NationalLibraryOfScotland/medical-history-of-british-india huggingface dataset:
7
APP. I. ]
its causes and characteristics.
I have now to point out that all of the so-called Calcutta plague
bacilli may be readily obtained from specimens of the blood of per-
fectly healthy animals under the influence of brief exposure to air,
even in scrupulously clean localities, such as my laboratory is, in
which the air is in no way exposed to any special chances of bacterial
contamination.
The data derived from the following experiment will probably be
sufficient to demonstrate this.
On the nth December 1896 about 30 C.C. of blood were drawn
from the jugular vein of a perfectly healthy fowl, under strictly aseptic
conditions, into two sterilised test tubes, A & B, stoppered, as usual,
with cotton-wool plugs. Half an hour later the plug was removed
from tube A and small quantities of blood were introduced into four
watch glasses exposed to the air for 20 minutes, and then set in an
incubator.
Cultivations from these specimens subsequently yielded growths,
which in all microscopical and macroscopical characters were identi-
cal with—
1st.— The only growth yielded by a cultivation of blood sent
to me by Dr. Cobb, and derived from an alleged case of
plague at Chinsurah.
2nd.—The growth from another type-cultivation obtained from
a man named Feney in the Station Hospital, and sent to
me by the Health Officer of Calcutta as a type-specimen
of plague bacilli.
3rd.—The large putrefactive bacillus which was present in the
original specimen of blood, and in the bouillon cultiva-
tion of the latter, sent to me by the Health Officer of
Calcutta as evidence that a man named Bipin, who died
in Calcutta early in November, had been suffering from
plague.
The tube B containing the other sample of blood was sealed up after
a few days to avoid evaporative loss, and showed no signs of decom-
position up to the 31st December, or three weeks after it had been
taken. Three cultivations of it were then made and yielded no growth
whatever. On the 3rd January portions of the clot were removed,
set in four watch glasses, exposed in the open laboratory for 20
minutes, and then placed in an incubator.
Decomposition rapidly set in in all of them, and they subsequently
yielded growths of the following characters :—
1st.—The large putrefactive bacillus, noted above as occurring
in the Health Officer's specimen of blood, and his culti-
vation of the latter from the case of Bipin.
8
Description of Plague :
[ APP. i.
2nd.—Small cocci and diplococci similar to those in the Health
Officer's cultivation of the same case, but more closely
agreeing in characters with those described as distinc-
tive of the plague bacillus, in that they were completely
bleached by treatment with solutions of iodine accord-
ing to the method of Gram.
3rd.—A schizomycete, which in all characters, macroscopical
and microscopical, was similar to that which was pre-
sent in a cultivation derived from the blood of the man
Cotta in Howrah, and supplied by the Health Officer as
a type-specimen of plague bacilli.
The evidence here is as complete as it well could be. The results in
the case of sample B show that the blood previous to exposure to air
contained no bacterial organisms, and those following even brief ex-
posure of portions of A and B to the chance of contamination equally
clearly demonstrate that various forms of schizomycetes, presenting all
distinctive characters of those alleged by the Health Officer to be
specific plague bacilli, are common aerial contents, and therefore liable
to appear in any cultivations of blood not conducted under strictly
aseptic conditions.
The 7 th February 1897.
No. 3.
Mr Haffkine's instructions for diagnosing plague.
1. Symptoms of disease:
Fever preceded sometimes by rigor. Trembling in the limbs.
Pain, swelling, and hardening of one or rarely several superfi-
cial glands, most frequently of one in the groin, more
rarely in a gland or glands of the armpit or of the cervical
region.
Delirium. Sometimes restless efforts at vomiting which may
or may not be successful ; constipation, or occasionally,
on the contrary, diarrhœa.
Comatose or semicomatose state of patient. Voice weak,
speech incoherent.
2. The most prominent of these symptoms is the affection of
superficial glands ; cases, when this symptom is absent, if there are
any, appear quite exceptionally.
3. In a case of plague a microscopic preparation, made with the
contents of the affected gland, will show numerous diplobacteria of
characteristic shape and size, described for the first time by Kitasato
in Hong-kong.
