NationalLibraryOfScotland/medical-history-of-british-india huggingface dataset:
4
Scientific Memoirs by
Number
of speci-
men.
Date at
which
pro-
cured.
Source.
General condition
which obtained.
Nature and treatment
of individual
samples.
Subsequent condition.
XII May
31st.
Bazaar . The milk siphoned
into distinct sam-
ples derived from
the upper and
lower strata.
I. Tube of the top
milk in the in-
cubator at 37°C.
June 1st. Reaction acid ; fully co-
agulated and separated.
2. Tube of the bot-
tom milk in the
incubator at
37ºC.
June 1st. Reaction acid, but not so
strongly so as that of the previous
sample : fully coagulated and separ-
ated.
XIII June
7th.
,,
. Milk treated as in
the previous spe-
cimen. Reaction
neutral.
1. Tube of the top
milk in the in-
cubator at 37°C.
June 8th. Acid ; fully coagulated
and separated.
2. Tube of the bot-
tom milk in the
incubator at
37°C.
XIV July
15th.
„
. Milk treated as
above. Reaction
faintly alkaline on
drying.
I. Tube of bottom
milk in incuba-
tor at 37°C.
July 16th. In soft coagulum, but not
separated.
July 17th Fully coagulated and
separated ; reaction strongly acid.
XV July
25th.
,,
.
Milk treated as
above. Reaction
neutral.
I. Tube as in the
previous case.
July 26th. Reaction acid ; in soft
coagulum, but not separated.
XVI July
29th.
„
. Treated as above.
Reaction neutral.
1. Tube as in the
previous case,
July 30th. Acid ; in soft coagulum,
afterwards undergoing separation.
XVII Aug.
3rd.
„
. Treated as above.
Reaction neutral.
I. Tube as in the
previous case.
August 4th. Not coagulated until it
was boiled, when coagulation and
separation occurred at once.
XVIII Aug.
15th.
„
. Treated as above.
Reaction faintly
alkaline.
I. Tube as in the
previous case,
August 15th. Reaction strongly acid ;
coagulated and partly separated,
with a little froth above.
This table shows the changes which specimens of milk of various qualities
normally tend to undergo if left without any special treatment. When first ob-
tained, the milk as a rule was neutral in reaction, in some cases faintly alkaline
on drying, and in one transitorily faintly acid. After an interval of 24 hours the
reaction in all ordinary samples of milk is acid and in most cases strongly so,
and, in association with this change in reaction, coagulation has occurred, in
most cases fully developed and with separation into coagula and fluid, but in
some not so far advanced and with an absence or incompleteness of separation.
In rare instances a certain amount of frothing is present, but as a rule the sur-
face of the fluid is merely covered with a layer of coagulated cream. The
occurrence of coagulation appears to be influenced by temperature, as in some
instances in which corresponding samples of milk were set simultaneously in
the open Laboratory and the Incubator at a temperature of 37°C., complete co-
agulation occurred more rapidly in the incubated specimens than in those ex-
Medical Officers of the Army of India.
5
posed to the ordinary atmospheric temperature, which, as a rule, was considerably
lower than that of the incubator, and in no instance was constantly at such a
high level during the entire period of 24 hours. In only one case was there
neither permanent acidity nor coagulation after the specimen had been under
observation for a period of 24 hours, and in this the milk had been obtained
under very special precautions. The specimen is that which is numbered VI in
the Table and was drawn direct into a sterilised beaker from which the plug of
cotton wool had just been withdrawn and at once transferred thence to a steri-
lised and plugged flask, and finally to a sterilised tube. The udder and 'teats of
the cow and the hands of the milkman had both been freshly washed, first with
a 5% solution of corrosive sublimate and then with spirit, immediately before the
milk was withdrawn. In spite of being kept constantly in the incubator, spon-
taneous coagulation had not occurred up to a period of 96 hours afterwards, and
only set in on boiling. The reaction too was quite exceptional, as it was only
transitorily acid after the lapse of 24 hours, and only faintly acid permanently
after 96 hours. The result clearly shows that the development of acidity and of
coagulation is determined, principally at, all events, by contamination of the fluid
during drawing and subsequent preservation.
TABLE II .—Results of Plate Cultivations of Unboiled Specimens of Milk from
various sources.
Number of
specimen.
