NationalLibraryOfScotland/medical-history-of-british-india huggingface dataset:
Medical Officers of the Army of India.
95
according as the micro-organisms, which are exposed to it, are in great or small
amount and according as they are virulent or attenuated bacteria. It is evident,
however, that inasmuch as a totally different specimen of spirillum blood would
have to be used for each observation, neither the number of spirilla nor their
virulence could be controlled in any series of experiments having as their object
an estimation, from day to day, of the amount of the anti-bacterial substances
present in the blood of a monkey after the crisis. This consideration quite
explains the somewhat anomalous results obtained by Bardach and others in
their attempts to make such an estimation.
We may now pass on to consider whether the theory enunciated above
can by analogy be brought forward as an explanation of the phenomena which
are characteristic of an attack of spirillum fever in the human subject.
The crisis which is seen in man is strictly analogous to that which occurs in
the monkey and subject, therefore, to the same explanation. The researches of
Gabritchewksy12 and Bardach 13 have demontstrated in vitro the presence of anti-
spirillar substances in the blood serum of patients during the whole of the apyrexial
period, viz., at a time when the spirilla are absent from the general blood
stream. Although there is no direct proof, it must be assumed that during this
apyrexial period the spirilla continue to live and multiply in the spleen and other
internal organs.' 4 The relapse, analogous to the second artificial infection of the
monkey, would, therefore, take place only after such time when the anti-bacterial
power of the blood had diminished to such an extent as to allow the spirilla to
again enter and cultivate themselves in that medium. Now inasmuch as the
spirilla continue to live and multiply during the apyrexia in the organism long after
the appearance of these bactericidal substances in the blood, the consideration that
these substances may be inoperative in vivo is brought before us. Such a con-
clusion would only be valid if it could be shown that the spirilla are really living
and cultivating themselves in a medium which contains an amount of bactericidal
substances equal to the quantity present in the circulating blood. For the co
m p l e t e r e s o l u t i o n o f t h i s q u e s t i o n i t i s o b v i o u s t h a t a q u a n t i t a t i v e a n d c o m p a r a t i v e
estimation of the amount of bactericidal substances present in the blood, and in
the organs in which the spirilla still live during the apyrexial period, would be
essential. The conditions necessary for such an investigation are seldom met
with and unfortunately in my experience no such opportunity has occurred.
It has been mentioned above that in the case of the monkey, spirilla are
present in the spleen for a short time after the crisis in a condition capable of
giving rise to infection in a new monkey, and that at this time they have com-
pletely disappeared from the blood. It is apparent, therefore, that some light might Z
96
Scientific Memoirs by
be thrown on the question under consideration by a comparative estimation of
the anti-bacterial substances contained in the splenic serum and in the blood
serum of monkeys immediately after the crisis. It is the result of such an in-
vestigation which I have now to present, and which I shall preface by giving
a short account of the technique employed.
The spleen, removed immediately after the crisis, was pounded in a mortar
with an equal weight of normal saline solution. This was then thrown on a filter
and a clear filtrate obtained. At the same time as the spleen was removed, blood
was collected from the ear into a capsule. This capsule was set aside for an hour
or two till a clear serum had exuded from the clot. The serum was then diluted
with equal parts of normal salt solution. By this means a blood serum of pra
ctically the same dilution as the splenic serum was obtained. For inasmuch as
the solid splenic substance amounts to only one-sixth of the total weight of the
spleen, the slight inaccuracy, which has been admitted into the experiments by
using for comparison blood serum of only one fold dilution, need not be taken
into account.
Both sera thus diluted were then mixed with equal parts of spirillum blood
taken from a monkey and prevented from clotting by the addition of citrate of
soda.15 A hanging drop preparation of each of these mixtures was put up.
The remainder was stored in capillary tubes.
All preparations were kept at the temperature of the laboratory, viz.—26°t
o 30° C.