9
APP. I. ]
its causes and characteristics.
4. To obtain the microbes with certainty a patient is to be select-
ed at the height of the symptoms, with glands largely increased, and
before any injections had been made into the glands. At the com-
mencement of the swelling, or in a convalescent patient, drop with-
drawn from the gland may fail to show the microbes.
5. For making a preparation, an ordinary or a grooved needle, a
spirit lamp, a microscopic slide and 5 per cent, carbolic lotion is to be
prepared. Then proceed as follows :—
Wash the skin just over the swollen gland with carbolic lotion.
Allow the lotion to dry in situ; pass the needle in the flame of a
spirit lamp and leave it to cool down. Steady the gland with the
fingers of left hand, so as to make it quite prominent, with the skin
tightly drawn over it ; and prick the gland through the skin. Press
out a drop of the whitish contents of the gland, withdraw the needle
and burn the part of it which has been introduced under the skin of
the patient, till it is red hot. Apply the slide to the drop of lymph
pressed out of the gland, and spread it as uniformly as possible over
about a half of the surface of the slide, avoiding any contact between
the infective material and the fingers or any other object.
6. Keep the slide in the hand till it is dry, and while holding it in
the hand by its free end, pass it rapidly in the flame five or six times,
with the preparation upwards, till the lower surface of the slide felt by
a finger is too hot to be left in contact with the finger without pain.
Excessive heating or charring will make the preparation unrecog-
nisable.
Disinfect the skin over the gland and apply a suitable dressing if
necessary.
7. In a certain number of cases, pricking the gland through the
skin fails to bring to the surface the characteristic material.
A more certain proceeding is to incise, with aseptic precautions,
the skin over the gland, and the gland itself, and to take directly
from the gland a drop of lymph on the point of a scalpel, spreading
it over a slide as directed above.
8. Another way consists in sucking up a drop from the gland by
means of a sterile Pasteur pipette, protected by a cotton plug.*
For this cauterise a small spot of the skin just over the gland.
Break the fine end of the pipette and pass the thin part of it rapidly
through the flame several times. Allow it to cool down and insert the
pipette into the cauterised part of skin, steadying the gland with the
fingers of left hand. By a boring motion introduce the tip of the
* Messrs. Kemp & Co., Bombay, have been instructed how to prepare such pipettes,
and have promised to sell boxes containing a dozen and half dozen of pipettes at the
cost price of a few annas a box.
b 2
10
Description of Plague :
[ APP. I.
pipette into the gland. The moment when this is reached will be
felt by the tissues offering less resistance to the penetration of
pipette.
Apply the mouth to the open end of pipette, and while constantly
moving the pipette to and fro, suck up a small quantity of fluid. The
best is the thick whitish contents of the gland blood to be avoided by
changing slightly the direction of the pipette or putting it deeper or
more superficially. Withdraw the pipette and blow out a drop of it
on the slide, spreading as above.
The pipette is to be burnt immediately after this, and all objects
used disinfected.
9. The use of a syringe is not advisable.
10. The slide is stained by any of the aniline dyes and examined
with an oil immersion lens, or sent for staining and examination to
the Bubonic Plague Research Committee, Petit Laboratory, Bom-
bay.
11. Any operation should be avoided with glands situated on the
abdomen above Poupart's ligament.
Reports by Mr. E. H. Hankin on experiments to test
the vitality of the plague bacillus.
No. 4.
No. 1600, dated Bombay Castle, the 26th February 1897.
From—A. M. T. JACKSON, Esq., I.C.S., Under Secretary to the Govern-
ment of Bombay, Revenue Department,
To—The Secretary to the Government of India, Home Department.
I am directed to forward, for the information of the Government
of India, copies of two reports, dated the 17th and 19th instant,
furnished by Mr. E. H, Hankin on researches he has made on
various articles of produce as regards their susceptibility to engender
or support the bubonic microbe. It will be seen that in his report,
dated the 19th instant, Mr. Hankin shows very clearly that the
various descriptions of grain and flour usually stored in Bombay con-
tain no trace oí the bubonic microbe, and his other report, dated the
17th idem, proves almost to demonstration that grain purposely in-
fected does not permit of the existence of the microbe for more than
about four to six days.
