NationalLibraryOfScotland/medical-history-of-british-india huggingface dataset:
8
Scientific Memoirs by
thick, yellowish strata, with a few more prominent masses. It had no perceptible
smell. No commas could be found, the growth apparently consisting entirely of
large, straight Bacilli
EXPERIMENT IV.—One hundred grammes of dry garden-earth were triturated
with about thirty grammes of fresh fæcal matter and the mixture set as usual in
a covered beaker. The fæcal matter was neutral in re-action. Forty-eight hours
later the earth appeared to be everywhere penetrated by white fungal hyphæ,
and on the surface a crop of young sporangia, of a species of mucor, had begun to
appear. Fifty cubic centimetres of neutral salt solution full of the commas from
two tube-cultivations of forty-eight hours' growth were now poured over it and
the beaker again set aside. Two days later a stratum of brownish alkaline
fluid, with a distinct thin scum, was still present on the surface. Some of this,
and some of the surface earth, were mixed with neutral salt solution and a plate-
cultivation of it set as usual. On the following day this had a sickly, somewhat
choleraic smell. The surface was covered by yellowish and bluish colonies, and
full of small interstitial ones. Six preparations were mounted, and in two of them
commas were present.
Six days after the addition of the commas to the earth another plate-
cultivation was started. Forty-eight hours later it was crowded with concrete
yellowish colonies, and also contained some large, diffused patches of yellowish
and bluish colour. The smell was non-choleraic and like that of the flowers
of Bassia latifolia. Twelve preparations were mounted, but none of them con
-tained commas.
Eight days after the commencement of the experiment another plate-cul
-tivation was inoculated from the earth. On the following day it had a faint
non-choleraic smell and was crowded with concrete colonies. Some diffuse
patches, of yellowish and bluish colour, were also present. Twelve preparations
were mounted from colonies which in any way resembled colonies of commas,
but none of them contained commas. Two days afterwards the cultivation had
a strong smell of bats. Another set of preparations was mounted, but, as before,
contained no commas.
EXPERIMENT V.—Twenty-five grammes of fresh fæculence was triturated
with seventy-four grammes of dry garden-earth. Water was then added and
the mixture thoroughly stirred up and boiled over a Bunsen flame until nearly
dry. It was then transferred to a clean glass beaker and the latter set in a
boiling water-bath for half an hour. On the next day the beaker was kept in the
bath for an hour, and the same process was repeated on the two following ones.
At this stage of the experiment a plate-cultivation was inoculated from the soil
by the usual method. On the following day it was found to be full of
yellowish defined, circular colonies. The smell was faint and non-choleraic. Pre
- p a r a t i o n s s h o w e d t h a t t h e c o l o n i e s w e r e a p p a r e n t l y a l l c o m p o s e d o f a s h o r t ,
thick, straight Bacillus. Twenty-four hours after the previous heating the beaker
Medical Officers of the Army of India.
9
was again heated in the bath, and finally for several hours over a Bunsen flame
On the following day the growth in two tube-cultivations of commas was mixed.
with some salt solution and poured over the earth.
Two days later a plate-cultivation was set as usual. On the following day
it contained immense numbers of colonies, large yellowish ones and minute blue
ones being apparently present in approximately equal numbers. The yellowish
colonies consisted of the form of Bacillus which appeared in the cultivation of
the earth previous to the additions of the commas, the blue ones of commas.
These were of rather large size and somewhat pale and granular. The smell of
the cultivation was bat-like.
Four days after the introduction of the commas another plate-cultivation
was set. On the next day it presented microscopic characters similar to those
present in the previous cultivation, the small blue colonies, however, being
relatively more abundant. The smell was a curious mixed one, the bat-like
odour being accompanied by a characteristic choleraic one. The results of
microscopic examination were identical with those in the previous case, the
large yellowish colonies being composed of large, straight Bacilli and the blue
ones of commas. On the following day the cultivation had a very strong,
unmixed odour of bats.