Date at
which
P r o c u r -
e d .
S o u r c e . Reaction.
Nature
of
cultivation.
Date at
which
set.
RESULT.
I Jan.
2nd.
Bazaar
. . .
Plate. Inoc-
ulated with
0.083 C. C.
Set in incu-
bator at 37°C.
Jan.
2nd.
January 3rd. Crowded with concrete colonies
in the proportion of about 100,000 per C. C.
The colonies were both superficial and inter-
stitial. In addition to these there were several
bluish, diffuse patches of growth, and two of
a bright greenish yellow tint. The schizomy
cete forms present included two kinds of
Micrococci ; a small Baccillus, the individual
rods with rounded ends and often united it
pairs; a slender straight Bacillus ; and
a limited number of larger rods, apparently of
Bacillus subtilis, which occurred among the
other forms and must apparently have been
due to a thin, diffuse, superficial growth
contaminating the specimens derived from
the distinct colonies. Smell of the cultivation
mouldy, non-choleraic.
II Jan.,
7th.
House-
supply.
...
Plate. Treat-
ed as in the
previous case.
Jan.
7th.
January 8th. A growth of colonies, similar to
those in the previous case, in the proportion of
20,000 per C. C.
6
Scientific Memoirs by
Number
of
specimen.
Date at
which
procure-
R d .
Source.
Reaction.
Nature
of
cultivation.
Date at
which
set.
RESULT.
III
Jan.
9th.
House-
supply.
. . . Plate. Treat-
ed as in the
previous spe-
emen.
Jan.,
9th.
January 10th. A growth of colonies in the
proportion of 48,150 per C. C. Almost all of
them appeared to consist of the short, oval,
often paired Bacilli described in the account
of the cultivation of specimen I. As in that
case scattered rods of larger size and appa-
rently due to diffuse growth of Bacillus
Subtilis were also present.
IV Jan.
16th.
, ,
.
Neutral .
, ,
. Jan.
16th.
January 17th. Crowded with concrete colonies
in the proportion of 132,000 per C. C. Also
a thin, diffuse, superficial growth visible at one
point. The defined colonies consisted of the
short oval Bacilli and the smaller form of
Micrococcus occurring in the cultivation of
specimen I. The diffuse growth consisted of
Bacillus subtilis.
V Mar.
1st.
, , . Faintly
alkaline.
, , . Mar.
1st.
March 2nd. Small colonies in the proportion
of 80,000 per C. C. One definite patch and
apparently a very thin, generally diffused film
of Bacillus subtilis. The small colonies con-
sisted of the common short oval Bacilli occur-
ring in the previous cultivations and of minute
straight Bacilli and large Micrococci similar
to those present in the cultivation of speci-
men I.
, , . Mar.
4th.
March 5th. Crowded with minute colonies of
short oval Bacilli and large Micrococcus.
VI Mar.
8th.
, , . Neutral .
, ,
.
Mar.
8th.
March 9th. Colonies present in the proper-
tions of 103,950 per C. C. These almost
entirely consisted of the large Micrococcus
occurring in several of the previous cultiva-
tions. As in other cases, a very limited,
superficial, diffuse growth of Bacillus subtili
appeared to be present, the stratum, how-
ever, being too thin to be recognisable.
, ,
.
Mar.
9th.
March 10th. Crowded with minute colonies of
the large Micrococcus and the oval Bacillus
occurring in previous cultivations.
VII Mar.
10th.
, ,
.
, ,
.
, ,
.
Mar.
10th.
March 11th. Colonies present in the propor-
tion of 51,000 per C. C. The greater number
of these consisted of the common lacteal
oval Bacilli. One large foliated patch of the
large Micrococcus.
VIII Mar.
12th.
, ,
.
, ,
.
, ,
.
Mar.
12th.
March 13th. Crowded with colonies in the
proportion of 340,000 per C. C. These con-
sisted of Bacillus subtilis, the common lacteal
oval Bacilli, large Micrococcus and slender
straight Bacilli. A certain number also were
apparently composed of some form of yeasty
growth of cells of some higher fungal species.
IX May
9th.
Zoolo-
gical
Garden.
, ,
.
Plate. Inoc-
ulated with
0.058 C. C.
Set in incu-
bator at 37°C.
May
9th.
