635 
of Edinhicrgh, Session 1883-84. 
The advantages just cited, as well as the experiments of Koch, 
were sufficient to induce me to also select the Anthrax hacillus for 
the purpose of ascertaining the relative value of disinfectants. 
Cultivation of Anthrax . — Dr Klein placed at my disposal a small 
supply of Anthrax hadlli in the spore state, and I had the advan- 
tage of studying his admirable paper * (Eleventh Annual Report of 
the Local Government Board), in which he has given full details 
of an improved method of cultivation. 
A large air-bath was set up, and all flasks, beakers, and pipettes 
heated for many hours to about 500° Eahr. Similarly the cotton- 
wool used was disinfected by a dry heat, the heat being carried to 
nearly the singeing point. 
As in Dr Klein’s method, pork as lean as possible, after suitable 
division, was boiled in water, the broth filtered through a sieve, and 
then the supernatant fat separated by means of a separating funnel. 
The broth was next accurately measured, and 10 c.c. or 50 c.c., titrated 
by d. n. soda, using as an indicator phenol-phthalein. In this way the 
precise acidity was known, and from the data thus obtained it was 
easy to exactly neutralise the remaining bulk. After neutralisation 
with soda, the broth was again boiled for some time, and then filtered 
hot into a previously sterilised flask. From this stock flask the 
cultivation flasks were charged with from 20 to 30 c.c. of the broth. 
The cultivation flasks had bulbs of about 70 c.c. capacity, the 
bottoms were flat, and the necks from four to five inches long, 
and narrow ; they were charged by means of a sterilised pipette, 
and then plugged for at least three inches with a cotton-wool plug. 
After plugging, the flasks were immersed in a water-bath, so that 
only about an inch of the neck was above the surface, and the 
water kept boiling for many hours. This method answered very 
well, and seemed to me an improvement on the usual plan of 
directly boiling the broth in the flask. After this process the flasks 
thus charged were placed in an incubator thirty at a time, and 
kept for three days and nights at the temperature of the blood. 
Those that remained clear were then considered sterilised, those 
showing the least turbidity were rejected, 
* “On the Relation of Pathogenic to Septic Bacteria, as illustrated by 
Anthrax Cultivation,” by E. Klein, M.D., F.R.S. Supplement containing 
the Report of the Medical Officer. 
