drug when introduced into the circulating blood is taken
into consideration, then " the effective dose in vivo " will be found to " correspond
fairly well with those in vitro " so that, according to these authors, the rational
conclusion is that " the therapeutic property of ' Bayer 205' is due to a direct
injury to the trypanosomes which renders them avirulent for the host and thus
readily destroyed and eliminated ".
The findings of Kligler and Weitzman are of some practical importance in that
they point to the possibility that, within certain limits, the dosage of " Bayer 205 "
might be increased to lengthen the intervals between two successive injections, and
this would, obviously, be a distinct advantage for field workers having to treat
prophylactically a large number of animals within a brief space of time. As a
matter of fact, the recent findings of Launoy and his co-workers [ 1930] in regard
to the prophylactic action of " Bayer 205 " against T. brucei infection would seem
to bring such a possibility within the bounds of practical realization, for these
workers found that when "Bayer 205" was used protectively on cats in doses
ranging from 0.0025 to 0.04 gramme, the period of resistance to infection increased
with the dose of the drug used, but not in arithmetical progression. In the case
of mice, the drug was used prophylactically in doses varying from 0.0001 gramme
to 0.0006 gramme, and within this range the protective action increased in a regular
PROPHYLAXIS AGAINST EQUINE SURRA
293
manner with the dose, each increase of 0.0901 gramme increasing the period of
resistance by about 5 days, although with large doses the progression was not so
regular.
Reference has already been made to the observations of Berg [1925 ] pointing
to the fact that "Bayer 205 ", when used alone, is valueless as a prophylactic agent
against T congolense infection. Berg, however, records having obtained satisfactory
results from the combined use of " Bayer 205 " and tartar emetic when tested upon
cattle exposed to natural infection in fly belts in South Africa. The animals were
injected in doses ranging from 2.5 to 10 grammes of " Bayer 205 " and with 1 to
1.5 grammes of tartar emetic, the two drugs being mixed and injected mainly
("in der Hauptsache")by the intravenous route. Perhaps it would be useful to
indicate here the actual method of preparing the two drugs for injection. One
gramme of tartar emetic is added to 40 c. c. of water in a clean flask, and at the
same time, the appropriate quantity of " Bayer 205 " is weighed and kept apart.
The mouth of the flask is closed with a piece of linen or wadding and the flask is
now placed in a vessel of cold water which is brought to the boil over fire. When
the mixture has been thus sterilized' for ½ hour, the flask is removed from the
jacket of water, the contents cooled to 60° C. and the previously weighed quantity
of " Bayer 205 " added to the solution. The results obtained from the prophylactic
use of the two drugs singly and in combination were as follows :—Out of 18 animals
injected with "Bayer 205" alone, 6 (or 33.5 per cent.) escaped infection; 13
animals received the combined injection, and of these 9 (or 69 per cent.) escaped
infection; of 3 animals treated with tartar emetic alone, 1 (or 33.5 per cent.) was
protected. 16 animals were used as controls, and all of these became infected. It
would, however, seem difficult to find any a priori explanation for the superiority
of the combined drugs over " Bayer 205 " alone, although, as suggested by Collier
[ 1926 ], it is possible that a " globulin Bayer" compound is formed when " Bayer
205 " is introduced into the circulating blood and that this compound is changed
by the addition of antimony. The method of treating with the combined drugs
would seem to merit trial under field conditions in India, since, if a combination
of the two drugs proves to be really superior to " Bayer 205 " alone, then the
former should doubtless be preferred in view of the rather prohibitive cost of
"Bayer 205".
REFERENCES.
Allen (1930). Observations on an Outbreak of Equine Surra with special Reference to Vectors,
and Means for Control. Vet. Journ., 86, 51-63.
Bakker (1925). Notes on Surra in Padang Sidempoean. Ned. Ind. Bladen V. Diergeneesk en
Dierenteelt, 37, 153-177. (Abstr. in Trop. Vet. Bull, 13, 76-78 ; 1925.)
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PROPHYLAXIS AGAINST EQUINE SURRA
295
Schmidt, & de Oliveira (1924). Mal do Cadeiras and scine Behandlung mit " Bayer 205 ". Arch. f.
schiffs-u. Tropen-Hyg., 28, 91-100. (Abstr. in Trop. Vet. Bull., 12, 113-114; 1924.)
Tejera (1924). Investigations in the Treatment of Horses affected with Trypanosomiasis in
Venezuela by " Bayer 205 ". Gac. Méd. de Caracas, 31, 177-180. (Abstr. in Trop. Vet. Bull.,
12, 114; 1924.)
Van Saceghem (1925). Le Traitement des trypanosomiases animales avec le 205 Bayer (Naganol).
Ann. Soc. Belge. Med. Trop., 5, 101-107. (Abstr. in Trop. Vet. Bull., 14, 50; 1926.)
