NationalLibraryOfScotland/medical-history-of-british-india huggingface dataset:
34
Scientific Memoirs by
While, therefore, the isotonic point for saline solutions is practically identical
for rat, snake, fowl, man and guinea-pig, the isotonic point, if we may use the ex-
pression for cobra-poison is very different. Dog's blood is exceedingly sensitive to
the poison ; hæmolysis in this animal occurring in very dilute solution, for instance,
in the strength 0. 5 c.c. = .0009 mgm. An examination of the table shows
us that there is no absolute relation between the hæmolytic reaction of the blood
of an animal to cobra-poison, and the resistance of the animal itself to the poison.
Thus while in this table cat's blood and guinea-pig's blood rank almost equally
low, as regards hæmolysis, yet the cat is far more resistant to the poison than the
guinea-pig. Again, on this scale, if it were taken as an indication of the resist-
ance of animals, we should rank a frog and a guinea-pig as equally resistant,
which, however, is far from being the case. These numbers must not, however,
be taken as absolute, as from time to time variations occur in the minimum
amounts of poison required to produce hæmolysis, These, however, are not
sufficiently large to negative the general relation shown above, and may be due
to changes in the poison, or in the blood, or in both.
It may be interesting here to point out the action of temperature on the
hæmolytic action which can be readily shewn by either of the above described
methods.
Thus.—
Table IV.
C . C .
C . P . = . 0 3 1 2 5 .
E x p . I .
Exp. II.
12
50
1.25
3.0
3.25
3.45
180 114
1.10
79
119
85
360.5
73
12
0
91
0
B. I C.C.=.03125.
18°
.
Standing over night
.
.
.
. .
Very incomplete Hæmolysis.
3 6 0 .
5
, ,
.
.
.
.
Complete Hæmolysis.
The action of antivenomous serum on the hæmolytic action of cobra poison.*
Antivenomous serum itself is hypertonic for human and guinea-pig's blood,
and the isotonic point is shown by the following table :—
Table V.
. 6 A n t i v e n o m o u s S e r u m
+ .
4 d i s t i l l e d w a t e r
.
.
.
.
No H
. 5 5 , ,
+ .45
,,
.
.
.
.
No H
. 5 , ,
+ .5
,,
.
.
.
.
H. incomplete.
. 4
,,
+ .6
,,
.
.
.
.
H. complete.
* The serum used in these experiments was that prepared at the Pasteur Institute, Lille, by Dr. A.
Calmette.
Medical Officers of the Army of India.
35
Consequently in these experiments a mixture of serum and water in the
proportion of 55:45 was used.
Taking then solutions of poison which readily hæmolyse, we proceeded to
try what might be the action of serum on this phenomenon. It was found that
the action could be completely arrested by using definite quantities of serum.
Table VI.
C.P. z c.c. = .2 mgm. in isotonic salt solution ; Human blood.
.5 c.c .C.P. . .
.
.
.
.
.
.
.
Complete H (in 1 hour).
.5 c.c. C.P. + .001 c.c. isotonic serum (antivenomous). . . Complete H.
.5 C.C. C.P. + .025 C.C.
, ,
, ,
.
.
Incomplete H.
.5 C.C. C.P. + .05 C.C.
, ,
, ,
.
.
Incomplete H.
.5 C.C. C.P. + .1 c.c
, ,
, ,
.
.
No H.
.5 C.C. C.P. + .2 C.C.
, ,
, ,
.
.
No H.
.5 C.C. C.P. + .3 C.C
, ,
, ,
.
.
No H.
.5 C.C. C.P. + .4 C.C.
, ,
, ,
.
.
No H.
.5 C.C. C.P. + .5 c.c.
, ,
, ,
.
.
No H.
The same results were obtained by varying the poison and keeping the
serum constant. Repeated observations showed that for the particular serum
used in these experiments .1 c.c. of isotonic serum is always sufficient to stop the
hæmolytic action of .5 c.c. of a solution of poison (1 c.c. = . 2 mgm.) on human
blood.
Further, if we take multiples of these numbers, the same relation holds
good.
Table VII.
C.P. 1 C.C.=.2 mgm.
.5 c.c. C.P. + .1 isotonic serum .
.
.
.
.
.