May 10th. Colonies of the common, large,
lacteal Micrococcus in the proportion of
25,245 per C. C.
Medical Officers of the Army of India.
7
Number of
specimen.
Date at
which
procur- Source.
Reaction.
Nature
of
cultivation.
Date at
which
set.
RESULT.
Plate. Treat-
ed as before.
May
11th.
May 13th. Absolutely crowded with colonies
of the common, large, lacteal Micrococcus.
X May
21st.
House-
supply.
Faintly
acid, be-
coming
neutral on
drying.
Plate. Treat-
ed as before.
May
21st.
May 22nd. Absolutely crowded with colonies
of the common, short, oval, lacteal Bacillus.
XI May
23rd.
,, . Neutral,
becoming
faintly al-
kaline on
drying.
, ,
.
May
23rd.
May 24th. Relatively very few colonies, the
proportion being only 3,400 per C. C. They
consisted of the common lacteal oval Bacilli
and large Micrococcus.. A very thin, diffuse
growth of Bacillus subtilis appeared also to
be present.
, ,
.
May
24th.
May 25th. Crowded with colonies of the com
mon lacteal oval Bacilli and large Mi-
crococci.
XII May
25th.
, ,
.
, ,
.
, ,
.
May
25th
May 26th. Crowded with colonies of the com-
mon lacteal oval, and small straight Bacilli,
and large Micrococcus. A thin, diffuse growth
of Bacillus subtilis appeared also to be pre
sent.
XIII May
271h.
Bazaar
, ,
.
, ,
.
May
27th.
May 28th. Number of colonies very small,
only giving a proportion of 8,670 per C. C.
They appeared to consist entirely of the
common large, lacteal Micrococcus. There
was also, seemingly, a very thin, diffuse
growth of Bacillus subtilis present.
XIV May
29th.
, ,
.
...
, ,
.
May
29th.
May 30th. Numerous colonies were present.
These consisted of large Micrococci and
common lacteal oval Bacilli, which, where
situated at the lower surface of the Agar-
Agar, had in many cases elevated it into bub-
bles, due to gaseous evolution.
XV May
31st.
, ,
.
, ,
.
Plate. Inoc-
ulated with
0.058 C. C.
Siphoned off
from bottom
of the milk.
May
31st.
June 1st.
Both plates alike. Containing
numerous colonies of the corn-
mon lacteal Schizomycetes.
Plate, Similar.
ly inoculated
from top of
milk.
, ,
.
, ,
.
XVI June
7th.
, ,
.
Neutral . Plate. Inoc-
ulated with
0.058 C. C.
of milk from
the top.
June
7th.
June 8th. Numerous colonies of the common
lacteal Schizomycetes.
Plate. Inoc-
ulated with
0.058 C. C.
from the bot-
tom.
, ,
.
June 8th. Colonies not nearly so abundant
as in the previous cultivation, but of the same
nature.
8
Scientific Memoirs by
Numbe o f
specimen.
Date at
which
procur- ed. Source. Reaction.
Nature
of
eultivation.
Date at
which
set.
RESULT.
XVII July
15th.
Bazaar Faintly
alkaline on
drying.
Plate. Inoc-
ulated with
0.03 C. C.
from the bot-
tom of the
milk.
July
15th.
July 16th. Crowded with colonies of common
lacteal Schizomycetes in the proportion of
340,965 per C. C. Small straight Bacill
and short oval ones the most prevalent forms
XVIII July
25th.
„ . Neutral . Plate. Inoc-
ulated as
above with
0.03 C. C.
„ . July 26th. Smell mouldy. Abundant colonies
of typical lacteal Micrococcus, oval and
small straight Bacilli.
XIX July
29th.
„ .
„
Plate. Inoc-
ulated as
above.
July
29th.
July 3oth. Numerous colonies in the propor-
tion of 51,172 per C. C. Most of them con-
sisting of the common lacteal large Micro-
coccus and short oval Bacilli. A few super-
ficial bluish colonies of small, straight Bacill
also present.
XX Aug.
15th.
„ . Faintly
alkaline.
„
. Aug.
15th.
August 16th. Colonies present in the propor-
tion of 39,847 per C. C. Consisting of com-
mon lacteal Micrococcus and small straigh
t B a c i l l i .