Observations of the hanging drop preparations were made at various inter-
vals after mixing, while from the capillary tubes specimens were periodically taken,
fixed on a slide, treated with acetic acid so as to dissolve out the hæmoglobin
and then stained with aniline gentian violet. The results obtained are detailed
as follows :—
Experiment 1.—Monkey S-13 suffered from an attack of spirillum fever of
two days' duration. Immediately after the crisis the spleen was removed and a
capsule of blood was collected from the car at the same time.
The spirillum blood used was taken from a monkey on the first day of fever.
This blood contained spirilla in considerable numbers. Preparations were made
in the manner described above and the following observations were recorded:—
( 1 ) Hanging drop preparations—
(a) Serum of spleen.— 30 minutes after mixing : spirilla very motile.
1 hour after mixing : spirilla still very motile.
2 hours after mixing : spirilla still very motile.
2 ½ hours after mixing: spirilla still very motile.
Medical Officers of the Army of India.
97
23 hours after mixing: some spirilla still very motile, others have a
sluggish movement, while some are quite motionless.
(b) Serum of blood.—30 minutes after mixing: some spirilla show slight
motility: many puckered-up, granular motionless forms.
1 hour after mixing: the same appearances as above.
1 ½ hours after mixing: no normal motile spirilla; some granular
forms which have a spasmodic shivering movement.
2 hours after mixing: no motility of any kind: some granular
spirillum- like bodies, but no normal spirilla.
23 hours after mixing: some granular spirillum-like bodies but no
normal spirilla.
(2) Stained preparations
—
(a) Serum of spleen.—
1 hour after mixing: a great number of typical spirilla, which stain evenly
and well.
2 hours after mixing: a great number of isolated spirilla; many small
clumps of spirilla: all spirilla take the stain well and regularly.
23 hours after mixing: a large number of isolated spirilla: several clumps
of spirilla, some large, some small: the spirilla generally take the
stain well, but there are some which stain very faintly.
(b) Serum of blood.—
1 hour after mixing: no typical normal spirilla: a number of dotted, gran
ular spirillum-like bodies which stain faintly and irregularly.
2 hours after mixing: some granular masses which, however, contain no
spiral forms: a few fine short badly stained bodies which might be
degenerated spirilla.
2 ½ hours after mixing: absolutely no appearance of spirilla or their re-
mains.
Experiment 2.—Monkey S-14 had a typical attack of spirillum fever of
2 days' duration. Shortly after the crisis the spleen was removed and at the
same time a sample of blood from the ear was collected in a blood capsule.
The spirillum blood used was taken from a monkey on the second day of fever.
This blood contained a very large number of spirilla.
In preparations made in the manner described above the following observa-
tions were recorded:—
(1) Hanging drop preparations—
(a) Serum of spleen.—
40 minutes after mixing: spirilla plentiful and very motile: no clumps.
Scientific Memoirs by
1½ hours after mixing: isolated actively motile spirilla: a few clumps
each made up of a small number of spirilla: no degenerate forms.
2½ hours after mixing: isolated actively motile spirilla: some clumps, in
which spirilla are still motile.
22 hours after mixing: no motile spirilla: a number of isolated motionless
spirilla: many clumps: no granular forms.
(b) Serum of blood.—
40 minutes after mixing: a very few isolated motile spirilla: several
clumps in which the spirilla still show motility.
1½ hours after mixing: no normal isolated spirilla: some granular pucker-
ed-up forms: many clumps in which the spirilla are slightly motile.
2½ hours after mixing: all motility completely ceased: only two isolated
spirilla which have granular appearance: many granular clumps.
22 hours after mixing: a number of granular clumps with granular
spirillum-forms at the margin: no isolated organisms: no motility:
no spirilla of normal appearance.
(2) Stained preparations—
(a) Serum of spleen.—
1 h o u r a f t e r m i x i n g : a g r e a t n u m b e r o f i s o l a t e d s p i r i l l a : s e v e r a l c l u m p s :
all spirilla, both isolated and clumped, take the stain regularly and
well.