Yakimov, and others (1927). An experiment on the Prophylactic Use of Naganol (" Bayer
205 ") in Camel Trypanosomiasis. Russian Journ. Trop. Med., 5, 43-46. (Abstr. in Trop. Vet.
Bull, 15, 97; 1927.)
Yakinoff, Amanchuloff, Arbuzoff, Schuravleff, and Clenomorskoff (1927). Versuch den prophylac¬
tischen Anwendung von Naganol (''Bayer 205") bei Trypanosomose der Kamele. Arch.f.
Schiffs—u. Trop. Hyg., 31, 536-541.
THE DIAGNOSIS OF "REDWATER" (PIROPLASMOSIS) IN
INDIAN CATTLE.*
BY
HUGH COOPER, M.R.C.V.S., I.V.S.,
Pathologist,
AND
P. R. KRISHNA IYER, G.M.V.C.,
Veterinary Inspector, Imperial Institute of Veterinary Research, Muktesar.
(Received for publication on 30th July 1931.)
In two articles contributed from this Institute to another Journal † a brief
account was given of the group of diseases known as the " Piroplasmoses " which
affect cattle in India. The object of the present article is to discuss the difficulties
that are not infrequently encountered in the diagnosis of the form of bovine piro-
plasmosis known as Tropical Redwater, caused by the protozoan parasite,
Babesia bigemina.
Tropical Redwater is a tick-borne disease and the transmitting tick in India is
believed to be Boophilus australis. The term " redwater " owes its origin to the
fact that the most striking symptom of the disease is the passing of red-coloured
urine (hæmoglobinuria) resulting from the destruction of red blood-corpuscles with
the consequent liberation of hæmoglobin. In countries, such as India, where the
disease is indigenous and widespread, cattle become infected as calves, at which age
they possess a high degree of natural resistance and thus acquire immunity by
virtue of becoming " carriers " of the parasite in a latent state of activity for the
rest of their lives. Clinical cases of acute and severe piroplasmosis as a primary
affection in adults occur in cattle imported from countries where the disease is
unknown, whilst in indigenous cattle the condition indicates an escape from infec-
tion early in life. The presence of these parasites in " carrier " animals, although
* Paper read at the 17th Indian Science Congress, Allahabad, 1930.
† 1. " Tick-borne Disease with some Remarks on the Diseases of Cattle caused by Protozoa ", by
Hugh Cooper, Agri. Jour. India, Vol. XXI, pp. 95—100 ; March 1926.
2. " The Piroplasmoses of Cattle in India", by Hugh Cooper, Agri. Jour. India, Vol. XXI,
pp. 313—317 ; July 1926.
( 296 )
DIAGNOSIS OF " REDWATER " (PIROPLASMOSIS)
297
ordinarily not attended with untoward symptoms, must be regarded as a source of
potential risk to them in view of the possibility of the resuscitation of these
organisms through the effects of intercurrent disease conditions, notably rinderpest,
or when the vitality of the host is depressed by some cause. Accounts of the
disease in technical literature are mainly based upon experience in other countries
and they only apply to India in the comparatively rare cases of primary infection
of highly susceptible adult cattle as mentioned above. As may be expected from
what has been said, in " carrier " animals the disease may manifest itself in varying
degrees of virulence, from the acute and severe form down to a mild resuscitation
of the piroplasms resulting in no more than a transient fever. The great variability
in clinical symptoms observed in the disease in India makes its detection frequently
difficult in practice, and since, as will be explained later, the existing methods of
diagnosis are far from adequate, it would seem probable that many true cases of
"redwater " have escaped the notice of workers in the field in the past.
The existing methods for the diagnosis of the disease are briefly as follows :—
I. IN LIVING ANIMALS.
1. By Microscopical examination of blood smears.—This is by far the best and
the only certain method of diagnosis. In properly stained films the piroplasms are
found inside red blood-corpuscles as two pear-shaped bodies joined at their tapering
extremities at a somewhat acute angle. Round forms, shaped in the form of signet
rings, are also not uncommon. Some amœbic forms with centrally placed chromatin
are also met with, whilst in very severe infections a few parasites are not infre-
quently seen which are free and outside the corpuscles. Recent researches carried
out at Muktesar have disclosed certain striking variations in the form of these
organisms in smears made post-mortem from animals dead of "redwater ". The
extent of modification in form undergone by the pear-shaped organisms is at times
so profound as to make it almost impossible to recognise them as piroplasms. In
the process of their disappearance the first change noticed is that the individuals
forming a pair get closer to each other at their broad ends, so as to become almost
parallel. They then contract in breadth and become much narrower than normal.
Later they become ovoid and finally circular, when they appear only as central
specks of chromatin surrounded by a thin halo of cytoplasm measuring not more
than a micron. These morphological changes were observed in their sequence in
a series of blood smears taken at two-hourly intervals from animals showing
numerous piroplasms, the examination of the smears having been started about
twelve hours prior to the death of the aninal and continued till about eighty hours
after death had occurred. The final stages of the parasite bear a striking resem-