This table records the results of cultivations of unboiled milk. These
show that the milk in common use in Calcutta contains a varying but always
very large number of schizomycete organisms. The numbers actually present
in those cases where an estimate was made by numeration of colonies present
within determined areas of plate-cultivations ranged from a proportion of only
3,400 per C. C. of milk to over 300,000 per C. C.
Determinations of this nature were made in regard to 14 specimens with
the following results :—
Case 1
2
100,000
20,000
Schizomycetes
„
per
„
C. C.
„
of milk
„
3 48,000
„ „ „ „
4 132,000
„ „ „ „
5 80,000
„ „ „ „
6 103,950
„ „ „ „
7 51,000
„ „ „ „
8 340,000
„ „ „ „
9 25,245
„ „ „ „
10
3,400
„ „ „ „
1 1
8,670
„ „ „ „
12 349,965
„ „ „ „
13
14
51,172
39 847
„ „ „ „ „ „ „ „
One noteworthy point is that the exceptional specimen of milk procured under
special precautions, and which did not undergo the normal acid fermentation and
coagulation, was not free from schizomycetes, nor did it even contain an excep-
10
Scientific Memoirs by
reveal them. It is very probable that growths of Bacillus subtilis of this
nature were present in other cultivations of the series than those of which it is
recorded, as the results of cultivations of boiled milk, as will be shown pre
-sently, clearly demonstrate that spores of Bacillus subtilis are of constant occur
-rence in ordinary samples .of milk. The schizomycetes, which may be fairly re
-garded as normal constituents of ordinary samples of milk in Calcutta, are thus
a large Micrococcus, an oval Bacillus, a small slender Bacillus, and finally
Bacillus subtilis.
Other forms, such as the small species of Micrococcus occurring in specimen
I of Table II, and one or two other Bacilli, sometimes occur in small numbers,
but their appearance is an exceptional, not a normal phenomenon. In the case
of one specimen, No. VIII of Table II, cultivation, in addition to colonies of
common lacteal schizomycetes, yielded others composed of dense masses of
yeast-like cells. The first four photographs in the plate show the proportions of
colonies present in cultivations of unboiled specimens of milk of twenty-four
hours' growth. The first three are from cultivations set immediately after the milk
was obtained and figure 3, when taken along with the two previous ones, shows
the relatively small numbers of colonies appearing in a cultivation of the speci
-men of milk obtained under special precautions to avoid contamination, as com
- p a r e d w i t h t h e n u m b e r s p r e s e n t i n c u l t i v a t i o n s o f s p e c i m e n s o f t h e c o m m o n m i l k
supply of Calcutta. The fourth figure, compared with the third,shows the excessive
multiplication of schizomycetes in specimens of milk when kept for any length
of time. Both cultivations to which the figures correspond were obtained from
the same sample of milk, but that illustrated in figure 3 was set immediately
after the sample was obtained, while that of figure 4 was not set until 48 hours
later. One point which came out very clearly in this series of cultivations was
the relative repression, and in many cases apparent complete suppression of
Bacillus subtilis in milk, which is associated with the presence of the other
schizomycetes normally present in the fluid and the acid fermentation with which
their presence and multiplication is associated. As will be shown in the fourth
Table, spores of Bacillus subtilis appear to be a constant content of ordinary.
specimens of milk, but the development of the species in all these cultivations
was inconspicuous, and in many instances no development at all seemed to
occur. The repressive influence of the other forms was specially indicated by
the fact that when growths of Bacillus subtilis did occur they were associated
with the first cultivations derived from any specimen, that is, at periods when
the numbers of the common lacteal schizomycetes were relatively small, as com-
pared with what they were subsequently.
Medical Officers of the Army of India. 1 1
TABLE III.—Changes occurring in Boiled Specimens of Milk from various sources
and under various conditions.
Number
of speci-
men.
Number
of corre-
sponding
specimen
of unboil-
ed milk in
Table I.
Date at
which p r o -
cured.
Source.
General
condition when
obtained.
Nature and treatment
of individual
samples.
Subsequent condition.
I
...
Jan.
9th.
House-
supply.
. . .
I. Boiled in a steri-
lised tube and
set in the incuba -
tor at 37°C.
January 10th. Remaining ap-
parently . unaltered ; quite fluid.
.