.
.
.
No H.
I C.C. C.P. + .2
,,
.
.
.
.
.
. .
.
.
No H.
2 c.c. C.P. + .4
, ,
.
.
.
.
.
.
.
.
.
No H.
3 c.c. C.P. + .6
, ,
.
.
.
.
.
.
.
.
.
No H.
These observations show accordingly that the antivenomous serum posses-
ses the power of neutralizing the hæmolytic action of cobra-poison in vitro in the
same manner as the antiricin neutralizes ricin in the test tube.
This action is, moreover, a specific one, as various other sera such as the
antidiphtheritic, antistreptococcus, antitetanus, antityphoid have no such action.
Correspondence between the antihœmolytic action in vitro and the protec-
tive action of the serum in corpore.
For a guinea-pig weighing 250—350 grms. mgm. of fresh poison is the
minimum certain lethal dose, death ensuing in from 5—8 hours. Now in Table
VI it is shown that the hæmolytic action of this quantity of poison is completely
counteracted by .1 c.c. of isotonic antivenomous serum. And injection of this
non-hæmolytic mixture is never fatal to the animal when incompletely neutralized
F 2
the animal may or may not die. The following Table VIII. summarizes a number
of experiments on this point :—
Table VIII.
(1) Mixtures of serum and poison which give
no hæmolysis, i.e., .1 mgm. cobra toxin.
+ .1 c.c. isotonic cobra antitoxin.
Animals taken
(subcutaneous injection).
Deaths.
12
0
(2) Mixtures of serum and poison which give
hæmolysis, i e., 1 mgm. toxin+less than
.1 c.c. isotonic antitoxin..
.
13
9
(3) Poison controls .1 mgm. cobra-toxin.
1 3
13
But on the other hand, if larger quantities of poison be taken, although
neutralized as regards hæmolysis, the animals died (5 out of 6).
Thus 3 c.c. C.P. (= .6 mgm.)+.6 c.c. isotonic serum is a non-hæmolytic
mixture. Yet it is rapidly fatal to the animal. So that the correspondence
only holds good for the dose already mentioned and does not obtain for mul-
tiples. An explanation of this fact may lie in the probable complex nature of
cobra-poison. L. J. Martin has recently drawn attention to the fact that many
snake-poisons including cobra-poison contain at least two proteid substances of
which one is coagulable and indiffusible, the other is incoagulable and diffusible,
and he believes that the coagulable proteid is a hæmolytic poison, whilst the
incoagulable one is principally a nerve cell-poison.
Now, if this be so, it is obvious that a single lethal dose mixed with the
quantity of serum necessary to neutralize the hæmolytic poison would contain
only a small quantity of the other poisonous substance, whilst six times this lethal
dose would contain 6 times as much of this other poisonous substance which
would remain active. Thus if a single lethal dose of poison contains two sub-
stances A and B of which A is present in a single lethal dose and B in sublethal
quantity, then six lethal doses would contain 6 A + 6 B. Now 6 A are
neutralized by the serum, while 6 B remain free and act fatally.
Briefly these experiments may be summarized as follows :—
(1) Cobra-poison is strongly h æmolytic in vitro.
(2.) This action is neutralized by anti-venomous serum, and the action of the
latter is specific.
(3.) For certain doses (.1 mgm.) the measure of this neutralization in vitro is a
measure of the neutralization in corpore for guinea-pigs.
(4.) The action is chemical and not cellular or vital.
NOTE.—In our late experiments the serum we have used possessed a less antih æmolytic power than
that of our original experiments. Whereas 0.1 c. c. of diluted serum only was necessary to neutralize 0.1 mgm.
of poison for this reaction ; of our later serum 0.4 c. c. of undiluted serum was necessary, i. e., nearly eight
times as much. If the serum then contains at least two substances, A and B, of which A is here in less quan-
tity than previously, then if the quantity of B has remained unchanged (or has increased), it is evident that
taking hæmolsis as our test reaction multiple doses will now contain greater quantities of B. And with
this serum we have found that multiples of 5.(i.e., .5. mgm, C. P.) produce no effect upon the animal when,
as previously, serum is added in sufficient quantity to prevent h æmolysis.
Medical Officers of the Army of India.
37
PART II.
