NationalLibraryOfScotland/medical-history-of-british-india huggingface dataset:
37
As soon as tetanus symptoms set in, both were taken out of the cold chamber
and kept at room temperature, when the disease became chronic, and remained
localised to the right leg (seat of spores) and side, and was severe for three weeks,
when both began to improve, and eventually recovered.
The control guinea pig inoculated with spores only, and kept at room tem-
perature remained well ; and another control inoculated with ¼ c.c. of the tetanus
culture from which the spores were prepared developed tetanus on the second day,
and died on the third day.
It is evident from these two experiments that cold has an influence in pro-
ducing conditions favourable for infection with "washed tetanus spores," and that
this influence is greater when quinine is given in addition. One would infer from
these experiments that it is possible for a latent infection to be stirred into activity
by cold or other depressing influences.
Table VI.
Experiments to test the influence of Cold on Guinea Pigs in the production of Tetanus
after the injection of "Washed Tetanus Spores."
No. of
experiment.
Date of
injection of
spores.
Time of
exposure
to cold.
Whether
quinine was
given, and if
so the amount.
Result.
REMARKS.
1
9th April
1910, right
hind ley.
9th to 12th
April, kept in
cold chamber
at about 38º
F.
No quinine . Tetanus, 12th April,
three days after
spores.
Severe and confined to
right leg and right side
of body. Lasted for
three weeks, when it gra-
dually cleared away.
2
Ditto
. 9th to 11th
April kept in
cold chamber
at about 38º
F.
½ gr. only, on
front of chest,
9th April.
Tetanus, 11th April,
two days after
spores.
Ditto ditto.
3 (Control of
spores.)
Ditto
. Not exposed
to cold.
No quinine . Remained well.
4 (Control of
tetanus cul-
ture from
which
spores
were pre-
pared.)
0.25 c.c. teta-
nus culture,
9th April.
Ditto
. Ditto
. Tetanus, 11th April.
Died, 12th April.
Symptoms local at outset,
then became general.
In No. 1 experiment, cold diminished the animal's powers of resistance to such an extent that "washed tetanus
spores" were able to grow and produce infection ; and in No. 2 experiment the ½ gr. of quinine hastened the
influence of cold by one day.
THE 7TH SERIES OF EXPERIMENTS.
This series was primarily intended to demonstrate the favouring influence of
quinine in producing tetanus in guinea pigs, when given hypodermically on front
of the chest, and for two days before washed tetanus spores were given hypoder-
mically in the hind leg. Owing to the accidental contamination of the culture
from which the spores were prepared the series proves nothing so far as quinine is
concerned, as the two control animals contracted tetanus the same time as the
four experimental ones.
38
The control anærobic cultivation made from the washed spores, in addition to
growing tetanus bacilli, grew another spore-bearing bacillus resembling malig-
nant œdema ; and a similar bacillus (in addition to tetanus bacilli) was isolated
from one of the controls and also from one of the experimental animals after death.
The results of this series are given as an illustration of the necessity of using
pure "washed spores," and the importance of never omitting control experiments.
The series also serves another purpose, viz., as an example of the fact that "washed
tetanus spores" mixed with other spores produce tetanus, when pure "washed
tetanus spores" would fail to do so.
Table VII.
The results of this series of experiments on Guinea Pigs are given as an illustration of
the importance of using pure cultures only when experimenting with "Washed
Tetanus Spores," and the necessity for control experiments. The culture used had
got accidentally contaminated with another spore-forming bacillus.
No. of
experiment.
Date of
injection of
spores.
Date of
injection of
quinine.
Quantity of
quinine
injected.
Results.
REMARKS.
1
25th February
1910.
23rd to 25th
February.
½ gr. daily for
two days.
Tetanus, 26th Febru-
ary. Died, 27th
February.
This series proves nothing
so far as quinine is con-
cerned, owing to the fact
that the two controls
contracted tetanus. In
all seven experiments
there was local swelling at
the seat of injection of
spores, and bacilli resem-
bling malignant œdema
present in stained speci-
mens. Both these and
tetanus bacilli were isola-
ted by cultivations from
two cases (one control
and one experiment).
2
Ditto
.
Ditto
.
Ditto
.
Ditto ditto.
3
Ditto
.
Ditto
.
Ditto
.
Ditto ditto.
4
Ditto
.
Ditto
.
Ditto
.
Ditto ditto.
5 and 6 (Con-
trols to
spores.)
25th February
1910.
Nil.
Nil.