Again boiled, January
10th and 11th.
The specimen throughout re-
mained seemingly unaltered,
save for a faint brownish tinge
which it acquired in connection
with the repeated boiling. Sub -
sequent to the 2nd boiling it
had a faint acid reaction. It
remained fluid for as long as it
was kept in the laboratory.
II
...
Jan.
16th.
„ Reaction neu-
tral.
I. Boiled in a steri-
lised tube and set
in the incubator
at 37°C.
January 17th. Remaining ap -
parently unaltered ; quite fluid.
Again boiled on Ja-
nuary 17th.
Subsequent to the second boiling
it remained unaltered during a
period of many weeks in which
it was retained in the labo-
ratory.
III
II Mar
8th,
„
Neutral . Treated as above . March 9th. Reaction strongly
acid ; fully coagulated and
separated into coagulum and
fluid.
IV
III Mar.
10th.
„
„
.
„
„
.
March 11th. Remaining fluid
and seemingly unaltered.
Re-boiled on March
11th.
March 13th. Remaining un-
altered.
V
IV Mar
12th.
„
„
. Treated as above . March 13th. Reaction neutral
quite fluid.
Boiled again on 13th
March.
March 14th. Seemingly un -
altered.
VI
V Mar.
15th.
Bazaar.
„ . Treated as above . March 16th. Coagulated and
separated into coagulum and
fluid; smell caseous; reaction
acid, becoming feeble on dry -
ing.
VII
VI May
9th.
Zoolo-
gical
Garden.
„ .
„
„
.
May 10th. Quite fluid ; reaction
neutral, becoming feebly alka -
line on drying.
Re-boiled once in the
next five days and
twice on the 6th day.
Remained unaltered throughout
as long as it was retained in the
laboratory.
C 2
12
Scientific Memoirs by
Number
of speci-
men.
Number
of corre -
sponding
specimen
of unboil-
ed milk in
Table 1.
Date at
which
p r o -
cured.
Source.
General
condition when
obtained.
Nature and treatment
of individual
samples.
Subsequent co ndition.
VIII
. . .
May
9th.
Zoologi-
cal Gar-
den.
Obtained at the
same time as
the previous
specimen,but
from a differ -
ent cow. The
udder was
not in this
case special-
ly prepared
and the calf
had to be al-
lowed to suck
ere the ani -
mal would
let down her
milk. It was
drawn direct
into a steri -
lised beaker
and transfer -
red to three
sterilised
flasks.
1. Boiled in a steri-
lised flask and
set in the incu-
bator at 37°C.
May 10th. Fully coagulated
and separated. Reaction acid.
2. Treated as above. May 10th. Coagulated, but
separation only beginning; co m -
plete separation occurring at
once of boiling.
3. „ „ . May 10th. Result as in sample
2.
IX
VII May
21st.
House-
supply.
Re-action faint-
ly acid, be-
coming neu-
tral on dry-
ing.
1. Boiled in a steri-
lised flask and
set in the incu-
bator at 37ºC.
May 22nd. Fully coagulated and
separated ; reaction acid, be-
coming very faintly so on dry-
ing.
2. Treated as above . May 22nd. In soft coagulum,
which on boiling underwent
complete separation ; reaction
acid, becoming very faint on
drying.
3. Boiled in a steri-
lised tube and
set in the incu-
bator at 37°C.
May 22nd. Fully coagulated
and separated ; reaction acid
faintly so on drying.
X
VIII May
23rd.
„
Neutral, be-
coming faint-
ly alkaline
on drying.
1. In sterilised flask
at 37°C.
2. ,, .
May 24th. In soft coagulum,
undergoing separation on
boiling.
3. In sterilised tube
at 37°C.
May 24th. Fully coagulated and
separated ; reaction acid, faint-
ly so on drying.
XI
IX May
25th.
„
Reaction neu-
tral, faintly
alkaline on
drying.
1. Boiled in a steri-
lised flask at 8
A.M., and 1 P.M.
During the in -
terval and sub -
sequently in in -
cubator at 37°C.
May 26th Undergoing full co-
agulation and separation on
being anew boiled.
2. Treated as above . May 26th. Results as above.
Medical Officers of the Army of India.
13
Number
of speci-
men.
Number
of corre-
sponding
specimen
of unboil-
ed milk in
Table I.