Seven days from the beginning of the experiment another cultivation was
started. Seventy-two hours later it had a strong, decidedly choleraic smell, and
was crowded with colonies. These, as before, consisted of large yellowish and
of small blue ones. The blue ones consisted of commas, both isolated and in
the vibrionic condition.
Ten days after the addition of the commas to the earth another plate was
set. It was examined after forty-eight hours and found to have a faint smell,
and to be full of colonies similar to those in the previous cultivation. As in it,
the blue colonies contained commas of slender form and only staining feebly.
On the twelfth day another cultivation was set. Twenty-four hours later
it was crowded with minute colonies and had a very faint, rather pleasant, vinous
smell. On the next day the smell remained unchanged no commas could be
found, the colonies seeming to consist of two forms of straight Bacilli only, one
presenting short, thick, isolated rods, the other slender ones, both separated
and associated in filaments.
On the fourteenth day another cultivation was set. Twenty-four hours
after it was absolutely crowded with colonies, the majority of which were of large
size and yellowish colour. The smell was disagreeable and bat-like. Small
slender commas were present in half the preparations, which were mounted.
Another cultivation was set on the fifteenth day. Two days later it had a
faint, choleraic smell, and was covered with bluish and yellowish colonies, of
various sizes. Five, out of six, preparations which were mounted contained
commas.
Scientific Memoirs by
On the twenty-first day another plate was inoculated. Two days later
it had a strong disagreeable smell, and was covered with large blue and yellow
colonies. Three out of four preparations contained commas.
'Twenty-eight days after the inoculation of the soil another plate was set.
Forty-eight hours later it had a strong but somewhat choleraic smell, and con-
tained numerous colonies, the majority of a yellowish tint, but a few bluish ones
also being present. The former consisted of large, thick, straight rods, the
latter of small commas.
On the forty-seventh day a final cultivation was set. In the course of
twenty-four hours it had become full of colonies, like those in the previous case,
and had acquired a faint, somewhat choleraic smell. The blue colonies, as before,
consisted of commas.
EXPERIMENT VI.—Some fresh fæcal matter was triturated with 100
grammes of dry garden-earth and set in a covered beaker as in the previous ex-
periment. Two days afterwards the growth of two tube-cultivations of commas
of four days' date was stirred up in sterilised salt solution and poured over the
earth. Nine days later the soil was still very moist, with a slightly fæculent
odour and a neutral re-action. A plate-cultivation of it was set. Numerous
schizomycete colonies and patches of fungal hyphæ appeared in it. It had no
perceptible odour and no commas could be found in it.
On the nineteenth day the surface of the soil was still quite moist, emitted
a faint stercoraceous odour, and was covered by a puffed-up sheet of fungal
mycelium, with radiant, whitish tufts of hyph. A plate-cultivation was set.
On the following day it was covered by a continuous stratum, of ochre colour,
which involved isolated colonies of several kinds. The smell was faint and dis-
agreeable but non-choleraic. No commas were to be found in it.
Taking the experiments as a whole, they indicate that comma-Bacilli, when
introduced into soil and water of very different qualities, so long as these retain
their natural condition, tend to disappear very rapidly.
The periods of survival are shown in the following tables :
—
Table I.—Periods of Survival of Commas in Water.
No. of Ex-
periment.
Quality of Water, &c.
Period.
No. I.
Fairly clean : unboiled . Disappearance in 4 days.
„ II.
, ,
, ,
.
, ,
,,
5 ,,
„ III.
Foul : unboiled
.
.
, ,
,,
4 ,,
„
„
IV.
V.
„
boiled
„
unboiled
.
, ,
, ,
,,
2 5
9
,,
10
Medical Officers of the Army of India.
Table II.—Periods of Survival of Commas in Soil.
No. of Ex-
periment.
Quality of Soil.
Period.
No. I.
Garden earth
. . Absent after 26 days.
"
II.
, ,
, ,
.
.
, ,
, ,
14 , ,
" III .
, , , ,
.
•
, ,
, ,
10 , ,
„
IV.
Garden earth and fæces .
, ,
, ,
6 , ,
„
V.
Garden earth and faces;
boiled.
Still present after 47 days.
„ VI.
Garden soil and fæces
unboiled.