2. I am to add that the results of Mr. Hankin's researches have
been telegraphed to Dr. Cleghorn at Venice with a request that they
may be communicated to the Sanitary Conference now sitting in that
city. Copies of the reports have also been sent to Dr. Cleghorn.
II
APP. I. ]
its causes and characteristics.
No.5.
Dated Bombay, the 17th February 1897.
From—E. H. HANKIN, Esq., Chemical Examiner and Bacteriologist,
North-Western Provinces and Oudh,
To—The Secretary to the Government of Bombay.
In accordance with Government Order No. 564/107-9, dated 2nd Feb-
ruary, I have the honor to submit the following report on the vitality of
the bubonic plague microbe in the chief grain exports from Bombay.
2. My experiments have been carried out with numerous specimens
of the following articles :—
Linseed.
Yellow rapeseed.
Brown
„
Tilseed or gingelly seed.
Groundnuts.
Castor seeds.
Poppy seeds.
Wheat (new hard red). /
,,
(another lighter variety),
Flour.
3. In order to test whether the microbe was still present in a liv-
ing condition in a specimen of previously infected grain, my method
has been to make an extract of this grain and to inject it into a
mouse. The great susceptibility of mice to bubonic plague and
to infectious diseases in general, while advantageous in one sense,
has been disadvantageous in another, in that the mice might have
died owing to the presence of other microbes than the bubonic in
the grain. In some cases the post mortem appearances have sug-
gested that this has occurred ; but, as it is impossible to give an abso-
lute proof that the bubonic microbe had nothing to do with the death
of the animal in these cases, I have in general regarded the sur-
vival of the mouse as the only adequate proof that the microbe in
question was no longer living in the previously infected grain.
4. My first experiment was carried out with the bubonic plague
microbe derived from cultures on agaragar. Two cultures, each
about a week old, were employed. Of those, one had been derived
from a patient in Bombay, the other had been recently isolated from
a rat found dead of the plague. The growth was scraped off the
surface of the jelly and well-shaken up in 15 cubic centimetres of
bouillon. The mixture was then placed in a burette. A sample of
the grain to be tested, weighing about a kilogram, was placed in a
previously sterilised stoppered foot-glass. One cubic centimetre of
the bouillon emulsion of the microbe was then poured into the foot-
glass, care being taken that it should fall into the centre of the grain.
The stopper was replaced, and the foot-glass was immediately vio-
lently shaken for about a quarter of an hour. At the end of this time
all traces of moisture from the bouillon that had been added had com-
pletely vanished. On shaking, a cloud of dry dust could be seen in the
air and in the foot-glass. In order to avoid inhaling this presumably
12
Description of Plague :
[ APP. I.
infected dust, the next part of the experiment was carried out in the
open air. The grain was poured from the foot-glass into the linen
bag in which I had received the specimen. From the bag a bulk of
about 40 cubic centimetres was poured into a measuring glass that
had been previously sterilised. This small bulk of grain was then
wetted with sterile bouillon and stirred up with a glass rod. About
a half to a quarter of this bouillon was sucked up into a pipette and
injected into a mouse. The white mice employed had generally been
in my possession for some time. Each lived in a separate cage. The
cage before being occupied had in all cases been sterilised in an
autoclave. The pipettes employed for the injections were always
disinfected by boiling in water previous to use. The skin of the
mouse at the point of inoculation was previously to the injection
rubbed with a mixture of carbolic acid, water, and alcohol.
5. The extracts made in this manner were as soon as possible
injected after the infection of the grain into mice with the following
results :—
Mouse treated with extract of linseed ... Died within 24 hours.
yellow rape-
seed
brown rape-
seed
tilseed
...
groundnuts
castor seed
poppy seed
wheat (new,
hard, red)
wheat (of an-
other kind)
flour
72
24
Survived.
Died within 48 hours.
