Evening bulletin. [volume] (Honolulu [Oahu, Hawaii]) 1895-1912, 1910-08-13


yet. nor are the prospects of these
being isolated very good, one OF
these being Of extremely slow growth
and the other containing spore
bearing bacillus as a contamination.
We have, however. a number of 9th
er tubes inoculated from other cases
which are not yet ready to be car
ried forward. but some Of these look
very promising. From these latter
we hope to secure additional strains
OF the gaclli. We believe from our
experience thus far that with in.
creaslng knowledge Of the details OF
the work, we may expect In future
to Isolate lepra bacilli with much
less difficulty than we at first expe-
rienced.
Our experience appears to indicate
that the aforementioned points are
essential TO success, but we Tecog-
nize that our experience is as yet
! too limited to state positively that
success can not be met with II one
should disregard these points:
Flrst-The kind OF amoeba one
employs is probably matter of In
difference, provided II is active
enough to migrate across Petri
dish before 1t encysts, otherwise Its
original symbia'nts can not be re.
placed by the symbiant of choice.
SecOnd-We employed -. coli, b.
typhi murium and S. cholerae as
! symbiants; We were only successful
when the latter organism was used.
Third.=We found II is essential TO
clear the salt agar media OF Clegg
with an egg. The egg, even after
coagulation and filtration. impar's
sufficient nutriment to the media to
sustain life Of the amoebae and
symbiant.
/ Fourth-Our incubators lowing
to the use of home made" gas)
nuctuated between "a c and 3S G.,
( but these minor variations appear
to have no unfavOrable influence on
4 growth.
Fifth--The experience of one of
| us tBrln"kerhonD working in the
! dry air Of Honolulu was that tho
media tubes should have an album
dance OF water OF condensation in
them, and should be capped to avoid
evaporation. In the experience OF
the writer working in the moist air
of Kalawao, this was not found TO
! be essential
Sixth--We only met with success
in our NEGOTIATIONS from rather
! early, untreated cases. Failure re-
suited from our late cases or early
cases that had been under Chaul-
moogra oil for a considerable period.
This, however. may have been only
COincident.
' Seventh--We only employed re-
cently-deposited, non-ulcerated nod
ules or the sera from such nodlues,
in making our inoculations. We be-
lieve the sera containing bacilll Is
| to be preferred TO the nOdule II.
self, rich media Of human tissue
appearing TO cause q lack of balance
between the number of amoeba and
the number of the symbiant spirilla
| present, and thus mitigates against
success.
Eighth-Clegg repeatedly inocu,
lated the same tube on three sue
cessiye days; we did not find this
practical, so endeavored to succeed
by single incubation. We found,
however, that such single inocula-
tions resulted in the amoeba destroy
ing the lepra bacilli by phagoeyto.
sis. To overcome this we allowed
the amoeba to first encyst, then
inO2ulated the tube with the leprous
material. It would appear that one
must resort to either one or the
other Of these methods, either the
tube must be enriched on successive
days to replace the bacilli lost, or
we must await a,stage OF the amoeba
when they are no longer capable of
phagocytosis.
Ninth-ln this, the ninth point,
our experience differs considerably
from that of Clegg. That gentle
man recommended the examination
OF the tubes about the tenth day
I for the purpose OF asaertaining
whether multiplication had occurred.
! This wc did In the early part Of
! our work and believe that our first
failures were largely We TO dis
turbing the tubes at this time. we