Tetanus, 26th Feb-
ruary. Died, 27th
February.
Both controls contracted
tetanus, which rendered
the series of experiments
useless.
7 (Control of
culture
from which
spores
were pre-
pared.)
¼ c.c. tetanus
culture, 25th
February.
Nil.
Nil.
Ditto ditto . Symptoms very acute, and
disease proved rapidly
fatal.
A cultivation made from the spores used for this series after heating for five minutes at a temperature of 75°C.
grew a mixture of tetanus and other spore-forming bacilli. The series proves that tetanus spores mixed with
other spores produce tetanus.
39
THE 8TH SERIES OF EXPERIMENTS.
The object of this series of experiments was to follow up the information,
gained from experiment 2, Table III (3rd series), in which tetanus bacilli were iso-
lated from the site of a quinine injection on front of the chest following the inocu-
lation of "washed tetanus spores" in the hind leg.
A tetanus culture was used for this series, and not "washed spores," as it
was necessary to produce tetanus without fail, in order to demonstrate whether
tetanus bacilli can be conveyed from the seat of infection on a distant part of the
body, such as the leg, to another part where quinine had been injected, such as the
front of the chest.
Seven guinea pigs were inoculated subcutaneously in the right hind leg
w i t h ¼ c . c . o f a p u r e t e t a n u s c u l t u r e , a n d o n t h e s e c o n d , d a y w h e n l o c a l t e t a n u s
was well marked in all, six of them were given ½ grain quinine subcutaneously
on the front of the chest. Soon after the quinine was given the disease became
severe and general, and they all died from generalised tetanus two days after-
wards.
The control animal which received the same amount of tetanus culture, but
no quinine, also died on the same day, but later on in the evening.
Immediately after death, anærobic cultivations were made from the subcuta-
neous tissues at the sites of injection of tetanus cultures and, quinine respectively;
and after incubating at a temperature 37° C. for a week, a good growth of tetanus
bacilli, showing well formed spores was present in each of the tubes from the seat
of injection of the tetanus culture in the leg, and in five out of six from the seat of
injection of quinine on front of the chest.
Every precaution was taken to sterilise the surface of the skin before cutting
into it to remove a small piece of subcutaneous tissue for inseminating the broth
tubes, and, separate sterilised instruments were used for the chest and leg, so there
was no possiblity of carrying over contaminations from one site to the other when
making the cultivations.
In five out of six experiments, infection was conveyed from the original site
of infection in the leg to the seat of quinine injection on the front of chest.
When these results are interpreted, it means that quinine given hypoder-
mically in the early stages of a tetanus infection starts a fresh focus of infection,
and a focus from which tetanus bacilli (which have not been injected into it) can
be isolated.
The question arises how did infection get conveyed from the leg to the chest
in these cases ? One can only surmise that early in a tetanus infection, the effi-
ciency of the phagocytes is severely taxed, and that some of them re-enter the cir-
culation carrying tetanus spores, and before they have been able to deal success-
fully with these spores they get stranded at the site where the quinine had been
40
injected, where they find dead tissue under anærobic conditions, the most suitable
conditions for the germination of tetanus spores.
A spore infected phagocyte entering the circulation would not be likely to get
stranded in healthy living tissue, and if it did, it would not find conditions
suitable for the spores to develop, and its phagocytic action would not be inter-
fered with.
As evidence in favour of this might be adduced the fact that after death in
cases of tetanus the bacilli are not found in anærobic cultivations made from
the internal organs or heart's blood. (See Tables IX, X, XII, and XIV.)
Table VIII.
Experiments on Guinea Pigs to prove whether hypodermic injections of Quinine in cases
of Tetanus infection produce local conditions suitable for the growth of Tetanus
bacilli .
No. of
experi-
ment.
Date and seat o f i n j e c t i o n
of tetanus
culture.
Date and seat
of injection
of quinine.
Result.
RESULT OF ANÆROBIC CULTI-
VATIONS MADE AFTER DEATH
FROM
REMARKS.
Seat of injec-
tion of
tetanus culture.
Seat of injec-
tion of
quinine.
1
25th August
1910, right
hind leg.
27th August
1910, ½ gr.,
chest.
Tetanus, 27th
August. Died,
29th August.
Growth of
tetanus bacil-
li.
Growth of
tetanus bacil-
li.
Tetanus local a t
first ; then became
severe and general
after quinine was
given. Infection
was conveyed from
the hind leg to
seat of quinine in-
jection on front of
chest.
2
Ditto
.
Ditto
.
