CULTURE MEDIA 45 



Blood-smeared Agar Medium. Make plain agar, put in tubes, sterilize in 

 autoclave and slant. When the agar solidifies, wash a finger with soap and hot 

 water, alcohol, i: 1000 bichloride sol., sterile water, then dry with sterile towel. 

 Prick the finger with a sterile needle, catch the blood in a sterile platinum loop 

 and smear it over the entire surface of the agar. 



Blood Agar Medium. Etherize a rabbit, shave and asepticize its chest, take 

 a IQ-CC. or 20-cc. sterile syringe and needle and withdraw blood from heart, 

 empty blood into a sterile bottle containing sterile glass beads and shake vigor- 

 ously until defibrinated. Bring temperature to 45C. in water bath. Have 

 tubes containing 10 cc. sterile, plain agar, liquefied, and at a temperature of 

 45C. With a sterile pipette add 5 cc. of blood to each tube of agar, mix blood 

 and agar and slant. 



Blood Alkali Agar Medium. Obtain ox blood in slaughter house. Collect 

 in small sterile bottles containing glass beads (100 cc. to 300 cc., cap bottles), 

 shake until defibrinated. 



Mix equal volumes of blood and normal sodium hydrate solution. Sterilize 

 in steam sterilizer 15 minutes each day for 2 days. 



Add i part of this alkaline blood to 7 parts sterile plain agar, place in tubes 

 and slant. Do not have cotton plugs any tighter than necessary. 



Loeffler's Blood-serum Medium. Blood serum (calf's or lamb's), 3 parts. 



Glucose bouillon, i part. 



Put in tubes, place in serum sterilizer. Heat at 57C. i hour each day for 

 6 days, then at r joC. i hour each day for 2 days. Or heat in Arnold steam 

 sterilizer at 8oC., i hour each day for 3 days. 



A Substitute for Ordinary Blood Serum. 1 "Add from 10 to 15 cc. of i per 

 cent, glucose bouillon to the white and yolk of one egg, make smooth mixture 

 in a mortar and place in tube. Inspissate and sterilize as for ordinary serum 

 slants." 



"When this medium is to be used for culturing tubercle bacilli, add about 

 i cc. of glycerin bouillon to each tube before final sterilization in the autoclave. 

 The cotton plugs should be paraffined to prevent drying of the slants in the 

 incubator." 



Egg Medium. Select a clean, fresh egg '(not more than 24 hours old) that 

 has a perfect shell, no cracks; shake to mix white and yolk, wash with warm 

 sterile water, then with warm i : 500 bichloride solution and again with sterile 

 water, wipe dry. To inoculate, pierce shell w r ith sterile needle and introduce 

 bacteria with platinum loop, seal opening with wax. If anaerobic culture is 

 desired, coat egg with paraffin. 



Dorset's Egg Medium. Selected clean fresh eggs with perfect shells. Wash 

 with sterile water, then with 5 per cent, phenol and finally with sterile water. 

 Break with a sterile knife and empty the eggs into a sterile flask. 



1 Stitt, "Practical Bacteriology," 3d edition, p. 25. 



