X, B, 5 Roberg: Bacterial Infections 331 
seared with a red-hot platinum point. It was then placed in 
5 per cent lysol for five minutes, washed three times in sterile 
salt solution, and the final washing in Dunham’s peptone was 
placed in a tube and incubated for twenty-four hours. This 
tube was perfectly clear and did not show any bacterial growth. 
The lysol-disinfected larva, when crushed in Dunham’s pep- 
tone, incubated for twenty-four hours, and plated on Dieudon- 
né’s medium, did not reveal the presence of cholera vibrios. 
f. Sand inoculated into Dunham’s peptone, incubated and 
plated on Dieudonné’s medium, did not reveal the presence of 
cholera vibrios. Several variously formed bacilli and cocci 
were present. 
g. As the results of e and f indicate that the vibrios had 
been outgrown, not only in the sand but also in the intestinal 
tract of the larve, it was necessary again to replenish the cholera 
vibrios by the addition of 24-hour-old broth cultures to the 
medium in which the larvze were developing. 
The larve and pupz were removed and placed in a new 
sterile stender dish containing sterile sand. This was again 
drenched with broth cultures of cholera vibrios. 
h. On April 16, as no flies had emerged, another transfer was 
made to a new sterile dish and again drenched with vibrios. 
7. On April 18, as no flies had emerged, they were again 
transferred and redrenched with vibrios. 
7. On April 20 one fly emerged. As I was not present at the 
time of emergence, it was not tested for vibrios contained in 
its intestines. Another transfer was made into a sterile dish 
and again drenched with vibrios. 
k. On the morning of April 21 six or seven flies had emerged. 
Four of the pupz which were soon to emerge into flies were 
removed and placed on a sterile moist filter paper and covered 
with a beaker. During the day these pupz were constantly 
watched and late in the afternoon one fly emerged. Immediately 
upon emergence the fly was chloroformed to prevent its escape 
or possible reinfection from cholera vibrios on the filter paper. 
This fly was placed in 5 per cent lysol for five minutes and 
washed three times in sterile salt solution. The final washing 
was in Dunham’s peptone, which was incubated for twenty-four 
hours and then plated on Dieudonné’s medium. Examination 
of the plate showed no cholera vibrios. 
The lysol-disinfected and washed fly was crushed in Dunham’s 
peptone solution, incubated for twenty-four hours, and streaked 
on Dieudonné’s medium. Examination of this plate revealed 
the presence of cholera vibrios. 