; Absent after 9 days.
The period during which they retain their vitality is very much shorter than
that in cases of pure cultivations, for in Calcutta successful cultivations may be
obtained from tubes up to a period exceeding sixty days. The fact of their
rapid disappearance from foul water if left in its natural state has been already
clearly demonstrated by Koch's celebrated observation on their occurrence in
the water of a tank in Calcutta during a period of epidemic prevalence of
cholera in the surrounding huts, i.e. during a period when fresh supplies of them
were liable constantly to gain access to the water—and their disappearance
shortly after the cessation of supply consequent on the termination of the out-
break.
Two exceptional cases are present, one in the set of experiments on water,
the other in that on the soil, but in both these the materials had not been left
under normal conditions previous to the access of the commas, but had been ex-
posed to the influence of temperatures high enough in the case of the water to
practically sterilise it, and in that of the soil to determine the destruction of
the filamentous fungi, and probably of large numbers of the schizomycete
organisms originally present in it. In both cases commas persisted longer than
in the other experiments, and in that of the soil the persistence was such as to
render the result practically identical with that in an ordinary pure tube-cultiva-
tion. The shorter period of persistence in the case of the boiled water may be
ascribed with probability to the much smaller amount and, therefore, more rapid
expenditure of nutritive material present. The proportion of nutritive material
present in a medium seems not only to determine the duration of vitality in the
commas, but also induces differences in the development of the individual
specimens. This is, of course, well known in connection with the appearance of
" involution" forms in nearly exhausted media, but, in addition to this, variations
in size wit bout alteration in formend to occur in many cases. There is frequently
C 2
Medical Officers of the Army of India.
15
possessed by the commas in their normal state.1 For example, while the
ordinary commas are killed by exposure to drying for a few hours, the Arthro-
spores are said to retain their vitality under similar circumstances for weeks,
and while commas are incapable of surviving the action of the normal gastric
juice the Arthrospores can survive it, and are therefore capable, without any
assistance, of inducing cholera in healthy animals to whose digestive canal they
have gained access.
The formation of Arthrospores, according to this, comes to replace the adju-
vant measures or the naturally developed dyspeptic conditions of the previous class
of experiments. In order, however, to show that the Bacilli alone are efficient
determinants of epidemic diffusion of cholera, it would still be necessary to show
that Arthrospores are normally formed under the ordinary conditions to which
the parent commas and spirochtal forms are exposed. Assuming that Hueppe
is perfectly correct in regard to the formation, nature, and properties of the
Arthrospores, there is as yet an absence of evidence to show that they are
formed under normal conditions. Their development has only been observed
under the influence of special conditions, and one of these apparently is that
the parent Bacilli were in pure cultivations.
There is a great tendency to assume that all phenomena presenting them-
selves in pure cultivations are to be taken as sufficient indices to the normal
behaviour of the special organism under observation in other circumstances.
But this, of course, by no means necessarily is the case, and there is no evi-
dence to show that in the case of the Comma-Bacilli any development of
Arthrospores would occur, even under the influence of exhaustion of medium
and special temperature, if the cultivations were no longer pure ones; or, in
other words, were the Bacilli exposed to a struggle for existence. On the
contrary, there is evidence indicating that under such circumstances there is
no tendency to the development of Arthrospores, but merely to rapid and final
disappearance of the Bacilli. Taking the case of the present experiments
on their behaviour in soil and water of various degrees of pollution—that is, in
media resembling those to which they must normally tend to gain access in
greatest numbers—the evidence was entirely in favour of rapid extinction and
against the development of any specially resistent forms. These results are
not isolated ones, for, although it has been affirmed that Comma-Bacilli are
specially inimical to certain saprophytic Schizomycetes, 2 the general consen-
sus of evidence and opinion must lead one to regard this as a very exceptional
phenomenon, and to believe that, generally speaking, and under normal cir-
cumstances, they are singularly incapable of holding their own in the struggle
for existence. When Comma-Bacilli are imported into any locality they do
1 Ueber die Dauerform der sogenannten Komma-bacillen—F. Hueppe. Deutschen Medicinischen Wochen.
schrift, 1885.