Ditto
.
Ditto
.
Ditto
.
Ditto.
3
Ditto
.
Ditto
.
Ditto
.
Ditto
.
Ditto
.
Ditto.
4
Ditto
.
Ditto
.
Ditto
.
Ditto
.
Ditto
.
Ditto.
5
Ditto
.
Ditto
.
Ditto
.
Ditto
.
Ditto
.
Ditto.
6
Ditto
.
Ditto
.
Ditto
.
Ditto
. No growth of
tetanus bacil-
li.
Symptoms same as
experiments Nos.
1 to 5.
7 ( C o n t r o l
of teta-
nus
culture
used.)
Ditto
.
Nil.
Ditto
.
Ditto
.
..
T e t a n u s l o c a l a t
outset ; then be-
came general.
In all these experiments the tetanus bacillus was recovered from the seat of injection of culture (hind leg) and
in out of five of the six cases in which quinine was given, tetanus bacilli were also isolated from the seat of injection
of the quinine (chest). The failure to isolate tetanus bacilli from the seat of quinine injection in No. 6 experi-
ment was very probably due to the fact that a hot glass rod was accidentally used to push down into a broth tube
the small piece of subcutaneous tissue used to inseminate it. The tetanus bacilli isolated from the seat of in-
jection of quinine were virulent for guinea pigs.
42
THE 10TH SERIES OF EXPERIMENTS.
The object of this series of experiments was to demonstrate what effects (if
any) quinine, morphia, lactic acid, and normal saline solution have in preparing
suitable local foci for the development of "washed tetanus spores," when either
of these materials are given subcutaneously, and followed 30 minutes later by the
injection of "washed tetanus spores" into the same site.
Six guinea pigs were taken and treated as follows:
—
1. Two received ½ gr. quinine disolved in ½ c.c. normal saline solution.
2. Two others received 1/20 grain morphia in ½ c.c. normal saline
solution.
3. One received ½ c.c. of a weak dilution of lactic acid (dilution made with
normal saline solution).
4. One control guinea pig received ½ c.c. normal saline solution.
The injections to these six guinea pigs were given subcutaneously in the
right hind leg. When 30 minutes had elapsed to allow for absorption to have
taken place, each received into the site of the previous injection ¼ c.c. of "washed
tetanus spores," and with the following results:
—
The two animals which received quinine developed acute tetanus on the
third day, and died next day.
The animal which received lactic acid developed acute tetanus on the
second day, and died next day.
The two animals which received morphia, and the one which received
normal saline solution remained well.
These experiments show that quinine and lactic acid, when injected hypoder-
mically produce sites suitable for the development of tetanus spores should they
ever reach these sites ; and that morphia and non-irritating solutions of normal
saline do not have this effect.
On the death of the three animals which received quinine and lactic acid,
anærobic cultivations were made from the subcutaneous tissues at the site of
injections in the leg, and also from the liver and the spleen, with the result that
tetanus bacilli were isolated from the leg in all three cases, but not from the
liver and spleen—a further proof that infection is not found in the internal healthy
organs in cases of acute tetanus.
43
Table X.
Experiments on Guinea Pigs to prove the influence of Quinine and Lactic Acid in
producing Tetanus, as contrasted with morphia and normal saline solution, when
" Washed Tetanus Spores " are used ; the spores being injected into the same site
as the quinine, lactic acid, morphia, or normal saline solution, but 30 minutes later.
No. of
experi- ment.
Date and seat of
injection of qui-
nine, or lactic
acid, or morphia,
or normal saline
solution.
Date and seat of
injection of spores.
Result.
RESULT OF ANÆROBI O
CULTIVATIONS MADE AFTER
DEATH FROM
REMARKS.
Seat of
inoculation. Liver. Spleen.
1 29th August
1910, ½ gr.
quinine, right
hind leg.
29th August
1910, into same
site as quinine,
but 30 minutes
later.
Tetanus, 1st
September. Died, 2nd
September.
Growth of
tetanus bacilli.
Negative Negative Quinine, and lactic acid, when
injected hypodermically,
produce sites favourable for
the development of " washed
tetanus spores " ; but nor-
mal saline solution and
morphia do not produce any
local favourable influence.
The quinine solution and
lactic acid did not produce
tetanus by dissolving the
tetanus spores, and liberat-
ing the toxine which they
contained. The 30 minutes'
time allowed for absorption
and the growth of tetanus
bacilli from the seat of
injection would go against
this view. In experiments
1, 2, and 3, although the dis-
ease was acute and rapid,
the infection remained local,
and the internal organs were
free from tetanus bacilli.