The influence of cobra-poison on the coagulation of the blood, and the
action of antivenomous serum upon this phenomenon in vitro.
The delay in clotting or complete absence of the same under the influence of cobra poison
can be readily shown by inducting blood direct from a vessel into the poison solution
as demonstrated by Dr. Cunningham.
TABLE I.
Guinea-pig : anæsthetized : cannula inserted in carotid.
Clotting time.
Next day.
I. 1 C. C. C. P.=20 m g m . + 1 c . c . B l o o d
. Black solution
.
. Fluid.
II. 1 c. c. C. P.=10 mgm. +1 c. c. Blood
. Black solution
.
. Fluid.
III. 1 c. c. C. P.= 5 m g m . + 1 c . c . B l o o d
. 1 h Black clot
.
. Solid jelly; no serum.
IV. 1 c. c. C. P.=2.5 m g m . + 1 c . c . B l o o d
. 18' Black
.
.
. Solid ; no serum.
V.
1c . c . C . P . = 1 . 2 5 m g m . + 1 c . c . B l o o d
. 9' Black
.
.
. Solid ; no serum.
VI.
1C . C . C .P
. = . 6 2 5 m g m . + 1
C . C .B
l o o d
.
...
. . .
VII.
1c . c . C .P
. = . 3 1 2 m g m . + 1
C . C .B
l o o d *
. 2'
2h deep red serum
.
...
VIII. 1 c. c. C. P.=.07 mgm. + 1 c. c. Blood*
. Bright red clot, red serum Semi-fluid clot.
IX. 1 c. c. C. P.=.02 mgm. +1 c. c. Blood* . Bright red clot, clear
serum.
Serum stained.
X. 1 c. c. C. P.= . 004 mgm.+1 c. c. Blood† . Bright red clot, clear
serum.
Serum stained.
XI. I c. c. C. P.=.002 mgm. +1 c. c. Blood
. Bright red, clear serum
Serum unstained.
XI I. 1 c. c . Saline 0.5 % +1 c. c. Blood
.
6' scarlet
.
.
. Contracted clot in serum.
* A fresh guinea-pig was used for VII, VIII, IX.
† A f r e s h g u i n e a - p i g .
The same effect can also be conveniently shown by mixing small quantities
of blood taken from the finger, with poison in small pieces of graduated
capillary tubing. The time when the blood can be inverted or when it can no
longer be blown out of the tube is taken as the clotting time. The solutions of
poison, or of poison and serum, are previously mixed and a small quantity intro-
duced into the tube ; an equal volume of 'blood is then sucked up from the
finger, the mixture rapidly made.
38
Scientific Memoirs by
TABLE II
.5 c. c. C. P.+1.25 mgm.
.
. .5 c. c. C. P. =1.25
mgm.
Antivenomous serum Saline 1%
.5 c. c. saline 1%
.
.
.
. 7 c. c antivenomous
serum.
Isotonic.
60' still fluid .
.
.
.
. (1 ) 12' firm
.
(1 ) 10'.
(2) 17' solid
.
(2) 15'.
60' still fluid and very small clot
(1) 3' . . . 10'
.
.
. 9'.
(2) 6' .
60' still fluid .
.
. . . 6'
. . . 6'
.
.
. 6½'.
24' still fluid
.
.
.
.
. 8'
.
.
1 6 '
. . . 5'.
60' still fluid and small clot
.
. 5½'
.
.
. 7'
.
.
. 10 '.
7½' another specimen
of blood.
60' still fluid and small clot
.
. 12'
. . . 10½'
.
.
. 8½.
(1) 10' second speci-
men.
(2) 18'
11½'.
60' still fluid, very small clot
.
. 12'
.
.
. 11½'
.
.
. 25' second :specimen.
60' still fluid, some clot
.
. 12'
.
.
. 7½'
.
.
. 12'.
NOTE.—When two figures are given in the table, the first signifies when the blood could be inverted, the second
when it was solid.
The table shows that a poison solution 1c. c.=.625 mgm. is able to keep
specimens of blood fluid for over an hour while with a mixture consisting of
.5 c. c. C. P.=1.25 mgm. +.7 isotonic serum (a mixture which gave only a trace
of hæmolysis) the clotting time was reduced to about the same as that of blood
diluted with saline.