2Bakteriolog. Untersuchung von choleraverdächtigen Fällen unter erschwerenden Umständen. Max Gruber,
1887.
20
Scientific Memoirs by Medical Officers of the Army of India.
venting the possibility of such importation needs no demonstration. But if it
be allowed that epidemics can only arise under the co-operation of certain favour
-ing local conditions, a hopeful field for efforts at prevention is at once opened out,
and with the practical recommendation that the improvements in local sanitation
which they necessarily imply will not have been absolutely fruitless, even should
they not at once arrive at their special aim in reference to cholera. On every
occasion on which quarantine or similar measures attempting to prevent impor
-tation are enforced, and cholera, in spite of them, succeeds in establishing itself,
even the most ardent believers in the benefit of such procedures must allow that
a futile waste of effort and money has taken place, but any improvement in local
conditions must remain a benefit, even though its immediate purpose be not
achieved.
CALCUTTA:
14th October, 1888.
Notes on the Life-history of Ravenelia sessilis, B. and Ravenelia
stictica, B. and Br.
BY
D. D. CUNNINGHAM, M.B.,
BENGAL MEDICAL SERVICE.
During the past few years I have, as opportunities presented themselves,
been accumulating notes on the life-history of the two species of Ravenelia
mentioned in the heading, and as these now are fairly complete save in regard
to one or two details, it appears to be worth while to publish them, more
especially in consideration of the very imperfect knowledge which has as yet
been arrived at in regard to the peculiarities of the genus. In so far as I am aware,
all that is yet known is that Ravenelia is a genus of the Uredineæ producing
uredospores and very peculiar teleutospores. In regard to the development of
the latter, however, there would appear to be no accurate information, and in re
-gard to the presence of any other forms of fructification no information of any
kind. In the present paper I hope to be able to show that in both species a
development, of what must apparently be regarded as spermogonia forms the
starting-point of each annual cycle of development, and that in Ravenelia sessilis
no less than five different forms of reproductive bodies are present,—spermatia,
two forms of uredospores, and two forms of teleutospores.
Both species are extremely common in the neighbourhood of Calcutta,
R. sessilis occurring on a large number of trees of Albizzia Lebbek, and
R. stictica appearing everywhere on the leaves of Pongamia glabra.
Taking R. sessilis first and the period of complete defoliation of the host,
which occurs in the beginning of the hot weather—as a rule during March—
as a starting-point, we find the following series of phenomena manifesting itself.
For the brief period during which leaves are entirely absent the only sites on the
trees of Albizzia Lebbek on which any traces of the parasite are to be found are
the dried-up and ripening pods, many of which remain adherent for a consider
-able period after the new crop of leaves has fully expanded. On these the
remains of large teleutosporic clusters, and sometimes of mixed ones containing
2 2
Scientific Memoirs by
both releutospores and uredospores are to be met with in varying numbers.
The number of spores which are present in them is, however, small. On the dried
dead pinnules beneath the trees, however, great numbers of teleutospores are
to be found, together with smaller numbers of uredospores. When the new crop
of leaves first makes its appearance and until the latter have fully expanded and
the flowers have unfolded in numbers, there is no indication of the presence of
the parasite. This state of things is, however, of very brief duration, and the
leaves speedily become spotted with patches of spermogonia (Fig. 13, Plate I).
Taking the phenomena presented by a tree in the year 1887 as an example,
the following was the sequence of events: On March the 11th the tree was
quite bare of leaves. On the 28th it was in full new leaf and just coming into
flower, but showed no traces of a new crop of Ravenelia sessilis. On April the
3rd the new leaves were universally spotted, due to the presence of groups of
spermogonia, and on the 17th the latter were beginning to dry up and blacken,
while pustules of uredospores were appearing here and there at the edges of the
groups.