1 3/4hours after mixing: same appearance as above.
2½4 hours after mixing: same appearance as above.
22 hours after mixing:a considerable number of isolated spirilla and
many clumps: all spirilla stain evenly and well.
(b) Serum of blood
.—
(c)
1 hour after mixing: a few isolated spirilla, much fewer than in splenic
preparation: some clumps of spirilla which stain normally: some
granular masses, which stain badly and in which no spirillum forms
can be made out.
1 3/4 hours after mixing: the same appearance as above.
2½ hours after mixing: the same appearance as above.
22 hours after mixing: a very few isolated spirillum-like bodies which
stain very irregularly: some small granular masses, which take the
stain very faintly and which contain no spiral forms.
Experiment 3.—Monkey S-12 suffered from an attack of spirillum fever
of 3 days' duration. Shortly after the crisis the spleen was removed. At the
same time a specimen of blood was taken from the ear.
Medical Officers of the Amy of India. 99
The spirillum blood was obtained from a monkey on the second day of fever.
This blood contained abundant spirilla.
Preparations were made as described above and the following observations.
recorded :—
(1) Hanging drop preparations
—
(a) Splenic serum.—
30 minutes after mixing: spirilla very abundant and very actively motile
no clumps.
1 ¼ h o u r s a f t e r . m i x i n g : a b u n d a n t i s o l a t e d s p i r i l l a s h o w i n g g r e a t m o t i l i t y :
many clumps in which spirilla are actively motile: no degenerated
forms.
2¼ hours after mixing: same appearance as above.
4 h o u r s a f t e r m i x i n g : a l a r g e n u m b e r o f i s o l a t e d a c t i v e l y s p i r i l l a :
some large clumps in which spirilla are still motile: other clumps.
made up of motionless and slightly granular spirilla.
22 hours after mixing: a few actively motile spirilla: a great many
clumps, in which many spirilla have a granular appearance.
(b) Serum of blood.—
30 minutes after mixing: some motile isolated spirilla: motility much less
than in splenic preparation: a great many clumps.
1¼ hours after mixing: very few slightly motile isolated spirilla: many
clumps in which the spirilla are quite motionless.
2¼ hours aftermixing: a few isolated spirilla showing slight movement:
many granular clumps: some clumps of apparently normal spirilla.
3 hours after mixing: all motility ceased: no isolated spirilla: many
clumps of granular motionless spirilla.
22 hours after mixing: no isolated spirilla: a few granular masses in
which some spirillum forms can be indistinctly made out.
(2) Stained preparations—
(a) Serum of spleen.—
5 0
m i n u t e s a f t e r m i x i n g : a l a r g e n u m b e r o f i s o l a t e d s p i r i l l a : v e r y m a n y
clumps, each composed of an enormous number of spirilla closely
fetled together: all spirilla, both isolated and in clumps, stain well
and regularly.
2 hours after mixing:
4½ hours after mixing:
same appearances as above.
I
22 hours after mixing:
A A
AA
100
Scientific Memoirs by
(b) Blood serum.—
50 minutes after mixing: a few isolated spirilla which take the stain well:
many clumps, each made up of only a few spirilla.
2 hours after mixing : a few isolated spirilla : no clumps seen.
4 ½ hours after mixing:
a very few spirilla which stain very faint-
ly and unevenly: some are isolated,
others in small clusters.
22 hours after mixing:
Experiment 4.—Monkey S-23 suffered from a typical attack of spirillum
fever of 3 days' duration. The spleen was removed shortly after the crisis,
and at the same time blood from the ear was collected in a capsule.
The spirillum blood was got from a monkey on the second day of fever. It
contained spirilla only in sparse numbers.
Preparations were made as in the other experiments, and the following ob-
servations recorded
:—
(1) Hanging drop preparations—
(a ) Serum of spleen.—
15 minutes after mixing: spirilla actively motile and isolated.
1 hour after mixing: no spirilla to be seen.
( b ) Serum of blood.—
15 minutes after mixing: no motility: a few motionless granular spirilla.