Finally, similar results were obtained by using citrated blood. To this were
added various quantities of poison and serum and two platinum loopfuls of
a 5% calcium chloride solution. C. P. 1 c.c.= . 2
m g m s .
TABLE III.
I. 0. 25 c. c. Blood+ 1.25 c. c. saline+call 2 .
.
.
.
.
.
10' nearly solid.
II. 0.25 c. c. Blood+ 0.5 c. c. C. P.+0.75 c. c. saline+call2
.
. 24' no clot.
III. 0.25 c. c. Blood+ (0.5 c. c. C. P. +.05 c. c. isotonic serum) +0.75 c. c. saline +call 2 30' small clot.
IV. 0.25 c. c. Blood +(0.5 c. c. C. P.+.075 c.c. „
„
)+0.75 c. c. saline+call 2 10' nearly solid.
V. 0. 25 c. c. Blood +(0.5 c. c. C. P.+ 0.1 c. c. „
, , )+0.6 c . c. saline+call 2 10' nearly solid.
VI. 0.25 c. c. Blood+(0.5 c. c. C. P.+ 0.2 c. c. „
„
)+ 0.55 c. c. saline+call 2 10' gelatinous clot.
VII. 0. 25 c. c. Blood +(0.5 c. c. C. P.+ 0.3 c. c. „
„
)+0.45 c. c. saline + call 2 20' gelatinous clot.
VIII. 0.25 c. c. Blood +(0.5 c. c. C. P .+ 0. 5 c. c. „
„ )+0.25 c. c. saline+call 2 20' gelatinous clot.
IX. 0'25 c. c. Blood+(0.5 c. c. C. P.+ 0.75 c. c. , ,
„ )+ call2 .
.
. 30' gelatinous clot.
X. 0.25 c. c. Blood+0.2 c. c. undiluted serum+ 1.05 c.c. saline+call2 .
. 45' nearly solid.
Medical Officers of the Army of India.
39
It will be seen that in Example II, no clot was obtained even after 24 hours,
while by the addition of o.I c.c. serum clotting was normal, viz., 10'. Further it
will be noted that the serum itself delays clotting somewhat.
The specificity of the action of the anti-venomous serum was easily shown
by either of these methods. Nor, on the other hand, had the anti-venomous
serum any action on the well known properties of leech extract and peptone
solutions in keeping blood fluid in vitro.
A few experiments were made with a view to determine whether the blood
of an animal artificially immunised had any greater resistance against this pro-
perty of cobra-poison than that of a normal blood. Thus a cat received Ioo
mgms. of cobra poison in the space of ten days. Sixteen days later, the blood
was tested with this result.
TABLE IV.
a saline+blood
—c. c.
clotting time about equal in all three tubes.
b C. P 2 mgm.+blood
—c.c.
c C. P 4 mgm+blood
—c, c.
That the blood possessed considerable resistance is evident from the fact
that as much as 4 mgm of cobra poison was unable to delay the clotting
time, much less to keep it permanently fluid. Coincidently with this, the serum
of this animal was shown to possess definite anti-toxic property as tested by
subcutaneous injection on guinea pigs.
C. P. I mgm.
. 340 gramme guinea-pig
. Died 24-36 hours.
C. P. I mgm+I c. c. cat's serum
..
,,
.
. No symptoms.
C. P..2 mgm.+ 1 c. c. serum.
,,
.
. No symptoms.
The variability in the time results, according to the method of determining
the clotting, have so far prevented us from using this reaction as a quantitative
test of the exact neutralization of this constituent of the poison by the anti-
venomous serum; but the qualitative reaction is fully shown by any of the
methods enumerated, and we see at any rate that the action of cobra-poison
upon shed blood is neutralized in vitro by the specific anti-toxin. And this
Dr. Cunningham has also demonstrated using a reaction other than hæmolysis
and coagulation.
On three new genera of plants from the Kachin Hills.
BY
MAJOR D. PRAIN, INDIAN MEDICAL SERVICE.
The year 1897 has been marked, so far as the records of the Calcutta
Herbarium are concerned, by the communication, for the first time, of botani-
cal specimens from the Kachin Hills in Upper Burma. The receipt of these
has been primarily due to the exertions of Captain Pottinger, R.A., who made
an adventurous journey into the unexplored country along the 'Nmai Kha from
March till June, and brought back with him a very interesting collection.
