102
Multiplication of B. anthracis, etc.
2. B, anthracis
Cultural
Ten goats, mostly from other experiments, and
14 guinea-pigs were used. With the goats, the
time of death after subcutaneous inoculation
of approximately the same amount of culture
was 31-85 hours (average 49). Within the hour
after death one femur was removed and the mar-
row cultured at once, the carcase was then placed
in a room at air temperature usually between 60
and 70ºF. for 6-24 hours when marrow from the
other femur was similarly cultured. In some
cases muscle was examined. With some of the
guinea-pigs the procedure was rather different;
both hind legs were removed, one was put in the
cold and the other at a higher temperature, usually
90ºF. After 8 or 24 or more hours the marrow of
each was cultured. In order to reduce the chance
of haemorrhage at the time of amputation the
operation was not performed for an hour or two
after death and ligatures were first applied to
the upper parts of the limbs. In all this work
in spite of every care the results were erratic,
most probably because the organism grows in
threads and colonies do not develop from single
organisms. It is, therefore, unnecessary to give
detailed results.
Microscopical
Since cultural examination gave indecisive
results, direct count was attempted of bacilli
and chains in the heart-blood of mice and guinea-
pigs immediately 'after death and after 6-12
hours storage of the carcase at 90°F. While in
the case of mice dying early, viz. within 24 hours
of inoculation, the chains had roughly doubled
in length during six hours at 90ºF.; similar evidence
could not be obtained with guinea-pigs. The
results as a whole indicate that multiplication of
the anthrax bacillus after death is not a, constant
feature.
3. Cl. chauvoei
The guinea-pigs and sheep, used for the experi-
ments, were inoculated intramuscularly in the
thigh with spore suspension or meat-liver culture,
with or without calcium chloride. In most cases
the sheep carcases were left in a shed at pre-
vailing air temperature, while guinea-pig carcases
were stored at 28° or 30ºC., the alimentary canal
in some cases being removed immediately after
death.
Cultural results
Preliminary experiments on 17 guinea
indicated that:
(1) At the time of death, unless this is un
delayed, the blackquarter orga
present in the blood, spleen and
marrow are in the vegetative form
animals which had been dead
hours, e.g. during the previous
spores might already have fo
in the bone-marrow.
(2) At the time of death most of the b
quarter organisms present in the
exudate are in the form of spores
cultures from the exudate are
infrequently contaminated with aer
(3) In carcases kept for 24-72 hours at 3
blackquarter bacilli in blood, s
and bone-marrow appeared to
multiplied and at least some of
had sporulated. One carcase was
into ice immediately after death
kept there for 44 hours when
multiplication was noted in the
marrow. A companion guinea-pig
at 28ºC. showed multiplication in
tissue. Multiplication in the
marrow may proceed slowly up
least the third day. The organ
also sporulates readily at 30ºC. in
taken immediately after death
allowed to clot under vaseline. 1
of the blood and gas product
beneath the seal indicates multip
tion [Hanna, 1897].
(4) In three of the animals it was proved
immunity tests that the organ
isolated from the blood after sto
of the carcase were blackqua
bacilli.
Observations of a similar nature were mac
17 sheep. While in sheep, as in guinea-
blackquarter organisms in the bone -marrow
the time of death are usually in the vegeta
form, some of them may have undergone spo
tion as early as three or "four hours after de
In two sheep, vegetative organisms alone
present in the bone-marrow six hours after de
whereas in both cases 22 hours after death
organisms had sporulated. In sheep cart
left at the prevailing air temperature, multipl
tion of this organism followed by sporula