6. Two days after the infection of the grain specimens were
again taken, extracted in exactly the same manner as before and
injected into another series of mice with the following results :—
Mouse treated with extract of linseed ... Died within 7 days.
yellow rape-
seed
brown rape-
seed
tilseed
groundnuts
castor seed
poppy seed
24 hours.
4 days.
Survived.
Died within 11 days.
48 hours.
48 „
13
APP. I. ]
its causes and characteristics.
Mouse treated with extract of wheat (first
variety) ...
wheat (second
variety) ...
flour
Survived.
[
Died within 48 hours.
7. At this time I found out that an extract of castor seeds to
which no bubonic microbes had been added was equally fatal as the
infected seeds to mice, and consequently was obliged to omit this
article from my experiments. The reason of this is that the castor
seeds contain a poison, known as ricin, which is intensely active on
subcutaneous injection. On stirring up the seeds with bouillon, as
described, caused a fatal dose to pass into solution. The mouse
injected with such an extract would certainly die whether or not the
bubonic microbe was present. If I made the extract of the seeds so
rapidly that a fatal dose of the poison would not be dissolved, I
could have no certainty that sufficient time had elapsed to allow the
microbes to pass into suspension in the bouillon.
Four days after the commencement of the experiment, extracts of
the grain were again made as before and injected into mice as
follows :—
Mouse treated with extract of linseed ... Died after 13 days.
yellow rape-
seed
brown
tilseed
...
groundnuts
poppy seed
wheat (first
variety)
wheat (se-
cond va-
riety) ...
flour ...
Survived.
9. Eleven days after the commencement of the experiments,
extracts were again injected. A change in procedure was made in
this case, in that the grains, instead of being merely stirred in a mea-
suring glass with a glass rod in order to produce the suspension in
the bouillon, were slightly ground up in a mortar until the mixture
14
Description of Plague :
[ APP. I.
of bouillon and debris was so thick that I could with difficulty suck
it up into the pipette. The results obtained were as follows :—
Mouse treated with extract of linseed ... Survived.
yellow rape-
seed
brown rape-
seed ... Di
tilseed ,..
groundnuts
poppy seed
wheat (first
variety) ...
wheat (second
variety)
flour ...
ed within 24 hours.
7 days.
24 hours.
24 ,,
24 „
24 ,,
24 ,,
10. Thirteen days after the commencement of the experiment,
extracts were again injected into mice with the following results. In
this as in later experiments the grain was extracted by grinding it in
a mortar with the bouillon :—
Mouse treated with extract of linseed ... Survived.
yellow rape-
seed
brown rape-
seed
tilseed
groundnuts
poppy seed
wheat (first
variety)
wheat (second
variety)
flour
.,,
Died after 24 hours.
Survived.
11. The mouse treated with the yellow rapeseed extract showed
no appearances microscopically or otherwise of having succumbed to
bubonic infection. There was nothing noteworthy at the seat of
inoculation in the back, but there was an extensive blood-stained
œdema on the underside of the neck, but appearances suggesting
that it had died from some accidental infection.
12. The above experiment indicates that the bubonic microbe
derived from pure cultures perishes within thirteen days after being
added to the abovementioned specimens of grain and seeds. The
question now arises whether the microbe would show a greater or
15
APP. I. ]
its causes and characteristics.
lesser degree of resistance when derived from the organs of ani-
mals dead of the disease. A reply to this question is given in the
following experiment.
13. The spleen of a rat and the spleen, liver, and œdema from a
mouse—both animals having died after plague inoculation—were
ground up in a sterile mortar with powdered glass and mixed with 10
cubic centimetres of bouillon. One cubic centimetre of the mixture
was added to half a kilogram of each of the specimens of grain imme-
diately and violently shaken as in the previous experiment. Ex-
tracts were at once injected into mice with the following results :—
Mouse injected with linseed
,,
,,
„ yellow rapeseed ...
„
„
„ brown
„
„
„
tilseed
„
„
„ groundnuts
,,
„
„ poppy seed
,,
,,
,, wheat (second
variety)
„
„
„ flour
Died within 48 hours.
Survived.
Died in 5 days.
,, ,, 6 ,,
,, ,, 6 ,,
Survived.
Died in 4 days.
