182 BACTERIA IN RELATION TO PLANT DISEASES. 
Harding inoculated Bact. campestre into animals with negative results. 
Harrison inoculated Bacillus solanisaprus into guinea-pigs and rabbits without positive 
results (see Vol. III, Basal Stem-rot of Potato). 
The following experiments with fishes in water containing bacteria of plant diseases 
were undertaken for me in 1905 by M. C. Marsh, bacteriologist of the Bureau of Fisheries, 
in Washington. I prepared the cultures myself. The summary is his: 
Bacillus aroideae.—About 25 liters Potomac tap water in glass aquarium jar, with constant flow 
of air in small bubbles delivering at the bottom. ‘Two sunfish, 1 small goldfish, and 1 mummichog 
were used with 14 cc. of a well-clouded 2-day bouillon culture of B. aroideae introduced on first day, 
February 17, 1905; March 5, 15 cc. of a 5-day culture added. March 13, after 26 days, all fishes 
alive and in good condition. 
March 5, injected largest of the above sunfish behind eye with about 0.5 cc. of a 5-day well- 
clouded bouillon culture of B. aroideae; producing great exophthalmia. Sunfish remained in jar 
containing B. aroideae mixed with water. March 13, fish alive, eye normal; time 8 days. 
Bacteria of carnation leaf spot, February 19, about 25 liters Potomac tap water in glass aquarium, 
aeration as above. One small black bass (5 inches), 2 sunfish, 1 mummichog. February 20, added 
two 20-day bouillon cultures of carnation bacteria, one 5-day bouillon culture, and one 20-day agar 
slant culture; February 28, added three 9-day bouillon cultures. 
March 13, after 21 days, all fish in good condition. 
Temperature of water 14.5° to 22° C. No change of water during experiments. Fishes fed 
very sparingly. 
In transmitting the above report Mr. Marsh made the following comment: 
I send you herewith a statement of the effect of the plant bacteria on fishes, from which it 
appears that the effect is mil. I did not make a direct inoculation with the carnation rot,on account 
of the result with the presumably more dangerous calla rot. It is not likely that these organisms 
would harm any fishes, though I was unable to try trout. The eye inoculation should have taken if 
there was any pathogenicity about the calla rot for fishes. 
INOCULATIONS OF BACT. TUMEFACIENS INTO FISH AND FROGs. 
In the spring of 1908, the writer made fourteen sets of inoculation experiments on fish 
and frogs with pure cultures of Bact. tumefaciens derived from tumors on the hothouse 
daisy (Chrysanthemum) to determine whether this organism would induce similar abnor- 
mal growths in cold-blooded animals, experiments on warm-blooded animals being con- 
sidered unnecessary because of the low maximum temperature of the organism (about 
36:6-C.), 
These inoculations were carried on in Washington in houses belonging to the Bureau 
of Fisheries with trout and roach kindly placed at my disposal from the stock tanks and with 
frogs bought from a Washington dealer. With exception of those used in Experiment IX, 
the trout were 8 to 10 inches long and were ordinary brook trout (Salvelinus fontanalis). 
The roach (Abramis chrysoleucus) were about 6 inches long. There were no checks on the 
roaches or frogs. The checks on the trout consisted of a school of about 100 fish of the same 
age and condition, and of which those I took had previously formed a part. These were in 
one of the ordinary exhibition tanks of the Bureau along with some rainbow trout. They 
were not checks in the strictest sense of the word because they were not wounded in any 
way. 
Inoculations of March 20, 1908.—These were made from four agar streak cultures 
48 hours old. None of them were hypodermic injections. 
I. Two trout. Each two needle-pricks in the eye-socket. 
II. Two trout. Each three needle-pricks in the region of the anus. 
III. Two trout. Each three or four needle-pricks in the fleshy fin (adipose). 
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