The spermogonia make their appearance as small groups of minute,
dull brown, circular, elevated points on somewhat discoloured and yellowish
areas of the surfaces of the pinnules (Fig. 13, Plate I). The discoloration,
as a rule, penetrates right through the thickness of the pinnules, and spermogonia
appear at corresponding points on both upper and under surfaces. The
size of the patches and the number of spermogonia which they contain
vary considerably in individual instances. In groups of average size the
spermogonial area has a diameter of about 0.5 mm., and is surrounded by a
discoloured zone about 0.25 mm. in breadth. The number of spermogonia
in a group varies from 1 or 2 up to 30, but, as a rule, 10 or 12 are present.
In most cases they are arranged around a central point in circular fashion,.
several concentric circles being present in the larger groups, but in some
instances the arrangement is irregular, or even almost linear. The number
of groups on individual pinnules varies very considerably. In one case as many
as 20 were present on the upper and 18 on the lower surface. Vertical sections
through the pinnules show that the spermogonial areas of the laminæ are much
thinner than surrounding ones, the patches being sunk below the level of the
rest of the surface both on the upper and under sides of the leaves.
These thin spermogonial areas are throughout so densely occupied by
fungal hyphæ that the normal structure of the pinnule is quite obscured.
They stain very deeply with Spiller's purple. In the deeper, central portions of
the, lamina the hyphæ are arranged horizontally, so as to form a dense stratified
mass. At the level of the epidermis the fungal tissue becomes more or less
vertical, and masses of cells pass outwards between and around the compressed
epidermal cells at particular points corresponding, to the bases of the
spermogonia. At the level of the outer surface of the epidermis dense masses
Medical Officers of the Army of India.
23
of erect twigs are given off, which gradually elevate the cuticle as they elongate.
The peripheral twigs of each mass elongate much more than the central ones,
and, growing up convergently around them, gradually form a definite bounding
layer, which, as time goes on, gradually acquires a brown colour (Fig. 12, Plate I).
The hyphæ of the included, cushion-like mass are slenderer than the peripheral
ones. They do not elongate so much, but terminate in pointed extremities, which
play the part of sterigmata and bear minute oval spermatia (Figs. 10, 11. Plate
I). As maturation advances a definite opening arises at the apex of each
spermogonium to permit of the discharge of the spermatia. Individual sper
-mogonia vary greatly in size. Three which were situated side by side in one
patch gave the following measurements, the first set of figures referring to
diameter and the second to height
—
1 . = . 0 7 5 9 x . 0 1 9 8 m m .
2. = .1320x.0330 „
3 . = . 0 6 6 0 x . 0 2 9 7 „
They are always very broad in proportion to their height. The spermatia mea
-sure about 3.25x2.75µ. After the spermogonia have matured they dry up and
remain as minute black points in the centre of the patches of uredosporic pustules,
which are next developed. Before they begin to dry up they usually present
a peculiar glistening appearance, apparently connected with exudation of fluid
from their orifices.
With the drying up of the spermogonia pustules of large uredospores
begin to appear in the peripheral zones of the patches. They differ both in
external appearance and in minute structure from the spermogonia. Instead of
appearing as minute circular bodies opening at the apex by a definite orifice,
they are of elongate form and open by mere irregular rupture. The latter
phenomenon is due to the fact that they have not anything corresponding to
the special bounding stratum of the spermogonia, and that the opening of the
pustules is effected by mere rupture of the cuticle of the leaf, which has been
gradually elevated by, and distended over, the growing mass of fungal tissue
represented by the spores and the bed from which they arise.
The uredosporic pustules gradually increase in number, and form one or
two rings around the central spermogonial areas (Fig. 14 a, Plate I). Individual
pustules sometimes become confluent, but the tendency to such an occurrence
is never so great here as in the case of teleutosporic ones. There is no diffu
-sion of infection over the general surface of this crop of leaves, each uredo
-sporic group of pustules is situated immediately around a previously developed
spermongonial one, and the number of groups of uredosporic pustules on any
pinnule is hence determined by that of spermogonial ones. The deeper tissues
of the leaf beneath the uredosporic areas are not nearly so much invaded by
mycelium as they are in the central spermogonial areas. In fact, as is the case
with the corresponding areas in the succeeding crop of leaves, it is often
very difficult to demonstrate the presence of mycelial elements in them at