CUL 77 VA TION OF BA CTERIA. 131 Glycerin Agar-agar.—For an unknown reason certain of the bacteria which will not grow upon the agar-agar as prepared above will do so if 3-7 per cent, of glycerin be added. Among these is the tubercle bacillus, which, not growing at all upon plain agar-agar, will grow well when glycerin is added—a fact discovered by Roux and Nocard. The glycerin may also be added to gelatin or any other medium. Blood Agar-agar was recommended by R. Pfeiffer for the cultivation of the influenza bacillus. It is ordinary agar-agar whose surface is coated with a little blood secured under antiseptic precautions from the finger-tip, ear-lobule, etc., of man, or the veins of one of the lower animals. Some bacteriologists prepare a hemoglobin agar-agar by spreading a little powdered hemoglobin upon the surface of the agar-agar. This has the disad- vantage that powdered hemoglobin is not sterile, and the medium must be sterilized after its addition. The blood agar-agar should be kept in the incubator a day or two before use so as to insure perfect sterility. Blood-serum.—The great advantage possessed by this medium is that it is itself a constituent of the body, and hence offers opportunities for the development of the parasitic forms of bacteria under the most natural con- ditions possible. It is the most difficult of all the media to prepare. The blood must be obtained from a slaughter- house in an appropriate receptacle, the best things for the purpose being tall narrow jars of about i liter capacity, with a tightly-fitting lid. The jars are sterilized by heat or by washing with alcohol and ether, are carefully dried, closed, and carried to the slaughter-house where the blood is to be obtained. As the blood flows from the severed vessels of the animal the jars are filled one by one. It seems advisable to allow the first blood to escape, as it is likely to become contaminated from the hair. By waiting until a coagulum forms upon the hair the danger of con- tamination is obviated. The jars when full are allowed to stand undisturbed until quite firm coagula form within