2
Ditto
. Ditto
. Ditto . Ditto . Ditto . Ditto .
3 29th August
1910, 1 c.c.
weak lactic
acid, right hind
leg.
29th August
1910, into same
site as lactic
acid, but 30
minutes later.
Tetanus, 31st
August. Died, 1st
September.
Ditto . Di to . Ditto .
4 29th August
1910, 1/20 gr.
morphia, right
hind leg.
29th August
1910, into same
site as morphia,
but 30 minutes
later.
Remained well.
5
Ditto
. Ditto
. Ditto.
6 (Control of the
pores used.)
29th August
1910, 1 c.c.
normal, saline
solution, right
hind leg.
29th August
1910, into same
site as normal
saline soultion,
but 30 minutes
later.
Ditto.
Quinine and lactic acid, when injected hypodermically, prepare sites favourable for the development of tetanus spores, but mor-
phia and normal saline solution do not produce this effect.
THE 11TH SERIES OF EXPERIMENTS.
This series of experiments was devised with the object of proving why it is
that tetanus so rarely follows hypodermic injections other than those of quinine,
due allowance being made for those exceptional cases in which tetanus germs were
inoculated along with the injected material.
Morphia is perhaps more often given as a hypodermic than any other drug,
and yet it is the rarest exception possible ever to hear of tetanus following its use.
The same may be said with regard to bacterial vaccines, sera, and such remedies,
which are always given hypodermically, and yet one seldom, if ever, hears of a
case of tetanus following their use, although they are now given all over the
world as a routine measure in the treatment of numerous human diseases, and
doubtless by many who have hazy notions as to what asepticism really means.
44
The two circumstances above all others which make quinine a more dangerous
weapon in the production of tetanus when given hypodermically are
-
(1) The local destruction of tissue at the seat of injection.
(2) The possible paralysing effect on phagocytic activity when large doses
are given.
The local destruction of tissue follows as a matter of course in all cases in
which quinine is given hypodermically ; and the paralysing effect on phagocytes
is a possibility when the drug is pushed and the patient's resistance lowered
by the effects of malaria, a high temperature, or other influences.
In continuation of the line of reasoning suggested by the results of the
experiments detailed in Table X, and with the object of testing experimentally
the effects of " washed tetanus spores " in the production of tetanus when mixed
with quinine and morphia respectively, and injected hypodermically, the following
experiments were carried out. Eight healthy guinea pigs of approximately the
same age and size were selected. Three were given ½ grain quinine dissolved in
c.c. normal saline solution, and mixed with ¼ c.c. " washed tetanus spores."
Three others were gi ven 1/20 grai n morphi a i n ½ c.c. normal sal i ne sol uti on, and
mi xed wi th ¼ c.c. " washed tetanus spores."
One was given ¼ c.c. " washed tetanus spores " only, and the eighth guinea
pig was given ¼ c.c. of the tetanus culture from which the spores were prepared.
The injections were in all cases given hypodermically in the hind leg, and with
the following results
:—
(1) The three animals which received the spores mixed with quinine all
developed tetanus ; one on the second day, one on the third day, and
the remaining one on the fourth day ; and death took place on the
fourth, ninth, and tenth days respectively after receiving the
injection.
After death anærobic cultivations were made from the sites of injection
of quinine mixed with spores, and tetanus bacilli were isolated in all
three cases.
(2) The three animals which received morphia mixed with spores remained
well, and not the slightest trace of local tetanus showed itself at any
time.
(3) The control animal which received " washed spores" only remained
well.
(4) The other control which received ¼ c.c. of the culture from which the
spores were prepared, contracted tetanus on the second day and
died next day.
It is evident from the results of this series of experiments that the chances of
contracting tetanus when tetanus spores are given hypodermically in a quinine
45
solution are much greater than when the spores are given with a hypodermic injec-
tion of morphia.
Table XI.
Experiments to test the effect of "Washed Tetanus Spores" when mixed with Morphia
or Quinine and injected into Guinea Pigs.
No. of
experiment.
Date of injection of mix-
ture of morphia, or
quinine and "washed
tetanus spores."
Seat of
injection.
Result.
REMARKS.
1
16th September 1910,
1/20 gr. morphia mixed
with washed tetanus
spores.
Right hind leg . Remained well.
2
Ditto ditto
. Ditto
.
Ditto
3
Ditto ditto
.
Ditto
.
Ditto.