(2) Stained preparations.—Specimens for these observations were not made
until an hour after the mixtures had been prepared. At this time no spirilla could
be found in either the splenic or blood serum specimens.
The above observations, although not mathematically precise, would appear
clearly to show that immediately after the crisis in monkeys, at a time when the
spirilla have disappeared completely from the blood, but are still present in a live
condition in the spleen, this latter organ is poorer in anti-bacterial substances
than the circulating blood.
Further, more light may be thrown on the subject by considering what results
have been obtained from similar investigations in other septicæmic diseases. In
the case of typhoid and Malta fevers the specific micro-organisms continue to
cultivate themselves in the body long subsequent to the appearance of the
specific agglutinins in the blood. Now the bacteria in these diseases are either
altogether absent from or present only in very small quantity in the circulating
blood, but are found to be present in greater amount in the spleen and other
internal organs. In patients dying during the course of these diseases, it is
possible to make a quantitative and comparative estimation of the amount of
agglutinins in the organs in which the bacteria are cultivating themselves and in
Medical Officers of the Army of India.
101
the blood serum. The result of such an investigation has been put on record by
Professor Wright and myself 16.
We made comparative estimations of the agglutinins in the serum of the
heart's blood and in the serum of the spleen, or of Peyer's patches, in four cases
dead of typhoid fever at different periods of the disease : further we made com-
parative estimations of the agglutinins in the serum of the heart's blood and in
the serum of the spleen in two cases dead of Malta fever. These observations
clearly established that the spleen in the case of typhoid and Malta fevers is much
poorer in agglutinating substances than the circulating blood. As a result of
these observations and other considerations stated in the publication referred to
above, we enunciated a theory as an explanation of the relapses which are so
common both in typhoid and in Malta fever. This theory may be briefly stated
as follows :—in both of these septicæmic diseases the bacteria become deposited
in the spleen and other internal organs. There they cultivate themselves forming
colonies which soon become protected from the bactericidal substances of the
lymph stream by the formation of a non-anti-bacterial envelope. As a result of
this the spleen will, as we have shown, contain, as a whole, less agglutinating
substances than the circulating blood. When the anti-bacterial substances in
the blood and lymph have increased to such an extent as to penetrate and
abolish the non-anti-bacterial envelopes which surround these colonies, the pro-
duction of toxins will be so diminished that the temperature will fall. If, however,
for some reason or other even a single colony escapes the full anti-bacterial
power of the lymph, owing, it may be, to its being shut off in a capillary which
has become blocked, or in some other part which was not freely washed by the
blood stream or by the lymph stream, the bacteria of this colony would go on
multiplying until the blood has become modified in such a manner as to bring
about a diminution of the anti-bacterial substances and thus render a relapse
possible.
These considerations, and the observations which I have put on record
above, go to show that a similar mental picture can be formed of the relapse
which is seen in every case of spirillum fever in man. Briefly then we may
enunciate the following theory. At the crisis some of the spirilla perish. It is
evident, however, that some must survive and from what we have seen to be
the bactericidal power of the blood and lymph, these must in some way or other
be protected from the full anti-bacterial power of these fluids. Under this
protection they would grow and multiply during the period of apyrexia. Toxin
would be produced, and in a short time so modify the blood as to diminish the
anti-spirillar substances to such an extent as to permit of the spirilla again
living in that medium. It is also probable that there may be a considerable
loss of the anti-bacterial substances by elimination. It is only by this latter
process that the blood can be thus altered before re-infection in the case of the
monkey.
The relapse is always of shorter duration and less severe than the primary
attack.
This is no doubt due to the fact that the organism at the beginning of the
relapse has already in its possession a considerable amount of anti-bacterial
substances and is therefore in a much better position to do battle with the spirilla
than it was at the commencement of the primary infection. The same explana-
tion may be brought forward to account for the shortness and mildness of the
second and third relapses when these occur.
Notes and references.