Captain Pottinger had further the kindness to enlist the sympathies of Lieute-
nant Cruddas, S.C., Commandant of the Battalion of Military Police at
Myitkyina, on behalf of the Herbarium. A native collector was sent to
Myitkyina; this man has been able, thanks to the assistance and opportunities
afforded him by Lieutenant Cruddas, to make large collections.
The Kachin Hill region forms the catchment area of the Upper Irrawaday,
and is thus wedged in between the well-marked phytogeographical areas
of South-West China on the one hand and of the Eastern-Himalaya on the other,
while it forms the meeting point of two equally distinct areas—that of
the Assam Ranges and that of the Shan Plateau—which to the south of
this region are separated by the Irrawaday Valley. On à priori grounds
one might expect that the Kachin Flora would exhibit a mixture in something
like equal proportions of the elements characteristic of all four areas mentioned.
Such, however, is by no means the case. The collections of 1897 have yielded
as many as 627 distinct Kachin plants, and of these only 195, or 31.1%, rather
less than a third of the flora, occur in China at all, while only three of the plants
met with in the Kachin Hills are peculiarly Chinese. In Eastern Indo-China,
the country to the east of the Irrawaday Valley, we find almost twice as many
Kachin plants, the number being 402, or 64.11%. The peculiarly Indo-
Chinese element is seven times as great as the peculiarly Chinese, for there
are 21 plants that are common only to Kachin and the country east of the
irrawaday. However, an even greater number of Kachin plants extend west-
ward to the more distant Himalayan region than eastward to the Shan Plateau,
the number being 416, or 66.34%. But while this is the case, the peculiarly
Himalayan element is far smaller than the peculiarly Eastern Indo-Chinese, as
only 8 plants occur in Kachin that are not found elsewhere except in the Hima-
laya. The real smallness of the Indo-Chinese influence is, however, very clearly
brought out when we consider the number of Kachin species that occur both
G
42
Scientific Memoirs by
in the Eastern Himalaya and in Assam without appearing east of the Irrawaday :
of this class there are 67 species, Io.68% of the whole Kachin flora. And
the affinity of the Kachin flora is seen to be most marked with the flora of the
Assam ranges from the fact that 522 species, or 83.25 of the Kachin flora,
extend to this particular region, and that 46 of these species, or 7% of the
Kachin flora, are peculiarly Assamese.
The comparative values of the influences exerted by the floras of the
four adjacent areas are, however, more accurately determined by using the
numbers indicated by each distributional arrangement that occurs, treating
the figures so obtained as numerators, employing as denominators the
number of adjacent areas involved in each case, and adding the fractions
relating to each area. The results are as follows :—Assam, 40.I%; Shan, 27%;
Himalaya, 22.4%, and China, I0.5%. The slightness of the Chinese and the
paramount nature of the Assamese influence on the Kachin flora is thus again
manifested.
As many as 53 of the forms reported appear to be endemic in the
Kachin region; this gives the considerable proportion of 8½%. Some of these
endemic forms are extremely interesting; among them are three, of which
descriptions are here given, that appear to constitute distinct generic types.
Cruddasia
prain.s
N a t . o r d . L E G U M I N O S Æ : T r a i b . P h a s e o l e œ .
Calycis
lobi acuti, 2 superiores in unum apice minute 2-dentatum connate,
cætera aequilata triangulares imo tamenlateralibus parum longiore. Vexillum
suborbiculatum, basi nec inflexum alae oblongo-ovatæ basi carinæ adhærentes;
carina cymbiformis erostris alis aequilonga, Stamen vexillare cæteris arcte
connatum, anthem uniformes. Ovarium sessile, 8-ovulatum; stylus filifor
mis incurvus sub stigmate terminali summo apice parce penicillatus ceterum
glaber. Legumen elongatum 2-valve planum coriaceum compressum, intus
inter semina tenuiter farctum. Semina plano-compressa, sub-orbicularia, hilo
ovato estrophiolata.—Caulis alte volubilis, foliis pinnatim 5-foliolatis, foliolis
anguste ovatis stipellatis. Stipules spinuloso-setace caducæ, basifixæ. Flores
purpurascentes in pedunculis elongatis fasciculato-racemosi, fasciculorum
rachide nodiformi. Bracteola et bracteae caducæ. Calyx vexilloque extus
sericeus. Species singula montium Kachin incola; C, insignis Prain in F ourn.
