Xew York Agkicultueal Expekimext 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 

 kei:)t in a moist chamber for 18 days with the same results. 



jSText, 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° Fahr, 

 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 jSTovember 1 six tubes of neutral beef-peptone agar were 

 inoculated with small pieces of the bro^vn, 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° Pahr. for one week but they produced no growth whatever. 



Finally, at the suggestion of Mr. IT. 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. c. 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. 



