New York AGRICULTURAL EXPERIMENT Station. 179 
finally rotted the rot started on the stem half instead of the spot- 
affected calyx half. At another time, two affected apples were 
kept in a moist chamber for 18 days with the same results. 
Next, an experiment was made to determine if the diseased 
tissue would produce any growth when placed in culture media. 
Four Petri dishes” containing potato agar slightly acidified with 
lactic acid were each inoculated at three different points with 
small pieces of the brown spongy tissue. This gave twelve points 
of inoculation with material from twelve different spots. The 
cultures were kept at the room temperature, about 70° Tahr. 
At the end of eight days one point of inoculation was overrun by 
a fungus which had gained admission to the culture by accident. 
The other eleven points of inoculation were entirely free from 
growth of any kind. 
On November 1 six tubes of neutral beef-peptone agar were 
inoculated with small pieces of the brown, spongy tissue taken 
from six different spots, and then poured into Petri dishes. At 
the end of eight days the only growth in the six dishes consisted 
of one fungus and two yeast colonies which were evidently 
intruders. We now tried cultures in an atmosphere devoid of 
oxygen. Six tubes were used — two of potato agar, two of beef- 
peptone agar and two of beef-peptone agar containing two per ct. 
of lactose. One tube of each kind was slightly acidified with malic 
acid and the other left neutral. The six tubes were inoculated 
with bits of spongy tissue from six different spots, thoroughly 
shaken and placed in a large bottle from which the oxygen was 
then removed by means of pyrogallic acid and potassium hydrox- 
ide solution. These cultures were kept at a temperature of about 
80° Fahr. for one week but they produced no growth whatever. 
Finally, at the suggestion of Mr. H. A. Harding, Station 
Bacteriologist, we tried apple peptone agar” as a culture medium. 
21 Two dishes acidified at the rate of one drop of 50 per cent. lactic acid to 
10 c. ec. agar, and the other two dishes at double this rate. 
22 Baldwin apple, 400 germs. Witte’s peptonum siccum, 10 germs. Agar, 
15 germs. Water, 1 liter. 