4
16th September 1910, ½
gr. quinine mixed with
washed tetanus spores.
Ditto
. Tetanus, 18th September.
Died, 20th September.
An anærobic cultivation
made from seat of in-
jection of quinine
mixed with spores
gave a growth of
tetanus bacilli.
5
Ditto ditto
.
Ditto
. Tetanus, 20th September.
Died, 26th September
Ditto ditto.
6
Ditto ditto
.
Ditto
. Tetanus, 19th September.
Died, 26th September.
Ditto ditto.
7 (Control of
spores
used.)
16th September, "washed
tetanus spores" only.
Ditto
. Remained well.
8 (Control of
culture
from which
spores
were pre-
pared.)
16th September, ¼ c.c.
tetanus culture only.
Ditto
. Tetanus, 17th September.
Died, 18th September.
Severe and rapid ; teta-
nus local at outset ;
then became general.
This series of experiments demonstrates that tetanus spores when mixed with quinine and injected into guinea
pigs produce tetanus, but when mixed with morphia do not produce tetanus.
THE 12TH SERIES OF EXPERIMENTS.
As all the foregoing experiments with quinine and tetanus spores had been
carried out on guinea pigs, it was deemed advisable to control a few of them by
similar experiments on monkeys.
The common brown monkey found in the hills in the neighbourhood of Kasauli
was used. The animals were full grown weighing about 12 to 14 lbs., in good condi-
tion, and had been in captivity for over five months.
This series included five monkeys treated as follows:
—
No. 1 Monkey. —Quinine was commenced at the same time that spores were
given.
On September 2nd, ½ c.c. "washed tetanus spores" were given hypodermi-
cally in the right hind leg, and three grains quinine hypodermically on front of chest.
The quinine was again repeated on the 3rd and 8th September, and in 4-grain doses.
46
Tetanus developed on September 19th, but the animal was ill for two days
before this. Lockjaw was present on the 19th September but not severe, and the
animal was able to eat and drink.
On. the 21st, there was difficulty of swallowing and the jaws could only be pressed
open a little and with difficulty.
The symptoms advanced slowly and become generalised. On the 26th the
animal was killed with chloroform.
Immediately after death, anærobic cultivations were made from the sub
-cutaneous tissues taken from seat of injection of spores, seat of injection of quinine,
and also from the liver, the spleen, and the heart's blood.
3 c.c. of the heart's blood were also injected subcutaneously into a guinea pig.
A growth of tetanus bacilli was obtained from the seat of injection of spores,
and seat of injection of quinine, but not from the liver, the spleen, or the heart's blood.
The guinea pig inoculated with 3 c.c. of the heart's blood remained well for
a week, when wasting set in, and it died on the 14th day, but showed no symp-
toms of tetanus at any time. An anærobic cultivation made from the site of injec-
tion of this monkey's blood into the guinea pig failed to grow tetanus bacilli.
No. 2 Monkey .—Quinine was commenced one day after spores were given.
On September 2nd, ½ c.c. " washed tetanus spores" was given hypodermi-
cally in the right hind leg.
On September 3rd, 4th and 8th, 4 grains quinine were given hypodermically
on front of the chest.
Tetanus developed on September 17th, and on that date there was slight
stiffness of the muscles of the neck and the jaws. On the 19th, lockjaw was well
marked, and there was great difficulty of swallowing, and the animal died next day
from acute generalised tetanus.
Immediately after death, anærobic cultivations were made from subcutane
- o u s t i s s u e t a k e n f r o m t h e s e a t o f i n j e c t i o n o f s p o r e s , a n d f r o m t h e s e a t o f i n j e c t i o n
of quinine, and also from the liver and spleen.
A growth of tetanus bacilli was obtained from the seat of injection of spores,
and seat of injection of quinine, but not from the liver or the spleen.
No. 3 Monkey.—Quinine was commenced one day before spores were given.
On September 2nd, 3 grains quinine were given hypodermically on the front
of the chest and next day ½ c.c. " washed tetanus spores " were given hypoder
- m i c a l l y i n t h e r i g h t h i n d l e g , a n d a t t h e s a m e t i m e 4 g r a i n s q u i n i n e o n f r o n t o f
chest. On September 8th it received another 4 grains quinine on front of chest.
Tetanus developed on September 15th, slight at first, but more marked on the 16th
when lockjaw was present.
On the 17th generalised tetanus was present, and the animal lay in its cage
bent forward (emprosthotonos) with shallow breathing, muscles of arms, legs,