Date at
which pro-
cured.
Source.
General
condition when
obtained.
Nature and treatment
of individual
samples.
Subsequent condition.
3. Boiled in a steri-
lised tube and
set in the incu-
bator at 37ºC.
May 26th. Fully coagulated and
separated.
4. Residuum in the
flask, clean but
non-sterilised, in
which the speci-
men was pro-
cured. Twice
boiled like I and
2, and left in the
open laboratory.
May 26th. Did not undergo
coagulation on being boiled
anew.
XII
X May
27th.
Bazaar. Reaction neu-
tral ; faintly
alkaline on
drying.
I. Boiled in a steri-
lised flask and
set in the open
laboratory.
May 28th. Not visibly coagu-
lated until reboiled, when coagu-
lation and separation occurred.
2. Boiled in a steri-
lised tube and
set in the incuba-
tor at 37°C.
May 28th. Fully coagulated and
separated; reaction acid, becom-
ing neutral on drying.
XIII
XI May
29th.
, ,
Obtained in a
sterilised flask
I. Boiled in a sterilised
flask and set in
the laboratory.
May 30th. In soft, loose coagu-
lum. Full coagulation and
separation taking place on
boiling.
2. A portion, after the
cream had been
allowed to rise and
partially skim-
med off, treated
as above.
3. Cream : boiled in a
sterilised tube and
set in the incuba-
tor at 37ºC.
May 30th. Fully coagulated
and separated.
4. Skimmed milk :
treated as 3.
5. Residuum in the
flask remaining
after the milk for
the previous speci-
mens had been
removed; boiled
and set in open
laboratory.
May 30th. Remaining unaltered
and not coagulating on being
boiled anew.
XIV
XII May
31st
, ,
Siphoned into
separate sam-
ples from the
top and bot-
tom.
1. Top-sample ; boiled
in a sterilised
tube and set in
the incubator at
37ºC.
June 1st. Fully coagulated and
separated : reaction acid.
Scientific Memoirs by
Number
of speci-
men,
Number
of corre-
sponding
s p e c i m e n o f u n b o i l -
e d m i l k i n
Table I.
Date at
which
pro
cured.
Source.
General
condition when
obtained.
Nature and treatment
of individual
samples.
Subsequent condition.
2. Bottom-sample ;
boiled in a steri-
lised tube and set
in the incubator
at 37°C.
June 1st. Quite fluid ; non-co-
agulant on boiling : reaction
feebly alkaline on drying.
3. Bottom-sample;
boiled in a steri-
lised flask and
set in the open
laboratory.
June 1st. Just as sample 2.
4. Nature and treat-
ment as in 3.
5. Residuum remain-
ing after the milk
for the previous
samples had been
drawn off: treated
as 3 & 4.
June 1st. Softly coagulated and
undergoing complete coagula-
tion and separation on boiling.
XV
XIII June
7th.
Reaction neu-
tral. Milk from
the bottom
siphoned off
into 34 steri-
lised tubes,
and milk
from the sur-
face into two
sterilised
tubes.
1. Thirty-three of the
sterilised tubes of
top-milk boiled at
10 A.M. and again
at 6 A.M. Set in
the incubator in
the interval and
after the second
boiling.
June 8th. All alike quite fluid and
neutral : non-coagulant on being
boiled anew. June 9th. Milk
still fluid ; contents of one tube
coagulating on being anew
boiled : all boiled again on
the 9th and kept in a boiling
water-bath for some hours both
on the 11th and 12th. One more
tube coagulated on the 13th.
All the rest, which have not yet
been employed in inoculation
experiments, remain fluid and
unaltered save slight browning
up to the present date, Septem-
ber 6th.
2. Residuum after re-
moval of the milk
for the tubes: boil-
ed for a long time
and left in the
open laboratory.
June 8th. Coagulating and se-
parating on boiling.
XVI
. . .
July
20th.
, ,
Neutral
. I. Milk from the bot-
tom ; boiled in a
sterilised tube and
set in the incuba-
tor at 37ºC.
July 21st. Quite fluid : neutral.
XVII
XV July
25th.
, ,
,,
.
1. Milk from the bot-
tom ; boiled in a
sterilised tube ;
in the incubator
at 37°C.
July 26th. Quite fluid.