130 HISTORY AND CAUSE OF THE COCONUT BUD-EOT. 



canary yellow of fermentation tubes with peptone and neutral red, 

 and reduced nitrates to nitrites. 



The writer of this paper likewise made attempts to isolate Bacillus 

 coli from the same diseased seedling used by Miss McCulloch. The 

 process was sunilar to that in experiment 1 and showed results as 

 follows : 



The agar plates which were poured from the diseased material showed in 24 hours 

 numerous round, white, raised, wet-shining colonies typical of Bacillus coli. Some 

 of the colonies were irregular in shape, even to radiate branching, and some were 

 bluish and iridescent in transmitted light, but these are variations often met with in 

 what passes for Bacillus coli. 



Transfers were made from the various colonies to plain beef-bouillon tubes and 

 thence to nitrate-bouillon tubes. After three days in the nitrate bouillon, tests were 

 made for the reduction of nitrates, and it was found that 6 out of the 13 tubes responded 

 to the test. Of these six, two were from round, white colonies; one from round, white, 

 iridescent; one from blue iridescent; one from radiate branched; and one from an 

 irregular blue iridescent colony. Those cultures which failed to show the reduction 

 were largely from round colonies. 



Fermentation tubes containing dextrose, peptone, and neutral red were inoculated, 

 and after two days at 37° C. showed the typical canary-yellow color in the closed 

 arm, together with an average of 35 millimeters of gas. Four out of seven of the tubes 

 tried responded to this test. From these four tubes transfers were made to Dolt's litmus- 

 lactose-glycerin agar slant tubes where all grew well, reddened the agar, and produced 

 good pink growths. 



Transfers were made from these same tubes in gelatin and placed in the thermostat 

 at 37° C, where an excellent growth took place. After 48 hours the tubes were 

 placed in an ice box and the medium soon became entirely solidified, showing that 

 no liquefaction had taken place. 



From these results it will be observed that the same conclusion may be derived 

 as from Miss McCulloch 's platings, that is, that Bacillus coli was isolated from the 

 diseased material obtained from an inoculation of Bacillus coli. 



Experiment No. 3. 



On April 14, 1910, two inoculations were made into coconut seed- 

 lings with cultures of Bacillus coli. 



The point of inoculation on the seedling was washed with a solution 

 of mercuric chloride before inoculation. 



On May 16, just 32 days afterwards, these two inoculations were 

 cut out and examined. They appeared as follows: 



(a) Dry, brown discoloration about upper part of hole. At extreme lower end were 

 water-soaked discolorations and slight signs of rot. At the lower part the tissues were 

 considerably split up. The edges of the cracks were yellowed, and in the cracks 

 were masses of what appeared to be bacteria, but little active motility was discernible. 



(b) Good water-soaked, brown, soft rot extending about 5 centimeters (PI. XII). 



Diseased material from these inoculations was carefully rinsed in 

 alcohol, soaked in mercuric chlorid, and rinsed in distilled water; 

 then, by means of sterile knives and forceps, small pieces were put 

 into test tubes containing beef bouillon and there thoroughly cut up. 



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