As. Soc., Beng., LXVII, 2, 287 .
This genus might with almost equal justice be referred to the
Galactieæ especially Galactic § Collœa or to the Diocleœ especially Pueraria.
It differs from both, however, as having the style bearded round the stigma, and
its 5-foliolate leaves give it a very distinctive facies. The genus is named in
Medical Officers of the Army of India.
43
honour of Lieutenant Cruddas, who has afforded much kind help in connection
with the investigation of the Kachin flora.
Pottengeria P r a i n
.
Nat. Ord. SAXIFRAGACEÆ: Trib. Escallonieœ.
Calycis tubus brevis late campanulatus basi ovarii adnatus lobis 5 ovatis
acutis persistentibus sinubus latis. Petala
Stamina 5 erecta
ad marginem disci perigyni affixa filamentis sursum subulatis, basi parum
explanatis ibique extus glandula mediana ornatis; antheræ.
Ovarium semisuperum. Capsula supera oblonga longitudinaliter parum
sulcata per stylos 3-partibilis stigmatibus coharentibus septicide 3-valvis,
placentis a marginibus carpellorum introflexis simulac secedentibus persisteni-
tibusque, singulis utroque margine semina circa I0 triente summo tantum fertili
gerentibus. Semina anguste fusiformia testa crustacea parum reticulate
utrinque parum producta; embryo cylindrica majuscula in axe albuminis
carnosi.—Folia alterna brevipetiolata, 5-nervia. Cymae multiflor axillares.
Species sirigula, montium Kachin incola; P. acuminata Prain in Fourn. As. Soc.
Beng. LXVII, 2, 291.
This plant constitutes the type of a very distinct genus near Itea which it
resembles in having a similarly partible style with a similarly half-superior ovary
and with the same septicidal dehiscence, but from which it differs markedly in hav-
ing 3 carpels in place of two and in the three filiform placentas being persistent.
The foliage recalls that of a Cinnamon or of some Melastomaceous plant and
is thus quite unlike anything else in the natural order to which the plant belongs.
The genus is dedicated to Captain Eldred Pottinger, R.A., to whom our
knowledge of the Kachin flora is primarily due.
Eurysolen Prain.
Nat, Ord. LABIATÆ Trib. Prasieœ.
Calyx tubuloso-campanulatus I0-nervus 5-dentatus dentibus 2 anticis
posticos 3 distincte excedentibus vix tamen 2-labiatus, intus glaber extus
parce puberulus, fructifer parum urceolatus dentibus erectis. Corollœ tubus
exsertus intus annulatus basi tenuis supra annulum antice gibbus; limbus
2 -
l a b i a t u s l a b i o p o s t i c o e r e c t o p a r u m c o n c a v o a p i c e r e t u s o , a n t i c o l o n g i o r e
patente 3-lobo lobis lateralibus minoribus medio majusculo. Stamina 4 di-
dynama anticis longioribus sub galea adscendentia distincte e tubo exserta ;
anther I-loculares ovato-reniformes explanat, iis staminum posticorum
G 2
G 2
44
Scientific Memoirs by Medical Officers of the Army of India.
parum minoribus filamentis omnibus pibs albis puberulis. Discusæ qualis.
Stylus apice 2-fidus lobis fere aequilongis acutis. Nuculae juniores extus læves
supra intusque papillosim glandulosæ.— Frutex scandens foliis ovato-
acutis basi cuneatis bracteis minutis. Flores parvi in spicas terminales axil-
laresque conferti. Species adhuc singula, emontibus Kachin recepta; Eurysolen
gracilis Prain in Ann. Roy. Bot. Gard., Calcutta, vol. IX, ined.
The genus is a very distinct one; it seems as if it might be best placed
among the Prasieae near Gomphostemma. The corolla tube is swollen in front
as in some species of Valeriana; the anthers are apparently one-celled from a
very early stage.