,, ,, 4 ,,
14. The fact that two mice in this series survived, and that the
others only succumbed after an unusual delay may be due to the fact
that the grains were extracted very rapidly with the bouillon and not
ground up in it as in the preceding and following experiments. Con-
sequently it is probable that fewer microbes than usual became sus-
pended in the bouillon extracts.
15. Two days later, extracts were again made and injected into
mice as follows :—
Mouse treated with extract of linseed
... Died within 24 hours.
yellow
rapeseed ...
brown.
rapeseed
tilseed
...
groundnuts
poppy seed
wheat
flour
24
24
Survived.
Died in 24 hours.
„ ,, 3 days.
Survived.
16. Four days after the commencement of the experiment, extracts
were again made and injected as follows :—
Mouse treated with extract of linseed
... Died in 5 days.
„
„
,,
yellow
rapeseed ... ,, ,, 3 ,,
16
Description of Plague:
[
APP. 1
Mouse treated with extract of brown rapeseed ...
tilseed
groundnuts
poppy seed
wheat
flour
Died in 2
Survived.
days.
17. The above mice that died shewed no appearances under the
microscope or inculturally suspicions of bubonic plague. Their
spleens were injected into three other mice which have remained in
good health.
18. Six days from the commencement of the experiment, extracts
were again injected into mice with the following; results :—
Mouse treated with extract of linseed
yellow
rapeseed
brown rape-
seed
tilseed
groundnuts
poppy seed
wheat
flour
... Survived.
... Survived.
19. Thus, in this experiment grain infected with the organs of
animals dead of the plague lost its infectious power completely within
six days.
20. The remaining point of the enquiry is the resisting power of
the microbes as it exists in secretions of the human patient. For this
purpose I chose the sputum in which the microbe has been shown by
Surgeon-Captain Childe, Professor of Pathology in the Grant Medical
College, to exist in large numbers and in a very virulent condition.
21. Sputum recently obtained from a patient was mixed with an
equal volume of bouillon. One cubic centimetre of the mixture was
immediately added as before to each of the specimens of grain. Owing
to an accident, the linseed could not be used in this experiment.
23. Extracts made immediately were injected into mice with the
following results :—
Mouse treated with extract of yellow rapeseed ... Died within 24 hours.
brown
„
tilseed
groundnuts
poppy seed
wheat
flour
... „
... ,,
... ,,
... ,,
... ,,
... ,,
24
24
24
24
48
24
17
APP. I. ]
its causes and characteristics.
23. Two days later, extracts were again made and injected. The
results were as follows :—
Mouse treated with extract of yellow rapeseed ... Died
brown
„
tilseed
groundnuts
poppy seed
wheat
flour
within 24 hours.
24
24
28
24
48
24
24. Four days after the commencement of the experiment, extracts
were again injected as follows : —
Mouse treated with extract of yellow rapeseed
„
,,
brown
„
„
„
tilseed
,,
,,
groundnuts
„
„
poppy seed
„
„
wheat
,,
„
flour
... Survived.
... Died within 24 hours.
... Survived.
...
„
... Died within 48 hours.
... „ 48
25. Six days after the commencement of the experiment, extracts
were made and injected with the following results :—
Mouse treated with extract of yellow rapeseed ...
brown
,,
tilseed
groundnuts
poppy seed
wheat
flour
...
...
...
...
...
...
Survived.
Died in 3 days.
20. No reason could be obtained from post mortem appearances
or cultures for thinking that the last-mentioned mouse had died of
bubonic plague. Further, the flour had shown no special capacity
for nourishing the bubonic microbe in my other experiments, so that it
would appear to be probable that the bubonic microbe had died out in
this case as in the other. Nevertheless, I am continuing my experi-
ments with flour and hope to make a separate report to you on the
subject in a few days.
27. Earlier experiments carried out with another kind of grain
(bajri) has shown me that the bubonic microbe in this case dies out
within two days.
28. In these experiments I have kept the grain damp, so that
eventually it became covered with mildew. In the experiments now
described, by keeping the grain in a dry condition in bags, I imitated
more closely the conditions that exist in commerce. Under such
b 3