(1) Transactions of the Royal Medical and Chirurgical Society of
London, Vol. LXIII, 1880:
Carter, Spirillum Fever: 1882.
(2) Loc. cit.
(3) The amount of citrate of soda necessary to prevent clotting of
monkey's blood was found to be 1 part to 200 parts of blood.
(4) Annal : de l'Inst. Pasteur : 1896, p. 630.
(5) Annal : de l'Inst. Pasteur : 1899, p. 363.
(6) See note 3.
(7) British Medical Journal, February 5th, 1898.
(8) In view of the fact that this blood had to be mixed with serum, the
proportion of citrate of soda employed was increased to 1 part in
1 0 0 p a r t s o f b l o o d .
(9) Annal : de l'Inst. Pasteur, Vol. V, p. 545.
(10) Virchow's Archiv, CIX. p. 176.
(11) Loc. cit.
(12) Loc. cit.
(13) Loc. cit.
(14) The theory that the spirilla form spores and survive in this condi-
tion in the internal organs during the period of apyrexia is not
supported by any observations, and has been abandoned by all
investigators.
(15) Vide note 8.
(16) Lancet: December 23rd, 1899.
Fig. 1.
Fig. 3.
Fig. 4.
Fig. 2.
A
B
L
Lith by A. C. Singha.
EXPLANATION OF PLATE I.
A.—Fig. 1. Root, showing the regular division into alternate white and black zones.
Root cap evident at lower extremity.
Fig. 2. Root which has been injured and is being continued in its main direction by
two branches directed downwards. A smaller branch is directed up
- w a r d s .
Fig. 3. Root showing irregular distribution of the white zones. Root cap well
shown.
Fig. 4. Root devoid of any zoned appearance, except in its branches, rising above
the mud. The upper dark-tinted portion is aerial, the lower lighter-tinted
part is sunk in the mud.
All four figures are natural size.
B.—Base of three stems of Phœnix paludosa Roxb. reduced ½.
On the Anatomy of the roots of Phœnix paludosa Roxb.
BY
A. T. GAGE, M.A., M. B., B.Sc.
LIEUTENANT, I.M.S.
Curator of the Herbarium attached to the Royal Botanic Garden, Sibpur, near Calcutta.
The roots of Phœnix paludosa Roxb. are thickly clustered around the base of
the stem as far up as from six inches to about two feet above the surface of the
swampy soil or water in which the palm grows.
They may to some extent be concealed beneath the remains of the fibrous
leaf bases, which may have to be broken away to reveal the roots. Usually, how-
ever, they are quite evident, and are seen in all stages of development on the
same stems. Some appear as little wartlike excrescences on the stem, others
may reach a length of two or three feet with their lower three-fourths sunk in
the mud. The majority of the roots, however, vary from three to six inches in
length, with a diameter varying from the thickness of a goose quill to that of the
little finger. The tip of each root is protected by a well developed root cap.
The roots arising from the trunk of the palm grow downwards till they pene-
trate into the mud, the root cap serving as a very efficient 'shield' during this
operation. As the resistance offered by soft mud cannot be very great, it
is possible that the root cap may protect the growing point against the attacks
of aquatic insects or fresh water crustaceans. All the roots, however, do not grow
downwards.
A glance at Plate I, B., discovers a considerable proportion of small roots
directed vertically upwards. These are branches arising from roots which have
penetrated well into the mud. They arise from the roots below the surface, and
curve upwards, assuming ultimately a practically vertical upward direction.
All branch roots do not grow vertically upwards. If an originally downward
directed root is so injured that further growth of it cannot proceed, one or more
branch roots are developed above the injured part and these carry on the down-
ward direction of the original root. See Plate I, A., figure 2 .
A large root which has penetrated about eight inches or a foot into the mud
shows a marked difference between its aerial and its subterranean portion. The
aerial portion is of firm texture and dark brown or almost black in colour, whilst
the buried portion is thicker, much paler in colour, and of a loose spongy consist-
ence.
BB